Bacillus thuringiensis (or Bt) is a Gram-positive, soil-dwelling bacterium, commonly used as a biological alternative to a pesticide; alternatively, the Cry toxin may be extracted and used as a pesticide. B. thuringiensis also occurs naturally in the gut of caterpillars of various types of moths and butterflies, as well as on the dark surface of plants.
|Spores and bipyramidal crystals of Bacillus thuringiensis morrisoni strain T08025|
|Bacillus thuringiensis |
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. There are however many crystal-producing Bt strains that do not have insecticidal properties.
Discovery and studyB. thuringiensis was first discovered in 1902 by Japanese biologist Shigetane Ishiwatari. In 1911, B. thuringiensis was rediscovered in Germany by Ernst Berliner, who isolated it as the cause of a disease called Schlaffsucht in flour moth caterpillars. In 1976, Zakharyan reported the presence of a plasmid in a strain of B. thuringiensis and suggested the plasmid's involvement in endospore and crystal formation. B. thuringiensis is closely related to B.cereus, a soil bacterium, and B.anthracis, the cause of anthrax: the three organisms differ mainly in their plasmids. Like other members of the genus, all three are aerobes capable of producing endospores. 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 the plasmid.
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, the alkaline pH of their digestive tract activates the toxin. Cry inserts into the insect gut cell membrane, forming a pore. The pore results cell lysis and eventual death of the insect.
Use in pest controlSpores and crystalline insecticidal proteins produced by B. thuringiensis have been used to control insect pests since the 1920s. 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. The Belgian company Plant Genetic Systems was the first company (in 1985) to develop genetically engineered (tobacco) plants with insect tolerance by expressing cry genes from B. thuringiensis.
B. thuringiensis-based insecticides are often applied as liquid sprays on crop plants, where the insecticide must be ingested to be effective. It is thought that the solubilized toxins form pores in the midgut epithelium of susceptible larvae. Recent research has suggested that the midgut bacteria of susceptible larvae are required for B. thuringiensis insecticidal activity.
Bacillus thuringiensis serovar israelensis, a strain of B. thuringiensis is widely used as a larvicide against mosquito larvae, where it is also considered an environmentally friendly method of mosquito control.
Genetic engineering for pest control
UsageIn 1995, potato plants producing Bt toxin were approved safe by the Environmental Protection Agency, making it the first pesticide producing crop to be approved in the USA. By 1996, Bt Maize, Bt Potato and Bt cotton were being grown by farmers in the USA.
Bt crops (in corn and cotton) were planted on 281,500 km² in 2006 (165,600 km² of Bt corn and 115900 km² of Bt cotton). This was equivalent to 11.1% and 33.6% respectively of global plantings of corn and cotton in 2006. Claims of major benefits to farmers, including poor farmers in developing countries, have been made by advocates of the technology, and have been challenged by opponents. The task of isolating impacts of the technology is complicated by the prevalence of biased observers, and by the rarity of controlled comparisons (such as identical seeds, differing only in the presence or absence of the Bt trait, being grown in identical situations). The main Bt crop being grown by small farmers in developing countries is cotton, and a recent exhaustive review of findings on Bt cotton by respected and unbiased agricultural economists concluded that "the overall balance sheet, though promising, is mixed. Economic returns are highly variable over years, farm type, and geographical location" .
Environmental impacts appear to be positive during the first ten years of Bt crop use (1996–2005). One study concluded that insecticide use on cotton and corn during this period fell by 35.6 million kg of insecticide active ingredient which is roughly equal to the amount of pesticide applied to arable crops in the EU in one year. Using the Environmental Impact Quotient (EIQ) measure of the impact of pesticide use on the environment, the adoption of Bt technology over this ten year period resulted in 24.3% and 4.6% reduction respectively in the environmental impact associated with insecticide use on the cotton and corn area using the technology.
AdvantagesThere are several advantages in expressing Bt toxins in transgenic Bt crops:
- The level of toxin expression can be very high thus delivering sufficient dosage to the pest.
- The toxin expression is contained within the plant system and hence only those insects that feed on the crop perish.
- The toxin expression can be modulated by using tissue-specific promoters, and replaces the use of synthetic pesticides in the environment. The latter observation has been well documented worldwide.
Health and safetyOverall, Bt-modified crops appear to be environmentally safe. The proteins produced by Bt have been used in sprays for agricultural weed control in France since 1938 and the USA since 1958 with seemingly no ill effects on the environment.
Bt toxins are considered environmentally friendly by many farmers[who?] and may be a potential alternative to broad spectrum insecticides. The toxicity of each Bt type is limited to one or two insect orders, and is nontoxic to vertebrates and many beneficial arthropods. The reason is that Bt works by binding to the appropriate receptor on the surface of midgut epithelial cells. Any organism that lacks the appropriate receptors in its gut cannot be affected by Bt.
