Enhanced resistance of transgenic tetraploid Isatis indigotica by Bt and CpTI to moths / 中草药

ABSTRACT

Objective Two insect resistance genes Bacillus thuringiensis(Bt) crystal protein gene CryIA(c) and cowpea trypsin inhibitor gene CpTI were introduced into tetraploid Isatis indigotica to enhance the resistance to moths.Methods I.indigotica was transformed with a plasmid,pGBI121S4ABC,containing CryIA(c) Bt and CpTI and the selectable gene(Npt-Ⅱ) driven by the CaMV35S promoter via Agrobacterium tumefaciens LBA4404 mediated transformation.Then PCR and Southern blotting assay were conducted followed by moth bioassay test.Results Co-transgenic rate of the two genes in tetraploid I.indigotica was 16.67%.The integration and expression of introduced genes in T0 regenerated transgenic plants were confirmed by Southern blotting assays.Insect bioassay test demonstrated transgenic lines had significant inhibition to moths,compared with wild type control.Conclusion Co-transformation of Bt and CpTI genes is an effective strategy to enhance the resistance to moths for tetraploid I.indigotica.