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Plant seed assemblies comprising fungal/bacterial antagonists

a technology of fungal/bacterial and seed assembly, which is applied in the field of plant seed assembly comprising a fungal/bacterial antagonist combination, can solve the problems of low yield of hybrids, no chemical or biological fungicide available, and loss of crops, so as to achieve effective control, prolong shelf life, and control the effect of root and stalk ro

Inactive Publication Date: 2013-09-26
NOVOZYMES BIOAG AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0014]One advantage of the invention is that root and stalk rot can be controlled with a composition that is not toxic to humans. Another advantage of the invention is that root and stalk rot can be controlled more economically than with chemical fungicides. Yet another advantage of the invention is that it provides a biocontrol agent or bio-pesticide with extended shelf life. Thus, a seed can be treated with the biocontrol agent and stored for a period of months and still host a viable biocontrol agent that will colonize the root when the seed is placed in the ground, germinates and grows. Furthermore, the disclosed biocontrol agent is competitive with natural soil microbes that occur in the rhizosphere while providing pathogen protection for the plant. A further advantage of the invention is that the combination of a fungal / bacterial antagonist is more effective in controlling fungal pathogens in the plant rhizosphere than either a fungal antagonist or a bacterial antagonist alone. Thus, the invention provides an easy-to-use, effective means of controlling plant pathogens that have been only been controllable by rotation management. A further advantage of the invention is that its use produces more consistent results than the use of either a fungal antagonist or a bacterial antagonist alone, as shown by the Working Examples presented herein. In fact, use of the antagonist combinations disclosed herein is shown to be functional when use of its individual constituent antagonists is not.
[0015]The compositions disclosed herein may be integrated into Integrated Pest Management (IPM) programs, the inventive compositions may be used in combination with other management systems. As an alternative to synthetic agents, biocontrol agents (bio-pesticides) offer the advantage of containing naturally derived constituents that are safe to both humans and the environment. Specifically, bio-pesticides offer such advantages as being inherently less toxic than conventional pesticides, generally affecting only the target pest and closely related organisms, and are often effective in very small quantities. For these reasons, bio-pesticides often decompose quickly and, therefore, are ideal for use as a component of Integrated Pest Management (IPM) programs.
[0016]The applicant has shown through a variety of laboratory and field trials that Bacillus amyloliquefaciens TJ 1000 and Trichoderma virens G1-3 are compatible with one another and that they act synergistically to consistently produce increased yield in plants. These results were presented in the parent application referenced above.
[0017]Field trials were conducted as part of the applicant's continuing research effort that tested other known Bacillus amyloliquefaciens strains and other known Trichoderma virens isolates. The purpose of testing was to determine whether the surprising synergism between a Bacillus amyloliquefaciens bacterium and a Trichoderma virens fungus disclosed in the parent application would be present between other strains and isolates of the same genus and species.
[0018]This testing by the applicant did result in the discovery of a synergistic activity between other isolates and strains of Trichoderma virens and Bacillus amyloliquefaciens. These results are presented in the final three working examples at the end of this document. The results show that other isolates of Trichoderma virens and other strains of Bacillus amyloliquefaciens do have synergistic properties. The applicant's research has also confirmed that the combination of T. virens G1-3 and Bacillus amyloliquefaciens TJ 1000 is superior to combinations comprising any other tested strains, but that synergies among other combinations do exist. These synergies have led the applicant to the conclusion that his patent rights should include combinations of all Trichoderma virens isolates and all Bacillus amyloliquefaciens strains.
[0019]In illustrative embodiments, the invention is an inoculum, a seed coated with the inoculum, a plant protected with the inoculum, a method of producing the inoculum and a method of protecting a seed or a plant with the inoculum. A further embodiment of the inoculum comprises a combination of a fungus and a bacterium. Preferably, the fungus is a species of Trichoderma and the bacterium is a species of Bacillus, preferably a spore-forming strain of Bacillus. More preferably, the fungus is Trichoderma virens and the bacterium is Bacillus amyloliquefaciens, although other combinations are also envisioned. Even more preferably, the fungus is Trichoderma virens G1-3 (ATCC 58678) or Trichoderma virens Gl-21 (an isolate that is commercially available from Thermo Trilogy Corporation) and the bacterium is Bacillus amyloliquefaciens TJ 1000 or 1 BE (ATCC BAA-390) or Bacillus amyloliquefaciens FZB24 (a strain that is commercially available from Earth Biosciences, Inc.).

Problems solved by technology

Early and late season stalk and root rot are major causes of crop loss.
There is no chemical or biological fungicide available that addresses the problem of late season root and stalk rot in corn.
Corn growers can also select hybrids that have better “standability,” but such hybrids usually have lower yields.
Unfortunately, the corn varieties with the highest yields are usually those most susceptible to late season root and stalk rot.
Furthermore, it has little or no shelf life unless refrigerated at 4 degrees Centigrade at a minimum of 20 percent moisture.

