[go: up one dir, main page]

WO1990010691A1 - Culture de tissus - Google Patents

Culture de tissus Download PDF

Info

Publication number
WO1990010691A1
WO1990010691A1 PCT/US1990/001333 US9001333W WO9010691A1 WO 1990010691 A1 WO1990010691 A1 WO 1990010691A1 US 9001333 W US9001333 W US 9001333W WO 9010691 A1 WO9010691 A1 WO 9010691A1
Authority
WO
WIPO (PCT)
Prior art keywords
nutrient
controlled release
medium
growth
salts
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/US1990/001333
Other languages
English (en)
Inventor
Carolyn J. Sluis
Alice U. Hahn
Rodney K. S. Kahn
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.)
GRACE-SIERRA HORTICULTURAL PRODUCTS Co
Original Assignee
GRACE-SIERRA HORTICULTURAL PRODUCTS Co
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 GRACE-SIERRA HORTICULTURAL PRODUCTS Co filed Critical GRACE-SIERRA HORTICULTURAL PRODUCTS Co
Publication of WO1990010691A1 publication Critical patent/WO1990010691A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0018—Culture media for cell or tissue culture
    • C12N5/0025—Culture media for plant cell or plant tissue culture

Definitions

  • This invention relates to an improvement in tis ⁇ sue culturing. More particularly, it relates to an improved method for delivering biologically active materi ⁇ als such as nutrients to tissue culture environments.
  • tissue culturing is a general term applied to micropropagation processes in which tissues or cells of higher animals or plants are grown artificially in a controlled environment. In these processes, tissues or cells, either as suspensions or as solids are maintained under conditions conducive for their growth and multiplication. These conditions include proper temperature, proper gaseous and liquid environment, and proper supply of nutrient.
  • Tissue culturing as applied to plants, is pres ⁇ ently viewed as an expensive method.
  • micropropagation represents one of the few means by which many forestry, plantation and other difficult-to-root spe ⁇ cies can be clonally reproduced
  • the high cost of tissue culture techniques has prevented broader application in the marketplace. Consequently, the appearance of clonal forests, fields and crops has not materialized.
  • the present invention seeks to address these problems and provide a cost effective way for delivering nutrients and/or essential minerals or other needed components to a tissue culture environment.
  • controlled release technology a technology primarily employed in drug delivery, pesticide delivery, and fertilizer delivery to potted plants and field crops, can advantageously maintain levels of nutrients, minerals, and other biologically ac- tive essential growth elements in a tissue culture medium at levels at or near optimum for sustained periods.
  • this invention provides an improved tissue-culturing process in which plant cells are grown in an aqueous-based gel matrix containing nutrients essential to the growth of the cells.
  • the improvement involves placing in that growth medium an insoluble or slowly soluble depot of one or more of the nutrients or other biologically active materials.
  • the depot has the property of being able to continuously release the one or more agents it contains into said growth medium over a prolonged period of time, thereby replacing the agents consumed by the growing cells and maintaining the agent level at or near optimum levels.
  • the present invention finds principal applica ⁇ tion with plant tissue cultures.
  • the nutrients which are delivered can include salts, organic growth regulators, as well as sugars and . other materials consumed by the growing plant.
  • the depots employed are generally made up of one or more bodies of nutrient and/or agent surrounded by a coating which limits the rate of dissolution or release of the body of nutrient or agent into the medium.
  • the sustained release nutrient sources are used in combination with an initial loading of wholly soluble nutrients and agents.
  • the initial loading serves to provide levels of nutrients and agents adequate for initial growth, and the sustained- release material adds nutrient and agents as they are consumed by the growing tissue.
  • Figure 1 is two graphs showing the changes in conductivity observed over a 6 week period in agar (A) and water (B) tissue culture media when six different levels of controlled release nutrients are added.
  • Figure 2 is a graph similar to Figure 1 showing the release of nutrient into a tissue culture medium containing an initial concentration of Murashige-Skoog's salts.
  • Figure 3 is a series of graphs showing the de ⁇ pletion of nutrient from a tissue culture environment when various plants are cultured.
  • Figure 4 is a series of graphs showing the sum of the depletion shown in Figure 3 plus the release of material released by the controlled release nutrient.
