WO2023161802A1

WO2023161802A1 - Methods and systems for controlling xylella fastidiosa in olive trees

Info

Publication number: WO2023161802A1
Application number: PCT/IB2023/051593
Authority: WO
Inventors: Luca Alessandro GRANDI; Michael Christian OEHL; Nicolas Schmitt; Dimitri Georges VERWEIRE
Original assignee: Invaio Sciences International GmbH
Current assignee: Invaio Sciences International GmbH
Priority date: 2022-02-22
Filing date: 2023-02-22
Publication date: 2023-08-31
Anticipated expiration: 2024-08-22

Abstract

The present disclosure relates generally to methods and systems of treating infected plants and/or controlling bacteria, fungi, virus and/or other pathogens that cause disease in plants, such as olive trees, and more specifically to methods and systems for controlling Xylella fastidiosa infection in olive trees.

Description

METHODS AND SYSTEMS FOR CONTROLLING XYLELLA FASTIDIOSA IN OLIVE TREES

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63/312,764, filed February 22, 2022, which is incorporated herein by reference in its entirety.

FIELD

[0002] The present disclosure relates generally to methods and systems of treating infected plants and/or controlling bacteria, fungi, virus and/or other pathogens that cause disease in plants, such as olive trees, and more specifically for controlling Xylella fastidiosa infection in olive trees.

BACKGROUND

[0003] Xylella fastidiosa is a plant pathogenic bacterium. Xylella fastidiosa can be divided into various subspecies that affect different plants. For example, Xylella fastidiosa subsp. has been found to cause “olive quick decline syndrome” (OQDS) in southern Italy and is spreading to central and North Italy as well as Spain, Portugal, France and possibly Greece. OQDS is a severe disease that can cause extensive leaf, twig and branch wilting, and is frequently accompanied by plant death. See M. Scortichini et al., Phytopathologia Mediterranea (2018), 57, 1, 48-72.

[0004] Xylella fastidiosa is a xylem-limited pathogen that is spread through the feeding activity of certain insect vectors. Infection by this pathogen causes blockage of the xylem vessels. See C.R. Girelli et al., Plants (2019), 8, 115, 1-17. Current methods to control this pathogen are generally not effective, in part because of the inability of bactericides to penetrate host xylem vessels where the pathogen is located.

[0005] For example, current methods to control Xylella fastidiosa include pruning and/or uprooting trees. However, this method can lead to huge cost and lost income, particularly since it can takes up to 10-20 years before a good level of production is obtained after replanting new trees. In addition, the trees may contribute to the cultural heritage of a region, as well as touristic attractiveness and economy of the region. Pruning has generally been found to be an ineffective solution because, within the same year, the symptoms typically spread to other areas in the tree.

[0006] Current methods may involve replanting with varieties that show resistance and/or tolerance o Xylella fastidiosa. However, this method can take decades to develop these varieties, plant them, and obtain yields comparable to the trees that need to be replaced without any certainty of success in the first instance. In addition, these new varieties are unlikely the same as those currently certified as Protected Designation of Origin or Protected Geographical Indication (also known as Indicazione Geografica Protetta in Italian), which may hinder the ability to sell at a certain premium by olive oil producers. Furthermore, mutations in the Xylella bacterium might break the resistance and/or tolerance overtime.

[0007] Other methods may involve foliar spraying of Dentamet®, a mixture of zinc and copper complex with hydracids of citric acid. However, a large volume of the formulation is typically needed to cover tree canopy. Applicant would like to improve upon such methods because only an estimated 10-30% of product actually land on the tree’s foliage. Such methods also have been observed to be less effective in older trees (for example, trees older than 100 years), and when symptoms are in advanced stages (for example, when more than 50% of the tree is affected or when 50% or less of the canopy foliage left).

[0008] Yet other methods may involve drilling a hole in the tree to administer the formulation. However, the practicality of such method is difficult to execute on a commercial scale and has a high risk of trunk damage, which may lead to rot and other negative effects associated with severely wounding the tree.

[0009] Further, providing a formulation through conventional methods, e.g., foliar spray or drenching, may (i) cause higher phytotoxicity, (ii) take longer period for certain residue to reach acceptable level from last treatment or administration of the formulation, and/or (iii) show low efficacy.

[0010] Thus, improved injection protocols and systems for controlling the pathogen controlling Xylella fastidiosa infection of an olive tree are needed. BRIEF SUMMARY

[0011] In one aspect, provided are methods for controlling Xylella fastidiosa infection of an olive tree. In some embodiments, the olive tree has a trunk. In some embodiments, the trunk has a circumference and a diameter. In some embodiments, the infected olive tree exhibits at least one symptom caused by Xylella fastidiosa. In some embodiments, the method comprises measuring the trunk circumference or the trunk diameter, or both. In some embodiments, the method comprises determining a symptom level of the infected olive tree. In some embodiments, the method comprises injecting the infected olive tree with an injection formulation comprising zinc and copper. In some embodiments, when the trunk circumference is less than about 140 cm, or the trunk diameter is less than about 45 cm, or both, the injection formulation comprises an initial concentration of the zinc (Z 1). In some embodiments, when (i) the trunk circumference is between about 140 cm and about 200 cm, or the trunk diameter is between about 45 cm and about 65 cm, or both, and (ii) the symptom level of the infected tree is less than or equal to about 50%, the injection formulation comprises the initial concentration of the zinc (Z 1). In some embodiments, when (i) the trunk circumference is between about 140 cm and about 200 cm, or the trunk diameter is between about 45 cm and about 65 cm, or both, and (ii) the symptom level of the infected tree is greater than about 50%, the injection formulation comprises about twice the initial concentration of the zinc (1.5Z1-2.5Z1). In some embodiments, when (i) the trunk circumference is greater than about 200 cm, or the trunk diameter is greater than about 65 cm, or both, and (ii) the symptom level of the infected tree is less than or equal to about 20%, the injection formulation comprises about the initial concentration of the zinc (Zl). In some embodiments, when (i) the trunk circumference is greater than about 200 cm, or the trunk diameter is greater than about 65 cm, or both, and (ii) the symptom level of the infected tree is between about 20% and about 50%, the injection formulation comprises about twice the initial concentration of the zinc (1.5Z1-2.5Z1). In some embodiments, when (i) the trunk circumference is greater than about 200 cm, or the trunk diameter is greater than about 65 cm, or both, and (ii) the symptom level of the infected tree is greater than about 50%, the injection formulation comprises about four times the initial concentration of the zinc (3.5Z1- 4.5Z1). In some embodiments, the initial concentration of the zinc is about 0.08% w/w to about 0.23% w/w. [0012] In some embodiments a method of treating an infected olive tree comprises, a) measuring the trunk circumference or the trunk diameter, or both; b) determining a symptom level of the infected olive tree; and/or c) injecting the infected olive tree with an injection formulation comprising zinc and copper. In some embodiments, the injection formulation comprises a concentration of zinc at about 0.08% w/w to about 0.23% w/w. In some embodiments of the aforementioned method of treating an infected olive tree, steps a), b), or c), or any combination thereof, is performed about 7 weeks before flowering, after fruit set, post-summer, and at the end of the season. In some embodiments, about 7 weeks before flowering is around March, after fruit set is around May and June, post-summer is around September, and the end of the season is around October.

[0013] In some embodiments, the method comprises injecting the infected olive tree with a low-dose injection formulation. In some embodiments, the low-dose formulation comprises about two thirds the initial concentration of the zinc. In some embodiments, injecting the infected olive tree with a low-dose injection formulation occurs about 3 weeks before flowering. In some embodiments, injecting the infected olive tree with a low-dose injection formulation occurs at or during pre-summer season. In some embodiments, about 3 weeks before flowering is around April. In some embodiments, the pre-summer season is around July.

[0014] In some embodiments, the method comprises assigning a trunk measurement range. In some embodiments, the trunk measurement range is selected from a set of reference trunk measurement ranges based on the circumference and/or diameter of the trunk. In some embodiments, the method comprises assigning a symptom level range. In some embodiments, the symptom level range is selected from a set of reference symptom level ranges. In some embodiments, the method comprises selecting an injection concentration. In some embodiments, the injection concentration is selected from a set of reference injection concentrations. In some embodiments, the method comprises selecting an injection concentration range. In some embodiments, the injection concentration range is selected from a set of reference injection concentration ranges. In some embodiments, selection of the injection concentration or the injection concentration range is based on the trunk measurement range and/or the symptom level range. In some embodiments, the method comprises applying an injection formulation to the infected olive tree. In some embodiments, the injection formulation comprises zinc and copper. In some embodiments, zinc is present in the injection formulation at the selected injection concentration. In some embodiments, zinc is present in the injection formulation within the selected injection concentration range.

[0015] In some embodiments, the set of reference trunk measurement ranges comprises a low trunk measurement range. In some embodiments, the set of reference trunk measurement ranges comprises a medium trunk measurement range. In some embodiments, the set of reference trunk measurement ranges comprises a high trunk measurement range. In some embodiments, the low trunk measurement range comprises a trunk circumference range of less than about 140 cm, or a trunk diameter of less than about 45 cm, or both. In some embodiments, the medium trunk measurement range comprises a trunk circumference range between about 140 cm and about 200 cm, or a trunk diameter between about 45 cm and about 65 cm, or both. In some embodiments, the high trunk measurement range comprises a trunk circumference range of greater than about 200 cm, or a trunk diameter of greater than about 65 cm, or both.

[0016] In some embodiments, the set of reference symptom level ranges comprises a low symptom level range. In some embodiments, the set of reference symptom level ranges comprises a medium symptom level range. In some embodiments, the set of reference symptom level ranges comprises a high symptom level range.

[0017] In some embodiments, an infected tree having a low symptom level range exhibits less than about 20% symptom level. In some embodiments, an infected tree having a medium symptom level range exhibits between about 20% and 50% symptom level. In some embodiments, an infected tree having a high symptom level range exhibits greater than 50% symptom level.

[0018] In some embodiments, the symptom level is measured based on one or more Xylella fastidiosa symptoms. In some embodiments, Xylella fastidiosa symptoms comprise scorching, discoloration, stunting, wilting, premature leaf abscission, shriveled fruit, premature fruit abscission, dieback, or plant death, or any combination thereof. In some embodiments, the symptom level is measured based on one or more Xylella fastidiosa symptoms affecting canopy of the olive tree.

[0019] In some embodiments, the reference injection concentrations of copper and/or zinc comprise a low injection concentration. In some embodiments, the reference injection concentrations of copper and/or zinc comprise a medium injection concentration. In some embodiments, the reference injection concentrations of copper and/or zinc comprise a high injection concentration. In some embodiments, the medium injection concentration of copper and/or zinc is about twice the low injection concentration of copper and/or zinc. In some embodiments, the high injection concentration of copper and/or zinc is about four times the low injection concentration of copper and/or zinc. In some embodiments, the low injection concentration of copper and/or zinc is about 0.08% to about 0.23% w/w. In some embodiments, the medium injection concentration of copper and/or zinc is about 0.16% to about 0.46%. In some embodiments, the high injection concentration of copper and/or zinc is about 0.32% to about 0.92%.

[0020] In some embodiments, the reference injection concentration ranges of copper and/or zinc comprise a low injection concentration range. In some embodiments, the reference injection concentration ranges of copper and/or zinc comprise a medium injection concentration range. In some embodiments, the reference injection concentration ranges of copper and/or zinc comprise a high injection concentration range. In some embodiments, the lower limit of the medium injection concentration range of copper and/or zinc is about twice the lower limit of the low injection concentration range. In some embodiments, the lower limit of the high injection concentration range of copper and/or zinc is about four times the lower limit of the low injection concentration. In some embodiments, the low injection concentration range of copper and/or zinc is about 0.08% to about 0.23%. In some embodiments, the medium injection concentration range of copper and/or zinc is about 0.16% to about 0.46% w/w. In some embodiments, the high injection concentration range of copper and/or zinc is about 0.32% to about 0.92% w/w.

[0021] In some embodiments, the low injection concentration is selected when the low trunk measurement range is assigned. In some embodiments, the low injection concentration is selected when the medium trunk measurement range is assigned and the low or medium symptom level range is assigned. In some embodiments, the low injection concentration is selected when the high trunk measurement range is assigned and the low symptom level range is assigned. In some embodiments, the medium injection concentration is selected when the medium trunk measurement range is assigned and the high symptom level range is assigned. In some embodiments, the medium injection concentration is selected when the high trunk measurement range is assigned and the medium symptom level is assigned. In some embodiments, the high injection concentration is selected when the high trunk measurement range is assigned and the high symptom level range is assigned.

