Genetic Transformation Of Bread Baking Quality Gene1Dx5in Xindong-26Wheat And Generations Detection

The artical make commercial wheat Xindong-26(Triticun aestivum L), as aresearch object, which is a salt-tolerant and winter wheat.Using the genetic engineer-ing, the linear core fragment of1Dx5was prepared by PCR, following the foreigngene was imported into receptor via pollen tube pathway to obtain transgenicwheat.with the method of SDS-PAGE and multiplex PCR, the expression situation ofexogenous gene in transgenic offspring were detected and analyzed, in order to newgermplasm of transgenic wheat are both strong salt-tolerance and gluten.this is notonly provide a new germplasm resources for food security and accelerat the pace ofthe production of high quality wheat, but also has a vital significant thatimprovement of wheat quality by molecular biology techniques and cultivation ofresource-efficient crops.The main findings are as follows:(1) comparation and analysis of1Dx subunit gene between wheat and relatedspecies: comparison and analysis of the1Dx type subunits of nucleic acid, proteinand secondary structure with the bioinformatics. The results showed that: in thenucleic acid level, the similarity is up to96.58%, there are90SNPs and resulting inthe replacement of53amino acids; the similarity is96.74%in the amino acid level,and there are seven deletion/insertion of amino acid peptide; in the prediction ofprotein secondary structure level, the similarity is high up to97.71%, the fourchanges of the secondary structural elements may be caused by SNP, repetitivesequences, insertions and deletions may affect the random curl the length ofsecondary structure which may affect the function of the biology of protein subunits.Analysis the differences of between each gene on the DNA and protein levelrevealed to some extent relationship in the molecular structure, biological functionand species evolutionary, reveal genetic transformation and detection of theexogenous gene. It is provide technical solution and theoretical guidance fordetection of exogenous gene in transgenic wheat.(2) Genetic transformation and detection of the exogenous gene: Firstly, pHMW-1Dx5and core fragment of1Dx5gene were imported to the receptor material viapollen tube pathway, respectively. Secondly, the HMW-GS composition of T0generation was deteced with method of SDS-PAGE. The results show that there arefive high molecular weight glutenin subunits in the T0generation of core fragment 1Dx5gene, include1Dx2,1Bx7,1By8,1Dy12, and a new protein subunit–BandXthat electrophpretic mobility faster than1Dx5, its frequency is0.2%. but electrophp-retic mobility of a new subunit lower than1Dx2in the T0generation of pHMW1Dx5,its frequency is close to0.15%.(3) Detceting and analysis of T1generation of pHMW1Dx5: The results showthat positive signal of the exogenous gene were found by PCR, it production is450bp indicated that the exogenous gene may integrated into the genome of receptors.the result of SDS-PAGE indicated that exogenous gene may be expressed andresulted in change of HMW-GS composition.(4) Detection and analysis of T1generation of core fragment1Dx5gene:detection and analysis through the establishment of the multiplex PCR system and bySDS-PAGE. The results show that Characteristic fragment of1Dx2and1Dx5intransgenic T0lines by Multiplex-PCR show that foreign gene had been inserted intothe genome of wheat cultivar. Secondly, a novel protein X was tested in HMW-GScompositions by SDS-PAGE, that may be ascribed to insertion of foreign gene.Security genetic transformation system is based on principle of gene capture, theseexplore new avenues for genetically modifide wheat.