Enhancing Cry1Ac toxicity by expression of the Helicoverpa armigera cadherin fragment in Bacillus thuringiensis.
Insect cadherin proteins localized in the midgut epithelium were identified as receptors for Bacillus thuringiensis insecticidal crystal proteins (Cry toxins). These cadherins facilitated toxin monomer oligomerization and mediated oligomer binding to secondary receptors. It has been reported that Manduca sexta, Helicoverpa armigera, Anopheles gambiae and Diabrotica virgifera cadherin toxin binding regions function as synergists for Cry1A, Cry4Ba and Cry3A toxicity against target insects. In the present study, the toxin binding region fragment of the H. armigera cadherin (hacad1) gene was cloned and fused with the promoter of the cry3Aa gene. The fusion gene pro3Aa-hacad1 and the cry1Ac gene were inserted into shuttle vector pHT304 and introduced into B. thuringiensis acrystalliferous strain BMB171 for coexpression (resulting in recombinant strain BMB1073). SDS-PAGE and mass spectrum analysis showed that BMB1073 could express HaCad1 and Cry1Ac proteins together. Bioassay results demonstrated that insecticidal activities against H. armigera and Spodoptera exigua could be increased 5.1-fold and 6.5-fold, respectively, by BMB1073 compared with the strain which can only express the Cry1Ac protein. Our discovery showed that coexpression of HaCad1 and Cry1Ac toxin in B. thuringiensis enhanced the insecticidal activity of Cry1Ac toward Lepidoptera insects. This finding also revealed a novel strategy for engineering strains and transgenic plants with higher insecticidal activity.
Peng
Donghai
D
State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, People's Republic of China.
Xu
Xiaohui
X
Ruan
Lifang
L
Yu
Ziniu
Z
Sun
Ming
M
D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics / Q000378:metabolism*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000633:toxicity*; D015820:Cadherins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004731:Endotoxins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000633:toxicity*; D015870:Gene Expression; D006460:Hemolysin Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000633:toxicity*; D007306:Insecticides / Q000737:chemistry / Q000633:toxicity; D009036:Moths / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D018411:Spodoptera / Q000254:growth & development
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D015820:Cadherins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; D011993:Recombinant Fusion Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis