[go: up one dir, main page]

WO2015111081A2 - Composition herbicide pour combattre la mauvaise herbe parthenium et souche correspondante - Google Patents

Composition herbicide pour combattre la mauvaise herbe parthenium et souche correspondante Download PDF

Info

Publication number
WO2015111081A2
WO2015111081A2 PCT/IN2015/000045 IN2015000045W WO2015111081A2 WO 2015111081 A2 WO2015111081 A2 WO 2015111081A2 IN 2015000045 W IN2015000045 W IN 2015000045W WO 2015111081 A2 WO2015111081 A2 WO 2015111081A2
Authority
WO
WIPO (PCT)
Prior art keywords
alternaria
parthenium
strain
ncim
inoculum
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/IN2015/000045
Other languages
English (en)
Other versions
WO2015111081A3 (fr
Inventor
Kumar Singh AJAY
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
PRAHARAJU LAXMINARAYANA
Original Assignee
PRAHARAJU LAXMINARAYANA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by PRAHARAJU LAXMINARAYANA filed Critical PRAHARAJU LAXMINARAYANA
Priority to AU2015208711A priority Critical patent/AU2015208711A1/en
Publication of WO2015111081A2 publication Critical patent/WO2015111081A2/fr
Publication of WO2015111081A3 publication Critical patent/WO2015111081A3/fr
Anticipated expiration legal-status Critical
Priority to AU2018267591A priority patent/AU2018267591B2/en
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom

