Analysis Of Resources Diversity Of ’Huangjincha’[Camellia Sinensis] And Study On CsICE1Gene Transformed Into Tobaccos

Tea plants were originated from Yunan province. In southern China ecological condition, warm and humid, heavily rainfall, tea plants prefer thermophilic to chill. Low temperature is an important abiotic stress constraint restricting geographical distribution. In recent year, extreme climate could seriously affect the quality and production of tea in many tea areas. So the mechanism of cold resistance becomes a hot topic in the research field of tea science.’Huangjincha’(Camellia Sinenesis) were unique and precious tea germplasm resource in Hnan province, they were found in recent year. Huangjincha have many characteristics, such as early sprouting, high yield, good quality and strong resistance.111tea strains from’Huangjincha’ population were analyzed by AFLP-sliver staining protocol for detecting the genetic diversity and relationship. The results provide the basis for the protection and utilization of’Huangjincha’ population from molecular level. Furthermore, functional analysis of cold regulated gene CsICE1, it set good foundation for mechanism of cold stress and tea breeding.The results of this thesis were listed as follows:1.20early maturing strains were selected and studied the main ingredients, in2011and2012spring. The results indicated that No.8has some characteristics, such as early maturing, high amino acid, high theanine, the low ratio of TP/TAA (tea polyphenol/total amino acid) that was fit for green tea. According to these features, No.8was preliminary forecasted suits green tea production.2.111tea strains from’Huangjincha’ population were analyzed by AFLP-sliver staining protocol for detecting the genetic diversity and relationship. AFLP fingerprinting of111samples with primer combinations E37M32, E41M33and E41M42revealed a total number of unambiguous229bands, of which186were polymeric and the polymorphism frequency was81.22%. The results showed that abundant diversities existed among samples of genomic DNA. The mean of effective number of alleles, Nei’s gene diversity and Shannon’s information index of111samples were1.42±0.35,0.25±0.18,0.38±0.25respectively. As analyzed by NTSYSpc2.1, the similarity coefficient of111samples ranged from0.65to0.99, and the mean was0.76. These tea strains were divided into8groups by UPGMA based on the mean of similarity coefficient, and AFLP dendrogram was established. In conclusion, the results provide the basis for the protection and utilization of ’Huangjincha’ population from molecular level.3. The expression level of cold regulated genes form huangjincha in4℃temperature were analyzed by RT-qPCR. According to these results, the three genes CsICE1, CsRAV and CsDHN2were speculated to constitutive expression. The other four CsCBF1, CsERF, CsRbcS and CsDHN1were speculated to inducible genes. All of them play key roles in cold stress for tea plant.4. The overexpression vector of CsICE1gene was constructed via agrobacterium tumefaciens binary vector, and it was transformed to agrobacterium GV3101.5. Transgenic tobaccos were obatained via leaf disks, and cold tolerances of T1s were analysis. The transformation of CsICE1form Huangjincha was set good foundations for enhance cold tolerance in tea plant via gene engineering.