Compositions and methods for controlling plant pests

Details Compositions and methods for controlling plant pests Compositions and methods for controlling plant pests

2001-11-27

2003-10-28

Tate, Christopher R. (Department: 1651)

Drug, bio-affecting and body treating compositions

Designated organic active ingredient containing

Peptide containing doai

C514S015800, C424S093460, C424S093462, C424S405000

Reexamination Certificate

active

06638910

ABSTRACT:

FIELD OF THE INVENTION
The present invention is in the field of biopesticides. More particularly, this invention relates to the finding that a novel strain of
Bacillus subtilis
, AQ713, can inhibit a broad range of fungal and bacterial plant diseases and also have activity against insects. The invention also relates to fungicidal, bactericidal, and insecticidal compositions comprising this novel Bacillus strain and the antibiotics and metabolites produced by this strain either alone, or in combination with other chemical and biological pesticides.
BACKGROUND OF THE INVENTION
For a number of years, it has been known that various microorganisms exhibit biological activity so as to be useful to control plant diseases. Although progress has been made in the field of identifying and developing biological pesticides for controlling various plant diseases of agronomic and horticultural importance, most of the pesticides in use are still synthetic compounds. Many of these chemical fungicides are classified as carcinogens by the EPA, are toxic to wildlife and other non-target species. In addition, pathogens may develop resistance to chemical pesticides (see, e.g., Schwinn et al., p. 244, ADVANCES IN PLANT PATHOLOGY: PHYTOPHTHORA INFESTANS, THE CAUSE OF LATE BLIGHT OF POTATO (Academic Press, San Diego 1991).
Every year 250-300 million dollars of chemical pesticides are used to control corn rootworm infestations. Many of these chemical pesticides are toxic to humans, wildlife and other nontarget species. Also some have been found in the ground water. New chemical insecticides cost 100 million to develop.
Biological control offers an attractive alternative to synthetic chemical fungicides. Biopesticides (living organisms and the naturally produced compounds produced by these organisms) can be safer, more biodegradable, and less expensive to develop.
Biopesticides developed from microorganisms are highly desired for integrated pest management programs in agriculture, public health and urban settings. One commonly used biopesticide is the gram positive bacterium
Bacillus thuringiensis
. Pesticidal
B. thuringiensis
strains are known to produce crystal proteins during sporulation, which are specifically toxic to certain orders and species of insects and nematodes (See, e.g., U.S. Pat. No. 4,999,192 and U.S. Pat. No. 5,208,017). Proteinaceous endotoxins produced by
B. thuringiensis
also act as insecticidal agents against corn rootworm and other beetles (e.g., U.S. Pat. No. 5,187,09; Johnson, T. J. et al. (1993),
J. Economic Entomology
, 86: 330-333).
B. thuringiensis
endotoxins have been shown to be effective as purified crystals, washed cell pellets, and expressed proteins. Warren et al. (WO 96/10083), discloses non-endotoxin proteins produced during the vegetative stage of
Bacillus cereus
and
B. thuringiensis
. These vegetative proteins, called Vip1 and Vip2 have potent activity against corn rootworm (northern and western) (Estruch et al. (1997),
Nature Biotechnology
15:137-141 and Mullins et al. (1997),
Appl. Environ. Microbiol
. 63, (in press).
One
B. thuringiensis
thermostable metabolites, termed beta-exotoxin has also been shown to have pesticidal properties. Burgjeron and Biache (1979),
Entomophaga
11:279-284 report a beta exotoxin that is active against Colorado potato beetle (
Leptinotarsa decemlineata
). In addition, the known
B. thuringiensis
betaexotoxins exhibits non-specific pesticidal activity, killing not only nematodes, but also flies, armyworms, mites, and corn rootworms. Sigma exotoxin has a structure similar to beta-exotoxin, and is active against Colorado potato beetle (Argauer et al. (1991)
J. Entomol. Sci
. 26:206-213). Alpha-exotoxin is toxic against larvae of
Musca domestica
(Cluthy (1980)
FEMS Microbiol. Lett
