葎草性别相关SRAP分子标记的鉴定

Identification of Sex Related SRAP Molecular Marker in Humulus scandens

葎草是一种研究雌雄异株植物性别分化分子机制的理想材料。本研究以葎草雌、雄基因池为模板,采用序列相关扩增多态性(SRAP)分子标记技术筛选与性别相关的分子标记,从256对引物组合中筛选出了50对可以稳定扩增出特异条带的引物组合,聚丙烯酰胺凝胶电泳结果显示,50对引物组合共扩增出671条带,247条呈现多态性,多态率为36.81%。后以葎草5个雄性单株、5个雌性单株为材料,采用琼脂糖凝胶电泳对引物组合me15+em11,me14+em16,me13+em12和me7+em10扩增结果再次进行验证,结果表明尽管扩增条带有所不同,但是雌、雄性特异条带是比较稳定的。对本研究中获得性别相关特异条带进行克隆、测序及染色体定位,将会有助于研究葎草性染色体演化的分子机制。

Humulus scandens is a model material for studying the molecular mechamisms of sex differentiation in dioecious plants.We used SRAP（sequence related amplified polymorphism） to screen male or female specific fragments in H.scandensby using male and female gene pool as template.Fifty pairs of primers showed a good amplification pattern after screening 256 pairs of primers.Polyacrylamide gel electrophoresis amplification patterns of SRAP showed that 671 bands were amplified using 50 primer combinations,of which 247 bands were polymorphic and the polymorphic ratio is 36.81%.With five single male plants and five single female plants as materials,we validated the amplification patterns of primer combinations me15 ＋em11,me14 ＋em16,me13 ＋em12 and me7 ＋ em10 again by agarose gel electrophoresis.Although the amplification patterns were more or less different between polyacrylamide gel electrophoresis and agarose gel electrophoresis,male or female specific bands were relatively stable.If cloning,sequencing and chromosomal locating the obtained sex-related DNA sequences on chromosome of H.scandens by using fluorescence in situ hybridization technique,it will be contributed to studying the molecular mechanisms of sex chromosome evolution in H.scandens.