油棕新品种遗传多样性的ISSR分析(摘要)(英文)

目的]从DNA分子水平上对油棕新品种进行遗传多样性的分析,旨在为这些新选育油棕种质的开发利用提供分子水平的理论依据。方法]采用改良的CTAB法提取油棕叶片总DNA。从加拿大哥伦比亚大学所设计的60个引物在部分模板中进行筛选,筛选出能扩增出多态性的引物23个用于样品的进一步分析。通过一定的摸索实验,最终确定用于油棕的各个组分的ISSR最佳反应体系。优化反应体系总体积为15.00μl,各个组分的具体用量为:MgCl2(25mmol/L)+10×Buffer2.25μl、dNTP(10mmol/L)0.30μl、Taq酶(5U/μl)0.15μl、引物(0.4μmol/μl)1.00μl、模板DNA(50ng/μl)1.00μl、ddH2O(超纯水)10.30μl。PCR扩增程序为:94℃预变性4min,然后94℃变性1min,52℃退火1min,72℃延伸1min,35个循环,72℃保温10min,4℃保温。采用2%琼脂糖凝胶电泳,EB染色检测扩增产物。ISSR数据的统计分析方法:每个样品的扩增产物电泳分离谱带在某一位点上按有或无记录,存在时赋值为"1",无带的记为"0"。将每个引物对所有样品的读带记录结果形成"0-1"矩阵。用NTSYS-pcversion2.1计算Jaccard’s相似系数,在此基础上用非加权类平均聚类法(UPGMA)进行聚类分析,绘制亲缘关系树状图。结果]用23个引物共扩增得到201条扩增条带,其中多态性条带占41.8%。根据ISSR遗传相似系数按UPGMA法聚类,以所有材料间的平均遗传相似系数0.838为阈值,可将其明显的划分为2大类:采自海南的R1～R12新种质材料与采自云南的V20新种质材料合为一类;采自云南其余的新种质材料为一类。用NTSYS-pc软件所得出的主成分分析图也能够明显的将海南和云南引进的新油棕种质材料区分开来,海南的R1～R12油棕与云南的V20油棕基本上归为一类;与聚类图不同的是,云南的新油棕种质V15、V16、V17、V19、V21、V22基本上归为一类,而品种V13、V14与V18则较远一些。结论]海南与云南新选育的油棕种质材料可能来源于不同的国家或地区。

ISSR Analysis on Genetic Diversity of New Oil Palm Germplasm

Objective]The aim of this study was to analyze the genetic diversity of new oil palm germplasm from DNA molecular level and provides a theoretical basis for the development and utilization of new oil palm germplasm. Method]The genetic diversity of 22 new oil palm germplasm from Hainan and Yunnan were analyzed by ISSR technique. Result]A total number of 201 bands were amplified with 23 ISSR primers,and polymorphic bands accounted for 41.8%. 22 oil palm varieties were divided into two groups by cluster analysis：Group I included new varieties of R1-R12 from Hainan and V20 from Yunnan,while group II included new varieties of V13-V22 from Yunnan,except V20. Conclusion]New breeding oil palm germplasm from Hainan and Yunnan may come from different countries or regions.