[0022] In some embodiments, the low injection concentration range is selected when the low trunk measurement range is assigned. In some embodiments, the low injection concentration range is selected when the medium trunk measurement range is assigned and the low or medium symptom level range is assigned. In some embodiments, the low injection concentration range is selected when the high trunk measurement range is assigned and the low symptom level range is assigned. In some embodiments, the medium injection concentration range is selected when the medium trunk measurement range is assigned and the high symptom level range is assigned. In some embodiments, the medium injection concentration range is selected when the high trunk measurement range is assigned and the medium symptom level is assigned. In some embodiments, the high injection concentration range is selected when the high trunk measurement range is assigned and the high symptom level range is assigned.

[0023] In some embodiments, a) assigning a trunk measurement range selected from a set of reference trunk measurement ranges based on the circumference and/or diameter of the trunk; b) assigning a symptom level range selected from a set of reference symptom level ranges; c) selecting an injection concentration from a set of reference injection concentrations or selecting an injection concentration range from a set of reference injection concentration ranges based on the trunk measurement range and the symptom level range; and/or d) applying an injection formulation comprising zinc and copper to the infected olive tree, wherein zinc is present in the injection formulation at the selected injection concentration or within the selected injection concentration range, i.e., steps a), b), c), or d), or any combination thereof is performed about 7 weeks before flowering, after fruit set, postsummer, and/or at the end of the season.

[0024] In some embodiments, the method comprises injecting the infected olive tree with a low-dose injection formulation. In some embodiments, the low-dose injection formulation comprises zinc and copper. In some embodiments, injecting the infected olive tree with a low-dose injection formulation occurs about 3 weeks before flowering. In some embodiments, injecting the infected olive tree with a low-dose injection formulation occurs at or during pre-summer season. In some embodiments, the zinc concentration of the low-dose injection formulation is about two thirds the low injection concentration. In some embodiments, the zinc concentration of the low-dose formulation is selected from a low-dose concentration range. In some embodiments, the lower limit of the low-dose concentration range is two thirds the lower limit of the low injection concentration range. In some embodiments, the upper limit of the low-dose concentration range is two thirds the upper limit of the low injection concentration range. In some embodiments, about 3 weeks before flowering is around April. In some embodiments, pre-summer season is around July.

[0025] In some embodiments, the circumference and/or diameter of the trunk is measured at a reference height from the ground. In some embodiments, the reference height is between about 40 cm and about 80 cm from the ground.

[0026] In some embodiments, the injecting of the injection formulation is performed using an injection system comprising an injection tool. In some embodiments, the injection tool is operatively connected to a fluid delivery unit. In some embodiments, the fluid delivery unit is configured to deliver the injection formulation.

[0027] In some embodiments, the injecting of the injection formulation comprises piercing the trunk of the olive tree using the injection tool of the injection system. In some embodiments, the injecting of the injection formulation comprises delivering at least a portion of the injection formulation from the fluid delivery unit through the injection tool into and no further than the active vasculature of the olive tree.

[0028] In some embodiments, the olive tree is suffering from olive quick decline syndrome. In some embodiments, the Xylella fastidiosa is Xylella fastidiosa subsp. pauca. In some embodiments, the injection formulation is distributed throughout the trunk and other parts of the olive tree. In some embodiments, the other parts of the olive tree comprise fruits. In some embodiments, the other parts of the olive tree comprise leaves. In some embodiments, the olive tree is at least about 30 years old. In some embodiments, the olive tree is at least about 100 years old.

[0029] In some embodiments, the delivery unit is a spring-loaded fluid delivery unit. In some embodiments, the delivery unit comprises a pressurized formulation cartridge.

[0030] In some embodiments, the method comprises replacing the fluid delivery unit with a second fluid delivery unit. In some embodiments, the method comprises delivering at least a portion of the injection formulation from the second fluid delivery unit through the injection tool into and no further than the active vasculature of the olive tree.

[0031] In some embodiments, the injection tool remains in the trunk of the olive tree over at least one growing season. In some embodiments, the injection tool remains in the trunk of the olive tree over multiple re-injections. In some embodiments, the trunk of the olive tree has bark. In some embodiments, the method comprises removing at least a portion of the bark prior to piercing the trunk.

DESCRIPTION OF THE FIGURES

[0032] The present application can be understood by reference to the following description taken in conjunction with the accompanying figures.

[0033] FIG. 1 depicts an example of a dosing matrix.

[0034] FIG. 2 depicts an example of an annual dosing program.

[0035] FIGS. 3A-3C depict an example of an injection tool that may be used with the methods and systems described herein.

[0036] FIGS. 4A-4D depict an example of a multi-port injection tool that may be used with the methods and systems described herein.

[0037] FIG. 5 depicts an example of a spring-loaded fluid delivery unit that may be used with the methods and systems described herein.

[0038] FIG. 6A depicts an example of a chassis that may be used with the methods and systems described herein.

[0039] FIG. 6B depicts an example of a canister that may be used with the methods and systems described herein.

[0040] FIG. 7A-7E depict olive trees with desirable and undesirable injection sites illustrated.

[0041] FIG. 8 A depicts delta DI plot for water injection.

[0042] FIG. 8B depicts delta DI plot for foliar treatment. [0043] FIGS 8C-8E depict delta DI plots for various injection rates.

[0044] FIGS. 9 A and 9B depict delta DI plots for different injection frequencies.

[0045] FIGS. 10A-10C depict delta DI plots for various initial DI conditions.

[0046] FIGS. 11 A and 1 IB depict a graph showing the amount of copper and zinc residue in the olive fruit.

DETAILED DESCRIPTION

[0047] The following description is presented to enable a person of ordinary skill in the art to make and use the various embodiments. Descriptions of specific devices, techniques, and applications are provided only as examples. Various modifications to the examples described herein will be readily apparent to those of ordinary skill in the art, and the general principles defined herein may be applied to other examples and applications without departing from the spirit and scope of the various embodiments. Thus, the various embodiments are not intended to be limited to the examples described herein and shown, but are to be accorded the scope consistent with the claims.

Injection formulations

[0048] In one aspect, provided are injection formulations. In certain embodiments, the injection formulation is water soluble. In some variations, the injection formulation comprises nutrients. In certain variations, the injection formulation comprises micronutrients.

[0049] In certain embodiments, the injection formulation comprises zinc, copper, or manganese, or any salt thereof, or any combination of the foregoing. In some variations, the injection formulation comprises zinc and copper, or any salt thereof. In other variations, the injection formulation comprises zinc and manganese, or any salt thereof.

[0050] In some embodiments, when the injection formulation is said to comprise a certain element or molecule, e.g., zinc, copper, manganese, or citric acid, the element or molecule may exist in the injection formulation in any form, e.g., as an ion, in an insoluble form, as part of a salt, or as part of a complex. In some embodiments, when the concentration of a certain element or molecule in a formulation is specified, the concentration should account for all of such element or molecule in the formulation regardless of what form they are in, e.g., as an ion, in an insoluble form, as part of a salt, or as part of a complex. [0051] In some embodiments of the foregoing, the injection formulation further comprises citric acid. In certain variations, the citric acid is complexed with the micronutrient(s) in the formulation. In one variation, the citric acid is in the form of hy dracids. In one variation, the injection formulation comprises a mixture of copper and zinc complexed with citric acid (e.g., under the form of hy dracids). In another variation, the injection formulation comprises a mixture of zinc and manganese complexed with citric acid (e.g., under the form of hy dracids). In yet other variations, the injection formulation comprises a mixture of zinc, manganese and copper complexed with citric acid (e.g., under the form of hy dracids). In other embodiments, other chelating agents may be used to bind to or complex with zinc or copper or manganese, e.g., aminopolycarboxylic acids such as ethylenediaminetetraacetic acid (EDTA).

[0052] In some embodiments, the injection formulation comprises a stock formulation or a commercially available formulation. In some embodiments, the injection formulation comprises a stock formulation diluted with water or other solvents or formulations. In some embodiments, the injection formulation comprises a commercially available formulation diluted with water or other solvents or formulations. In some embodiments, stock formulations comprise commercially available formulations. Examples of commercially available formulations suitable for use in the methods herein comprise Dentamet® (DiAgro) and Bio-D (DiAgro).

[0053] In some variations, commercially available formulations or stock formulations may be diluted and/or further formulated for use in the methods described herein. For example, in some embodiments, the injection formulation comprises one part Dentamet® and four parts water. In certain variations, the injection formulation comprises Dentamet® diluted in water so that a balance between obtaining a sufficient amount to get activity while avoiding phytotoxicity in the tree is achieved.

[0054] In some embodiments, the injection formulation comprises about 5% of a stock formulation or a commercially available formulation, meaning that the stock formulation or the commercially available formulation is diluted 20-fold. In some embodiments, the injection formulation comprises about 5%, 10%, or 20% of a stock formulation or a commercially available formulation. In some embodiments, the injection formulation comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of a stock formulation or a commercially available formulation. In some embodiments, the injection formulation comprises a stock formulation or a commercially available formulation in the amount less than any of the following: 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the injection formulation comprises a stock formulation or a commercially available formulation in the amount greater than any of the following: 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the injection formulation comprises a stock formulation or a commercially available formulation with the upper limit of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, and an independently selected lower limit of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, wherein the lower limit is less than the upper limit.

[0055] In some embodiments, the stock formulation comprises between about 7% and 20% zinc sulfate heptahydrate. In some embodiments, the stock formulation comprises less than about 15% sulfuric acid. In some embodiments, the stock formulation comprises less than about 10% malic acid. In some embodiments, the stock formulation comprises less than about 10% citric acid monohydrate. In some embodiments, the stock formulation comprises between about 2% and about 4.5% tribasic copper sulfate.

[0056] In some variations, the stock formulation comprises between 1.59% and 4.55% zinc. In some variations, the stock formulation comprises less than about 9.14% citric acid. In some variations, the stock formulation comprises between about 0.3% and about 0.7% copper. In some embodiments, the stock formulation comprises between 1% and 5% zinc. In some embodiments, the stock formulation comprises less than about 10% citric acid. In some embodiments, the stock formulation comprises between about 0.1% and about 1% copper.

[0057] In some embodiments, the stock formulation comprises about 0.1%, 0.2%, 0.3%,

0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%,

6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% zinc. In some embodiments, the stock formulation comprises less than about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% zinc. In some embodiments, the stock formulation comprises more than about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% zinc. In some embodiments, the zinc concentration in the stock formulation has an upper limit of about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%, and an independently selected lower limit of about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%, where the lower limit is less than the upper limit.

[0058] In some embodiments, the stock formulation comprises about 0.1%, 0.2%, 0.3%,

6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% copper. In some embodiments, the stock formulation comprises less than about 0.1%, 0.2%, 0.3%, 0.4%,

0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%,

7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% copper. In some embodiments, the stock formulation comprises more than about 0.1%, 0.2%, 0.3%, 0.4%,

7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% copper. In some embodiments, the copper concentration in the stock formulation has an upper limit of about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%, and an independently selected lower limit of about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%, where the lower limit is less than the upper limit.

[0059] In some embodiments, a low-dose injection formulation is a diluted injection formulation. In some embodiments, a low-dose injection formulation is an injection formulation diluted with water or any suitable solvent or formulation. In some embodiments, a low-dose injection formulation comprises an injection formulation or a low-injection- concentration injection formulation further diluted by 2/3, meaning that 2/3 of the low-dose injection formulation is the injection formulation or the low-injection-concentration injection formulation. In some embodiments, a low-dose injection formulation comprises an injection formulation or a low-injection-concentration injection formulation further diluted by about 29/30, 14/15, 9/10, 13/15, 5/6, 4/5, 23/30, 11/15, 7/10, 2/3, 19/30, 3/5, 17/30, 8/15, 1/2, 7/15, 13/30, 2/5, 11/30, 1/3, 3/10, 4/15, 7/30, 1/5, 1/6, 2/15, 1/10, 1/15, or 1/30. In some embodiments, a low-dose injection formulation comprises about 3.3% Dentamet®. In some embodiments, a low-dose injection formulation comprises about 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, or 20% Dentamet®.