Definitions

  • the present invention provides a novel herbicidal isolate of Alternaria sp. or cell- and spore-free filtrate or crude filtrate or a crude suspension or partially pure obtained therefrom, useful for the control of Parthenium weeds.
  • the present invention also discloses herbicidal compositions comprising fungal isolates formulated in a growth medium for maintaining the viability of the product when the biological control composition is applied to weed.
  • the present invention also discloses methods of screening fungal isolates to determine if they exhibit biocontrol activity.
  • the present invention also discloses a whole genome study for detecting Alternaria sp. isolates that exhibit biocontrol activity to Parthenium weeds.
  • Parthenium is regarded as one of the worst weeds because of its invasiveness, potential for spread, and economic and environmental impacts, and it is noxious because it is highly adaptable to almost all type of environmental conditions, can invade all types of land, also causes high losses in the yield of field crops and direct contact with plant or plant parts for long time causes dermatitis sometimes it may lead to death of person, fever and asthma.
  • Parthenium is hazardous to human health. Health impacts on humans include contact dermatitis, skin irritation, nausea, giddiness and respiratory problems like bronchitis and asthma, eye irritation, and sinusitis (hay fever). Sesquiterpene lactones specially Parthenin found in almost all the plant parts are causing these hazards. Parthenium causes acute toxicity in cattle and milk becomes bitter tasting due to the presence of parthenin, which is also hepatotoxic. If it is present in animal diet then causes dermatitis with pronounced skin lesions and a significant amount (10-50%) of P. hysterophorus in the diet can kill cattle and buffalo. In India, P. hysterophorus causes a yield decline of up to 40% in agricultural crops.
  • allelochemicals, phenolics and, sesquiterpene lactones mainly parthenin
  • it inhibits the germination and growth of plants including pasture grasses, cereals, vegetables and other plant species.
  • it depletes nutrients from and releases toxic substances into the soil.
  • Parthenium pollen can also inhibit fruit set in beans, eggplant, peppers, tomatoes, and other plants. Because their extremely large no. in the field area inhibits the pollens of crop plant to disperse and pollinate.
  • Parthenium seeds Contamination of crop plant seeds with Parthenium seeds restricts their sale and export. By acting as host for insects such as cotton mealybug and disease-causing organisms such as tomato leaf curl virus, it promotes spreading of these diseases in the field. It is commonly called as congress grass, carrot weed, white top etc.
  • Some of the local names by which Parthenium is known are chatak chandani, broom brush, gajari and safedtopi. It is among the top ten worst weeds of the world and has been listed in the global invasive species database.
  • Ploughing the weed in before the plants reach the flowering stage and establishing pastures or other plants may be effective.
  • the manual removal is usually neither very effective nor economical, because of the rapid regrowth requiring repeated removals for season- long control.
  • Several cultural practices also used such as preventing introduction of Parthenium seeds by keeping clean the equipments, livestock, animal feed, people, and vehicles, preventing physical spread of the seeds by cultivators, shoes, tires, machinery.
  • Parthenium weed can be suppressed by growing competitive crops (fodder sorghum sunflower and maize) or self-perpetuating competitive plant species like Cassia sericea, C. tor a, Tagetuserecta (marigold), and Ab tilon indicum, Croton bonplandianus and C. sparsiflorus, Cassia auriculata, in non-crop areas which will compete with the weed and reduce its population.
  • competitive crops for example, dry sorghum sunflower and maize
  • self-perpetuating competitive plant species like Cassia sericea, C. tor a, Tagetuserecta (marigold), and Ab tilon indicum, Croton bonplandianus and C. sparsiflorus, Cassia auriculata, in non-crop areas which will compete with the weed and reduce its population.
  • crop rotation using marigold (Tagetes spp.) during rainy season instead of the usual crop, is found effective in reducing parthenium infestation in
  • Parthenium management through using natural herbicides from fungi is a new kind of bioherbicide as an alternative to chemical herbicide.
  • Mycoherbicides can be divided into fungus preparation herbicide and fungus derived herbicide by virtue of the effective components from the pathogen itself or its phytotoxin.
  • some phytotoxin screened from pathogenic weeds showed potential herbicidal activity.
  • Some experts suggest these phytotoxin be developed as new kind of Bioherbicide as an alternative chemical herbicide.
  • Phytotoxin used for mycoherbicide can be fungal derived product.
  • the pathogen which produces phytotoxins as a microbial herbicide must fit certain requirements: (i) Be reproduced by biological techniques, (ii) grow fast after spraying or be capable of killing weeds within definite time, (iii) suit industrial production and (iv) suitable for packaging, transport and use. Because the fungal toxin used in mycoherbicides usually occur naturally in the areas where they are utilized, they tend to be less harmful to the environment than chemical herbicides. The fungal toxin are often more selective in their mode of action so the risk of damage to other plants is reduced. Mycoherbicides are, as a rule, less toxic to people and animals than chemical herbicides.
  • the present invention provides a herbicidal composition suitable for controlling Parthenium sp., the composition comprising a herbicidal agent from an Alternaria strain.