  • Figure 5 is a graph comparing the cumulative release of controlled release nutrient into a tissue culture medium with the cumulative uptake of nutrient by . three species of plants in a tissue culture.
  • Figure 6 is a graph showing relative growth observed with Ficus benjamina when various levels of controlled release nutrient are added.
  • Figure 7 is a graph showing the relative biomass of Boston fern observed when various levels of controlled release nutrient are added.
  • Figure 8 is a graph showing the relative growth of potatoes observed when various levels of controlled release nutrient are added.
  • the typical environments in which the present invention are employed comprise an aqueous gel medium in which cells or living tissue are artificially grown.
  • these environments are small separately enclosed sterile regions such as closable jars, culture dishes and the like.
  • the environment includes an aqueous growth medium. This is, in the case of plants, typically an agar-based medium which contains substantial quantities of sugars such as sucrose, glucose, and coconut milk and essential minerals and the like.
  • the environment also includes needed gases such as CO- or oxygen and, in the case of plants, a source of light.
  • United States Patent 3,819,960 is incorporated herein by reference and includes a description of the various materials including minerals, metal salts, growth hormones, amino acids and vitamins which may be advantageously present in the growth environ ⁇ ment.
  • United States Patent 4,038,778, also incorporated by reference shows other culturing conditions.
  • the agents delivered to the growth environment using the present invention are most typically water-soluble macronutrients such as potassium salts, nitrogen containing salts, and phosphorous containing salts.
  • macronutrients such as potassium salts, nitrogen containing salts, and phosphorous containing salts.
  • Micronutrients such as salts or other soluble compounds of iron, zinc, manganese, copper, nickel, molybdenum and boron can also be added.
  • Organic materials can also be added including vitamins, growth factors and growth regulators and the like.
  • the invention could be used to deliver all the macro and micronutrients to a tissue-culturing medium.
  • the invention can be used in combination with known nutrient sources at or below their usual levels.
  • a typical known plant nutrient package is Murashige-Skoog's salts which are typically used at from 0.4 to 1.0 and preferably 0.4 to 0.8 times its normal use levels. (See Revue Physiologia Plantarur ⁇ 15:473 for details on Murashige-Skoog's salts.)
  • a typical salt mixture is used to supply macro and micronutrients and the controlled release material is used to supply macronutrients as they are consumed by the growing plants.
  • Representative nutrients in tissue culture media include the following.
  • Macronutrients potassium and ammonium nitrates, magnesium sulfate, calcium chloride, and monopotassium phosphate (Murashige-Skoog's macroelements).
  • Micronutrients include manganese, zinc and cop ⁇ per sulfates, potassium iodide, boric acid, calcium chloride and molybdenum-sodium oxide (Murashige-Skoog's microelements) .
  • NiCl 2 (6H 2 0) 0.030 Iron may also be present such as at 27.3 mg/1 of FeS0.(7H 2 0).
  • Cobalt is another typical constituent such as at 0.02 mg/1 of CoCl 2 (2H 2 0).
  • Typical vitamins and growth factors can include:
  • Nicotinic acid 1.0 mg/1
  • Vitamin B fi 1.0 mg/1
  • Vitamin B. 1.0 mg/1
  • the initial culturing media will contain these microelements and up to these levels of macroelements with the controlled release material replenishing macroelements and optionally microelements as they are consumed.
  • the invention can employ a controlled release material capable of delivering and maintaining the appropriate levels of N, P and K.
  • the controlled release material can deliver N, P, K, Ca and Mg. In yet another embodiment, the controlled release material can deliver N, P, K, Ca and Mg and at least one of Cu, Co, Mo, I, Zn, B, Mn, and Fe.
  • the present invention can be used to deliver vitamins, hormones, essential nutrients and the like.
  • nutrients are delivered to tissue culturing environments by the use of controlled release systems. These systems are made up of a depot of nutrient and means to prevent the immediate release of the nutrient into the tissue culture environ ⁇ ment of use.
  • the nutrients are water soluble solids or, at times, liquids.
  • the environment of use is aqueous.
  • Any release mechanism which will prolong the release of the nutrients over a period of not less than 7 days can be used, with preferred systems prolonging and extending the release period to between 7 and 360 days and especially between 21 and 150 days.