. 8:1-7). Gamma-exotoxins are various proteolytic enzymes, chitinases and proteases. The toxic effects of gamma exotoxins are only expressed in combination with beta-exotoxin or delta-endotoxin. Forsberg et al. (1976) “
Bacillus thuringiensis
: Its effects in Environmental Quality,” National Research Council of Canada. Stonard et al. (1994)
ACS Symposium Series
551:25 report a water-soluble secondary metabolite active against corn rootworm in the supernatant of a
Bacillus cereus
strain.
There are no documented strains of
Bacillus subtilis
with broad spectrum insecticidal activity.
Screening programs have identified certain Bacillus spp. (Bacillus spp. includes
B. subtilis, B. cereus, B. mycoides, B. thuringiensis
) strains that exhibit antifungal activity. (See e.g. Stabb et al. (1990)
Applied Environ. Microbiol
. 60: 4404-4412). These strains have been shown to produce zwittermicin-A and or kanosamine (Milner et al. (1996)
Appl. Environ. Microb
. 62: 3061-3066), two antibiotic agents that are effective against the soil borne disease damping off, caused by
Phytophthora medicaginis, P. nicotianae, P. aphanidermatum
or
Sclerotinia minor
(See Stabb et al., supra). Zwittermicin-A is a water soluble, acid stable linear aminopolyol molecule (see, He et al, (1994)
Tetra. Lett
. 35 (16) 2499-2502.
U.S. Pat. No. 5,049,379 to Handelsman et al. describes how zwittermicin-A produces damping off in alfalfa and soybeans. When the seed was coated with
B. cereus
ATCC 53522, the pathogenic activity of root rot fungus is inhibited. Similarly application of spore-based formulations of certain
B. cereus
strains to soybean seeds or the soil surrounding the seeds has been shown to improve soybean yield at field sites. (See, Osburne et al. (1995)
Am. Phytopathol. Soc
. 79(6): 551-556). Methods of applying biopesticides are well known in the art and include, for example, wettable powders, dry flowables, microencapsulation of effective agents, liquid or solid formulations of antibiotic fractions from suitable cultures. (See e.g., U.S. Pat. No. 5,061,495 to Rossall or U.S. Pat. No. 5,049,379 to Handelsman).
Smith et al. (1993)
Plant Disease
77(2) 139-142 report that the activity of the soil-borne fungus,
Pythium aphanidermatum
, that causes cottony cucumber leak can be suppressed using zwittermicin-producing
B. cereus
strain UW85. Leifert et al. (1995)
J. Appl. Bacteriol
. 78: 97-108 report that the production of anti-Botrytis and anti-Alternaria antibiotics by two Bacillus strains,
B. subtilis
CL27 and
B. pumilis
CL 45. The whole broth and cell-free filtrates were active against Botrytis and Alternaria in in vitro tests and were active against Botrytis in in vivo small plant tests on Astilbe. Leifert et al. (1997) U.S. Pat. No. 5,597,565 disclose
B. subtilis, B. pumilis
, and
B. polymyxa
that are particularly effective at inhibiting post harvest disease causing fungi. They also disclose the presence of antibiotics produced in the cell-free culture filtrate and their activity at different pH values, but they do not identify these compounds.
Rossall (1994) U.S. Pat. No. 5,344,647 discloses
Bacillus subtilis
strains with broad anti-fungal activity. Sholberg et al. (1995)
Can. J. Microbiol
. 41: 247-252, Swinburne et al. (1975)
Trans. Brit. Mycol. Soc
. 65: 211-217, Singh and Deverall (1984)
Trans. Br. Mycol. Soc
. 83: 487-490, and Ferreira, et al. (1991)
Phytopathology
81: 283-287. Baker et al. (1983)
Phytopathology
73: 1148-1152 disclose the use of Bacillus spp. and
Bacillus subtilis
as biocontrol agents of fungal plant pathogens. Baker et al. (1983)
Phytopathology
73: 1148-1152 also report on an antifungal
Bacillus subtilis
for use on plant pathogens. Pusey et al. (1988)
Plant Dis
. 72: 622-626, Pusey and Robins (U.S. Pat. No. 5,047,239), and McKeen et al. (1986)
Phytopathology
76: 136-139 disclose control of post harvest fruit rot using
B. subtilis
. McKeen et al, supra, have shown that antibiotics similar to the low molecular weight iturin cyclic polypeptides contribute to this fungicidal activity of
B. subtilis.
Liu et al. (1995) U.S. Pat. No. 5,403,583 disclose a
Bacillus megaterium
, ATCC 55000 and a method to control the fungal plant pathogen,
Rhizoctonia solani
. Islam and Nandi (1985)
Journal of Plan

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