  • Alternaria sp AGPH04 was deposited in the patent collection of the IMTECH under the terms of the Budapest Treaty and has been assigned accession number MTCC 5973 and identified as Alternaria alternata. It is also deposited in NCIM under accession number 1371.
  • the present invention relates to a method for controlling growth of Parthenium weed comprising contacting the Parthenium plants with the herbicide composition according to the invention.
  • the invention relates to use of an Alternaria sp AGPH 04 for producing herbicidal agent effective for controlling growth of Parthenium plants.
  • the present invention also discloses process of production and characterization of naturally occurring host specific phytotoxic compound that is excreted by the Alternaria sp. AGPH04.
  • the invention also relates to study of sequence homology and phylogenic analysis of strain showed 98% nucleotide similarity with other similar genus isolate.
  • the present invention also relates to whole genome study for identification of novel probe and primer pair sequence for use in detecting isolates that exhibit bio-control activity. This is first report of whole genome study of mycoherbicidal strains Alternaria sp AGPH04 against Parthenium weed.
  • Fig 1 illustrates the macroscopic morphology of Alternaria in accordance with present invention
  • Fig 2 illustrates the microscopic morphology of Alternaria in accordance with present invention
  • Fig. 3 illustrates phylogenetic relationship of Alternaria sp. AGPH#04 to selected species from the genera Alternaria based on 18S rRNA genes in accordance with present invention:
  • Fig. 4 Schematic diagram for extraction of phytotoxic compound from AGPH04
  • Fig. 5 illustrates typical HPLC, CI 8 reverse phase column elution Profile of crude purified Toxin in accordance with certain embodiments of present invention
  • Fig 7 and Fig 8 illustrate base quality score distribution, left (Rl) and right (R2) end of the paired end read for the fungal sample AGPH04in accordance with present invention
  • Fig 9 and Fig 10 illustrate base composition distribution of left and right end of the paired end read sequence for the fungal sample AGPH04in accordance with present invention
  • Fig 1 1 and Fig 12 illustrate The GC distribution of left and right end of the paired end read sequence for the fungal sample AGPH04in accordance with present invention
  • Fig 13 illustrates BLASTX E Value distribution for the fungal sample AGPH04in accordance with present invention
  • Fig 14 illustrates BLASTX similarity score distribution for the fungal sample AGPH04in accordance with present invention
  • Fig 15 illustrate annotated result of top 15 organisms found in BLASTX for the fungal sample AGPH04 in accordance with present invention
  • Fig 16 illustrate top 13 terms in biological function category identified using Gene ontology annotation for sample AGPH04 in accordance with present invention
  • Fig 17 illustrate top 13 terms in molecular function category identified using Gene ontology annotation for sample AGPH04in accordance with present invention
  • Fig 18 illustrate top 13 terms in cellular component category identified using Gene ontology annotation for sample AGPH04in accordance with present invention
  • Table 1 BLAST results of ITS- 1, 5.8 S, and ITS-2 rDNA sequence data of AGPH#04
  • Table 2 Thermal Stability of Phytotoxin
  • Table 5 Effects of metabolites produced by Alternaria on excised leaves of Parthenium
  • Table 6 Effects of Alternaria AGPH#04 on Shoot cut of Parthenium
  • the present invention relates to Parthenium control compositions comprising living fungal cells, Cell free broth, crude metabolites or purified compound of plant pathogens effective for the control of Parthenium weeds.
  • control methods of weeds, plant diseases and insects substantial developments have taken place in approximately the last 100 years owing to developments in chemistry.
  • chemical control methods have replaced traditional cultivation control methods.
  • the rapid progress of synthesized organic pesticides in the middle of the 19th century led to increased field crop yields, improved quality, farming labor saving, etc., so that the world food production was improved rapidly.
  • An object of this invention is to provide novel weed control means capable of taking over the position of synthesized herbicides.
  • the present inventors have found certain strains of the fungus Alternaria sp. as pathogens effective for the control of Parthenium.
  • the present invention therefore provides a weed control composition comprising living Alternaria sp.
  • the pathogens, Alternaria sp., useful in the practice of this invention exhibit a specific pathogenicity to Parthenium.
  • the use of these microorganisms as mycoherbicide makes it possible to control Parthenium, a troublesome weed, without adversely affecting adjacent some economic crops.
  • the pathogens which are useful in the practice of this invention have been chosen from a variety of naturally occurring microorganisms and therefore are free of the potential problem of environmental pollution by synthesized organic herbicides and can be used safely. Namely, Alternaria sp.
  • Pathogens useful in the practice of this invention were found by subjecting to pure isolation pathogens, which had been collected from lesions of naturally-infected Parthenium, and then selecting, from the thus-isolated pathogens, those being pathogenic to Parthenium but non-pathogenic to economic crops like chilies, tomato and brinjal.
  • Pathogens useful in the practice of this invention were selected by conducting both herbicidal activity and pathogenicity tests on Parthenium and chilies, tomato and brinjal plants with respect to strains isolated from naturally-infected Parthenium.
  • As a result of a morphological and 18s rRNA identification of strains it was found to be classified as Alternaria sp named as AGPH#04. It is deposited to NCIM with accession number NCIM 1371.
