Study On The Transformation Of Wheat By Biolistic Particle With Rnai Genes Related To Resistance To Fusarium Haed Blight

Wheat is one of the most important food crops in the world,its industry plays an important role in the agricultural production,Fusarium head blight(FHB)is a global climate disease caused by Fusarium graminarum,which not only causes incalculable economic losses,but infects the quantity of wheat,if we feed grain accumulation of toxin,it can cause a threat to the health of human and animals,which affect the wheat yield and quality greatly.Taking the advantange of genetic engineering technology,assisted in FHB traditional resistant breeding,we can broaden the the FHB resistance germplasm resources,so as to accelerate the breeding process effectively.In this study we use transgenic technology mediated by particle bombardment,and transform the exogenous resistant genes into wheat.the aim is to provide research materials for the research of wheat FHB resistance improvement and lay a foundation for the study of resistance mechanism.This study import RNAi(RNA interference)gene constructs which from Fusarium graminarum into Xiangmai 76(X76)via particle bombardment,including the chitin synthase gene construct(Chs3b?Chs7),glucan synthase gene construct(gls),sterol 14 alpha methylation enzyme gene construct(CYP51).we get different materials of gene combinations by the identification,we also breed and identify the resistance transgenic wheat got by our laboratory precancerous study,the main results are as follows:1.The selection and identification of transformation wheat with the chitin synthase gene 7 RANi(dubiubi-chs7As3As4).We choose PMI/mannose as selected system,we get two lines of T0 generation form different callus,named JA7/JA8,PCR identification form selfing offspring showed,the gene can inherit stably by generation,and tend to be homozygous in T3,the result of greenhouse single flower inoculation in T3 showed that strain JA7 increased significantly in resistance of FHB which compared with X76,the same with field single flower inoculation of T4 generation,compared with the normal transgenic plants and transgenic plants,the plant height,spike length,spike grain number of some strain JA8 increased,but spike grain weight does not showed significant difference.2.The selection and identification of transformation wheat with the chitin synthase gene 3 RNAi(dubicmps-chs3b-As2-chs3bAs3,dubi35s chs3b-As5-chs3b-Asl)?sterol enzyme gene 14 alpha demethylation gene RNAi construct(dubi35ss-CYP51BAC-DONS),we get 1 strain which identificatied with all purpose gene,dubi35ss-CYP51BAC-DONS gene loses in T2,the separation of gene is larger,we find no plants of target gene and selection marker gene separation.3.The selection and identification of transformation wheat with the chitin synthase gene and glucan synthase gene RNAi construct,we get two different combinations,1)dubicmps-chs3b-As2-chs3b As3,dubi35s chs3b As5-chs3b Asl and dubiubi gls3-gls6 co-transformed into X76,we get 4 strains,named from LA1 to LA4,from 3 callus,PCR detection of T1 generation showed that the plants only have dubicmps-chs3b-As2-chs3b As3,T2 is homozygous.2)we also get 1 strain from the combination of chitin synthase gene dubi35s 6*chs3b5-2and glucan synthase gene dubiubi gls3-gls6,named LG1.LG2,the current generation is T0.4.The selection and identification of transformation wheat with glucan synthase gene RNAi construct(dubiubi-gls2 gls6,dubiubi gls3-gls6),with the same method mentioned above,we get 2 plants,named DG1,DG2,the current generation is T1,compared with offspring genes present trend of other batch conversion material,T1 generation gene is homozygous,and has two purpose gene at the same time,compared with non-transformed plants,this grow well,have more tillers,full grain,good agronomic characters.5.The breeding and identification of transformation wheat with resistent gene.1)The transformation wheat with protein kinase gene dubiubi PkcAs3-Chs7As4,Amt-Blf receptor is Xiangmai 76,greenhouse vaccination in T4 and field vaccination in T5,strain RA1,RA3b showed significantly resistance to FHB,and performed stablily.2)The transformation wheat with Chitinase gene UEch42,antibacterial peptide gene Ubi-AFP2,the receptor is Yangmai 158,strain WB22 showed significant resistance compared with control in T4,the same with the previous generation,it prove that the FHB resistance of transgene strains could inheritance stably.3)The breeding and identification of materials taking by high generation contain chs7As3As4 gene,greenhouse single flower inoculation showed that DA3,DA4 strains had significant resistance to FHB in T4.