紫花苜蓿品种鉴定的RAPD分子标记技术研究

试验分别以中苜一号(Medicago sativa L. cv.Zhongmu No.1)、肇东(M.sattva L. cv.Zhodong)、甘农三号(M.sativa L. cv.Gannong No.3)、公农一号(M.satzva L. cv.Gongnong No.1)紫花苜蓿品种的混合DNA(由各品种60个单株的DNA等量混合)为模板筛选RAPD特异性引物,分析比较品种鉴定的特异性谱带.结果表明:从120条随机引物中筛选出S1、S78、S89和S¨5共4条随机引物,可以通过特异性分子标记位点反映品种间的差异;另外,筛选出的RAPD标记引物对各品种60个单株DNA的测试显示品种内部存在特异性标记的单株比例较高;通过RAPD分子标记引物的筛选、标记程序的完善,确定品种特异性标记谱带,可以进行紫花苜蓿(Medicago sativa)品种的鉴定.

Study on the Verification of Alfalfa Cultivars by RAPD Molecular Markers

Alfalfa,a perennial legume forage,is important for the establishment of cultivated grassland and production of hay and silage.Superior cultivars of alfalfa have been distributed to large areas of northern China. Four alfalfa cultivars: Zhongmu No.1,Zhaodong,Gannong No.3,and Gongnong No.1 were selected as research materials.The present experiment screened primers which could verify cultivars by amplifying specific nucleotide sequence in DNA (mixed by 60 individuals) by RAPD-PCR.The results show that 120 primers were screened and four primers: S1,S78,S89,S115,could mark specific nucleotide sequence;electrophoresis bands were clear and specific.These different alfalfa cultivars could be identified by combination of four primers.The PCR amplified effect of RAPD in individual plants of each cultivar indicated that the rate of individuals with specific marker was high and steady.