  • ⁇ In selecting a release mechanism it should be kept in mind that the degree of precision required is not extreme and that in most cases one can safely trade off precise control of release kinetics for cost savings. As will be obvious, in the areas of commercial plant produc-
  • cost control is of utmost importance.
  • One useful and often cost effective release mechanism is to disperse the particles of nutrient in a solid body of slowly soluble material such as a slowly soluble polymer.
  • Slowly degradable esters such as the
  • orthoesters or slowly hydrolyzable materials can be used as an erodible matrix.
  • Another release mechanism which can be employed involves the use of a semipermeable layer surrounding one or a plurality of depots of the nutrients which layer
  • 2c permits the passage of water into the depot with the subsequent gradual dissolution of the soluble salts (nutrients) and thus the generation of osmotic pressure within the depots.
  • This osmotic pressure can be used to drive the nutrients out of the depots either through
  • 3Q preprovided microholes or through holes formed in situ by the pressure build up.
  • a seemingly limitless family of materials are available to use as these osmotic membranes and include cellulose acetate, various resins and the like.
  • a representative listing of typical osmotic 35 membrane materials is included in United States Patent 3,845,770, which is incorporated herein by reference.
  • Other representative disclosures of controlled release coatings can be found in United States Patents 4,019,890; 3,342,577; 4,369,055 and 3,264,089, all in ⁇ corporated herein.
  • a good system is the Osmocote system employed by Sierra Chemical Co. This is a heat-cured resin system described in United States Patents 4,657,576 and 3,223,518.
  • the Osmocote " resin system involves a co- polymer of dicyclopentadiene and alkyd resin from soy bean oil.
  • the basal medium contained Murashige-Skoog's (1962) inorganic salts (M/S salts) and vitamins, 100 mg/1 m-inositol, and 30 g/1 sucrose.
  • Hormones were added as required by the various plant spe ⁇ cies for shoot proliferation. The pH was adjusted to 5.7 prior to addition of 0.7% agar.
  • the media were autoclaved in 1 1 bottles at 120 C and 20 psi for 17 minutes and dispensed into various sterile containers in a HEPA-filtered laminar flow hood before gelling. Most of the work was done in plastic containers known in the trade as sundae cups, some in standard 25 x 100 mm petri dishes, some in a plastic containerized systems commercially sold as "sandwich boxes", and some in Magenta boxes. In all cases the medium volume was kept constant at 60 ml per container.
  • Plants were inoculated onto medium by insertion with forceps and left to grow for 4-6 weeks in culture rooms kept at 23 C, using a 16 hour photo period and cool white fluorescent lights at 400-800 f.c.
  • Osmocote brand osmotic release sustained delivery greenhouse soil fertilizer was used as the sustained release nutrient source.
  • This material which was used as 1-2 mm prills and as microprills, included nitrogen as ammonium, 9 .3% ; nitrogen as nitrate, 7.7%; phosphorous expressed as P- j O,-, 6%; and potassium expressed as K Raven0, 12% (all by weight). This material had the characteristic of giving a sustained release of these nutrients over a period of up to at least about 120 days.
  • the sustained release material also contained, in some cases, a mixture of other minerals.
  • the added mineral ormula ⁇ tion had 1.5% w calcium
  • controlled release material was about 40% nutrient and 60% inerts and controlled release coating.
  • the initial strength of the medium could be reduced for two reasons: (1) to accommodate any "quick" release from broken or imperfect prills, and (2) to start the plants off at a lower salt level and build it up over time.
  • the initial strength of the M/S salts was reduced from full strength to half strength in initial experiments. Some plants cannot tolerate such low levels and 0.7-0.8 times the typical M/S salts were commonly used in those cases. 2. Conductivity Measurements.
  • Conductivity measurements were used as a quick and reproducible way to monitor the release of nutrients to the growth environ ⁇ ment.
  • the conductivity meter used was a Chemtrix 103 instrument with automatic temperature compensation, calibrated daily against KC1 standard solutions. It was determined that completely filling the probe tip with agar medium excluding air pockets gave the same reading as measuring the same amount of salts in aqueous solution. The hollow tip of the probe was pushed through the agar to fill it completely, care taken to exclude air or plant debris. Conductivity is temperature dependent so care was taken that the agar was not too warm or too cool during measurement.
