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Molecular Mechanisms of Resistance to Bacillus Thuringiensis Cry1Ac Toxin in Plutella Xylostella (L.)

Author: QinYue
Tutor: ZhangYouJun
School: Chinese Academy of Agricultural Sciences
Course: Agricultural Entomology and Pest Control
Keywords: Diamondback moth Cry1Ac Fitness Cadherin Gene Expression
CLC: S433
Type: Master's thesis
Year: 2011
Downloads: 44
Quote: 1
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Abstract


Bacillus thuringiensis Bacillus thuringiensis arising insecticidal crystal proteins (Insecticidal crystal proteins, of ICPs), target insects with a highly selective, but the friendly people and other non-target biological and environmental security in the world to get a wide range of applications in plays a huge role in pest control and other chemical pesticides, long-term resistance to Bt selection pressure under pests will also have. It is reported that there are a variety of insects to the Bt toxin resistance, but the diamondback moth Plutella xylostella still is so far the only pest resistance to Bt formulation in the field. Clear diamondback moth to Bt-resistant mechanism, provide some theoretical foundation of the Plutella xylostella Bt resistance management, eliminate the election of indoor resistant diamondback moth, diamondback moth biological characteristics observed, fitness research and cloning and expression of the diamondback moth larvae body class cadherin (Cadherin-like Protein, CAD), specific results are as follows: 1. resistant and eliminate the election by two years, the resistant diamondback moth populations Cry1Ac toxin anti- level from 750 times to 1,500 times more. Observe the 4 ° C treatment pupae (7d) and allergy hybrid fecundity of the diamondback moth, 4 ° C treatment pupae can significantly reduce the fecundity of the diamondback moth, can see resistant populations (Cry1AcR) from the hybridization experiments, anti- the cross population fecundity (SS ♂ × Cry1AcR ♀) was significantly lower than the sensitive populations (SS) and orthogonal populations (SS ♀ × Cry1AcR ♂) fecundity inferred low temperature on the pupal processing and hybrid female resistance levels may affect the fecundity of the diamondback moth. 2. Construct life tables of the diamondback moth, compared resistant and susceptible diamondback moth populations range of biological parameters. The results show that, compared with sensitive populations SS, the resistant populations Cry1AcR pupae weight was significantly reduced, a significant reduction in fecundity, sex ratio, hatching and pupation rate significantly lower, simultaneously Cry1AcR population intrinsic rate of increase, the net the finite rate of increase of the growth rate and within weeks were reduced, resistant strains of the relative fitness of susceptible strains only 0.57. Shows a significant adverse diamondback moth resistant population growth and reproductive capacity. Cloned diamondback moth SS Cry1AcR two group classes cadherin gene, the full-length cDNA was 5148bp, encoding 1716 amino acids. Confront the two populations of sensitive cadherin-like compared to 98.3% nucleotide similarity, the amino acid similarity of 98.8%, compared with sensitive populations SS the resistant population Cry1AcR 87 bases and 21 amino acids different. The homology analysis that cadherin-homology of the genes and other insects. 4. Using Bac-to-Bac system of CAD expression will build CAD recombinant baculovirus with no connection of exogenous gene expression vector as control transfected Sf9 cells, and then to collect the CAD gene and empty vector transfected cells as well as There are no transfected normal cells protein. The collected protein by SDS-PAGE electrophoresis detection, the results showed that only cells transfected CAD recombinant baculovirus capable of specifically express approximately 190 kD protein product with a theoretical expected value match, indicating that the expression of CAD obtained.

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CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Plant pest and its control
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