US 2015O135371 A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0135371 A1 LeVeau et al. (43) Pub. Date: May 14, 2015 (54) SYNERGY-BASED BIOCONTROL OF PLANT Related U.S. Application Data PATHOGENS (60) Provisional application No. 61/902,046, filed on Nov. (71) Applicant: The Regents of the University of 8, 2013. California, Oakland, CA (US) Publication Classification (72) Inventors: Johannes Henricus Josephus LeVeau, (51) Int. Cl. Davis, CA (US); Hung Kim Doan, AOIN 63/00 (2006.01) Davis, CA (US) (52) U.S. Cl. CPC ...................................... A0IN 63/00 (2013.01) (21) Appl. No.: 14/537,620 (57) ABSTRACT Methods and compositions are provided for control of patho 22) Filed1C Nov.OV. 10,U, 2014 genicen1C fungalTungal Or OOmVCetOuSy 1infecti nect1On. Patent Application Publication May 14, 2015 Sheet 1 of 5 US 2015/O135371 A1 FIG. 1 Patent Application Publication May 14, 2015 Sheet 2 of 5 US 2015/O135371 A1 FIG 2 Patent Application Publication May 14, 2015 Sheet 3 of 5 US 2015/O135371 A1 Ca33 at Serenades Ca35 and o: - - - --- Ca35 and Seenace & Serenade , * Cat35 and Serenade : Cat35 and Serenade re FIG 3 Patent Application Publication May 14, 2015 Sheet 4 of 5 US 2015/O135371 A1 8. Ca35 and Serenade -> 8 ::::::::::::::::::::: F.G. 4 Patent Application Publication May 14, 2015 Sheet 5 of 5 US 2015/O135371 A1 FIG. 5 US 2015/O 135371 A1 May 14, 2015 SYNERGYBASED BOCONTROL OF PLANT the part of said plant contacted with bacteria of the genus PATHOGENS Collimonas or a product thereof is the seed of said plant. In Some embodiments, the part of said plant contacted with CROSS-REFERENCES TO RELATED bacteria of the genus Bacillus or a product thereof is the seed APPLICATIONS of said plant. In some embodiments, the part of said plant 0001. This application claims priority to U.S. Provisional contacted with bacteria of the genus Collimonas or a product Application 61/902,046, filed Nov. 8, 2013, the contents of thereof is the foliage of said plant. In some embodiments, the which are hereby incorporated in their entirety for all pur part of said plant contacted with bacteria of the genus Bacillus poses. or a product thereof is the foliage of said plant. 0007. In some embodiments, the contacting of a) com BACKGROUND OF THE INVENTION prises contacting the plant with bacteria of the genus Colli monas and a product thereof. In some embodiments, the 0002 Fungal and oomycetous pathogens can cause sig contacting ofb) comprises contacting the plant with bacteria nificant damage to a wide variety of commercially important of the genus Bacillus and a product thereof. In some embodi plant varieties including crops and ornamental plants. Such ments, the contacting of a) comprises contacting the plant pathogens can kill plants, reduce yield, reduce plant strength with bacteria of the genus Collimonas and a product thereof, (e.g., decrease resistance to lodging), cause symptoms of and the contacting ofb) comprises contacting the plant with mineral deficiency, and predispose plants to infection by other pathogens. As such, there is great interest in developing bacteria of the genus Bacillus and a product thereof. compositions and methods for control of fungal and oomyc 0008. In some embodiments, the fungal or oomycetous etous pathogens. infection comprises an infection by a fungal pathogen from a 0003) Fungal and oomycetous pathogens are typically class selected from the group consisting of Plasmodiophoro controlled by use of synthetic chemicals (e.g., fungicides). mycetes, Oomycetes, Chytridiomycetes, Zygomycetes, However, effective synthetic chemicals can be dangerous, Ascomycetes, Basidiomycetes, Deuteromycetes, Sordari toxic, and expensive. In some countries, certain fungicides or omycetes, and combinations thereof. In some cases, the fun anti-Oomycetous chemicals have been restricted or banned gal or oomycetous pathogen is from the class Oomycetes. In for these and other reasons. Therefore, there is great interest Some embodiments, said fungal or oomycetous infection in developing fungal control methods and compositions that comprises infection by a fungal pathogen from a genus do not rely on synthetic chemical fungicides or anti-Oomyc selected from the group consisting of Fusarium, Geotrichum, etous compounds, or reduce the use of Such chemicals. Aspergillus, Alternaria, Botryosphaeria, Colletotrichum, 0004 Biocontrol agents are a promising candidate for Magnaporthe, Verticillium, Cryphonectria, Botrytis, reducing or eliminating the need for Such chemicals. Biocon Monilinia, Sclerotium, Rhizoctonia, and combinations trol agents are typically microorganisms, such as bacteria, or thereof. In some embodiments, said fungal or oomycetous one or more products thereof, that are applied to a plant or a infection comprises infection by an Oomycete from a genus plant propagation material (e.g., Soil) to control a pathogen. selected from the group consisting of Pythium, Phytophthora, However, there are a limited number of commercially avail and combinations thereof. able biocontrol agents. For these and other