  • Desiccated medium was observed to have a higher conductiv ⁇ ity than fresh medium. Often at the end of a culture period the medium had a higher conductivity than fresh medium. Often, the ending conductivity can be quite low showing that plants in tissue culture were able to use up much of the salt in the medium.
  • Figure 1 shows the increase in conductivity observed when controlled release nutrients are released into agar and water and illustrates how, by the practise of the present invention, nutrient is delivered to tissue culture media at a relatively constant rate over at least an 8 week period.
  • the volume was scaled down to fit in 25 x 150 mm test tubes to increase replication.
  • Object 1 To Be Able to Get Controlled Release Material in Sterile Form.
  • tissue culture it must be obtained in sterile form, i.e., autoclaved, dry heat sterilized, irradiated, fumigated, or chemically treated.
  • this material can be autoclaved separately from the medium quite satisfactorily.
  • autoclaved in water a great deal of the nutrient in the material is released.
  • prills from autoclaving, but it seems to be minor.
  • Moisture from the steam dries out over about a week if the prills are left to stand.
  • Controlled release material was added to the surface of water agar (nonsterile but containing 50 mg/1 Benlate) at levels of 260, 520, and 1040 mg/60 ml medium.
  • the medium was kept in the culture room at 75-85 F and conductivity measurements were taken weekly for up to 12 weeks .
  • the release rate was linear over time and fairly proportional over the doses of controlled release material tested (Figure 1A) .
  • Controlled release material was added to 60 ml DI water containing half strength Murashige-Skoog salts (0.5 M/S) to test whether external salt concentration af ⁇ fected release rate (Figure 2).
  • the release rate was substantially linear and identical to the release rate seen with the agar media or plain water, showing that controlled release rate is relatively independent of external salt or external water potential.
  • prills were- sorted "to ' •- exclude most of the irregular prills, culling 25-30% of the product in order to reduce the uncontrolled release.
  • prills were weighed into test tubes for autoclaving. When the prills were applied to medium, a spatula was used to dislodge prills stuck in the tube. Some prill breakage does occur, and this would bias the lower-dose treatments more than the higher doses.
  • Autoclaved prills were a different color from nonautoclaved prills even after sitting on agar or in water for weeks. The coating took on a darker brown color after autoclaving. However, there was no large or consistent effect of autoclaving on release rate in spite of the color difference.
  • autoclaved prills were designated 'A' and nonautoclaved prills 'NA'.
  • the controlled release nutrient used had a 17-9- 13 formulation.
  • the nutrients come from ammonium nitrate, ammonium phosphates, calcium phosphates, and potassium sulfates. This is a mixture of 60% 15-15-15 and 40% 21-7- 14 fertilizers.
  • the microprills of the controlled release materials weigh 2-3 mg each so in a 260 mg treatment there are about 100 prills. This number assures that about 60 prills are 15-15-15 and about 40 prills are 21-7-14. With larger prills and fewer prills/dose, there is less assur ⁇ ance of the proper mix of the two materials. This problem is not an issue with a single-component type fertilizer. The other reason for microprills is better distribution spread on or mixed throughout the medium.
  • Dose rates were chosen as follows: Level 1 was set at 260 mg which is equal to the weight of salt in 60 ml of M/S medium (4.3 g/L) . This level was referred to as IX. The other levels were 2X (520 mg) and 4X (1040 g) . The best dose is determined empirically—that which best enhances plant growth.
  • Nutrient depletion curves were generated from 3 crops (Ficus benjamina, Syngonium “White Butterfly", and Boston fern) ( Figures 3A, 3B and 3C) . There were 4-6 dif ⁇ ferent groups of each of these species grown in rotation such that one group was subcultured each week. Basically, medium conductivity was measured weekly, four dishes with duplicate readings per dish. Coefficients of variation between duplicate readings are commonly 1% and between repetitions of the same treatment about 10%. Culture vigor can be variable depending on explant size, prior history, and extent of endophytic bacteria. Consequently, greater variation can be expected between groups than within the same group measured week to week.