  • the deposition were converted to depositions under the requirements of the Budapest Treaty, IMTECH to get accession number MTCC 5973.
  • the weed control compositions of this invention which possess the specific pathogenicity against Parthenium only, can use selected strains of Alternaria sp.
  • cultured living fungal cells can be used directly as they are, or after culturing the cells, the culture resulting from germ-free filtration to use a metabolite thereof.
  • the mycoherbicidal compositions of the invention are prepared by dispersing the cultures in suitable medium at an application rate of active agent, preferably ranging from aboutlOml of formulated culture broth in one litre of water.
  • Water is a suitable medium for dispersing toxin-containing cultures. It is suitable to use formulations of toxin from crude fungal inocula or fractions thereof, such as cell-free filtrates, thereby obviating the need to isolate the pure compound.
  • the cell free filtrate, crude filtrate as well as the partially purified toxin is effective in controlling weeds. Therefore, the cell free culture filtrate may be used as such, if desired, in the herbicidal composition, thereby obviating the need for any purification steps.
  • formulations of culture broth are definitely the most preferred and suitable.
  • Effective microbial weed control compositions and microbial origin weed control compositions can be produced by mass-culturing the Alternaria sp. and efficiently obtaining spores, both under asceptic conditions. When applied to the weed, these weed control compositions possess selective herbicidal activities against Parthenium only and show substantially no pathogenicity against economic crops and other weeds. They therefore have highly-selective herbicidal activities, and are free of potential problem of environmental contamination and thus can be used safely.
  • Pathogens Alternaria sp. Useful in the practice of this invention will hereinafter be specifically described by the following examples.
  • the present invention provides a herbicidal composition for controlling Parthenium sp., the composition comprising a herbicidal agent or phytotoxin from an Alternaria strain.
  • the Alternaria strain preferably is a strain having the characterizing features of Alternaria strain AGPH#04 as deposited at IMTECH under accession number. MTCC 5973.
  • a characterizing feature of this strain is its 18s rRNA sequence. Sequence analysis showed 92% nucleotide similarity of the Fungi to Alternaria species but also indicated nucleotide variation of this fungus from other known Genus (Figl).
  • AGPH#04 has surprising features in respect of herbicidal activity, in particular against Parthenium sp.
  • the Alternaria strain having the characterizing features of Alternaria strain AGPH#04 most preferably is Alternaria sp.
  • the herbicidal agent may be a hyphal or a spore inoculum, such as a conidial inoculum.
  • the term inoculum referring to an agent comprising fungal biomass.
  • the inoculum may be viable or non-viable.
  • a non-viable inoculum may be obtained by subjecting a viable inoculum to an inactivating treatment as is known in the art. Heat treatment may be suitably used for inactivating an inoculum.
  • Hyphal and/or (conidia) spore inocula may be obtained from solid or liquid fermentations.
  • a hyphal inoculum may be selected as a hyphal suspension.
  • a (conidia)spore inoculum may be selected from a spore suspension.
  • the herbicidal agent may be culture broth (fermentation broth).
  • culture broth refers to the liquid in which the fungus was grown.
  • PDA medium may be used for fungal growth.
  • Fungal growth may commence between 4 and 9 days, such as between 5 and 8 days, preferably between 6 and 7 days, more preferably about 7 days.
  • Culture broth may be used as it results from a liquid fermentation and may contain fungal biomass such as hyphal and/or (conidio) spore biomass.
  • the culture broth is at least partially purified.
  • the term "at least partially purified" comprises partially purified and may be substituted for this term.
  • a partially purified culture broth results from performing a number of purification steps that increase the contents of certain compounds of the culture broth (while removing others).
  • a partially purified culture broth remains a mixture of compounds.
  • a partially purified culture broth may be cell free culture filtrate (CFCF) for example obtained by filtering-off the fungal biomass with any known means such as microfiltratibn.
  • CFCF cell free culture filtrate
  • a partially purified culture broth may further be obtained by (organic) solvent extraction of certain compounds from the culture broth or from a CFCF obtained therefrom.
  • the solvent is selected from hexane, Ethyl acetate, chloroform, methanol or mixtures therefrom.
  • the selected solvent preferably comprises hexane or Ethyl acetate and preferably is hexane or ethyl acetate. Most preferably hexane is used.
  • a potent herbicidal agent is obtained as is further detailed in the examples.
  • the invention therefore further relates to a herbicidal agent obtainable by organic solvent extraction, in particular a hexane or ethyl acetate extraction, of CFCF of the Alternaria strain used.
  • the solvent extract may have the elution profile presented in figure 2, or a similar profile, on a HPLC, CI 8 Reverse phase column. The use of hexane for solvent extraction is most preferred.
  • the herbicidal agent according to the invention will comprise a number of secondary metabolites. "A number of within the present invention should be construed as meaning one or morej such as a plurality, for example 2-10 such as, 3-9, 4-8, 5-7, or 6. Secondary metabolites in the herbicidal agent may be further purified. According to certain embodiments the secondary metabolites are purified to > 80% purity, such as >85%, >90%, >95%, >99% up to 99.9 % purity.
