Mechanism of Insecticidal Action: Use of Spores and Proteins in Pest Control

Bacillus thuringiensis
(or Bt) is a Gram-positive, soil-dwelling bacterium, commonly used as

a biological pesticide. B. thuringiensis also occurs naturally in the gut of caterpillars of various types
of moths and butterflies, as well on leaf surfaces, aquatic environments, animal feces, insect-rich
environments, and flour mills and grain-storage facilities.[1][2]
During sporulation, many Bt strains produce crystal proteins (proteinaceous inclusions), called δ-
endotoxins, that have insecticidal action. This has led to their use as insecticides, and more recently
to genetically modified crops using Bt genes, such as Bt corn.[3] Many crystal-producing Bt strains,
though, do not have insecticidal properties.[4]
Mechanism of insecticidal action

Upon sporulation, B. thuringiensis forms crystals of proteinaceous insecticidal δ-endotoxins (called
crystal proteins or Cry proteins), which are encoded by cry genes. In most strains of B. thuringiensis,
the cry genes are located on a plasmid (cry is not a chromosomal gene in most strains).
Cry toxins have specific activities against insect species of the orders Lepidoptera (moths and
butterflies), Diptera (flies and
mosquitoes), Coleoptera (beetles), Hymenoptera (wasps, bees, ants and sawflies) and nematodes .
Thus, B. thuringiensis serves as an important reservoir of Cry toxins for production of biological
insecticides and insect-resistant genetically modified crops. When insects ingest toxin crystals, their
alkaline digestive tracts denature the insoluble crystals, making them soluble and thus amenable to
being cut with proteases found in the insect gut, which liberate the toxin from the crystal. The Cry
toxin is then inserted into the insect gut cell membrane, paralyzing the digestive tract and forming a
pore. The insect stops eating and starves to death; live Bt bacteria may also colonize the insect
which can contribute to death. The midgut bacteria of susceptible larvae may be required for B.
thuringiensis insecticidal activity.
In 1996 another class of insecticidal proteins in Bt was discovered; the vegetative insecticidal
proteins (Vip). Vip proteins do not share sequence homology with Cry proteins, in general do not
compete for the same receptors, and some kill different insects than do Cry proteins.
In 2000, a novel functional group of Cry protein, designated parasporin, was discovered from
noninsecticidal B. thuringiensis isolates.[20] The proteins of parasporin group are defined as B.
thuringiensis and related bacterial parasporal proteins that are not hemolytic, but capable of
preferentially killing cancer cells.[21] As of January 2013, parasporins comprise six subfamilies (PS1 to
PS6).[22]
Use of spores and proteins in pest control[edit]

Spores and crystalline insecticidal proteins produced by B. thuringiensis have been used to control
insect pests since the 1920s and are often applied as liquid sprays.[23] They are now used as
specific insecticides under trade names such as DiPel and Thuricide. Because of their specificity,
these pesticides are regarded as environmentally friendly, with little or no effect on
humans, wildlife, pollinators, and most other beneficial insects, and are used in organic farming;
[24]
however, the manuals for these products do contain many environmental and human health
warnings,[25][26] and a 2012 European regulatory peer review of five approved strains found, while data
exist to support some claims of low toxicity to humans and the environment, the data are insufficient
to justify many of these claims.[27]
New strains of Bt are developed and introduced over time[28] as insects develop resistance to Bt,[29] or
the desire occurs to force mutations to modify organism characteristics[30] or to use homologous
recombinant genetic engineering to improve crystal size and increase pesticidal activity[31] or broaden
the host range of Bt and obtain more effective formulations.[32] Each new strain is given a unique
number and registered with the U.S. EPA[33] and allowances may be given for genetic modification
depending on "its parental strains, the proposed pesticide use pattern, and the manner and extent to
which the organism has been genetically modified".[34] Formulations of Bt that are approved for
organic farming in the US are listed at the website of the Organic Materials Review Institute (OMRI)
[35]
and several university extension websites offer advice on how to use Bt spore or protein
preparations in organic farming.[36][37]
Use of Bt genes in genetic engineering of plants for pest

control
The Belgian company Plant Genetic Systems (now part of Bayer CropScience) was the first
company (in 1985) to develop genetically modified crops (tobacco) with insect tolerance by
expressing cry genes from B. thuringiensis; the resulting crops contain delta endotoxin. The Bt
tobacco was never commercialized; tobacco plants are used to test genetic modifications since they
are easy to manipulate genetically and are not part of the food supply.
In 1995, potato plants producing CRY 3A Bt toxin were approved safe by the Environmental
Protection Agency, making it the first human-modified pesticide-producing crop to be approved in the
USA,[43][44] though many plants produce pesticides naturally, including tobacco, coffee plants, cocoa,
and black walnut. This was the 'New Leaf' potato, and it was removed from the market in 2001 due
to lack of interest. For current crops and their acreage under cultivation, see genetically modified
crops.
In 1996, genetically modified maize producing Bt Cry protein was approved, which killed the
European corn borer and related species; subsequent Bt genes were introduced that killed corn
rootworm larvae.
The Bt genes engineered into crops and approved for release include, singly and stacked:
Cry1A.105, CryIAb, CryIF, Cry2Ab, Cry3Bb1, Cry34Ab1, Cry35Ab1, mCry3A, and VIP, and the
engineered crops include corn and cotton.[47][48]:285ff
Corn genetically modified to produce VIP was first approved in the US in 2010.[49]
In India, by 2014, more than seven million cotton farmers, occupying twenty-six million acres, had
adopted Bt cotton.[50]
Monsanto developed a soybean expressing Cry1Ac and the glyphosate-resistance gene for the
Brazilian market, which completed the Brazilian regulatory process in 2010.[51][52]

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Bacillus thuringiensis

(or Bt) is a Gram-positive, soil-dwelling bacterium, commonly used as

Mechanism of insecticidal action

Use of spores and proteins in pest control[edit]

Use of Bt genes in genetic engineering of plants for pest

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