  • Endpoint conductivity can give a clue as to the proper length of the subculture interval. It appears that when plants are growing rapidly, they take up more salts, rather than if more salts are taken up, the plants grow more rapidly. The key, then, is to get plants to grow rapidly and keep them supplied with proper levels of salts. Second, the mineral uptake appears to increase rapidly during the first two weeks and decrease there ⁇ after. In the case of Ficus, the decrease may have more to do with the total depletion of the medium than a physiological response. Mineral uptake was estimated from the slope of the depletion curve.
  • Uptake Towards the end of sub ⁇ culture when salts .are no longer being taken up, the slope (uptake) approaches zero. Uptake is maximal when the conductivity of the medium is falling most rapidly. The variation is due to both sampling error and to extraneous factors affecting culture performance. Peak uptake seems to be correlated with the period of shoot multiplication which is followed by shoot elongation and leaf expansion. This may indicate a greater requirement for salts during the first half of the subculture interval. Alternatively, the duration of the multiplication phase might be limited by available salts.
  • the controlled release nutrient employed does not appear to be phytotoxic. It was added to plant cultures at three levels covering a 4-fold range and no injurious response, except due to salinity was noted. In fact, repeated enhancement of growth with added controlled release material shows that either there was no phytotoxicity or that any toxic effect was masked by sup ⁇ plying mineral elements . In the release kinetics experiments, and when taking endpoint conductivities, the pH tended to be buffered about 4.8-5.5, perhaps because of the phosphate in the material. Prills in direct contact with the plant (i.e., about the base, or when sprinkled on leaves, in leaf axils) did not produce any acute localized lesions at the point of contact.
  • the controlled-release nutrient can be used to supply nutrients to match plant needs in tissue culture as shown in Figures 4 and 5.
  • End point conductivity graphs show addition of between 260 and 520 mg of nutrient per 60 ml .over the subculture period maintains conductivity near the initial value. (Note: 260 mg of nutrient in this volume equals the nutrient levels achieved with standard levels of Murashige-Skoog salts.) This may be indicative of an optimal dose rate.
  • the uptake of salts by plants changes week by week whereas the release from the controlled release material is linear and constant. Although medium conductivity was not measured weekly in experiments, it can be estimated from the plant uptake curves and the nutrient release curves ( Figure 4).
  • the resulting salt profile is the sum of plant uptake and nutrient release.
  • conductivity of the medium tends to rise a little since the release rate exceeds plant uptake.
  • conductivity either rises only slightly or falls (depending on dose) when plant uptake peaks.
  • the uptake rate declines, the medium conductivity increases gain.
  • the controlled release product is best suited to crops that steadily take up salts with no sharp peaks or that have broader peaks of uptake.
  • a controlled release product can e custom formulated and blended to give second order release kinetics in order to accommodate peak uptake.
  • controlled release material c was added to media at both IX initial medium strength and ⁇ 1X initial medium strength. There were two reasons for doing this: 1) to accommodate any "quick" release from broken or imperfect prills and 2) to start the plants off at a lower salt level and build it up over time. When it g was determined that prill sorting could eliminate a great deal of the "quick" release, it became more desirable to use sorted prills since the mineral balance of M/S salts would be less altered.
  • One of the early premises was that the M/S formulation was optimized for "batch" cultures and 5 that initial strength of the medium was set higher than optimal in order to sustain growth over a 4-8 week period.
  • M/S is said to be a "high salt" medium for tissue culture. If so, then newly explanted material might do better on lower initial medium strength provided that nutrients are continuously supplied. Also, newly explanted or small pieces of tissue might be less tolerant of high salt levels whereas older, larger tissue pieces might tolerate higher salt levels (perhaps depending on surface area/ volume) . Then again, as indicated above, salt uptake seems to peak during weeks 2-3. Perhaps, peak uptake reflects peak need. Also, M/S may not necessarily have been set at a supraoptimal level; it might be set at a point of diminishing returns. Maybe some plants grow faster at 2-3X M/S but not so much faster as to economi ⁇ cally justify using more than IX M/S.
  • the safest course may be to maintain constant medium strength near starting levels. That way, there is a chance of eliminating a lag without creating another problem by adapting the plant to higher salt levels.
