Cloning Of L-GalLDH And DHA CDNA Fragments From Actinidia Deliciosa Fruit And RAPD Analysis On Germplasm Resources In Genera Actinidia

China is the center of origin and distribution of genus Actinidia and there are various genotypes,which provide broad breeding materials for creating new cultivars.The superposition in geographical distribution and the natural hybridization resulted in emergence of many transitional biotypes,which make the boundary of interspecies and interspecies taxonomic unit confuse.The varieties in A. chinensis and A. deliciosa were extensively cultivated in kiwifruit production.Cultivating valuable kiwifruit cultivars still be a great challenge,gene clone and molecular markers are widely applied in breeding of kiwifruit,and reach some achievements.L-GalLDH and DHAR play important role in the biosynthesis accumulation and reproduction of AsA. For understanding the mechanism of AsA accumulation,we cloned L-GalLDH and DHAR.Huayou is a new kiwifruit cultivar.For deeply grasping genetic and biological characteristics of Huayou,not only biological characteristics were investigated and compared among 3 Actinidia cultivars,but also analysis Huayou by RAPD and do the cluster analysis in the software of DPS(6.55).Meanwhile,52 materials of Actinidia germplasm provided by Zhouzhi county were analysised by RAPD.The aim of the study is to give some theory bases of making use of the germplasm and selecting the parents of hybridization.The main results were as follow:1. 857 bp and 594 bp cDNA encoding L-galactono-1,4-lactone dehydrogenase(GalLDH)and dehydroascorbate reductase(DHAR) fragments were cloned from fruit of Actinidia deliciosa. In GenBank. sequences analysis showed that 88% GalLDH gene identity in nucleotide sequence to that of Rosa roxburghii Tratt ,and 99% DHAR gene identity in nucleotide sequence to apple.2. Most biological characteristics of flower,fruit,leaf,branch and lenticels etc.on Huayou are identical to Hongyang,some characteristics of fruit set are identical to Qinmei,and characteristics of phenophase are between Hongyang and Qinmei .the result of RAPD shows Huyou is a new cultivar which has the same characteristics to A. deliciosa,And has the same to A. chinensis,and most characteristics belong to A. chinensis,few belong to A. deliciosa. cluster analysis shows Huayou first become cluster with 5 A. chinensis cultivars,finally become cluster with 5 A. deliciosa cultivars. The genetic distance between Huayou and 5 A. chinensis cultivars is from 0.3143 to 0.4667,but 5 A. deliciosa cultivars is from 0.4194-0.8333.3. Of the 90 random primers screened,30 primers exhibited sufficient polymorphic band patterns total of 246 RAPD bands were generated by 30 selected arbitrary primers,among which 212 bands were polymorphic.The result of cluster analysis showed that A. chinensis and A. deliciosa have closer relationship.A few wild kiwifruit species'genetic distance is more far,some cluster with A. deliciosa, some cluster with A. chinensis,these indicated they have more genetic diversity.Among most A. deliciosa cv.in this study have closer relationship,indicating they have similar genetic background.