Method used

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  • Plant seed assemblies comprising fungal/bacterial antagonists
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Examples

Experimental program
Comparison scheme
Effect test

first greenhouse working example

[0073]Greenhouse testing was conducted to determine the effectiveness of the disclosed biocontrol agents. Treated and untreated corn seeds were grown in soil infested with seven percent Fusarium infested wheat seed. In this testing, the following treatment codes were used:

[0074]CONTROL—Nothing on the seed

[0075]TJ 1000—Bacillus amyloliquefaciens TJ1000 or IBE

[0076]TJ 0300—Trichoderma virens G1-3

[0077]TJ 1300—50 / 50 combination of Trichoderma virens G1-3 and Bacillus amyloliquefaciens TJ1000 or 1BE

[0078]TJ 1310—competitive culture of Trichoderma virens G1-3 and Bacillus amyloliquefaciens TJ1000 or 1BE, resulting in a 70 / 30 ratio of Trichoderma to Bacillus

[0079]The results of greenhouse testing are presented in Table 0. The rating scale used was 9=worst plant protection and 1=best plant protection. Seed treated with biocontrol organisms grown in competitive culture showed an increase in plant protection over seed treatments with the same biological control organisms grown in non-compet...

second greenhouse working example

[0085]Greenhouse Methods: All test cultures were grown in MYE (three percent Molasses, 0.5 percent Yeast Extract) broth for five days. Bacteria were grown up individually (non-competitive) and with T. virens G1-3 (competitive culture). T. virens G1-3 was also grown in a non-competitive culture for testing. T. virens G1-3 and test bacteria grown in non-competitive culture were also applied to the same seed to test the effectiveness of non-competitive culture versus competitive culture. Corn seeds were treated to give a final concentration of 1×109 bacteria / fungal spores (may also be referred to a Colony Forming Units or CFU) per acre. Seed treatment was done with a Gustafson Benchtop Seed Treater, Model BLT. Seeds were grown in soil infested with seven percent Fusarium-infested wheat seed. After four weeks, plant heights were taken as well as plant biomass. Plant heights were taken by measuring from the soil line to the tallest leaf, biomass of the plants was taken by cutting the pla...

trial working example

Combinations Field Trial Working Example

[0099]Materials and Methods: A field trial was conducted using the corn variety NK 3030Bt to using the following biological treatments of the seed at a rate of approximately 106 CFU per seed. The seed was planted at a seeding rate of 25,000 seeds per acre in 30-inch rows in a randomized, replicated block. Each entry was replicated four times. The pathogen levels were natural populations at a location near Groton, S. Dak. The entries were as follows:

[0100]Control: Maxim Seed treatment (Maxim is a trademark of Syngenta Crop Protection)

[0101]TJ 1000—Bacillus amyloliquefaciens TJ 1000 or 1 BE

[0102]TJ 0300—Trichoderma virens G1-3

[0103]TJ 1300—50 / 50 combination of B. amyloliquefaciens TJ1000 or 1BE and T. virens G1-3

[0104]TJ 1310—Coculture 30 / 70 combination of B. amyloliquefaciens TJ1000 or 1 BE and T. virens G1-3

[0105]TJ 66 / 300—50 / 50 combination of Bacillus lentimorbus and T. virens G1-3

[0106]Results: The trial produced significant yield response o...

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Abstract

A seed treated with a fungal / bacterial antagonist combination and a seed assembly comprising a seed and a fungal / bacterial antagonist combination. The fungal / bacterial antagonist combination comprises a Trichoderma virens fungal antagonist and a Bacillus amyloliquefaciens bacterial antagonist for controlling plant pathogens as a biocontrol agent, bio-pesticide or bio-fungicide. In preferred embodiments, the invention produces an increase in plant yield. Control of early and late season stalk and root rot caused by fungi such as Fusarium, Phythium, Phytophthora and Penicillium in tomatoes, peppers, turf grass, soybeans, sunflower, wheat and corn is achieved.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a divisional of U.S. patent application Ser. No. 12 / 228,696, filed Aug. 16, 2008, which is a divisional of U.S. patent application Ser. No. 10 / 940,036, filed Sep. 13, 2004, now U.S. Pat. No. 7,429,477, which is a continuation-in-part of U.S. patent application Ser. No. 10 / 067,185, filed Feb. 1, 2002, now U.S. Pat. No. 6,808,917, which claims the benefit of U.S. Provisional Application No. 60 / 265,998, filed Feb. 2, 2001; the disclosures of which applications and patents are incorporated by reference as if fully set forth herein. The application also incorporates by reference the disclosure of U.S. Patent Application Publication No. US 2005-0096225 A1 as if fully set forth herein.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]The U.S. Government has a paid-up license in this invention and the right in limited circumstances to require the patent owner to license others on reasonable terms as provided...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01N63/04A01N63/02A01N63/22A01N63/38
CPCA01N63/00A01C1/06A01N63/04A01N63/02A01N2300/00A01N63/22A01N63/38
Inventor JOHNSON, THOMAS D.
Owner NOVOZYMES BIOAG AS
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