  • the macroelements are so called because of the large quantities utilized in plant growth. Micronutrients are required for proper growth but are taken up in small quantities.
  • the controlled release nutrient can supply macro-and minor elements which are likely to depleted in a closed system such as the tissue culture vessel. For this reason, it is not absolutely necessary that the quality or analysis of the fertilizer salts used in controlled release product be the same as in M/S salts.
  • the added materials are being introduced to the system only to sup- plement an M/S based medium, not to substitute for it.
  • When growth in vitro depletes major salts it become less important which salts are supplied than whether salts are allowed to become major limiting factors. While it is possible to formulate controlled release material to duplicate the relative proportions of M/S salts, it may be determined that this is not actually necessary or economi ⁇ cally justified in some cases.
  • Controlled release treated cultures were generally larger than those without treat ⁇ ment. The effect seemed greater at 3 density than at 5 density (opposite of what was expected) . The difference observed with added nutrient disappeared by week 8. 'Three-density' clumps outgrew those at 5 density after the fourth week. Conductivity readings showed that the higher density used up medium salts faster than the lower density, as expected.
  • the supply of controlled release nutrient elevated medium conductivity above that normally present at each time point. However, the 260 mg rate failed to maintain medium strength near the initial value. A 520 mg rate might have been more appropriate for the 5 density treatments. It is likely that light or space had become more limiting at week 8 than mineral supply.
  • the Boston fern produced about 18 g
  • Ficus benjamina produced about 30 g
  • potato produced about 10 g fresh weight.
  • the variation in carrying capacity is puzzling. But consider the initial weight of the explants: 2-3 g for Boston fern, 3-5 g for Ficus benjamina, and 0.2-0.5 g for potato. This makes about a 6-9 fold increase for Boston fern, a 6-10 fold increase for Ficus benjamina, and a 20-50 fold increase for potato.
  • controlled release fertilizer did not seem to increase the carrying capacity of the system. It did appear to promote faster growth in a given period of time, provided other limiting factors were not overriding it. It is possible that as limiting factors are removed, controlled release nutrient will have an even larger effect on plant growth. As noted, a significant ingredient to longer subculture interval is lower plant density. At lower density, the onset of limiting factors is delayed, permit ⁇ ting controlled release material to enhance the rate of growth. Hence, controlled release material is a factor in obtaining the highest possible multiplication in order to decrease labor and material costs in plant micropropagation.
  • the prills were readily sterilizeable, using a variety of techniques, without altering their properties.
  • the release mechanism was operative in pure water or in gels, such as agar. Release rates were constant irrespec ⁇ tive of the presence or absence of external salts. Sort ⁇ ing to remove nonspherical, broken or- uncoated prills eliminated most of the early, uncontrolled release which was initially observed.
  • explants can deplete tis ⁇ sue culture media of salts to less than 10% of the original amount. This depletion is dependent on length of subculture interval and correlates with the amount of tis- sue growth, i.e., if plants are growing vigorously, they will use more salts.
  • the number of prills needed to deliver controlled-release fertilizers was not excessive. Ap ⁇ plication of the proper dose of fertilizer prills needed to maintain conductivity near initial levels, resulted in a light to medium coverage of the media surface with prills.
  • the pH was buffered at about 5. With controlled release material addition, one can approximate chemostasis with regard to NPK nutrition and pH control.
  • the longev- ity of the coated product can be 70-90 days—enough to cover a 12 week subculture if desired. This has obvious implications for crops that produce storage organs (such as bulbs, corms, or tubers) .
  • Explant utilization of medium salts is variable. Factors such as species, prior growth history, and initial explant fresh weight can influence the amount of salt de ⁇ pletion. Where depletion is higher, the addition of controlled release material should have greater effect, and where depletion is lower, it may have only a small effect. There did not seem to be any phytotoxicity related to the prills or their contents. Diffusion did not seem to be a major limiting factor especially since plants take up salts over a period of several weeks.