There is clear evidence from laboratory settings that Bt toxins can affect non-target organisms. Usually, but not always, affected organisms are closely related to intended targets. Typically, exposure occurs through the consumption of plant parts such as pollen or plant debris, or through Bt ingested by their predatory food choices. Nevertheless, due to significant data gaps, the real-world consequences of Bt transgenics remains unclear.
Not all scientific reports on Bt safety have been positive. A 2007 study funded by the European arm of Greenpeace, suggested the possibility of a slight but statistically meaningful risk of liver damage in rats. While small statistically significant changes may have been observed, statistical differences are both probable and predictable in animal studies of this kind,(known as Type I errors), that is, the probability of finding a false-positive due to chance alone. In this case, the number of positive results was within the statistically predicted range for Type I errors.
The observed changes have been found to be of no biological significance by the European Food Safety Authority. A 2008 Austrian study investigating the usefulness of a long-term reproduction mouse model for GM crop safety reported that Bt-treated corn consumption in mice appeared to be correlated with reduced fertility via an unknown biochemical mechanism.
Limitations of Bt cropsevolutionary pressure for pests resistant to that toxin. Already, a Diamondback moth population is known to have acquired resistance to Bt in spray form (i.e., not engineered) when used in organic agriculture. The same researcher has now reported the first documented case of pest resistance to biotech cotton.
One method of reducing resistance is the creation of non-Bt crop refuges to allow some non-resistant insects to survive and maintain a susceptible population. To reduce the chance that an insect would become resistant to a Bt crop, the commercialization of transgenic cotton and maize in 1996 was accompanied with a management strategy to prevent insects from becoming resistant to Bt crops, and insect resistance management plans are mandatory for Bt crops planted in the USA and other countries. The aim is to encourage a large population of pests so that any genes for resistance are greatly diluted. This technique is based on the assumption that resistance genes will be recessive.
This means that with sufficiently high levels of transgene expression, nearly all of the heterozygotes (S/s), the largest segment of the pest population carrying a resistance allele, will be killed before they reach maturity, thus preventing transmission of the resistance gene to their progenies. The planting of refuges (i. e., fields of non-transgenic plants) adjacent to fields of transgenic plants increases the likelihood that homozygous resistant (s/s) individuals and any surviving heterozygotes will mate with susceptible (S/S) individuals from the refuge, instead of with other individuals carrying the resistance allele. As a result, the resistance gene frequency in the population would remain low.
Nevertheless, there are limitations that can affect the success of the high-dose/refuge strategy. For example, expression of the Bt gene can vary. For instance, if the temperature is not ideal this stress can lower the toxin production and make the plant more susceptible. More importantly, reduced late-season expression of toxin has been documented, possibly resulting from DNA methylation of the promoter. So, while the high-dose/refuge strategy has been successful at prolonging the durability of Bt crops, this success has also had much to do with key factors independent of management strategy, including low initial resistance allele frequencies, fitness costs associated with resistance, and the abundance of non-Bt host plants that have supplemented the refuges planted as part of the resistance management strategy.
Insect resistanceIn November 2009, Monsanto scientists found that the pink bollworm had become resistant to Bt cotton in parts of Gujarat, India. In four regions, Amreli, Bhavnagar, Junagarh and Rajkot the crop is no longer effective at killing the pests. This was the first instance of Bt resistance that was confirmed by Monsanto anywhere in the world. Monsanto confirmed field resistance of the worm to the Cry1Ac first generation Bollgard cotton, which expresses a single Bt gene.
Secondary pestsChinese farmers have found that after seven years of growing BT cotton the populations of other insects other than bollworms, such as mirids, have become significant problems.  Similar problems but with mealy bugs have been reported in India.
Lepidopteran toxicityThe most publicised problem associated with Bt crops is the claim that pollen from Bt maize could kill the monarch butterfly. This report was puzzling because the pollen from most maize hybrids contains much lower levels of Bt than the rest of the plant and led to multiple follow-up studies.
It appears that the initial study was flawed by faulty pollen-collection procedure; researchers fed non-toxic pollen mixed with anther walls containing Bt toxin. The weight of the evidence is that Bt crops do not pose a risk to the monarch butterfly.
Wild maize genetic contaminationA study in Nature reported that Bt-containing maize genes was contaminating maize in its center of origin. Nature later "concluded that the evidence available is not sufficient to justify the publication of the original paper." However, there still remains a controversy over the highly unorthodox retraction on the part of Nature. In 1998, Chapela, one of the original paper's authors spoke out against Berkeley accepting a multi-million dollar research grant from the Swiss pharmaceutical company, Novartis.
A subsequent large-scale study, in 2005, failed to find any evidence of contamination in Oaxaca. However, further research confirmed initial findings concerning contamination of natural maize by transgenic maize.
However, further studies, such as that published in Molecular Ecology in 2008, have shown some small-scale (about 1%) genetic contamination (by the 35S promoter) in sampled fields in Mexico. One meta-study has found evidence for and against Bt contamination of maize, concluding that the preponderance of evidence points to Bt maize contamination in Mexico.