Treatment Protocol

[0060] In some aspects, provided are methods for controlling Xylella fastidiosa infection of an olive tree. In some embodiments, the olive tree has a trunk. In some embodiments, the trunk has a circumference. In some embodiments, the trunk has a diameter. In some embodiments, the infected olive tree exhibits at least one symptom caused by Xylella fastidiosa. In some embodiments, the olive tree is infected. In some embodiments, the olive tree is not infected. In some embodiments, the methods described herein are used only for olive trees with one or more Xylella fastidiosa symptoms. In some embodiments, the methods described herein are not applicable to trees beyond a certain dimension. For example, in some variations, the methods described herein are not applicable to trees with a circumference greater than about 300 cm.

[0061] In some embodiments, the method comprises measuring the trunk circumference. In some embodiments, the method comprises measuring the trunk diameter. In some embodiments, the method comprises measuring the trunk circumference, the trunk diameter, or both.

[0062] In some embodiments, the trunk circumference or the trunk diameter is measured at a reference height from the ground. In some embodiments, the reference height is measured from the ground. In some embodiments, the reference height is between about 40 cm to about 80 cm. In some embodiments, the reference height is about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 cm. In some embodiments, the reference height is lower than about any of the following: 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 cm. In some embodiments, the reference height is higher than about any of the following: 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 cm. In some embodiments, the reference height has an upper limit of about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 cm, and an independently selected lower limit of about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 cm, wherein the lower limit is less than the upper limit.

[0063] In some embodiments, the method comprises determining a symptom level of the olive tree. In some embodiments, the termination of a symptom level is based on one or more Xylella fastidiosa symptoms. In some embodiments, Xylella fastidiosa symptoms may comprise scorching, discoloration, stunting, wilting, premature leaf abscission, shriveled fruit, premature fruit abscission, dieback, and plant death. In some embodiments, the symptom level is measured based on one or more Xylella fastidiosa symptoms affecting canopy of the olive tree. In some embodiments, the symptom level is measured based on the percentage of the canopy affected by one or more Xylella fastidiosa symptoms.

[0064] In some embodiments, the method comprises injecting the olive tree with an injection formulation described herein.

[0065] In some embodiments, when the trunk circumference is less than Cl, or the trunk diameter is less than DI, or both, the injection formulation comprises an initial concentration or low injection concentration of the zinc, an initial concentration or low injection concentration of the copper, or an initial concentration or low injection concentration of a stock formulation or a commercially available formulation.

[0066] In some embodiments, when (i) the trunk circumference is between Cl and C2, or the trunk diameter is between DI and D2, or both, and (ii) the symptom level of the infected tree is less than or equal to S2, the injection formulation comprises an initial concentration or low injection concentration of the zinc, an initial concentration or low injection concentration of the copper, or an initial concentration or low injection concentration of a stock formulation or a commercially available formulation.

[0067] In some embodiments, when (i) the trunk circumference is between Cl and C2, or the trunk diameter is between DI and D2, or both, and (ii) the symptom level of the infected tree is greater than S2, the injection formulation comprises a medium injection concentration of the zinc, a medium injection concentration of the copper, or a medium injection concentration of a stock formulation or a commercially available formulation. [0068] In some embodiments, when (i) the trunk circumference is greater than C2, or the trunk diameter is greater than D2, or both, and (ii) the symptom level of the infected tree is less than or equal to SI, the injection formulation comprises an initial concentration or low injection concentration of the zinc, an initial concentration or low injection concentration of the copper, or an initial concentration or low injection concentration of a stock formulation or a commercially available formulation.

[0069] In some embodiments, when (i) the trunk circumference is between C2 and C3, or the trunk diameter is between D2 and D3, or both, and (ii) the symptom level of the infected tree is less than or equal to SI, the injection formulation comprises an initial concentration or low injection concentration of the zinc, an initial concentration or low injection concentration of the copper, or an initial concentration or low injection concentration of a stock formulation or a commercially available formulation.

[0070] In some embodiments, when (i) the trunk circumference is greater than C2, or the trunk diameter is greater than D2, or both, and (ii) the symptom level of the infected tree is between SI and S2, the injection formulation comprises a medium injection concentration of the zinc, a medium injection concentration of the copper, or a medium injection concentration of a stock formulation or a commercially available formulation.

[0071] In some embodiments, when (i) the trunk circumference is between C2 and C3, or the trunk diameter is between D2 and D3, or both, and (ii) the symptom level of the infected tree is between SI and S2, the injection formulation comprises a medium injection concentration of the zinc, a medium injection concentration of the copper, or a medium injection concentration of a stock formulation or a commercially available formulation.

[0072] In some embodiments, when (i) the trunk circumference is greater than C2, or the trunk diameter is greater than D2, or both, and (ii) the symptom level of the infected tree is greater than S2, the injection formulation comprises a high injection concentration of the zinc, a high injection concentration of the copper, or a high injection concentration of a stock formulation or a commercially available formulation.

[0073] In some embodiments, when (i) the trunk circumference is between C2 and C3, or the trunk diameter is between D2 and D3, or both, and (ii) the symptom level of the infected tree is greater than S2, the injection formulation comprises a high injection concentration of the zinc, a high injection concentration of the copper, or a high injection concentration of a stock formulation or a commercially available formulation.

[0074] In some embodiments, when the trunk circumference is greater than C3, or the trunk diameter is greater than D3, or both, the tree is not injected with an injection formulation for treating Xylella fastidiosa infection.

[0075] In some embodiments, Cl is about 140 cm. In some embodiments, C2 is about 200 cm. In some embodiments, C3 is about 300 cm. In some embodiments, DI is about 45 cm. In some embodiments, D2 is about 65 cm. In some embodiments, D2 is about 95 cm. In some embodiments, SI is about 20%. In some embodiments, S2 is about 50%. In some embodiments, Cl is about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, or 400 cm. In some embodiments, C2 is about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, or 400 cm. In some embodiments, C3 is about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, or 400 cm. In some embodiments, C2 is greater than Cl. In some embodiments, C3 is greater than C2. In some embodiments, DI is about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, or 145 cm. In some embodiments, D2 is about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, or 145 cm. In some embodiments, D3 is about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, or 145 cm. In some embodiments, D2 is greater than DI. In some embodiments, D3 is greater than D2. In some embodiments, SI is about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, and S2 is about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, wherein S2 is greater than S 1.

[0076] In some embodiments, the olive tree presents an irregular structure, such as complex structure, significant trunk splitting, or significant trunk cavities. In some embodiments, the method comprises determining whether the olive tree presents a significant irregular structure. In some embodiments, when the olive tree is determined to have a significant irregular structure, the methods provided herein are not applicable. In some embodiments, when the olive tree is determined to have a significant irregular structure, the injection formulation or concentration is selected independent of the trunk circumference or the trunk diameter. In some embodiments, when the olive tree is determined to have a significant irregular structure, the injection formulation is selected based on the symptom level. In some embodiments, when (i) the olive tree presents a significant irregular structure and (ii) the symptom level of the tree is less than SI, the injection formulation comprises the initial concentration of zinc, copper, stock formulation, or commercially available formulation. In some embodiments, when (i) the olive tree presents a significant irregular structure and (ii) the symptom level of the tree is between SI and S2, the injection formulation comprises the medium injection concentration of zinc, copper, stock formulation, or commercially available formulation. In some embodiments, when (i) the olive tree presents a significant irregular structure and (ii) the symptom level of the tree is greater than S2, the injection formulation comprises the high injection concentration of zinc, copper, stock formulation, or commercially available formulation.

[0077] FIG. 1 provides an exemplary dosing matrix where Cl is 140 cm, C2 is 200 cm, C3 is 300 cm, DI is 45 cm, D2 is 65 cm, D3 is 95 cm, SI is 20%, and S2 is 50%, where “1” in the matrix corresponds to 5% Dentamet® (i.e., 20-fold dilution) in the injection formulation, “2” in the matrix corresponds to 10% Dentamet® (i.e., 10-fold dilution) in the injection formulation, and “3” in the matrix corresponds to 20% Dentamet ® (i.e., 5-fold dilution) in the injection formulation.

[0078] Dosing with the strategy provided in the dosing matrix of FIG. 1 or the method provided herein can (i) reduce or minimize phytotoxicity, (ii) reduce or minimize the time required to reach the residue limit (e.g., for copper or zinc) from last treatment or injection, and/or (iii) increase or maximize control of the Xylella fastidiosa infection.

[0079] In some embodiments, the initial concentration or low injection concentration of zinc is between about 0.08% and about 0.23%. In some embodiments, the initial concentration or low injection concentration of copper is between about 0.015% and about 0.035%. In some embodiments, the initial concentration or low injection concentration of copper is about 0.010%, 0.015%, 0.020%, 0.025%, 0.030%, 0.035%, 0.040%, 0.045%, 0.050%, 0.055%, 0.060%, 0.065%, 0.070%, 0.075%, 0.080%, 0.085%, 0.090%, 0.095%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, or 5%. In some embodiments, the initial concentration or low injection concentration of zinc is about

0.010%, 0.015%, 0.020%, 0.025%, 0.030%, 0.035%, 0.040%, 0.045%, 0.050%, 0.055%,

0.060%, 0.065%, 0.070%, 0.075%, 0.080%, 0.085%, 0.090%, 0.095%, 0.1%, 0.2%, 0.3%,

0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, or 5%.

[0080] In some embodiments, the initial concentration or low injection concentration of a stock formulation or a commercially available formulation is about 5%, meaning that the stock formulation or the commercially available formulation is diluted 20-fold for the injection formulation. In some embodiments, the initial concentration or low injection concentration of a stock formulation or a commercially available formulation is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.

[0081] In some embodiments, the medium injection concentration is about twice the initial concentration or low injection concentration. In some embodiments, the medium injection concentration is about 1.5 to 2.5 times the initial concentration or low injection concentration. In some embodiments, the medium injection concentration is higher than the initial concentration or low injection concentration. In some embodiments, the medium injection concentration is about 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 times higher than the initial concentration or low injection concentration. In some embodiments, the medium injection concentration of zinc or copper is about 0.010%, 0.015%, 0.020%, 0.025%, 0.030%, 0.035%, 0.040%, 0.045%, 0.050%, 0.055%, 0.060%, 0.065%, 0.070%, 0.075%, 0.080%, 0.085%, 0.090%, 0.095%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25%. In some embodiments, the medium injection concentration of a stock formulation or a commercially available formulation is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0082] In some embodiments, the high injection concentration is about twice the medium injection concentration. In some embodiments, the high injection concentration is about 1.5 to 2.5 times the medium injection concentration. In some embodiments, the high injection concentration is about four times the initial concentration or low injection concentration. In some embodiments, the high injection concentration is about 3.5 to 4.5 times the initial concentration or low injection concentration. In some embodiments, the high injection concentration is higher than the medium injection concentration. In some embodiments, the high injection concentration is about 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 times higher than the medium injection concentration. In some embodiments, the high injection concentration of zinc or copper is about 0.010%, 0.015%, 0.020%, 0.025%, 0.030%, 0.035%, 0.040%, 0.045%, 0.050%, 0.055%, 0.060%, 0.065%, 0.070%, 0.075%, 0.080%, 0.085%, 0.090%, 0.095%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the high injection concentration of a stock formulation or a commercially available formulation is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0083] In some embodiments, the method comprises assigning a trunk measurement range selected from a set of reference trunk measurement ranges based on the circumference, diameter, or any physical properties of the trunk. In some embodiments, the set of reference trunk measurement ranges comprises one or more reference trunk measurement ranges. In some embodiments, the set of reference trunk measurement ranges comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more reference trunk measurement ranges. In some embodiments, the reference trunk measurement ranges in the set are nonoverlapping. In some embodiments, the trunk measurement range is assigned such that the circumference or the diameter of the tree trunk falls within the trunk measurement range. For example, if the reference trunk measurement ranges comprise less than 140 cm, between 140 cm and 200 cm (inclusive), and greater than 200 cm with respect to the trunk circumference, then an olive tree with a tree trunk circumference of 120 cm (e.g., measured at a certain reference height from the ground) would be assigned “less than 140 cm” as the trunk measurement range.