  • compositions and methods of the invention include the provision of application of cell free culture filtrate or their metabolites to produce a novel herbicidal agent.
  • Herbicide compositions may be prepared as a liquid formulation by suspending the broth, partially purified broth, such as cell free broth, crude or purified metabolites in an agriculturally acceptable carrier for application to the weed or the location where it is growing.
  • any agriculturally acceptable carrier can be used whether it is liquid or solid as long as it can be employed in agricultural or horticultural formulations and is preferably biologically inert.
  • Exemplary agriculturally acceptable liquid carriers include, but are not limited to, water, surfactants, vegetable oils, and mineral oils.
  • the agriculturally acceptable carrier for a liquid formulation is water, and the herbicide comprises a cell free broth.
  • Herbicide compositions can also be prepared as granular formulations, flowable formulations, or wettable powder formulations by mixing with an agriculturally acceptable carrier, which is then applied to weed.
  • Suitable agriculturally acceptable solid carriers include mineral powders, such as clay, talc, bentonite, calcium carbonate, diatomaceous earth and white carbon; vegetable flours such as soybea flour and starch, and some polymers such as polyvinyl alcohol and polyalkylene glycol.
  • a surfactant such as a Tween, for example Tween 20, or Tween 80 is used.
  • a surfactant such as a Tween, for example Tween 20, or Tween 80 is used.
  • Tween 20 for example Tween 20
  • Tween 80 is used.
  • an oil such as a mineral oil, for example paraffin oil, or a vegetable oil, such as coconut oil is used.
  • Antibiotics may also be included Antibiotics (antimicrobials) may be selected from Streptomycin, Ampilox, Azithromycin may be selected from these antibiotics. .
  • In general formulating agents can be added in an amount of 0.2-0.9 % w/w, such as 0.3-0.8%, 0.4-0.7%, 0.4-0.6%, such as about 0.5% w/w to the herbicidal agent.
  • Antimicrobials may be added in an amount of 0.05-0.5% w/w such as 0.06-0.4 %, 0.07-0.3%, 0.08-0.3%, 0.09-0.3 %, 0.1-0.3 % such as about 0.2% w/w to the herbicidal agent.
  • a further aspect of the invention relates to a method of controlling weeds, in particular Parthenium sp.
  • the method includes applying the herbicide composition to the Parthenium plants.
  • the herbicide composition can be applied by spraying a solution of j fungal herbicidal agents, such as metabolites, at weed in an amount sufficient to coat the leaves of the weed.
  • One application may be sufficient to reduce current growth; however, repeat applications may be necessary if regrowth of the plant occurs from resistant or below ground structures.
  • the method of the present invention may be used in addition to or in conjunction with other control measures.
  • the method is for suppressing or preventing (controlling) growth of Parthenium sp. and the method comprises contacting the Parthenium plants, preferably Parthenium hysterophorus plants, with the a composition of the invention. Details of the technical features of the composition of the invention have been discussed above. As the skilled person will understand the composition of the invention will be used in an effective amount sufficient to bring an effect, in particular control (reduction or inhibition) of growth of Parthenium plants. It is within the ambit of the knowledge and skill of the skilled person to determine effective amounts. According to certain embodiments effective amounts may be in the range of 10ml of formulated cell free culture broth to 1 litre of water.
  • the contacting of the Parthenium plants with the composition is achieved by applying the composition on the plants for example by spraying. In other embodiments of the present invention, the contacting of the Parthenium plants with the composition is achieved by mixing the composition in soil where Parthenium is or may be present as seeds and/or roots.
  • a further aspect of the invention relates to the use of an Altemaria strain, preferably a strain having the characterizing features of Altemaria strain AGPH#04 as deposited at IMTECH under accession number MTCC 5973 for producing a herbicidal agent effective for controlling growth of Parthenium sp, in particular Parthenium hysterophorus.
  • the technical features of the herbicidal agent have been discussed above in connection to the discussion of the composition of the invention. Similar to what is discussed for the composition of the invention, the herbicidal agent may be selected from a hyphal inoculum, a spore inoculum, preferably a conidial inoculum, culture broth, preferably at least partially purified culture broth metabolites.
  • a further aspect of the invention relates to a herbicidal agent obtainable by a method comprising solvent extraction, preferably hexane or ethyl acetate extraction, of cell free culture filtrate (CFCF) of a Altemaria strain, preferably a strain having the characterizing features of Altemaria strain AGPH#04 as deposited at IMTECH under accession number MTCC 5973.
  • the Altemaria strain is strain AGPH#04 as deposited at IMTECH under accession number MTCC 5973 or a strain derived therefrom.
  • the CFCF is provided, preferably by culturing the selected strain in a liquid culture and producing a CFCF from the culture broth.
  • a (organic) solvent extraction preferably hexane or ethyl acetate extraction is performed on said CFCF.
  • the solvent extraction may be executed according to any known methods.
  • the strain was grown at 28°C ⁇ 1°C under in one litre flasks containing suitable medium.
  • the sterile media was inoculated with 1 ml of a spore suspension of fungus, shaken gently, and incubated as standing cultures for 14 days.
