Abstract:

By optimizing DNA extraction method, annealing temperature, amplification of cycle number and Mg2+ concentration, we established an optimal ISSR-PCR reaction system for Phalaenopsis in this study. 8 primers were used to generate 193 brands of DNA fragments, the average percentage of polymorphic bands was 92.2%, and the range of amplified fragments was 300~2 500 bp.  This result indicated that the tested species possessed abundant genetic polymorphism. The 16 species of Phalaenopsis can be divided into 2 groups at L=0.222 5, one stand for Japanese/Taiwan species, and the other is for species newly cultured by the Company. The potential hybrid combinations of different species were suggested according to the affinity distance determined by molecular marker.  The establishment of ISSR molecular marker system in Phalaenopsis has provided theoretical basis for analyzing genetic relationship of Orchidaceae ornamental plants.

Key words: ISSR, Phalaenopsis hybrid, genetic relationship, molecular breeding

摘要：

从DNA提取、退火温度、扩增循环数、镁离子浓度等方面对ISSR-PCR反应体系的条件进行了优化,筛选出8个引物,共扩增出193条片段(多态性条带比例为92.2%),所扩增的条带数分布范围为300~2 500 bp,表明供试品种间具有丰富的遗传多态性。聚类分析结果表明：来源于不同地域的16个品种在遗传距离L=0.222 5处可分为2个类群；来源于日本、台湾的品种分别聚类为一簇,公司新育成的品种为一簇。根据分子标记确定的亲缘性远近提出了可能的杂交组合。蝴蝶兰ISSR分子标记优化体系的建立为兰科类观赏植物的亲缘关系分析提供了理论依据。

关键词: ISSR;蝴蝶兰;亲缘关系;分子育种