Research On EST-SSR Markers And Two Genes Responsed To GA In Ramie

Ramie fiber is a kind of important natural materials.However,compared with cotton fibers,ramie fiber has some shortcoming such as rough and tickling feeling.In this study, excavate and identify molecular markers and gene which related to the development of ramie fiber was carried out by molecular markers and transgenic breeding theories and methods of biological technology..This paper based on Ramie transcriptome sequencing results to explore genes related to fiber development respectively from the aspects of the development function marking and identification of functional genes:①Develop EST-SSR markers of ramie and filter the polymorphism EST-SSR markers.Take EST-SSR markers to explore the molecular markers associated with the fineness quality traits of ramie fiber.②Two ESTs respectively homologous with a-amylase and E3gene were sreening from GA treatment expression spectrum.On the basis of related research progress, these EST may be associated with fiber development.Clone the full length of the two genes,then build its expression vector to further arabidopsis functional verification to identify the role in the process of ramie fiber development.1)8392SSR locis with more than two two nucleotides were found by scanning all ESTs of the ramie transcriptome and exploring distribution characteristics.the amplification of612site EST-SSR primer locus which include139polymorphism EST-SSR primers locus are completed and500allelic variations are checked out.lt detect3.59number of alleles in per primer sites.The average Polymorphism Information Content (PIC) is0.4615. level of linkage disequilibrium measured by R2((Squares Allele-Frequency Correlations))is far less than by D’(Standardized Disequilibrium Coefficients) by TASSEL software, This indicated that linkage disequilibrium between ramie markers was mainly caused by restructuring,and it also indicated that the contribution to linkage disequilibrium of mutations is small. The "k" that represent the subgroup numbers of association group was4. by evaluating the group structure, Association analysis results showed that locus25348 associated with fiber fineness characters and that locus25348was an ethylene response factors by EST information annotation.2) The full length coding region of Bn-a-amylase was cloned.The length of the sequence which encoded765amino acids is2298bp, with predicted pI and molecule weight of this protein encoded by gene were4.95and36.81KD.The carboxyl end of protein contain two conservative domain structure.The results of subcellular localization analysis showed that the Bn-a-amylase located in cytoplasm and there is no signal peptide and the membrane structure domain.The full length coding region of BnSINAT2which is924bp was cloned.307amino acids were encoded by this gene and predicted pI and molecule weight of this protein encoded by gene were7.779and34.466KD.Two conservative domain structure were found in the protein. The results of subcellular localization analysis showed that the BnSINAT2located in cytoplasm and there is no signal peptide and the membrane structure domain. And we build the expression vector pRI101-Bn-a-amylase and pRI101-BnSINAT2for two genes respectively.