  • the culture filtrates were collected by successive passage through Whatman No.l filter paper, and a sartorius membrane filter (0.20 ⁇ ).
  • the culture filtrate which has been passed through the sartorius membrane filter. Cell free culture filtrate extracted with chloroform to get to solvent extract.
  • the spore's concentration was adjusted with haemocytometer to 2.5x10 6 spores/ml.
  • the potential of isolates were assessed using detached leaf bioassay or shoot cut bioassay method.
  • the strains showing LAD Leaf area damage between 80 to 100%, has taken for second trails.
  • Pure cultures of the recovered fungi were prepared from single conidium and maintained on half strength PDA slants as stock cultures.
  • the isolated fungi were identified to the genus based on their conidial morphology and growth characteristics on various growing media. Further identification and characterization were done on isolate that was confirmed to be highly pathogenic to P. hysterophorus by applying Koch's postulates. Cultures grown on PDA as well as sporulating on infected plants in the greenhouse were used for taxonomic characterization of the conidia.
  • Alternaria isolate was isolated from infected Parthenium leaves plants exhibiting symptoms of leaf disease. During survey to Village Adavi Ramanpalam, PenuballiMandal, Khammam District, Andhra Pradesh, Parthenium leaves infected with pathogenic fungus Alternaria sp AGPH#04 is a case in point to develop as herbicide. The isolates were grown on the PDA medium and stock cultures of these were maintained at AGBIO Systems Laboratory and also deposited in the NCIM, Pune, India and given an accession number NCIM 1371. This strain has also been deposited at IMTECH, Chandigarh, India under the Budapest treaty under accession number. MTCC 5973. The strain has identified as Alternaria alternata.
  • the colony is flat, downy to woolly and is covered by grayish, short, aerial hyphae in time.
  • the surface is greyish white at the beginning which later darkens and becomes greenish black or olive brown with a light border.
  • the reverse side is typically brown to black due to pigment production.
  • the macroscopic figures of Alternaria sp AGPH04 is mentioned in Fig 1.
  • Alternaria sp. has septate, dark hyphae.
  • Conidiophores are also septate and sometimes have a zigzag appearance. They bear simple or branched large conidia (8-16 x 23-50 ⁇ ) which have both transverse and longitudinal septations. These conidia may be observed singly or in acropetal chains and may produce germ tubes. They are ovoid to obclavate, darkly pigmented, muriform, smooth or roughened. The end of the conidium nearest the conidiophore is round while it tapers towards the apex. This gives the typical beak or club-like appearance of the conidia.
  • the microscopic figures of Alternaria sp. is mentioned in Fig 2.
  • EXAMPLE 2 The microscopic figures of Alternaria sp. is mentioned in Fig 2.
  • Alternaria sp AGPH#04 are potential, highly hosts specific mycoherbicide for control of the Parthenium weed (Table 1 & Fig 3).
  • the fungal isolates were characterized based on partial DNA sequence of the internal transcribed spacer regions of the nuclear ribosomal RNA gene.
  • the ITS sequence was obtained using the deposited database of ITS. Both isolates ITS sequence obtained was not 100% identical to any ITS sequence deposited in Gene Bank. The 95% similarity has been shown by Alternaria alternata AF21879. This unique ITS sequence of the two isolates provides strong evidence to consider these pathogens as new from speicalis that will facilitate and encourage the introduction and acceptance by regulatory authorities for practical field application.
  • Table 1 BLAST results of ITS- 1, 5.8 S, and ITS-2 rDNA sequence data of AGPH#04.
  • Alternaria sp was grown at 28° C ⁇ 1°C under in one litre flasks containing 250 ml modified Richard medium with 1% host leaves extract.
  • the sterile media was inoculated with 1 ml of a spore suspension of Alternaria sp, shaken gently, and incubated as standing cultures for 14 days.
  • the culture filtrates were collected by successive passage through Whatman No. l filter paper, and a Sartorius membrane filter (0.20 ⁇ ).
  • the sterile filtrates were stored at 4°C depending on assay or fractionation.
  • HPLC High Performance Liquid Chromatography
  • CFCF of AGPH#04 was subjected to different temperature treatment viz. 40, 50, 60, 100 and 120°C. Each treatment was carried out for 15 mins. The phytotoxic activity of each treatment was assessed using the shoot cut bioassay. Each treatment was carried out in triplicate and CFCF at room temperature served as control and uninoculated medium served as second control.
  • Parthenium seeds were planted in pot. The plants were watered as needed. The photoperiod was 14 h. The fungal inoculum was applied using a sprayer to run-off. Control groups received a filtrate of autoclaved distilled water. Parthenium plants were used in these experiments. Following inoculation, plants were incubated on greenhouse benches under conditions as described above. Three replicates of weed were used for each treatment. The experiment was repeated four times. Symptom development was monitored daily. Results were observed at the beginning and the end of the experiments. All treated plant showed wilting and complete necrosis. The results are shown in Table 4. In Parthenium seedling, the damage resulting from the crude, cell-free filtrates and partially purified toxin were identical, including visibility of necrosis of the leaves of the weeds. Sterilized distilled water was used as controls for the method.