Landscapes

  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

On ajoute des éléments nutritifs à libération régulée, à des environnements de culture de tissus afin de remplacer et d'ajouter les éléments nutritifs essentiels consommés par les tissus en croissance.
PCT/US1990/001333 1989-03-10 1990-03-12 Culture de tissus Ceased WO1990010691A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US32242189A 1989-03-10 1989-03-10
US322,421 1989-03-10

Publications (1)

Publication Number Publication Date
WO1990010691A1 true WO1990010691A1 (fr) 1990-09-20

Family

ID=23254807

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1990/001333 Ceased WO1990010691A1 (fr) 1989-03-10 1990-03-12 Culture de tissus

Country Status (1)

Country Link
WO (1) WO1990010691A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991002071A3 (fr) * 1989-08-09 1992-05-14 Dekalb Plant Genetics Procedes et compositions de production de plantes monocotyledones fecondes ainsi que de leurs cellules transformees de maniere stable
US5484956A (en) * 1990-01-22 1996-01-16 Dekalb Genetics Corporation Fertile transgenic Zea mays plant comprising heterologous DNA encoding Bacillus thuringiensis endotoxin
US5508468A (en) * 1990-01-22 1996-04-16 Dekalb Genetics Corporation Fertile transgenic corn plants
US5550318A (en) * 1990-04-17 1996-08-27 Dekalb Genetics Corporation Methods and compositions for the production of stably transformed, fertile monocot plants and cells thereof
US6946587B1 (en) 1990-01-22 2005-09-20 Dekalb Genetics Corporation Method for preparing fertile transgenic corn plants
US6960709B1 (en) 1993-08-25 2005-11-01 Dekalb Genetics Corporation Method for altering the nutritional content of plant seed
EP2055781A2 (fr) 1996-11-29 2009-05-06 Third Wave Technologies, Inc. Endonucléases FEN-1, mélanges et procédé de division
US7615685B2 (en) 1990-01-22 2009-11-10 Dekalb Genetics Corporation Methods of producing human or animal food from stably transformed, fertile maize plants

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3223518A (en) * 1961-08-18 1965-12-14 Archer Daniels Midland Granular fertilizer having a plurality of coatings and the process of making
US4019890A (en) * 1973-12-28 1977-04-26 Chissoasahi Fertilizer Co., Ltd. Method for producing coated fertilizer
US4369055A (en) * 1978-01-09 1983-01-18 Chissoasahi Fertilizer Co., Ltd. Coated granular fertilizer capable of controlling the effect of temperature upon dissolution-out rate
US4537860A (en) * 1982-12-08 1985-08-27 Monsanto Company Static cell culture maintenance system
US4657576A (en) * 1984-11-16 1987-04-14 Sierra Chemical Company Granular fertilizer composition having controlled release and process for the preparation thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3223518A (en) * 1961-08-18 1965-12-14 Archer Daniels Midland Granular fertilizer having a plurality of coatings and the process of making
US4019890A (en) * 1973-12-28 1977-04-26 Chissoasahi Fertilizer Co., Ltd. Method for producing coated fertilizer
US4369055A (en) * 1978-01-09 1983-01-18 Chissoasahi Fertilizer Co., Ltd. Coated granular fertilizer capable of controlling the effect of temperature upon dissolution-out rate
US4537860A (en) * 1982-12-08 1985-08-27 Monsanto Company Static cell culture maintenance system
US4657576A (en) * 1984-11-16 1987-04-14 Sierra Chemical Company Granular fertilizer composition having controlled release and process for the preparation thereof

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
COMMUNICATIONS IN SOIL SCIENCE AND PLANT ANALYSIS, (New York, New York), Volume 16(8), issued 1985, (YEAGER et al.); "Response of four woody ornamental Species to superphosphate and controlled-release fertilizers", pages 853-863, see the entire document. *
COMMUNICATIONS IN SOIL SCIENCE AND PLANT ANALYSIS, (New York, New York), Volume 17(1), issued 1986, (PILL et al.): "Tomato seedling growth in peat and peat-like blocks amended with hydrophilic polymer", pages 45, see the entire document. *
HORTSCIENCE (St. Joseph, Mo), Volume 20(6), issued December 1985, (DOUGHTY et al.), "Comparison of three Slow-release fertilizers in the production of seedling dwarf date palm", pages 1088-1089, see the entire document. *
HORTSCIENCE (St. Joseph, Mo), Volume 20(6), issued December 1985, (DOUGHTY et al.); "Evaluation of Liquid and Slow Release Fertilizer Applications in the Production of Dwarf Date Palm", pages 1090-1091, see the entire document. *
HORTSCIENCE (St. Joseph, Mo), Volume 22(5), issued October 1987, (NEUMAIER et al.); "Effect of light and fertilizer Rate and Source on flowering, growth, and quality of hibiscua", pages 902-904, see the entire document. *
HORTSUENCE (St. Joseph, Mo), Volume 18(2), issued April 1983, (POOLE et al.); "Establishment and Growth of In virtro-cultured Dieffenbachia", pages 185-187, see especially Experiments 3 and 4. *

