马槟榔DNA提取及其RAPD反应体系的优化  

Extration of Genomic DNA of Capparis masaikai Lévl and Optimization of the RAPD Reaction System

作　　者：于旭东[1] 张超[2] 顾文亮[1] 胡新文[1] 郭建春[3] 

机构地区：[1]海南大学农学院,海口570228 [2]海南大学园艺园林学院,儋州571737 [3]中国热带农业科学院热带生物技术研究所,海口571101

出　　处：《基因组学与应用生物学》2011年第1期117-121,共5页Genomics and Applied Biology

基　　金：国家自然科学基金(30760025);国家自然科学基金(31060123)共同资助

摘　　要：马槟榔是我国特有的仅分布在热带、亚热带地区的珍稀濒危野生果树,马槟榔种子中所富含的植物甜蛋白马宾灵(Mabinlin)在食品甜味剂领域有着广阔的应用前景。为了对我国马槟榔野生种质资源进行遗传多样性分析,本研究建立并优化了马槟榔基因组DNA的RAPD技术最优反应体系。结果表明,采用SDS-CTAB方法提取的马槟榔基因组DNA满足RAPD试验要求;RAPD最优反应体系(25μL)为:引物的终浓度为1.0μmol/L,dNTPs的终浓度为0.2mmol/L,模板DNA的终浓度为1600ng/L,TaqDNA聚合酶用量为40U/L。本研究通过对马槟榔DNA的提取及RAPD反应体系的优化,旨在为下一步对我国马槟榔野生种质资源进行遗传多样性分析提供基础。Capparis masaikai Lévl is an rare and endangered specie of wild fruit tree which only distributes in tropical and subtropical regions of China. The plant sweet protein Mabinlin which is rich in the seeds of Capparis masaikai Lévl has a broad application prospects in the food sweeteners industry. In order to analysis the genetic diversity of the wild Capparis masaikai Lévl germplasm, this research aimed to screen out optimum RAPD reaction system for genomic DNA of Capparis masaikai Lévl. The results showed that the improved SDS-CTAB DNA extraction method was used to extracted the genomic DNA of the leaves of Capparis masaikai Lévl. The single factor experiment was adopted to select the optimization the dNTPs concentration, template DNA concentration, Mg2＋ concentration, Taq enzyme concentration and the primer concentration. The results shows that the optimum reaction system （25 μL） for RAPD amplification of the Capparis masaikai Lévl DNA was listed as follows： 1.0 μmol/L primer, 0.2 mmol/L dNTPs, 1 600 ng/L DNA and 40 U/L Taq enzyme. All the results of this investigation could be used for further analysis of the species diversity of the wild Capparis masaikai Lévl germplasm.

关 键 词：马槟榔 RAPD 反应体系 种质 

分 类 号：Q943.2[生物学—植物学]