  • Detached Parthenium leaves were used to test the, biological activities of cell free filtrates, crude filtrate and partially purified toxin. Detached leaves were placed on moistened filter paper inside 9-cm diameter sterile Petri plates. The inocula of crude filtrates, cell-and spore-free filtrates, and the partially purified phytotoxin were applied to the leaves with micropipettes. Ten leaves were used for each treatment. Control leaves received distilled water. The plates were sealed with parafilm and incubated at Room temperature under 12 h light condition. The phytotoxic effects on the treated excised leaves were evaluated visually for damage for 5 days. The result of this test is depicted in Table S.
  • Formulation refers to the process of blending of secondary metabolite or microbe with inert carriers to alter their physical characteristics to enhance its shelf life and field performance.
  • Formulation involves the use of formulants which include: adjuvants, surfactants, wetting agents, spreaders and preservative.
  • adjuvants increases the efficacy of post emergence herbicides by increasing the wettability of the target surface as they reduce surface tension. Adjuvants are also known to enhance penetration. In order to investigate the compatibility of the phytotoxin produced by the test fungus with various formulants were tested.
  • Table 8 Testing of various formulations of AGPH#04 by Seedling bioassay. 24 hpt 48 hpt 72 hpt.
  • Plant fungal pathogen in the genus Altemaria infects a remarkable range of host plants and is major causes of agricultural yield losses.
  • Altemaria sp DNA was extracted by Scigenom developed protocol and the sequencing library was prepared using Illumina paired end DNA sample prep kit. Sequencing was performed using Illumina Genome Analyser. Short reads were assembled de novo using velvet and assembly quality was improved by a pipeline including two alternate assemblers Edena and Minimus. Alternaria sp AGPH#04 (NCIM 1371) was sequenced by a whole genome shotgun approach using the Ilium ina Genome Analyzer. Fastq quality check, De novo assembly of genome study. Gene prediction, ORF annotation and Blastx search performed.
  • Fastq quality check involved checking of quality parameters for the sequence obtained from sequencer.
  • Base quality score distributions, average base content per read and GC distribution in the reads were performed for an input fastq file.
  • the base quality score distribution, the quality of left (Rl) and right (R2) end of the. paired end read for the sample is shown in fig 7, 8.
  • Base composition distribution of left and right end of the paired end is shown in fig 9, 10.
  • the x-axis represents sequencing cycle and y axis represents nucleotide percentage.
  • bias in the base composition towards the beginning of reads. Biasing in sequence composition is generally observed in NGS experiments.
  • the GC distribution of left and right end of the paired end read sequence were performed (Fig 11 , 12).
  • the x-axis represents average GC content in the sequence and y axis represents total percentage of reads.
  • the average GC content of the reads in the sample followed a normal distribution.
  • the fastq files were trimmed before performing Denovocontigs assembly. Scigenom scientists have trimmed three bases from the beginning and two bases from the end of the reads. Summary of the trimmed reads are provided in Table 10. Table 10: Trimmed read summary for samples AGPH04
  • the BLASTX search E value distribution is provided in Fig 13. Around 89% of the ORFs found using BLASTX for the sample had confidence level of at least le-50, which( '
  • BLASTX similarity score distribution for sample is shown in Fig 14. 83% of the predicated ORFs found using BLASTX had similarity of more than 60% at protein level with the existing protein at NCBI database in Sample AGPH04.
  • Majority of the top BLASTX hit belong to Pyrenophorateres f teres.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Pest Control & Pesticides (AREA)
  • Biotechnology (AREA)
  • Agronomy & Crop Science (AREA)
  • Plant Pathology (AREA)
  • Virology (AREA)
  • Mycology (AREA)
  • Dentistry (AREA)
  • Wood Science & Technology (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne le développement d'un nouveau mycoherbicide pour combattre Parthenium hysterophorus. La présente invention concerne également des isolats fongiques d'Alternaria sp AGPH04 utilisant des spores en tant filtrat exempt de cellules et de spores, un filtrat brut, ou une suspension brute de la culture et éventuellement d'autres additifs. On a observé que la souche fongique est particulièrement active contre Parthenium. La souche fongique a été déposée auprès de NCIM en tant que collection de culture privée et s'est vu attribuer le numéro NCIM 1371. La souche a été également déposée dans la collection de brevets de l'IMTECH dans le cadre du Traité de Budapest et s'est vu attribuer le numéro MTCC 5973. L'homologie de séquence et l'analyse phylogénétique de la souche ont montré 98 % de similitudes avec un isolat d'autres genres similaires. La présente invention concerne également l'étude du génome entier pour l'identification d'une nouvelle séquence de paires de sonde et d'amorce servant à détecter les isolats qui manifestent une activité de lutte biologique. La présente invention concerne également un procédé de production et de caractérisation de composé phytotoxique spécifique à un hôte d'origine naturelle excrété par Alternaria sp AGPH04.
PCT/IN2015/000045 2014-01-27 2015-01-23 Composition herbicide pour combattre la mauvaise herbe parthenium et souche correspondante Ceased WO2015111081A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU2015208711A AU2015208711A1 (en) 2014-01-27 2015-01-23 An herbicidal composition for controlling Parthenium weed and strain thereof
AU2018267591A AU2018267591B2 (en) 2014-01-27 2018-11-20 An Herbicidal Composition for Controlling Parthenium Weed and Strain Thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IN326CH2014 IN2014CH00326A (fr) 2014-01-27 2014-01-27
IN326/CHE/2014 2014-01-27

