摘要
利用正交设计和完全随机试验优化蒺藜苜蓿(Medicago truncatula Gaertn)SSR标记的PCR反应体系。结果表明,适合于一年生苜蓿(Medicago L.)遗传分析的SSR技术体系为:10μL反应体系中TaqDNA聚合酶为1U,dNTP浓度为0.20mmol/L,Mg2+浓度为2.0mmol/L,引物浓度为1.5umol/L,模板DNA用量为20ng/uL;利用该反应体系将2个蒺藜苜蓿亲本和杂交种有效鉴别;利用SSR标记对19个一年生苜蓿种质进行SSR-PCR扩增,用8%的非变性聚丙烯酰胺凝胶电泳检测,不同种质间DNA谱带多态性丰富,通过UPGMA方法聚类分析可以明确区分19个一年生苜蓿种质。
SSR reaction conditions on annual medics were optimized. The model legume, Medicago truncatula. , was used to set up a SSR-PCR amplification system for the annual medics. The results show that the optimal SSR-PCR reaction system for annual medics was 1 U TaqDNA polymerase, 0. 20 mmol/L dNTP, 2. 0 mmol/L Mg^2＋, 1. 5 μmol/L primer, and 20 ng/μL DNA in the total volume of 10 μL. Through above PCR system, SSR fragments of 19 annual medics＇ germplasms were obtained and detected. Polymorphism between different lines was abundantly detected by 8% fro polyacrylamide gel. Germplasms of 19 annual medics were definitely differentiated using cluster analysis based on the UPMGA.
出处
《草地学报》
CAS
CSCD
2007年第5期429-436,共8页
Acta Agrestia Sinica
基金
甘肃省教育厅科学技术研究项目(0507)
甘肃农业大学科技创新项目(200518)
草业生态系统教育部省部共建重点实验室和国家自然科学基金(30671486)
关键词
一年生苜蓿
蒺藜苜蓿
SSR反应
正交设计
杂交种鉴定
种质鉴定
Annual medic
Medicago truncatula Gaertn.
SSR reaction
Orthogonal design
Hybridization identification
Germplasm identification