[0084] In some embodiments, the set of reference trunk measurement ranges comprises a low trunk measurement range. In some embodiments, the set of reference trunk measurement ranges comprises a medium trunk measurement range. In some embodiments, the set of reference trunk measurement ranges comprises a high trunk measurement range. In some embodiments, the set of reference trunk measurement ranges comprises an excess trunk measurement range. In some embodiments, the low trunk measurement range comprises a trunk circumference range of less than Cl, or a trunk diameter of less than DI, or both. In some embodiments, the medium trunk measurement range comprises a trunk circumference range between Cl and C2, or a trunk diameter between DI and D2, or both. In some embodiments, the high trunk measurement range comprises a trunk circumference range of greater than C2, or a trunk diameter of greater than D2, or both. In some embodiments, the high trunk measurement range comprises a trunk circumference range of between C2 and C3, or a trunk diameter of between D2 and D3, or both. In some embodiments, the excess trunk measurement range comprises a trunk circumference range of greater than C3, or a trunk diameter of greater than D3, or both.

[0085] In some embodiments, the set of reference trunk measurement ranges comprises mtm ranges separated by (mtm - 1) boundary dimensions selected from any suitable dimensions, e.g., about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, and 300 cm. For example, when the set of reference trunk measurement ranges comprises 3 circumference ranges separated by about 140 cm and about 200 cm, the first reference trunk measurement range is less than about 140 cm, the second reference trunk measurement range is between about 140 cm and about 200 cm, and the third trunk measurement range is greater than about 200 cm.

[0086] In some embodiments, the method comprises assigning a symptom level range selected from a set of reference symptom level ranges based on one or more Xylella fastidiosa symptoms. In some embodiments, the set of reference symptom level ranges comprises one or more reference symptom level ranges. In some embodiments, the set of reference symptom level ranges comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more reference symptom level ranges. In some embodiments, the reference symptom level ranges in the set are non-overlapping. In some embodiments, the symptom level range is assigned such that the symptom level of the tree falls within the symptom level range. For example, if the reference symptom level ranges comprise less than 20%, between 20% and 50% (inclusive), and greater than 50%, then an olive tree with a symptom level of 10% (percent canopy affected by one or more Xylella fastidiosa symptoms) would be assigned “less than 20%” as the symptom level range.

[0087] In some embodiments, the set of reference symptom level ranges comprises a low symptom level range. In some embodiments, the set of reference symptom level ranges comprises a medium symptom level range. In some embodiments, the set of reference symptom level ranges comprises a high symptom level range. In some embodiments, an infected tree having a low symptom level range exhibits less than SI symptom level. In some embodiments, an infected tree having a medium symptom level range exhibits between SI and S2 symptom level. In some embodiments, an infected tree having a high symptom level range exhibits greater than S2 symptom level.

[0088] In some embodiments, the set of reference symptom level ranges comprises m_si ranges separated by (m_si - 1) boundary symptom levels selected from any suitable symptom levels, e.g., about 0%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, and 100%. For example, when the set of reference symptom level ranges comprises 3 symptom level ranges separated by about 20% and about 50%, the first reference symptom level range is less than about 20%, the second reference symptom level range is between about 20% and about 50%, and the third symptom level range is greater than about 50%.

[0089] In some embodiments, the method comprises selecting an injection concentration from a set of reference injection concentrations or selecting an injection concentration range from a set of reference injection concentration ranges based on the assigned trunk measurement range and/or the symptom level range. In some embodiments, the set of reference injection concentrations comprises one or more reference injection concentrations. In some embodiments, the set of reference concentrations comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more reference injection concentrations. In some embodiments, the set of reference injection concentration ranges comprises one or more reference injection concentration ranges. In some embodiments, the set of reference concentration ranges comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more reference injection concentration ranges. In some embodiments, the reference injection concentration ranges in the set are non-overlapping. In some embodiments, a reference injection concentration ranges in a set may or may not overlap with other reference injection concentration ranges.

[0090] In some embodiments, a reference concentration is about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,

75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%. For example, if there are 3 reference concentrations in a set, they may be 5%, 10%, and 20%, respectively.

[0091] In some embodiments, a reference concentration range has a lower limit of about

0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%,

0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%,

1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.5%, 4%,

4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%,

15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,

75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, a reference concentration range has an upper limit of about 0.01%, 0.02%, 0.03%, 0.04%,

0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%,

0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%,

2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%,

8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%

30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%,

98%, 99%, or 100%. In some embodiments, a reference concentration range has a lower limit of about 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%,

0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%,

1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.5%,

4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%,

14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,

70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%, and an independently selected upper limit of about 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%,

0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%

1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%,

2.8%, 2.9%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%,

10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%

50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%, where the lower limit is less than the upper limit. For example, if there are 3 reference concentration ranges in a set, they may be between 0.1% to 0.25%, between 0.2% and 0.5%, and between 0.3% and 1%, respectively. In some embodiments, these ranges may be overlapping as in this example. In some embodiments, only some of the ranges may be overlapping. In some embodiments, none of the ranges may be overlapping.

[0092] In some embodiments, the reference injection concentrations comprise a low injection concentration, a medium injection concentration, and a high injection concentration as described herein. In some embodiments, the low injection concentration is about 0.08% to about 0.23%. In some embodiments, the medium injection concentration is about 0.16% to about 0.46%. In some embodiments, the high injection concentration is about 0.32% to about 0.92%. In some embodiments, the low injection concentration is about 5%. In some embodiments, the medium injection concentration is about 10%. In some embodiments, the high injection concentration is about 20%.

[0093] In some embodiments, the set of reference injection concentration ranges comprise a low injection concentration range, a medium injection concentration range, and a high injection concentration range. In some embodiments, the low injection concentration range is about 0.08% to about 0.23%. In some embodiments, the medium injection concentration range is about 0.16% to about 0.46%. In some embodiments, the high injection concentration range is about 0.32% to about 0.92%.

[0094] In some embodiments, a reference injection concentration range is about ±0.001%, ±0.002%, ±0.003%, ±0.004%, ±0.005%, ±0.01%, ±0.02%, ±0.03%, ±0.04%, ±0.05%, ±0.1%, ±0.15%, ±0.2%, ±0.25%, ±0.3%, ±0.35%, ±0.4%, ±0.45%, ±0.5%, ±0.55%, ±0.6%, ±0.65%, ±0.7%, ±0.75%, ±1%, ±2%, ±3%, ±4%, ±5%, ±6%, ±7%, ±8%, ±9%, ±10%, ±15%, ±20%, ±25%, ±30%, ±35%, or ±40% with respect to a reference injection concentration described herein. For example, when 5% can be selected or recommended as an injection concentration, 5%±2% can be selected or recommended as an injection concentration range.

[0095] In some embodiments, the lower limit and/or the upper limit of the medium injection concentration range is about twice the lower limit and/or the upper limit of the low injection concentration range. In some embodiments, the lower limit and/or the upper limit of the high injection concentration range is about twice the lower limit and/or the upper limit of the medium injection concentration range. In some embodiments, the lower limit and/or the upper limit of the medium injection concentration range is about 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 times higher than the lower limit and/or the upper limit of the low injection concentration range. In some embodiments, the lower limit and/or the upper limit of the high injection concentration range is about 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 times higher than the lower limit and/or the upper limit of the medium injection concentration range.

[0096] In some embodiments, the method comprises applying an injection formulation to the olive tree. In some embodiments, the injection formulation comprises zinc, wherein zinc is present in the injection formulation at the selected injection concentration or within the selected injection concentration range. In some embodiments, the method comprises applying an injection formulation comprising copper to the olive tree, wherein copper is present in the injection formulation at the selected injection concentration or within the selected injection concentration range. In some embodiments, the method comprises applying an injection formulation to the olive tree, wherein the injection formulation comprises a stock formulation or a commercially available formulation at the selected injection concentration or within the selected injection concentration range. In some embodiments, the concentration can be selected from any of the concentrations or concentration ranges provided herein.

[0097] In some embodiments, a low injection concentration is selected when a low trunk measurement range is assigned. In some embodiments, a low injection concentration is selected when a medium trunk measurement range is assigned and a low or medium symptom level range is assigned. In some embodiments, a low injection concentration is selected when a high trunk measurement range is assigned and a low symptom level range is assigned. In some embodiments, a low injection concentration is selected when an excess trunk measurement range is assigned and a low symptom level range is assigned.

[0098] In some embodiments, a medium injection concentration is selected when a medium trunk measurement range is assigned and a high symptom level range is assigned. In some embodiments, a medium injection concentration is selected when a high trunk measurement range is assigned and a medium symptom level is assigned. In some embodiments, a medium injection concentration is selected when an excess trunk measurement range is assigned and a medium symptom level is assigned.

[0099] In some embodiments, a high injection concentration is selected when a high trunk measurement range is assigned and a high symptom level range is assigned. In some embodiments, a high injection concentration is selected when an excess trunk measurement range is assigned and a high symptom level range is assigned.

[0100] In some embodiments, a low injection concentration range is selected when a low trunk measurement range is assigned. In some embodiments, a low injection concentration range is selected when a medium trunk measurement range is assigned and a low or medium symptom level range is assigned. In some embodiments, a low injection concentration range is selected when a high trunk measurement range is assigned and a low symptom level range is assigned. In some embodiments, a low injection concentration range is selected when an excess trunk measurement range is assigned and a low symptom level range is assigned.

[0101] In some embodiments, a medium injection concentration range is selected when (i) a medium trunk measurement range is assigned and a high symptom level range is assigned. In some embodiments, a medium injection concentration range is selected when a high trunk measurement range is assigned and a medium symptom level is assigned. In some embodiments, a medium injection concentration range is selected when an excess trunk measurement range is assigned and a medium symptom level is assigned.

[0102] In some embodiments, a high injection concentration range is selected when a high trunk measurement range is assigned and a high symptom level range is assigned. In some embodiments, a high injection concentration range is selected when an excess trunk measurement range is assigned and a high symptom level range is assigned.

[0103] In some embodiments, when the olive tree presents an irregular structure, trunk measurement does not affect selection of injection concentration or injection concentration range. In some embodiments, a low injection concentration or a low injection concentration range is selected when (i) the olive tree presents an irregular structure and (ii) a low symptom level range is assigned. In some embodiments, a medium injection concentration or a medium injection concentration range is selected when (i) the olive tree presents an irregular structure and (ii) a medium symptom level range is assigned. In some embodiments, a high injection concentration or a high injection concentration range is selected when (i) the olive tree presents an irregular structure and (ii) a high symptom level range is assigned.

[0104] In some embodiments, no injection concentration or injection concentration range is selected when an excess trunk measurement range is assigned or when the tree presents a significant irregular structure. Injection programs

[0105] In some embodiments, injecting the injection formulation or any of the methods described herein are performed 4 times a year. In some embodiments, injecting the injection formulation or any of the methods described herein are performed 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times a year. In some embodiments, injecting the injection formulation or any of the methods described herein are performed more than 1 time, more than 2 times, more than 3 times, more than 4 times, more than 5 times, more than 6 times, more than 7 times, more than 8 times, more than 9 times, or more than 10 times a year. In some embodiments, injecting the injection formulation or any of the methods described herein are performed less than 2 times, less than 3 times, less than 4 times, less than 5 times, less than 6 times, less than 7 times, less than 8 times, less than 9 times, or less than 10 times a year.

[0106] In some embodiments, injecting the injection formulation or any of the methods described herein are performed around March, around May or June, around September, and around October. In some embodiments, injecting the injection formulation or any of the methods described herein are performed around January, around February, around March, around April, around May, around June, around July, around August, around September, around October, around November, around December, or any combination thereof.

[0107] In some embodiments, injecting the injection formulation or any of the methods described herein are performed about 7 weeks before flowering. In some embodiments, injecting the injection formulation or any of the methods described herein are performed about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks before flowering. In some embodiments, about 5, 6, 7, 8, or 9 weeks before flowering is around March. In some embodiments, about 5, 6, 7, 8, or 9 weeks before flowering is around early March.

[0108] In some embodiments, injecting the injection formulation or any of the methods described herein are performed after fruit set. In some embodiments, injecting the injection formulation or any of the methods described herein are performed about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days after fruit set. In some embodiments, about 1, 2, 3, 4, 5, 6, 7, 8,

9, 10, 11, 12, 13, or 14 days after fruit set is around May or around June. 1, 2, 3, 4, 5, 6, 7, 8,

9, 10, 11, 12, 13, or 14 days after fruit set is around late May or around early June. [0109] In some embodiments, injecting the injection formulation or any of the methods described herein are performed post-summer. In some embodiments, post-summer is around September. In some embodiments, post-summer is around early September.