Publications (2)

Publication Number Publication Date
WO2015111081A2 true WO2015111081A2 (fr) 2015-07-30
WO2015111081A3 WO2015111081A3 (fr) 2015-11-19

Family

ID=52823745

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IN2015/000045 Ceased WO2015111081A2 (fr) 2014-01-27 2015-01-23 Composition herbicide pour combattre la mauvaise herbe parthenium et souche correspondante

Country Status (3)

Country Link
AU (2) AU2015208711A1 (fr)
IN (1) IN2014CH00326A (fr)
WO (1) WO2015111081A2 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113278532A (zh) * 2021-05-21 2021-08-20 山东农业大学 一株极细链格孢及其代谢产物和应用
CN114907990A (zh) * 2022-05-02 2022-08-16 中国海洋大学 一种链格孢菌及其用于去除废硅藻土中蛋白质的应用
CN116784145A (zh) * 2023-07-26 2023-09-22 四川省自然资源科学研究院(四川省生产力促进中心) 一种银胶菊的生态防控方法
CN118931735A (zh) * 2024-08-30 2024-11-12 西南林业大学 一株棉链格孢菌swfu-mm002及其应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
None

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113278532A (zh) * 2021-05-21 2021-08-20 山东农业大学 一株极细链格孢及其代谢产物和应用
CN114907990A (zh) * 2022-05-02 2022-08-16 中国海洋大学 一种链格孢菌及其用于去除废硅藻土中蛋白质的应用
CN114907990B (zh) * 2022-05-02 2023-07-14 中国海洋大学 一种链格孢菌及其用于去除废硅藻土中蛋白质的应用
CN116784145A (zh) * 2023-07-26 2023-09-22 四川省自然资源科学研究院(四川省生产力促进中心) 一种银胶菊的生态防控方法
CN118931735A (zh) * 2024-08-30 2024-11-12 西南林业大学 一株棉链格孢菌swfu-mm002及其应用

Also Published As

Publication number Publication date
IN2014CH00326A (fr) 2015-08-14
AU2018267591B2 (en) 2020-09-10
WO2015111081A3 (fr) 2015-11-19
AU2018267591A1 (en) 2018-12-06
AU2015208711A1 (en) 2016-09-15

Similar Documents

Publication Publication Date Title
AU2016258913B2 (en) Designed complex endophyte compositions and methods for improved plant traits
Tagele et al. Effectiveness of multi-trait Burkholderia contaminans KNU17BI1 in growth promotion and management of banded leaf and sheath blight in maize seedling
JP4883814B2 (ja) 植物病害に対する防除能を有する微生物、及び該微生物を用いた植物病害の防除剤
Ramos et al. Diversity and abundance of Beauveria bassiana in soils, stink bugs and plant tissues of common bean from organic and conventional fields
AU2018267591B2 (en) An Herbicidal Composition for Controlling Parthenium Weed and Strain Thereof
US11390844B2 (en) Use of compositions containing Streptomyces melanosporofaciens AGL225 in controlling plant diseases
KR102604427B1 (ko) 트리코더마 롱기브라키아툼 균주를 포함하는 식물병 방제용 조성물 및 이를 이용한 식물병 방제 방법
KR101708325B1 (ko) 식물기생선충에 대해 살선충 활성을 가지는 아스퍼질러스 나이거 f22 균주 및 이의 용도
CN111040976A (zh) 一株解淀粉芽孢杆菌及其应用
Tra et al. Bacterial endophytes from Chukrasia tabularis can antagonize Hypsipyla robusta larvae
CN104593266B (zh) 一种番茄内生真菌交织枝顶孢及其在番茄根结线虫病生防中的应用
CN101735956B (zh) 一种真菌代谢产物及其作为除草剂的用途
CN101919412B (zh) 一种炭疽菌菌株及含有其孢子的生物除草试剂
CN108441443A (zh) 一株防治植物线虫的菌株及其应用
KR101953835B1 (ko) 식물의 병 저항성을 증진시키는 아스페르질루스 테레우스 균주 및 이의 용도
AU2018267590B2 (en) A Novel Mycoherbicidal Composition for Suppressing Water Hyacinth
KR102556901B1 (ko) 담수에서 분리한 항균력 및 식물생장촉진능을 가지는 에드니아 속 nnibrfg15114 균주 및 이의 용도
KR102397132B1 (ko) 항균력 및 식물생장촉진능을 가지는 트라이코더마 아스퍼레룸 nnibrfg4324 균주 및 이의 용도
JP7468842B2 (ja) 植物病害防除用の農薬組成物及びそれを用いる植物病害防除方法
CN113925064A (zh) 耐硼赖氨酸芽孢杆菌在抑制火龙果软腐病菌生长方面的应用
McLean Biological control of onion white rot using Trichoderma harzianum
Hernández-Hernández et al. In vitro characterization of rhizobacteria and their antagonism with fungi that cause damping off in chili
Chan-Cupul et al. Trichoderma spp. condiospores production and its application with Streptomyces spp. for the management of Mycosphaerella fijiensis in banana
OMUKETI Diversity, Antagonistic Potential of Endophytes, Phytochemicals and Antimicrobial Activity of Selected Agroforestry Trees Against Xanthomonas Campestris Pv. Musacearum and Cercospora Zeae-Maydis
MEGHANA MOLECULAR DIVERSITY ANALYSIS AND ANTAGONISTIC POTENTIAL OF PLANT GROWTH PROMOTING MICROORGANISMS FOR THE MANAGEMENT OF EMERGING DRY ROOT ROT OF CHICKPEA CAUSED BY Rhizoctonia bataticola (Taub.) Butler.

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15715461

Country of ref document: EP

Kind code of ref document: A2

ENP Entry into the national phase

Ref document number: 2016/2532

Country of ref document: KE

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2015208711

Country of ref document: AU

Date of ref document: 20150123

Kind code of ref document: A

122 Ep: pct application non-entry in european phase

Ref document number: 15715461

Country of ref document: EP

Kind code of ref document: A2