摘要
为筛选与毛叶山桐子性别相关的分子标记,本研究优化了毛叶山桐子ISSR-PCR反应体系,并利用优化后的体系逐一筛选100条ISSR引物,然后通过聚丙烯酰胺凝胶电泳进行多态性检测.实验确立出最佳的25μL反应体系为：55ng DNA模板,1 U Taq酶,2μL dNTPs（2.5mmol/L）,2μL引物（10μmol/L）,2.5μL 10×Taq Buffer（含Mg2＋）,超纯水补足总25μL.利用该体系逐一筛选ISSR引物,在多次重复后只有UBC841引物扩增出的差异性条带可以稳定存在.PAGE电泳结果显示,UBC841扩增产生一条差异性条带,大小在250-300bp之间,存在于毛叶山桐子雌株中,但后期的测序发现该条带可能是由大小相近的条带组成的混合条带.虽然如此,引物UBC841仍可以作为毛叶山桐子性别相关的分子标记.
In order to screen the sex-related ISSR molecular marker in Idesiapolycarpa Maxim.var.vestita Diels,the inter-simple sequence repeat-polymerase chain reaction（ISSR-PCR）system was established and optimized.And we found the best ISSR-PCR reaction system with total volume of 25μL was55 ng DNA template,1U Taq polymerase,2μL dNTPs tendency（2.5mmol/L）,2μL primers（10μmol/L）,2.5μL 10 xTaq Buffer（including magnesium ion）and then adding ultrapure water to make up total25μL.With this system we discovered only one ISSR primer-UBC841,which could amplify one stable female-special fragment with size in the range of 250-300bp.By sequencing the size of female-special fragment,we found it was a mixed bands which were consist of many similar sizes.Even so,the primer UBC841 still can be used for sex-related markers to identify the gender of Idesia polycarpa Maxim.var.vestita Diels.
出处
《四川大学学报（自然科学版）》
CAS
CSCD
北大核心
2016年第2期465-470,共6页
Journal of Sichuan University(Natural Science Edition)
基金
四川省科技计划项目(2011HH 0006)
四川省支撑课题(2011JZ 0002)
关键词
毛叶山桐子
性别鉴定
雌雄异株
ISSR-PCR
Idesia polycarpa Maxim.var.vestita Diels
sex identification
dioecism
ISSR-PCR