[0110] In some embodiments, injecting the injection formulation or any of the methods described herein are performed at the end of the season. In some embodiments, end of the season is around October. In some embodiments, end of the season is around mid-to-end of October.

[oni] In some embodiments, the method comprises injecting the olive tree with a low- dose injection formulation. In some embodiments, the low-dose injection formulation comprises zinc and copper. In some embodiments, zinc or copper concentration of the low- dose injection formulation is about two thirds the initial concentration or the low injection concentration. In some embodiments, the low-dose injection formulation comprises about 3.3% Dentamet®.

[0112] In some embodiments, injection during the reproductive phase before flowering is avoided or performed with a low-dose injection formulation. In some embodiments, the method comprises injecting a low-dose injection formulation to the olive tree about 3 weeks before flowering. In some embodiments, the method comprises injecting a low-dose injection formulation to the olive tree about 1, 2, 3, 4, or 5 weeks before flowering. In some embodiments, about 1, 2, 3, 4, or 5 weeks before flowering is around April. In some embodiments, about 3 weeks before flowering is around early April.

[0113] In some embodiments, injection during seasons of high drought or heat stress is avoided or performed with a low-dose injection formulation. In some embodiments, the method comprises injecting a low-dose injection formulation to the olive tree in the presummer season. In some embodiments, the pre-summer season is around July. In some embodiments, the pre-summer season is around mid- July.

[0114] In some embodiments, injection close to harvest is avoided. In some embodiments, last annual injection is performed at least about 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or 10 weeks before harvesting.

[0115] FIG. 2 provides an exemplary injection schedule for injection formulations and low-dose injection formulations described herein. This exemplary injection schedule lays out total six injections, two of which (indicated as “Treatment A”) are for injection of a low-dose injection formulation, and the other four are for regular injection of injection formulation according to the methods described herein.

Olive Tree & Diseases

[0116] In some embodiments, the olive tree is suffering from olive quick decline syndrome. In some embodiments, the Xylella fastidiosa is Xylella fastidiosa subsp.

In some embodiments, the injection formulation is distributed throughout the trunk and other parts of the olive tree. In some embodiments, the other parts of the olive tree comprise fruits. In some embodiments, the other parts of the olive tree comprise leaves. In some embodiments, the olive tree is at least about 30 years old. In some embodiments, the olive tree is at least about 100 years old. In some embodiments, the olive tree is at least about 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, or 100 years old.

[0117] In some embodiments, this disclosure provides methods for enhancing or maintaining plant health, such as in olive trees. In some such embodiments, this disclosure provides methods for treating diseased plants and/or methods for controlling bacteria, fungi, viruses and/or other pathogens which cause disease in plants, such as in olive trees. In further such embodiments, this disclosure provides methods for treating olive trees whose xylem has been invaded by disease-causing bacteria, fungi, viruses, and/or other pathogens, for controlling the bacteria, fungi, virus and/or other pathogens causing the disease, and for preventing diseases by preventing sufficient colonization of the tree by the disease causing pathogens such as bacteria, fungi, and viruses.

[0118] In some embodiments, this disclosure provides methods of treating olive quick decline syndrome (“OQDS”). In some embodiments, the disclosure provides methods for controlling Xylella fastidiosa, which is a xylem-limited plant bacteria thought to cause the referenced disease. In certain embodiments, the disclosure provides methods for enhancing or maintaining the health of olive trees. In some embodiments, the disclosure provides methods for treating OQDS in olive trees. In some variations, the disclosure provides methods for controlling Xylella fastidiosa subsp. pauca in olive trees. In other variations, the disclosure provides methods for controlling Xylella fastidiosa subsp. multiplex in olive trees.

[0119] In some embodiments, controlling Xylella fastidiosa in olive trees using the systems, devices and methods herein includes reducing the bacterial concentration (titer) in the vascular system. In some variations, controlling Xylella fastidiosa in olive trees using the systems, devices and methods herein includes reducing the bacterial concentration (titer) in the vascular system by strengthening the tree’s natural defense system. In certain embodiments, the systems, devices and methods herein can provide a treatment that leads to suppression of the disease to a level where recovery of olive production occurs. In some variations, bacterial titer refers to the bacterial concentration in the vascular system of the infected tree. Bacterial titer may be measured using any suitable methods and techniques known in the art. For example, in one variation, bacterial titer is measured through quantitative PCR.

[0120] In some embodiments, recovery of olive trees infected by Xylella fastidiosa includes trees that show new green shoots and/or sprouts after treatment. In certain embodiments, the treatment may lead to growth of new green shoots and/or sprouts, which in turn may lead to new branches with green leaves. In some variations of the foregoing, controlling Xylella fastidiosa in olive trees using the systems, devices and methods herein results in an overall greener canopy with increased photosynthetic capacity. In certain variations, phenotypic observations may include estimating of vegetative index and photographic documentation of re-growth.

[0121] In other embodiments, controlling Xylella fastidiosa in olive trees using the systems, devices and methods herein includes at least partially or fully restoring xylem functionality of the infected olive trees. In certain embodiments of the foregoing, this may restore the tree’s productive capacity and overall tree health including the metabolomic profile of the tree. In some variations, metabolomic profile of the tree may be used to measure the tree health.

[0122] In yet other embodiments, controlling Xylella fastidiosa in olive trees using the systems, devices and methods herein includes at least partially or fully restoring yield capacity. In some variations, yield over the tree lifecycle is increased as compared to untreated control trees.

[0123] In certain embodiments, the method comprises delivering a formulation comprising one or more nutrients into an olive tree. In certain embodiments the method comprises precision delivery (also referred to as “precision injection”) of a formulation into the olive tree. Precision delivery refers to delivering the formulation only or substantially only into a target location in the olive tree. For example, in some embodiments, the target location is the active vasculature of the tree. In certain embodiments, the method comprises injecting an injection formulation into and no further than the active vasculature of the tree. In some variations, the active vasculature of the tree is the xylem and/or the phloem. In one variation, the active vasculature is active xylem (such as sapstream) and phloem. In further embodiments, precision delivery involves delivering the formulation into the active vasculature of the olive tree while minimizing damage to the tree relative traditional forms of injection drilling systems. In yet other embodiments, precision delivery involves using a system that can be configured to deliver formulation into and no further than the active vasculature of a tree.

[0124] In certain embodiments, the method comprises injecting an injection formulation comprising Dentamet® (DiAgro) into an olive tree, for example into the active vasculature of the tree using precision delivery devices and systems, such as those referenced herein. In some such embodiments, the methods comprise precise injection of an injection formulation comprising Dentamet® into the tree. In certain embodiments, the methods comprise injecting an injection formulation, for example precise injection of an injection formulation, comprising Dentamet® into the tree, for example into the active vasculature of the tree prone to disease caused by Xylella fastidiosa.

[0125] In some further embodiments, the olive tree is suffering from olive quick decline syndrome. In some variations, the infected olive tree has an infection level of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%; or between 10% and 90%, between 20% and 80%, between 25% and 75%, between 30% and 60%, or between 40% and 50%; or about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, or about 80%.

[0126] In other embodiments, the methods provided include injecting olive trees with no or low symptoms, including olive trees with less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%, or between 0% and 5% infection level. [0127] In certain variations of the foregoing, “infection level” of an olive tree is characterized by percent canopy loss. In one variation, the level of infection is characterized by the amount of canopy dried out and/or level of infected dried out canopy pruned away.

[0128] In certain embodiments, the olive tree is at least about 30 years old, at least about 50 years old, at least about 100 years old, at least about 150 years old, or at least about 250 years old.

[0129] In some embodiments, this disclosure also provides systems and devices for delivering injection formulations to the interior of the tree. In some embodiments, the systems comprise an injection tool operatively connected to a fluid delivery unit, wherein the injection tool is configured for precision delivery of the injection formulation to a target location inside the tree. In some embodiments, the systems are configured for precision delivery of an injection formulation into the active vasculature of an olive tree. In some embodiments, the fluid delivery unit further comprises the formulation. In other embodiments, the system comprises an injection tool, a fluid delivery unit, and a source of source of formulation in fluid communication with the fluid delivery unit.

[0130] In some embodiments, Xylella fastidiosa symptoms comprise scorching, otherwise known as leaf scorching, leaf bum, or sun scorch. In some embodiments, scorching comprises browning of the plant tissue. In some embodiments, scorching comprises darkening of the Xylem vessels of the plant. In some embodiments, Xylella fastidiosa symptoms comprise discoloration or leaf discoloration. In some embodiments, discoloration comprises turning of leaves to yellow or brownish color. In some embodiments, Xylella fastidiosa symptoms comprise stunting. In some embodiments, stunting comprises dwarfing of the plant. In some embodiments, Xylella fastidiosa symptoms comprise wilting. In some embodiments, Xylella fastidiosa symptoms comprise premature leaf abscission. In some embodiments, premature leaf abscission leads to bare petioles. In some embodiments, premature leaf abscission comprises premature dropping of leaves. In some embodiments, Xylella fastidiosa symptoms comprise poor or shriveled fruit. In some embodiments, Xylella fastidiosa symptoms comprise premature fruit abscission. In some embodiments, premature fruit abscission comprises premature dropping of the fruit. In some embodiments, Xylella fastidiosa symptoms comprise dieback. In some embodiments, dieback comprises dying of the plant branches from the tip inward. In some embodiments, Xylella fastidiosa symptoms comprise plant death. [0131] In some embodiments, the plant is an olive tree suffering from one or more of olive quick decline syndrome, olive leaf/peacock spot, anthracnose, Cercospora leaf spot, olive knot/tuberculosis, and Verticillium wilt. In certain embodiments, provided are methods for controlling Colletotricum spp. , Pseudocercospora cladosporioides, Xylella fastidiosa (including, e.g., Xylella fastidiosa supsp. pauca), Spilocea olivine/Cycloconium oleaginum, Pseudomonas savastanoi (including, e.g., Pseudomonas savastanoi pv. Savastanoi) and Verticlium species (including V. dahliae, V. albo-atrum, V. longisporum, V. nubilum, V. theobromae and V. tricorpus) in olive trees.

[0132] In some variations, provided are methods for treating one or more of: Olive Quick Decline Syndrome, Olive leaf/Peacock spot, Olive knot/tuberculosis, and Anthracnose in olive trees. In some variations, provided are methods for controlling one or more of: Xylella fastidiosa, Spilocea olivine, Pseudomonas savastanoi, and Colletotricum spp. in olive trees.

In certain variations of the foregoing, the methods comprise injecting into a plant an injection formulation comprising zinc and copper, such as Dentamet® (DiAgro).

[0133] In certain embodiments, the methods for controlling the pathogens described herein may include any one or more of the following: reducing bacterial titer (concentration) of the pathogen; inducing and/or triggering the plant-based defense mechanisms; xylem functional recovery; recovery of metabolomic profile to pre-infection state in the plant; growth of crops; reducing crop (e.g., fruit) drop; increasing yield and/or restablishing productivity; inducing new sprouting to re-establish green leaf canopy; preventing and/ or delaying infection and symptoms; and recovery of plant health.

Injection System

[0134] In some embodiments, the injecting of the injection formulation is performed using an injection system comprising an injection tool operatively connected to a fluid delivery unit, wherein the fluid delivery unit is configured to deliver the injection formulation. In some embodiments, the injecting of the injection formulation comprises piercing the trunk of the olive tree using the injection tool of the injection system. In some embodiments, the injecting of the injection formulation comprises delivering at least a portion of the injection formulation from the fluid delivery unit through the injection tool into and no further than the active vasculature of the olive tree. [0135] In some embodiments, an injection system is used to deliver the injection formulation to an olive tree. In some variations, the injection system comprises: an injection tool operatively connected to a fluid delivery unit. In certain variations, the injection tool comprises: a base having at least one inlet; and a body comprising at least one distribution reservoir, and at least one outlet. In some embodiments, the injection system comprises: an injection tool, a fluid delivery unit, and a source of active ingredient (including, for example, nutrients) formulated as a liquid.

[0136] In some variations, the body is shaped to pierce the tree, such as the trunk of the tree. In certain variations, the body is in the shape of a blade. In certain variations, the body has a cutting edge at the tip of the body, and the width of the cutting edge is narrower than width of the body in the area connected to the base.