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991002071A3 (fr) * 1989-08-09 1992-05-14 Dekalb Plant Genetics Procedes et compositions de production de plantes monocotyledones fecondes ainsi que de leurs cellules transformees de maniere stable
US5554798A (en) * 1990-01-22 1996-09-10 Dekalb Genetics Corporation Fertile glyphosate-resistant transgenic corn plants
US5508468A (en) * 1990-01-22 1996-04-16 Dekalb Genetics Corporation Fertile transgenic corn plants
US5538877A (en) * 1990-01-22 1996-07-23 Dekalb Genetics Corporation Method for preparing fertile transgenic corn plants
US5538880A (en) * 1990-01-22 1996-07-23 Dekalb Genetics Corporation Method for preparing fertile transgenic corn plants
US5484956A (en) * 1990-01-22 1996-01-16 Dekalb Genetics Corporation Fertile transgenic Zea mays plant comprising heterologous DNA encoding Bacillus thuringiensis endotoxin
US5780708A (en) * 1990-01-22 1998-07-14 Dekalb Genetics Corporation Fertile transgenic corn plants
US6946587B1 (en) 1990-01-22 2005-09-20 Dekalb Genetics Corporation Method for preparing fertile transgenic corn plants
US7064248B2 (en) 1990-01-22 2006-06-20 Dekalb Genetics Corp. Method of preparing fertile transgenic corn plants by microprojectile bombardment
US7615685B2 (en) 1990-01-22 2009-11-10 Dekalb Genetics Corporation Methods of producing human or animal food from stably transformed, fertile maize plants
US5550318A (en) * 1990-04-17 1996-08-27 Dekalb Genetics Corporation Methods and compositions for the production of stably transformed, fertile monocot plants and cells thereof
US6960709B1 (en) 1993-08-25 2005-11-01 Dekalb Genetics Corporation Method for altering the nutritional content of plant seed
US7547820B2 (en) 1993-08-25 2009-06-16 Dekalb Genetics Corporation Method for altering the nutritional content of plant seed
EP2055781A2 (fr) 1996-11-29 2009-05-06 Third Wave Technologies, Inc. Endonucléases FEN-1, mélanges et procédé de division

Similar Documents

Publication Publication Date Title
AU608743B2 (en) Hydrated, pregerminated seeds in gel capsules
O'neill et al. Elevated atmospheric CO2 effects on seedling growth, nutrient uptake, and rhizosphere bacterial populations of Liriodendron tulipifera L.
EP1096849B1 (fr) PROCEDE DE PRODUCTION ET D'ENSEMENCEMENT ET DE PROPAGATION ULTERIEURS $i(EX VITRO) D'EMBRYONS SOMATIQUES DE PLANTES PRE-GERMEES
EP3747267B1 (fr) Nouvelle souche methylobacterium sp. , compositions la comprenant et son utilisation en tant que biostimulant et bactérie fixant l'azote endophyte
Woodford et al. Preliminary results obtained with an apparatus for the study of salt uptake and root respiration of whole plants
WO1990010691A1 (fr) Culture de tissus
CN107047319B (zh) 一种银杏的组织培养方法及生根培养基
CN111492917A (zh) 一种高效鉴定水稻幼苗耐盐性能的方法
CN116904366A (zh) 一种嗜线虫沙雷氏菌、微生物包衣制剂及其应用
EP0275682A1 (fr) Procédé de multiplications de plantes bulbeuses
Leonard Method of Testing Legume Bacterial Cultures and Results of Tests of Commercial Inoculants in 1943
AU2021103083A4 (en) Method for identifying phosphorus activation efficiency of peanut germplasm under hydroponics
CN118909878B (zh) 一株碱蓬微杆菌7-6及其应用
US5543321A (en) Colored plant culture media
CN117660261A (zh) 一个增强水稻耐盐能力的根际促生菌及其应用
Song et al. Rapid assessment of acid phosphatase activity in the mycorrhizosphere and in arbuscular mycorrhizal fungal hyphae
CN118440837A (zh) 一株高产iaa且耐逆性优异的促生菌奇异虚构芽孢杆菌ypr-35及其应用
Tikhomirov et al. Light intensity and production parameters of phytocenoses cultivated on soil-like substrate under controled environment conditions
GB2152531A (en) Germination and sustained growth of orchid seeds
Woomer et al. A revised manual for rhizobium methods and standard protocols available on the project website
CN107099480A (zh) 一种发酵制备鸡血液体肥料的方法
Burton Legume inoculant production manual
RU2157065C2 (ru) Способ семенного размножения eustoma russelianum
CN109868228A (zh) 一株近玫色锁掷孢酵母及其应用
FUJIWARA et al. PHYSIOLOGICAL STUDIES OF PLANT ROOTS 1. INFLUENCES OF SOME ENVIRONMENTAL CONDITIONS ON THE GROWTH OF ISOLATED ROOTS OF RICE PLANT AND WHEAT

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB IT LU NL SE