苎麻细胞质雄性不育“三系”ISSR特异片段克隆和序列分析

Cloning and Sequencing for ISSR Specific Fragments in Cytoplasmic Male Sterile,Maintainer and Restorer Lines of Ramie(Boehmeria nivea)

利用ISSR分子标记技术对苎麻细胞质雄性不育"三系"mtDNA进行多态性分析;在选用的38个IS-SR引物中,有6个引物的扩增产物在不育系、保持系和恢复系之间存在差异。对这些特异性片段进行克隆和序列测定,结果表明:片段21-MS全长956bp,包含一个525bp的完整编码区,共编码174个氨基酸。片段31-M/R全长778bp,包含一个404bp的不完整编码区,共编码134个氨基酸;其核苷酸和氨基酸序列与已报道的多种植物中的番茄红素β-环化酶基因分别存在71～76和73～77的同源性。

The polymorphic analysis of mtDNA from ramie cytoplasmic male sterile （MS）, maintainer （M） and restorer （R） lines （three lines） were performed by using ISSR technique. Thirty-eight ISSR primers were used for testing mtDNA polymorphism and different amplified bands were observed in six primers in ramie ＂three lines＂, these specific bands were cloned and sequenced. Among those bands, the fragment 21-MS （GenBank accession number： EU122345） had 956 bp in total length and a 525 bp intact coding region that encoded a polypeptide consisting of 174 amino acids. Fragment 31-M/R （GenBank accession number： EU122344） had 778 bp in total length and a 404 bp incompletely coding region that encoded a polypeptide consisting of 134 amino acids; this fragment had 71%- 76 % and 73 %- 77 % of homology in nucleotide and amino acid sequences with lycopene beta-eyclase allozyme gene in many other plants.