[0137] In certain variations, the body comprises: at least one outlet that receives the injection formulation from the at least one inlet, and at least one distribution reservoir that retains the injection formulation proximate to adjacent tissue of the tree. In certain variations, the fluid delivery unit is configured to store and deliver the injection formulation. In certain variations, the fluid delivery unit comprises a pressurized container (e.g., a pressurized canister).

[0138] In some embodiments, the method comprises: piercing the trunk of an olive tree using the injection tool of the injection system; and delivering at least a portion of the injection formulation from the fluid delivery unit through the injection tool to the vasculature of the olive tree. In some variations, the injection formulation is delivered pneumatically or hydraulically.

[0139] In some embodiments, the injection formulation is precisely delivered. In some variations, the injection formulation is delivered into and no further than the active vasculature of the tree when the injection tool is inserted into the trunk of the tree. In one variation, the injection formulation is delivered into and no further than the xylem, or the phloem or both of the tree when the injection tool is inserted into the trunk of the tree.

[0140] In other embodiments, precisely delivering the injection formulation comprises inserting the injection tool into and no further than the active vasculature of the tree. In certain variations, precisely delivering the injection formulation comprises inserting the body of the injection tool into and no further than the active vasculature of the tree. In one variation, precisely delivering the injection formulation comprises inserting the injection tool such that the distribution reservoir is positioned in and no further than the active vasculature of the tree.

[0141] In some variations, the methods deliver at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% of the injection formulation into to the active vasculature of the tree. In one variation, the methods deliver at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% of the injection formulation into the xylem and/or phloem of the tree. In some variations of the foregoing, the methods deliver the injection formulation into to the active vasculature of the tree in an average maximum time of less than 10 minutes, or less than 5 minutes.

[0142] In certain embodiments, the method comprises injecting injection formulation into the vasculature through one or more sites on the trunk of the tree. In embodiments where the formulation is injected through multiple injection sites, a plurality of the injection systems described herein may be used. In some embodiments where the formulation is injected through multiple injection sites, the system comprises multiple injection tools operatively connected to a single fluid delivery unit.

[0143] In some variations, the method further comprises removing at least a portion of the bark around the injection site, e.g., prior to piercing the trunk.

[0144] The methods described herein generally provide one or more commercial advantages over the methods currently known in the art to treat Xylella fastidiosa. For example, advantages include one or more of a faster return to the production yields preinfection, fast response (e.g., curing), lower volumes of formulation needed, less loss of formulation to the environment, less damage to the tree, response in old trees including trees older than 100 years, response in trees with significant disease symptoms (e.g., with 50% or less remaining canopy foliage and faster administration to the trees).

[0145] In some variations, methods for enhancing plant health and methods for treating diseased plants comprises enhancing the health of older trees or treating diseased older trees. In some such embodiments, the trees are olive trees at least 100 years old or more. In other variations, methods for treating diseased plants comprises treating plants suffering from severe cases of a disease. In certain such variation, the plants are olive trees suffering from more severe cases of Xylella fastidiosa, such as olive trees with 50% or less canopy foliage.

[0146] In embodiments, the injection systems comprise an injection tool, a fluid delivery unit, and an injection formulation source. In operation, the injection tool is operatively connected to the fluid delivery unit such that injection formulation flows from the source through the injection tool into the plant. In some embodiments, the source of injection formulation is independent of the fluid delivery unit. In other embodiments, the source of injection formulation is integral with the fluid delivery unit. Certain embodiment of injection systems suitable for use in the methods described herein are described in further detail below.

Injection Tool

[0147] In some embodiments, the injection tool includes a body, at least a portion of which is designed to be lodged into the trunk of a tree. The body has a channel system (e.g., having one or more channels) through which the injection formulation can flow. In some variations, the liquid formation enters the injection tool through one or more inlets, and exits the injection tool through one or more outlets through which the injection formulation is delivered to the interior of the tree. In some embodiments, the lodged portion of the body is sized and shaped to reduce or minimize damage to the target tree when inserted into the tree, while maintaining efficient functionality of the injection tool in delivering the desired dosing of the injection formulation over the desired time period directly to the sapwood and not the heartwood of the trunk of the tree. In other embodiments, the lodged portion of the body is sized and shaped to reduce damage to the target tree when inserted into the tree, as compared to traditional drilling injection system.

[0148] In some variations, exemplary injection tools are depicted in FIGS. 3A-3C and 4A-4D. With reference to FIGS. 3A-3C, depicted is an exemplary injection tool 6001 having a base 6010 and a body 6020. The body 6020 includes a cutting element. For example, in one variation, the body 6020 includes a cutting edge along the front face 6021, directed distally away from the base 6010. In some variations, the outlets 6027 and distribution reservoirs 6028 are within the body 6020. As shown in the side view of FIG. 3C, the body 6020 increases in thickness from the distal portion 6041 toward the proximal portion 6043 and the base 6010. [0149] In some embodiments, the base 6010 optionally includes a ribbed outer structure

6012, such as attachment cleats, to facilitate grasping of the base 6010 and to securely connect the injection tool 6001 with a fluid delivery unit. In some variations, at the transition between the base 6010 and the body 6020, a step is provided. The step forms an abutting face

6013. The abutting face 6013 extends relative to (e.g., away from) the body 6020. During insertion of the injection tool 6001, the abutting face 6013 contacts the tree and arrests further advancement of the injection tool 6001 into the tree. In some variations, a larger abutting face 6013 facilitates use with smaller and less robust trees having a comparably soft shell or boundary. The relatively large abutting face distributes forces from insertion over the correspondingly large face 6013 and thereby minimizes trauma to the tree. In certain variations, the abutting face 6013 further provides an enclosing face for the injection tool 6001 for establishing a robust coupling with the tree.

[0150] As shown in FIGS. 3B and 3C, the base 6010 includes an inlet 6011, which receives the injection formulation from the fluid delivery unit. The injection formulation travels through a main channel 6025, and is released through the outlets 6027 into the distribution reservoirs 6028. As shown in FIGS. 3 A and 3B, outlets 6027 open transversely into the respective distribution reservoirs 6028.

[0151] The injection formulation is delivered from the outlets 6027 transversely, for instance relative to the longitudinal body axis 6040 and the corresponding insertion direction 6030, into the distribution reservoirs 6028. The distribution reservoirs 6028 retain the injection formulation in residence proximate to adjacent plant tissues. In the example shown in FIG. 3B, the outlets 6027 extend proximally toward the base 6010 and transverse relative to the insertion direction 6030 of the injection tool 6001.

[0152] Because of the relatively small profile of the injection tool 6001, the injection tool 6001 is readily inserted and installed in comparably small trees or less robust trees having a softer plant material (e.g., tissues or the like). For instance, the injection tool 6001 is configured for softened striking or manual pressing of the injection tool 6001 into the tree.

[0153] As shown in FIG. 3B, the injection tool 6001 is inserted along an insertion direction 6030 corresponding to the longitudinal body axis 6040 of the injection tool 6001. The body 6020 of the injection tool 6001 spreads the tree material aside as the injection tool 6001 is inserted into the tree. Spreading of the tree material minimizes trauma to the tree material, and in some examples facilitates enhanced uptake of formulations.

[0154] As further shown in FIG. 3B, the outlets 6027 extend in outlet direction 6032 toward the distribution reservoirs 6028. The outlet direction 6032 is transverse to the insertion direction 6030 (and the longitudinal body axis 6040). For example, in some variations, the outlet direction 6032 is misaligned with the insertion direction 6030 (and the longitudinal body axis 6040) with an angle of 125 degrees or the like. The transverse orientation of the outlets 6027 isolates the outlets 6027 from tree material otherwise introduced into the outlet channels with insertion. Further, the distribution reservoirs 6028 facilitate positioning of the outlets 6027 within the body profile, for instance recessing the outlets 6027 from an exterior of the body profile.

[0155] With reference to FIGS. 4A-4D, depicted is another exemplary injection tool 6101 having a base 6110 and a body 6120. The base 6110 and the body 6120 share similar features as described above for base 6010 and body 6120. For example, the body 6120 may include a cutting edge along the front face of the body. The body further includes outlets that distribute the injection formulation into distribution reservoirs 6128 within the body 6120.

[0156] However, unlike the injection tool 6001 depicted in FIGS. 3A-3C, injection tool 6101 has a base 6110 that includes two inlets 6111, which receives the injection formulation from the fluid delivery unit. The injection formulation travels through a main channel, and is released through the outlets into the distribution reservoirs 6128.

[0157] In some variations, the injection tool may have a plurality of inlets operatively connected to a fluid delivery unit. In certain variations, the injection tool has two, three or four inlets operatively connected to a fluid delivery unit.

[0158] With reference again to FIGS. 4 A, 4B and 4D, the base 6010 optionally includes a ribbed outer structure, such as attachment cleats, to facilitate grasping of the base 6010 and to securely connect the injection tool 6001 with a fluid delivery unit.

[0159] In some embodiments, the injection systems described herein comprising the exemplary injection tools depicted in the figures do not require drilling a hole or installing a valve in the trunk of the tree before injecting the injection formulation. Fluid Delivery Unit

[0160] In some embodiments, the fluid delivery unit and the source of the injection formulation are integrated into a formulation cartridge, such as a pressurized container. In certain variations, the formulation cartridge is a pressurized canister. In operation, the injection formulation flows from the fluid delivery unit through the injection tool into the tree. WO 2020/021041, which is hereby incorporated by reference, provides additional embodiments and variations of the injection systems and components thereof, including in the figures therein.

[0161] In some embodiments, the injection systems or components thereof used in the methods described herein are as depicted in the figures. In some embodiments, the systems are configured to administer injection formulation comprising one or more active ingredients (including, for example, nutrients) to a tree or a part thereof. In certain embodiments, such systems are mounted onto a post portion of a tree, for example to a trunk of the tree.

[0162] In some embodiments, the methods provided herein include installing an injection tool in the trunk, stem, root or limb of a tree, operatively connecting the injection tool to a fluid delivery unit, and activating the fluid delivery unit to initiate the flow of fluid from the fluid delivery unit through the injection tool and into the tree. In other embodiments, two or more injection tools are installed into one or more of the stem, trunk, roots, limbs or the like of a tree to minimize trauma to the tree (e.g., by minimizing the size of a unitary hole in the tree or spacing the tools apart along the tree). In some such embodiments, the two or more injection tools are operatively connected to the same fluid delivery unit. In other such embodiments the two or more injection tools are operatively connected to independent fluid delivery unit.

[0163] In some variations, the fluid delivery unit comprises a spring-loaded fluid delivery unit. In certain variations of the foregoing, the spring-loaded fluid delivery unit is configured to operate at a pressure between 1.5-3 bar. In other variations, the fluid delivery unit comprises a fluid delivery unit comprising a pressurized container (e.g., a pressurized canister). Examples of suitable fluid delivery unit include the variations depicted in FIGS. 5 and 6A.

[0164] With reference to FIG. 5, depicted is an exemplary spring-loaded fluid delivery unit 9900. Base 9912 holds two springs 9908 within syringes 9910. A piston with a rubber seal divides the injection formulation from the spring chamber. Attached to each syringe body 9910 is a tube 9904 connected to a t-shaped connector 9902. The injection tip (not depicted in FIG. 5) is connected to the connector 9902. The spring-loaded fluid delivery unit 9900 can be filled through connector 9902.

[0165] In other exemplary embodiments, the spring-loaded fluid delivery unit may have a base holding one or multiple springs within one or multiple corresponding syringes. The design of the spring-loaded fluid delivery unity may vary based on the pressure, volume, time or other appropriate parameters to deliver the injection formulation. For example, in some variations, multiple springs (such as a dual spring) may be employed in the fluid delivery unit to allow for injection of a higher volume of the injection formulation. In other variations, a single spring with a larger syringe may be used, but may affect pressure range employed to inject the injection formulation.

[0166] With reference to FIG. 6A, depicted is an exemplary chassis-style injection system comprising a system housing for integrating various components of the injection system, including an injection tool. Injection system 9800 includes a chassis 9802 that has a delivery interface connecting to the injection tool 9806. The delivery interface includes, but is not limited to, passages, channels, tubing, reservoirs or the like that interconnect the formulation cartridge (not depicted in FIG. 6 A) and the injection tool 9806. The delivery interface extends to the injection tool and fluidly communicates the formulation to the distribution reservoirs of the injection tool. A flange 9880 may engage the formulation cartridge, resulting in activating the cartridge and maintaining it in place. In some embodiments, the position of the flange is adjustable to accommodate different length canisters and/or to permit activation at a desired time. As further depicted in FIG. 6A, in some embodiments, at least a portion of chassis includes a flexible portion 9882, for example to mitigate damage to the tip during installation. In one variation, in the exemplary injection system depicted in FIG. 6A, the chassis 9802 can further include an anchor 9890 to further facilitate coupling with the tree. For instance, a belt, strap or the like may be passed through the anchor 9890 to hold the injection system 9800 in place along the tree.

