The Genetic Transformation And Insect Bioassay Of Nine Cotton Endogenous Candidate Insect-resistance Gene

Cotton is one of the most important crop which has a great economic value and a practical value.Its fiber is an important raw material,and its cotton seed oil can also be used as energy source.Therefore,cotton occupies an important place in the national economy in the future.Whitefly is one of the main pests in cotton's production which has a typical of sucking mouth parts with which it can feeding with cotton's phloem SAP in the process of harm plants.At the same time it can also as a media to Spread of disease and in the actual production of the cotton which does a great deal of harm to the plant.In This study,we choose nine genes which cotton may resistant whitefly to do transgenic function verification.Eight genes of them are extracted by transcriptome analysis of cotton response to insect Bemisia tabaci data which probably be genes of cotton Interacting with whitefly.Compared Gene sequence with Arabidopsis thaliana,the function of those gene probably are: six receptor protein kinase genes or receptor protein genes :PR(phytosulfokin receptor 1),RLP(receptor like protein 6),LRR(Leucine rich repeat(LRR)family protein),PLK-a(cysteine rich RLK25),RLK-D(cysteine-rich RLK25),LPK(S-locus lectin protein kinase family protein),A XTH(XTH22),A CHA(Achitinase A).In addition,there is another one obtained by the method of homologous comparison which is Cotton endogenous gene homologous to Rice gene BPH3.Through RNAi,Overexpression and gene editing techniques,we design and build at a total of 13 expression vectors including: 10 RNAi vectors,two over expression vectors and one Crispr vector.Through genetic transformation technique technology,86 lines were genetically modified materials,and Southern-blotting were used to detect the number of the positive plants copy.Some of the genetically modified lines were plant in the ground,and by analyzed T1 generation of genetically modified RNAi lines,we cannot find obvious differenceswe with the traits such as plant type,flower.It have little affect in the overall agronomic traits Comparing with Jin668 control and negative control,except plant height.Some genetic interventionstrain insect-resistant traits differences compared with Jin668.The creation of these materials and identification insect-resistant study further.In the process of differentiation,BPH over-express vector cannot be differentiated after the transformation.So,we predict that endogenous BPH gene may speed up the formation of callus and the cell of differentiation.Through genetic transformation experiment,we can initial guess that cotton endogenous BPH gene may be associated with cell differentiation,but this guess also need to do more relevant experiments in the further validation.