Studies On The ISSR Molecular Makers Of Resistance To Sclerotinia Trifoliorum In Medicago Sativa L.

Along with south of our country the animal husbandry development,the high quality forage grass deficient has become its bottleneck.In recent years,the south has introduced the alfalfa which is known as "king of the forage grass".With cultivated area's expansion,alfalfa plant disease was serious day by day in recent years,specially south of our country the warm and damp climate,has provided the existing environment for the alfalfa Sclerotinia trifoliorum.The alfalfa Sclerotinia trifoliorum has become primary factors that restricts alfalfa production south of our country. Therefore,the research to ISSR Molecular Makers of Resistance to Sclerotinia trifoliorum in Medicago sativa L.is very essential and is urgent.The traditional breeding method wastes the time and the money,the molecular mark auxiliary choice(MAS)is judges the goal gene through the analysis and the goal gene close chain-like member mark whether to exist,may accelerate the goal gene greatly the shift and the use,eliminates the disadvantageous related character design and the cultivation ideal variety early,enhances the heredity breeding and the disease-resistant breeding efficiency.This research uses the molecular mark auxiliary choice to carry on the disease-resistant breeding,attempts to discover the ISSR member mark link with anti-Sclerotinia trifoliorum, accelerates anti-source screening and the resistant gene appraisal,will build the foundation for present's breeding work.The findings are as follows:After several year field experiments and the exsomatize leaf blade inoculation,screens high anti-5,high sensitivity 7,altogether 12 adult plants as experiment material.Compared with polyacrylamide gel electrophoresis and agarose gel electrophoresis,the agarose gel electrophoresis is more suitable to use in this experiment ISSR-PCR product examination.Constructs anti-,susceptible gene pond with the BSA law,47 ISSR primers were screened by garose gel electrophoresis,37 primers be able to amplify the band,and 10 showed polymorphism between anti- and susceptible gene pond.Further regarding this 10 primers the confirmation,the primer P825 to be able to amplify 1500bp in 5 disease-resistant plants the special belt,but 7 susceptible plants can not amplify this special banding;The primer P827 to be able to amplify 1380bp in 5 disease-resistant plants the special belt, but 7 susceptible plants can not amplify this special banding.Tentative determines these 2 marks to be possible linkage with the alfalfa anti-Sclerotinia trifoliorum gene.