[0167] With reference to FIG. 6B, a pressurized formulation cartridge 9810 is depicted. The exemplary cartridge 9810 includes a formulation container 9854 including a formulation reservoir therein having a quantity of the injection formulation. A cartridge cap 9856 encloses the formulation container 9854. A cartridge discharge port 9858 extends from the formulation cartridge 9810. Optionally, in certain variations, the cartridge discharge port includes an opening feature configured to transition from a closed configuration to an open configuration. The opening feature includes, but is not limited to, a valve, membrane or the like that is opened prior to coupling with the chassis of the injection system.

[0168] In some embodiments, once the injection tool is inserted into the trunk of the olive tree, the injection tool may remain untouched and in place over multiple re-injections. In certain embodiments, the method further comprises: replacing the fluid delivery unit with a second fluid delivery unit; and delivering at least a portion of the injection formulation from the second fluid delivery unit through the injection tool into and no further than the active vasculature of the olive tree. In some variations, the injection tool remains in the trunk of the olive tree over at least one growing season, at least two growing seasons, or at least three growing seasons.

Injection positions

[0169] Olive trees can have multiple shapes and sizes, depending on age, life history, and variety. Selecting the exact injection point(s) can be challenging. The goal is to increase the chances that the product reaches the whole canopy by placing the tip optimally. Even distribution of the product in the whole tree can reduce the chance of phytotoxicity events.

[0170] In some embodiments, when there are multiple branches from a main trunk, it is preferable to inject through the main trunk rather than through the branches, as illustrated by FIG. 7A. In this figure, an exemplary injection position with higher likelihood of efficient and/or even distribution of the injection formulation is indicated with a circle (O), and an exemplary injection position with lower likelihood of efficient and/or even distribution of the injection formulation is indicated with a cross (x).

[0171] In some embodiments, when there are multiple branches from a main trunk, it is preferable to inject through a part of the main trunk that is roughly equal distance from two or more branches, rather than through a part of the main trunk that is closer to a particular branch, as illustrated by FIG. 7B. In this figure, an exemplary injection position with higher likelihood of efficient and/or even distribution of the injection formulation is indicated with a circle (O), and an exemplary injection position with lower likelihood of efficient and/or even distribution of the injection formulation is indicated with a cross (x). [0172] In some embodiments, for more efficient and/or even distribution of the injection formulation, injection through damaged structures, for example, as indicated with a cross (x) in FIGS. 7C and 7D, should be avoided. Instead, for more efficient and/or even distribution of the injection formulation, injection through a part of the tree without such damages, for example, as indicated with a circle (O) in FIG. 7E, would be more appropriate.

[0173] In some embodiments, trunk circumference or trunk diameter is measured at or near the position where the injection formulation will be injected (i.e., injection point) or at or near a desirable injection point, for examples, the area indicated with a circle (O) in FIG. 7 A, 7B, or 7E. In some embodiments, the injection formulation is injected at or near the position where trunk circumference or trunk diameter was measured. In some embodiments, the reference height for measuring trunk circumference or trunk diameter is substantially the same as the height of the injection point.

[0174] In some embodiments, for more efficient and/or even distribution of the injection formulation, (i) injecting into damaged trunk parts should be avoided; (ii) injecting close to damaged trunk parts should be avoided; (iii) injection point should be equal distance from multiple branches, as much as possible; (iv) when a trunk is homogeneous and does not present significant splits or particular shapes, inject between about 40 cm to about 80 cm from the soil; and (v) when a tree has significant splits near or right from the soil or particular shapes or damages, inject into the side or spot that is directly linked to the main part of the canopy.

ENUMERATED EMBODIMENTS

[0175] The following enumerated embodiments are representative of some aspects of the invention.

1. A method for controlling Xylella fastidiosa infection of an olive tree, wherein the olive tree has a trunk, wherein the trunk has a circumference and a diameter, and wherein the infected olive tree exhibits at least one symptom caused by Xylella fastidiosa, the method comprising: a) measuring the trunk circumference or the trunk diameter, or both; b) determining a symptom level of the infected olive tree; and c) injecting the infected olive tree with an injection formulation comprising zinc and copper, wherein: when the trunk circumference is less than about 140 cm, or the trunk diameter is less than about 45 cm, or both, the injection formulation comprises an initial concentration of the zinc (Z 1); when (i) the trunk circumference is between about 140 cm and about 200 cm, or the trunk diameter is between about 45 cm and about 65 cm, or both, and (ii) the symptom level of the infected tree is less than or equal to about 50%, the injection formulation comprises the initial concentration of the zinc (Z 1 ); when (i) the trunk circumference is between about 140 cm and about 200 cm, or the trunk diameter is between about 45 cm and about 65 cm, or both, and (ii) the symptom level of the infected tree is greater than about 50%, the injection formulation comprises about twice the initial concentration of the zinc (1.5Z1-2.5Z1); when (i) the trunk circumference is greater than about 200 cm, or the trunk diameter is greater than about 65 cm, or both, and (ii) the symptom level of the infected tree is less than or equal to about 20%, the injection formulation comprises about the initial concentration of the zinc (Z 1); when (i) the trunk circumference is greater than about 200 cm, or the trunk diameter is greater than about 65 cm, or both, and (ii) the symptom level of the infected tree is between about 20% and about 50%, the injection formulation comprises about twice the initial concentration of the zinc (1.5Z1-2.5Z1); and when (i) the trunk circumference is greater than about 200 cm, or the trunk diameter is greater than about 65 cm, or both, and (ii) the symptom level of the infected tree is greater than about 50%, the injection formulation comprises about four times the initial concentration of the zinc (3.5Z1-4.5Z1).

2. The method of embodiment 1, wherein the initial concentration of the zinc is about 0.08% to about 0.23%.

3. The method of embodiment 1 or 2, wherein steps a)-c) are performed about 7 weeks before flowering, after fruit set, post-summer, and at the end of the season.

4. The method of embodiment 3, wherein about 7 weeks before flowering is around March, after fruit set is around May and June, post-summer is around September, and the end of the season is around October. 5. The method of any one of embodiments 1 to 4, further comprising injecting the infected olive tree with a low-dose injection formulation comprising about two thirds the initial concentration of the zinc, about 3 weeks before flowering and pre-summer season.

6. The method of embodiment 5, wherein about 3 weeks before flowering is around April and the pre-summer season is around July.

7. A method for controlling Xylella fastidiosa infection of an olive tree, wherein the olive tree has a trunk, and wherein the trunk has a circumference and a diameter, the method comprising: a) assigning a trunk measurement range selected from a set of reference trunk measurement ranges based on the circumference and/or diameter of the trunk; b) assigning a symptom level range selected from a set of reference symptom level ranges; c) selecting an injection concentration from a set of reference injection concentrations or selecting an injection concentration range from a set of reference injection concentration ranges based on the trunk measurement range and the symptom level range; and d) applying an injection formulation comprising zinc and copper to the infected olive tree, wherein zinc is present in the injection formulation at the selected injection concentration or within the selected injection concentration range.

8. The method of embodiment 7, wherein the set of reference trunk measurement ranges comprises a low trunk measurement range, a medium trunk measurement range, and a high trunk measurement range.

9. The method of embodiment 8, wherein: the low trunk measurement range comprises a trunk circumference range of less than about 140 cm, or a trunk diameter of less than about 45 cm, or both; the medium trunk measurement range comprises a trunk circumference range between about 140 cm and about 200 cm, or a trunk diameter between about 45 cm and about 65 cm, or both; and the high trunk measurement range comprises a trunk circumference range of greater than about 200 cm, or a trunk diameter of greater than about 65 cm, or both.

10. The method of any one of embodiments 7 to 9, wherein the set of reference symptom level ranges comprises a low symptom level range, a medium symptom level range, and a high symptom level range.

11. The method of embodiment 10, wherein: an infected tree having a low symptom level range exhibits less than about 20% symptom level; an infected tree having a medium symptom level range exhibits between about 20% and 50% symptom level; and an infected tree having a high symptom level range exhibits greater than 50% symptom level.

12. The method of embodiment 11, wherein the symptom level is measured based on one or more Xylella fastidiosa symptoms comprising scorching, discoloration, stunting, wilting, premature leaf abscission, shriveled fruit, premature fruit abscission, dieback, and plant death.

13. The method of embodiment 11 or 12, wherein the symptom level is measured based on one or more Xylella fastidiosa symptoms affecting canopy of the olive tree.

14. The method of any one of embodiments 7 to 13, wherein the reference injection concentrations comprise a low injection concentration, a medium injection concentration, and a high injection concentration.

15. The method of embodiment 14, wherein: the medium injection concentration is about twice the low injection concentration; and the high injection concentration is about four times the low injection concentration.

16. The method of embodiment 14 or 15, wherein the low injection concentration is about

0.08% to about 0.23%. 17. The method of embodiment 16, wherein: the medium injection concentration is about 0.16% to about 0.46%; and the high injection concentration is about 0.32% to about 0.92%.

18. The method of any one of embodiments 7 to 13, wherein the reference injection concentration ranges comprise a low injection concentration range, a medium injection concentration range, and a high injection concentration range.

19. The method of embodiment 18, wherein: the lower limit of the medium injection concentration range is about twice the lower limit of the low injection concentration range; and the lower limit of the high injection concentration range is about four times the lower limit of the low injection concentration.

20. The method of embodiment 18 or 19, wherein the low injection concentration range is about 0.08% to about 0.23%.

21. The method of embodiment 20, wherein: the medium injection concentration range is about 0.16% to about 0.46%; and the high injection concentration range is about 0.32% to about 0.92%.

22. The method of any one of embodiments 14 to 17, wherein: the low injection concentration is selected when (i) the low trunk measurement range is assigned, (ii) the medium trunk measurement range is assigned and the low or medium symptom level range is assigned, or (iii) the high trunk measurement range is assigned and the low symptom level range is assigned; the medium injection concentration is selected when (i) the medium trunk measurement range is assigned and the high symptom level range is assigned or (ii) the high trunk measurement range is assigned and the medium symptom level is assigned; and the high injection concentration is selected when the high trunk measurement range is assigned and the high symptom level range is assigned.

23. The method of any one of embodiments 18 to 21, wherein: the low injection concentration range is selected when (i) the low trunk measurement range is assigned, (ii) the medium trunk measurement range is assigned and the low or medium symptom level range is assigned, or (iii) the high trunk measurement range is assigned and the low symptom level range is assigned; the medium injection concentration range is selected when (i) the medium trunk measurement range is assigned and the high symptom level range is assigned or (ii) the high trunk measurement range is assigned and the medium symptom level is assigned; and the high injection concentration range is selected when the high trunk measurement range is assigned and the high symptom level range is assigned.

24. The method of any one of embodiments 7 to 23, wherein steps a)-d) are performed about 7 weeks before flowering, after fruit set, post-summer, and at the end of the season.

25. The method of any one of embodiments 14 to 17 and 22, further comprising injecting the infected olive tree with a low-dose injection formulation comprising zinc and copper about 3 weeks before flowering and pre-summer season, wherein the zinc concentration of the low-dose injection formulation is about two thirds the low injection concentration.

26. The method of any one of embodiments 18 to 21 and 23, further comprising injecting the infected olive tree with a low-dose injection formulation comprising zinc and copper about 3 weeks before flowering and pre-summer season, wherein the zinc concentration of the low-dose formulation is selected from a low-dose concentration range, wherein the lower limit of the low-dose concentration range is two thirds the lower limit of the low injection concentration range, and the upper limit of the low-dose concentration range is two thirds the upper limit of the low injection concentration range.

27. The method of embodiment 25 or 26, wherein about 3 weeks before flowering is around April and pre-summer season is around July. 28. The method of any one of embodiments 1 to 27, wherein the circumference and/or diameter of the trunk is measured at a reference height from the ground.