reasons, there 0009. In some embodiments, said bacteria of the genus remains a need to further develop methods and compositions Bacillus comprise Bacillus subtilis. In some embodiments, containing biocontrol agents for the control of pathogenic said bacteria of the genus Bacillus comprise Bacillus subtilis fungi and Oomycetes. var. amyloliquefaciens. In some embodiments, said bacteria of the genus Bacillus comprise Bacillus subtilis QST 713. In BRIEF SUMMARY OF THE INVENTION Some embodiments, said bacteria of genus Collimonas com prise a species selected from the group consisting of Colli 0005. In a first aspect, the present invention provides a monas arenae, Collimonas fingivorans, and Collimonas method of cultivating a plant resistant to a fungal or oomyc pratensis. In some embodiments, said bacteria of genus Col etous infection comprising the steps of: a) contacting said limonas comprise C. arenae Cal35. In some embodiments, plant with bacteria of the genus Collimonas, or a product said bacteria of genus Collimonas comprise C. arenae Cal35 thereof, and b) contacting said plant with bacteria of the genus and said bacteria of the genus Bacillus comprise Bacillus Bacillus, or a product thereof, wherein said plant shows a greater resistance to the infection or exhibits reduced symp subtilis QST 713. toms of fungal or oomycetous infection relative to a non 0010. In some embodiments, said bacteria of the genus contacted plant. In some embodiments, the plant is cultivated Collimonas or a product thereof are in the form of a liquid in Soil or other planting media that does not otherwise contain Suspension. In some embodiments, said bacteria of the genus bacteria of the genus Collimonas. In some embodiments, said Collimonas are in a liquid Suspension at a concentration of plant is a plant selected from the group consisting of tomato, approximately 1x10° cells per milliliter. In some embodi potato, Sweet potato, cassava, beets, ginger, horseradish, rad ments, said bacteria of the genus Bacillus or a product thereof ish, ginseng, turnip, any root or tuber crop, pepper, eggplant, are in the form of a liquid Suspension. In some embodiments, ground cherry, tomatillo, okra, other fruiting vegetables, said bacteria of the genus Bacillus are in a liquid Suspension cucumber cantaloupe, melon, muskmelon, squash, water with a concentration of approximately 1x10 or 1x10 colony melon and other cucurbit plants. forming units per gram. 0006. In some embodiments, the part of said plant con 0011. In some embodiments, said contacting of a) or b) tacted with bacteria of the genus Collimonas or a product comprises a root dip. In some embodiments, said contacting thereof is a root of said plant. In some embodiments, the part of a) or b) comprises a soil drench. In some embodiments, of said plant contacted with bacteria of the genus Bacillus or said contacting of a) and b) comprises a root dip. In some a product thereof is a root of said plant. In some embodiments, embodiments, said contacting of a) and b) comprises a soil US 2015/O 135371 A1 May 14, 2015 drench. In some embodiments, said contacting of a) and b) are endosperm, and seed coat) and fruit (the mature ovary), plant independently selected from the group consisting of root dip, tissue (e.g. vascular tissue, ground tissue, and the like) and soil drench, and spray. cells (e.g. guard cells, egg cells, trichomes and the like), and 0012. In a second aspect, the present invention provides a progeny of same. The class of plants that can be used in the plant cultivated by any of the foregoing methods. In some method of the invention is generally as broad as the class of embodiments, the present invention provides a plant in con higher and lower plants amenable to cultivation, including tact with bacteria from the genus Bacillus or a product angiosperms (monocotyledonous and dicotyledonous thereof, and bacteria from the genus Collimonas, or a product plants), gymnosperms, and ferns. It includes plants of a vari thereof. In some embodiments, the plant is cultivated in soil ety of ploidy levels, including aneuploid, polyploid, diploid, that does not otherwise contain bacteria of the genus Colli haploid and hemizygous. Specific embodiments of plants FiOS useful for the methods and compositions of the present inven 0013. In a third aspect, the present invention provides a tion include but are not limited to commercial food crops, and composition of matter comprising: a) bacteria of the genus energy crops. Collimonas or a product thereof, and b) bacteria of the genus 0019. The invention has use over broad range of plants, Bacillus or a product thereof. In some embodiments, the including species from the genera Anacardium, Arabidopsis, composition comprises bacteria of the genus Collimonas and Arachis, Asparagus, Atropa, Avena, Brassica, Citrus, Citrul a product thereof. In some embodiments, the composition lus, Capsicum, Carthamus, Cocos, Coffea, Cucumis, Cucur comprises bacteria of the genus Bacillus and a product bita, Daucus, Elaeis, Fragaria, Glycine, Gossypium, Helian thereof.