Bacillus mycoides strain for controlling insects

Details Bacillus mycoides strain for controlling insects Bacillus mycoides strain for controlling insects

1998-08-19

2001-04-03

Lankford, Jr., Leon B. (Department: 1651)

Drug, bio-affecting and body treating compositions

Whole live micro-organism, cell, or virus containing

Bacteria or actinomycetales

C435S041000, C435S252600, C504S116100

Reexamination Certificate

active

06210665

ABSTRACT:

FIELD OF THE INVENTION
This invention is in the field of biopesticides. More particularly, the present invention describes a novel, pesticidal strain of
Bacillus mycoides
that is active against insects such as corn rootworm, e.g.,
Diabrotica virgifera, D. longicornis, D. undecimpunctata
and aphids such as the green peach aphid,
Myzus persicae
. The novel
Bacillus mycoides
strain also produces a metabolite in the supernatant of a whole broth culture that is useful as a biocontrol agent in the treatment and prevention of insect infestation of plants.
BACKGROUND OF THE INVENTION
Every year 250-300 million dollars of chemical pesticides are used to control insect infestations. Many of these chemical pesticides are toxic to humans, wildlife and other nontarget species. In addition, some of these pesticides have been found in ground water. New chemical pesticides cost $100 million to develop.
Biological control offers an attractive alternative to synthetic chemical pesticides. Biopesticides (living organisms and the naturally-occurring compounds produced by these organisms) can be safer, more biodegradable, and less expensive to develop.
One commonly used biopesticide is the gram-positive bacterium
Bacillus thuringiensis
. Pesticidal
B. thuringiensis
strains are known to produce crystal proteins during sporulation that 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. 5,187,091, and Johnsonetal. (1993)
J. Economic Entomology
, 86: 330-333).
B. thuringiensis
endotoxins have been shown to be effective pesticides in the form of 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, designated Vip1 and Vip2, have potent insecticidal 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-metabolite designated 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
beta-exotoxins exhibit 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 exhibits pesticidal activity against Colorado potato beetle (Argauer et al. (1991)
J. Entomol. Sci
. 26:206-213). Alpha-exotoxin is toxic to larvae of Musca domestics (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 exhibiting pesticidal activity against corn rootworm in the supernatant of a
Bacillus cereus
strain.
There are no documented strains of
Bacillus mycoides
that produce metabolites exhibiting pesticidal activity against insects such as corn rootworms and aphids. Moreover, there are no known water-soluble metabolites produced by
Bacillus mycoides
with pesticidal activity against insects such as corn rootworms and aphids.


REFERENCES:
patent: 4999192 (1991-03-01), Payne et al.
patent: 5187091 (1993-02-01), Donovan et al.
patent: 5208017 (1993-05-01), Bradfisch et al.
patent: WO 96/10083 (1996-04-01), None
Argauer et al., “Evidence for a novel insecticidally active exotoxin produced by the HD 116 strain ofBacillus thuringiensisvar.morrisoni1”J. Entomol. Sci.26(2):206-213 (1991).
Burgjeron and Biache, “Alimentation au laboratoire dePerillus bioculatusFABR.avec des larves deLeptinotarsa decemlineatasay intoxiqu&igr;es par la toxine thermostable deBacillus thuringiensis berlinger”Entomophaga11(3):279-284 (1966).
Estruch et al., “Transgenic plants: An emerging approach to pest control”Nature Biotechnol.15:137-141 (1997).
Johnson et al., “Insecticidal activity of EG4961, a novel strain ofBacillus thuringiensistoxic to larvae and adults of southern corn rootworm (coleoptera: Chrysomelidae) and colorado potato beetle (coleoptera: Chrysomelidae)”J. Econ. Entomol.86(2):330-333 (1993).
Lüthy, “Insecticidal toxins ofBacillus thuringiensis”FEMS Microbiol. Lett.8:1-7 (1980).
Marrone et al., “Improvements in laboratory rearing of the southern corn rootworm,Diabrotica undecimpuncta howardibarber (coleoptera: Chrysomelidae), on an artificial diet and corn”J. Econ. Entomol.78(1):290-293 (1985).
Miller, “Single derivatization method for routine analysis of bacterial whole-cell fatty acid methyl esters, including hydroxy acids”J. Clin. Microbiol.16(3):584-586 (1982).
Stonard et al., “Microbial secondary metabolites as a source of agrochemicals”Amer. Chem. Soc. Symposium Series551:25-36 (1994).
Yu et al., “TheBacillus thuringiensisvegatative insecticidal protein vip3A lyses midgut epithelium cells of susceptible insects”Appl. Environ. Microbiol.63(2):532-536 (1997).

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