29. The method of embodiment 28, wherein the reference height is between about 40 cm and about 80 cm from the ground.

30. The method of any one of embodiments 1 to 29, wherein the injecting of the injection formulation is performed using an injection system comprising an injection tool operatively connected to a fluid delivery unit, wherein the fluid delivery unit is configured to deliver the injection formulation.

31. The method of embodiment 30, wherein the injecting of the injection formulation comprises: piercing the trunk of the olive tree using the injection tool of the injection system; and delivering at least a portion of the injection formulation from the fluid delivery unit through the injection tool into and no further than the active vasculature of the olive tree.

32. The method of any one of the preceding embodiments, wherein the olive tree is suffering from olive quick decline syndrome.

33. The method of any one of the preceding embodiments, wherein the Xylella fastidiosa is Xylella fastidiosa subsp. pauca.

34. The method of any one of the preceding embodiments, wherein the injection formulation is distributed throughout the trunk and other parts of the olive tree.

35. The method of embodiment 34, wherein the other parts of the olive tree comprise fruits.

36. The method of embodiment 34, wherein the other parts of the olive tree comprise leaves.

37. The method of any one of the preceding embodiments, wherein the olive tree is at least about 30 years old.

38. The method of any one of the preceding embodiments, wherein the olive tree is at least about 100 years old. 39. The method of any one of embodiments 30 to 38, wherein the delivery unit is a spring-loaded fluid delivery unit.

40. The method of any one of embodiments 30 to 39, wherein the delivery unit comprises a pressurized formulation cartridge.

41. The method of any one of embodiments 30 to 40, further comprising: replacing the fluid delivery unit with a second fluid delivery unit; and delivering at least a portion of the injection formulation from the second fluid delivery unit through the injection tool into and no further than the active vasculature of the olive tree.

42. The method of any one of embodiments 30 to 41, wherein the injection tool remains in the trunk of the olive tree over at least one growing season.

43. The method of any one of embodiments 30 to 42, wherein the injection tool remains in the trunk of the olive tree over multiple re-injections.

44. The method of any one of embodiments 31 to 43, wherein the trunk of the olive tree has bark, and the method further comprises: removing at least a portion of the bark prior to piercing the trunk.

EXAMPLES

[0176] The presently disclosed subject matter will be better understood by reference to the following Examples, which are provided as exemplary of the invention, and not by way of limitation.

Example 1: Efficacy Studies

[0177] FIGS. 8A to 8E, 9A, 9B, and 10A to 10C show delta DI scores plotted under various injection and treatment conditions.

Example 2: Residual Copper and Zinc Content in Olive Fruit

[0178] FIGS. 11 A and 1 IB show residual copper and zinc content in the olive fruit >32 days after final injection or treatment with various injection or treatment conditions in two separate trials. Copper levels in all injection or treatment conditions in both trials remained below 30 ppm, i.e., below the residue limit.

[0179] Copper and zinc levels were similar between water injection, foliar treatment, and injection at different dose or frequency.

Claims

CLAIMS What is claimed is:

1. A method for controlling Xylella fastidiosa infection of an olive tree, wherein the olive tree has a trunk, wherein the trunk has a circumference and a diameter, and wherein the infected olive tree exhibits at least one symptom caused by Xylella fastidiosa, the method comprising: a) measuring the trunk circumference or the trunk diameter, or both; b) determining a symptom level of the infected olive tree; and c) injecting the infected olive tree with an injection formulation comprising zinc and copper, wherein: when the trunk circumference is less than about 140 cm, or the trunk diameter is less than about 45 cm, or both, the injection formulation comprises an initial concentration of the zinc (Z 1); when (i) the trunk circumference is between about 140 cm and about 200 cm, or the trunk diameter is between about 45 cm and about 65 cm, or both, and (ii) the symptom level of the infected tree is less than or equal to about 50%, the injection formulation comprises the initial concentration of the zinc (Z 1 ); when (i) the trunk circumference is between about 140 cm and about 200 cm, or the trunk diameter is between about 45 cm and about 65 cm, or both, and (ii) the symptom level of the infected tree is greater than about 50%, the injection formulation comprises about twice the initial concentration of the zinc (1.5Z1-2.5Z1); when (i) the trunk circumference is greater than about 200 cm, or the trunk diameter is greater than about 65 cm, or both, and (ii) the symptom level of the infected tree is less than or equal to about 20%, the injection formulation comprises about the initial concentration of the zinc (Z 1); when (i) the trunk circumference is greater than about 200 cm, or the trunk diameter is greater than about 65 cm, or both, and (ii) the symptom level of the infected tree is between about 20% and about 50%, the injection formulation comprises about twice the initial concentration of the zinc (1.5Z1-2.5Z1); and when (i) the trunk circumference is greater than about 200 cm, or the trunk diameter is greater than about 65 cm, or both, and (ii) the symptom level of the infected tree is greater than about 50%, the injection formulation comprises about four times the initial concentration of the zinc (3.5Z1-4.5Z1) .

2. The method of claim 1, wherein the initial concentration of the zinc is about 0.08% to about 0.23%.

3. The method of claim 1 or 2, wherein steps a)-c) are performed about 7 weeks before flowering, after fruit set, post-summer, and at the end of the season.

4. The method of claim 3, wherein about 7 weeks before flowering is around March, after fruit set is around May and June, post-summer is around September, and the end of the season is around October.

5. The method of any one of claims 1 to 4, further comprising injecting the infected olive tree with a low-dose injection formulation comprising about two thirds the initial concentration of the zinc, about 3 weeks before flowering and pre-summer season.

6. The method of claim 5, wherein about 3 weeks before flowering is around April and the pre-summer season is around July.

8. The method of claim 7, wherein the set of reference trunk measurement ranges comprises a low trunk measurement range, a medium trunk measurement range, and a high trunk measurement range.

9. The method of claim 8, wherein: the low trunk measurement range comprises a trunk circumference range of less than about 140 cm, or a trunk diameter of less than about 45 cm, or both; the medium trunk measurement range comprises a trunk circumference range between about 140 cm and about 200 cm, or a trunk diameter between about 45 cm and about 65 cm, or both; and the high trunk measurement range comprises a trunk circumference range of greater than about 200 cm, or a trunk diameter of greater than about 65 cm, or both.

10. The method of any one of claims 7 to 9, wherein the set of reference symptom level ranges comprises a low symptom level range, a medium symptom level range, and a high symptom level range.

11. The method of claim 10, wherein: an infected tree having a low symptom level range exhibits less than about 20% symptom level; an infected tree having a medium symptom level range exhibits between about 20% and 50% symptom level; and an infected tree having a high symptom level range exhibits greater than 50% symptom level.

12. The method of claim 11, wherein the symptom level is measured based on one or more Xylella fastidiosa symptoms comprising scorching, discoloration, stunting, wilting, premature leaf abscission, shriveled fruit, premature fruit abscission, dieback, and plant death.

13. The method of claim 11 or 12, wherein the symptom level is measured based on one or more Xylella fastidiosa symptoms affecting canopy of the olive tree.

14. The method of any one of claims 7 to 13, wherein the reference injection concentrations comprise a low injection concentration, a medium injection concentration, and a high injection concentration.

15. The method of claim 14, wherein: the medium injection concentration is about twice the low injection concentration; and the high injection concentration is about four times the low injection concentration.

16. The method of claim 14 or 15, wherein the low injection concentration is about 0.08% to about 0.23%.

17. The method of claim 16, wherein: the medium injection concentration is about 0.16% to about 0.46%; and the high injection concentration is about 0.32% to about 0.92%.

18. The method of any one of claims 7 to 13, wherein the reference injection concentration ranges comprise a low injection concentration range, a medium injection concentration range, and a high injection concentration range.

19. The method of claim 18, wherein: the lower limit of the medium injection concentration range is about twice the lower limit of the low injection concentration range; and the lower limit of the high injection concentration range is about four times the lower limit of the low injection concentration.

20. The method of claim 18 or 19, wherein the low injection concentration range is about 0.08% to about 0.23%.

21. The method of claim 20, wherein: the medium injection concentration range is about 0.16% to about 0.46%; and the high injection concentration range is about 0.32% to about 0.92%.

22. The method of any one of claims 14 to 17, wherein: the low injection concentration is selected when (i) the low trunk measurement range is assigned, (ii) the medium trunk measurement range is assigned and the low or medium symptom level range is assigned, or (iii) the high trunk measurement range is assigned and the low symptom level range is assigned; the medium injection concentration is selected when (i) the medium trunk measurement range is assigned and the high symptom level range is assigned or (ii) the high trunk measurement range is assigned and the medium symptom level is assigned; and the high injection concentration is selected when the high trunk measurement range is assigned and the high symptom level range is assigned.

23. The method of any one of claims 18 to 21, wherein: the low injection concentration range is selected when (i) the low trunk measurement range is assigned, (ii) the medium trunk measurement range is assigned and the low or medium symptom level range is assigned, or (iii) the high trunk measurement range is assigned and the low symptom level range is assigned; the medium injection concentration range is selected when (i) the medium trunk measurement range is assigned and the high symptom level range is assigned or (ii) the high trunk measurement range is assigned and the medium symptom level is assigned; and the high injection concentration range is selected when the high trunk measurement range is assigned and the high symptom level range is assigned.

24. The method of any one of claims 7 to 23, wherein steps a)-d) are performed about 7 weeks before flowering, after fruit set, post-summer, and at the end of the season.

25. The method of any one of claims 14 to 17 and 22, further comprising injecting the infected olive tree with a low-dose injection formulation comprising zinc and copper about 3 weeks before flowering and pre-summer season, wherein the zinc concentration of the low- dose injection formulation is about two thirds the low injection concentration.

26. The method of any one of claims 18 to 21 and 23, further comprising injecting the infected olive tree with a low-dose injection formulation comprising zinc and copper about 3 weeks before flowering and pre-summer season, wherein the zinc concentration of the low- dose formulation is selected from a low-dose concentration range, wherein the lower limit of the low-dose concentration range is two thirds the lower limit of the low injection concentration range, and the upper limit of the low-dose concentration range is two thirds the upper limit of the low injection concentration range.

27. The method of claim 25 or 26, wherein about 3 weeks before flowering is around April and pre-summer season is around July.

28. The method of any one of claims 1 to 27, wherein the circumference and/or diameter of the trunk is measured at a reference height from the ground.

29. The method of claim 28, wherein the reference height is between about 40 cm and about 80 cm from the ground.

30. The method of any one of claims 1 to 29, wherein the injecting of the injection formulation is performed using an injection system comprising an injection tool operatively connected to a fluid delivery unit, wherein the fluid delivery unit is configured to deliver the injection formulation.

31. The method of claim 30, wherein the injecting of the injection formulation comprises: piercing the trunk of the olive tree using the injection tool of the injection system; and delivering at least a portion of the injection formulation from the fluid delivery unit through the injection tool into and no further than the active vasculature of the olive tree.

32. The method of any one of the preceding claims, wherein the olive tree is suffering from olive quick decline syndrome.

33. The method of any one of the preceding claims, wherein the Xylella fastidiosa is Xylella fastidiosa subsp. pauca.

34. The method of any one of the preceding claims, wherein the injection formulation is distributed throughout the trunk and other parts of the olive tree.

35. The method of claim 34, wherein the other parts of the olive tree comprise fruits.

36. The method of claim 34, wherein the other parts of the olive tree comprise leaves.

37. The method of any one of the preceding claims, wherein the olive tree is at least about 30 years old.

38. The method of any one of the preceding claims, wherein the olive tree is at least about 100 years old.

39. The method of any one of claims 30 to 38, wherein the delivery unit is a spring- loaded fluid delivery unit.

40. The method of any one of claims 30 to 39, wherein the delivery unit comprises a pressurized formulation cartridge.

41. The method of any one of claims 30 to 40, further comprising: replacing the fluid delivery unit with a second fluid delivery unit; and delivering at least a portion of the injection formulation from the second fluid delivery unit through the injection tool into and no further than the active vasculature of the olive tree.

42. The method of any one of claims 30 to 41, wherein the injection tool remains in the trunk of the olive tree over at least one growing season.

43. The method of any one of claims 30 to 42, wherein the injection tool remains in the trunk of the olive tree over multiple re-injections.

44. The method of any one of claims 31 to 43, wherein the trunk of the olive tree has bark, and the method further comprises: removing at least a portion of the bark prior to piercing the trunk.