中国石蒜属遗传多样性与亲缘关系的SSR分析

Analysis of Genetic Diversity and Inter-specific Relationships of Lycoris (Amaryllidaceae) from China by SSR-PCR

利用SSR-PCR分析中国石蒜属植物的遗传多样性与种间亲缘关系,从46对SSR引物选出18对扩增清晰、多态性好的引物对石蒜属15个种和外类群中国水仙进行了PCR扩增,采用非变性聚丙烯酰胺凝胶电泳和改良的银染法染色,结果表明供试材料具有高度的遗传多样性,多态性位点百分率达97.63%。通过UPGMA法将供试20份材料聚成4类。第I类包括安徽石蒜、长筒石蒜、黄长筒石蒜、忽地笑、中国石蒜、乳白石蒜,其中安徽石蒜与长筒石蒜的亲缘关系很近;第II类包括香石蒜、鹿葱、换锦花、红蓝石蒜和玫瑰石蒜;第III类由石蒜、矮小石蒜、稻草石蒜和江苏石蒜组成;第IV类为外类群中国水仙。分子聚类结果不仅支持乳白石蒜、鹿葱、红蓝石蒜、江苏石蒜、稻草石蒜的杂交起源观点,而且与形态学分类、物种的染色体倍性、核型存在较高的一致性,即除乳白石蒜外,染色体基数接近或等于11、核型含有11t或11t倍数的物种聚为一大类;染色体基数不等于11、核型为6M+10T的种聚成了另一大类。SSR-PCR容易区分同种不同产地的安徽石蒜或不同花色的长筒石蒜,外形十分相似的石蒜和矮小石蒜,忽地笑和中国石蒜。本研究认为,将安徽石蒜作为一个独立的种不太合适,它可能是长筒石蒜的变种或是以长筒石蒜为亲本的杂交种。

Genetic diversity and inter-specific relationships of Lycoris were analyzed by SSR-PCR technique.46 SSR primer pairs were employed to amplify genomic DNA of Lycoris.18 out of 46 SSR primer pairs which showed high polymorphism were used to detect genetic diversity and inter-specific relationships among 15 Lycoris species and Narcissus tazaetta var.chinensis as an outgroup in this study.The results indicated that the genetic diversity is high,and their percentage of polymorphic bands is up to 97.63% using the non-denaturing polyacrylamide gel electrophoresis and modified argentation.The 20 accessions were divided into four major groups by UPGMA clustering method.Group I included L.anhuiensis,L.1ongituba,L.longituba var.flava,L.aurea,L.chinensis,and L.albiflora.Group II included L.incarnata,L.squamigera,L.sprengeri,L.haywardii and L.rosea.Group III consisted of L.radiata,L.radiata var.pumila,L.straminea and L.houdyshelii.The N.tazetta L.var.chinensis was as an independent group IV.The molecular cluster result of SSR marker not only supports the likelihood hybrid origin of L.albiflora,L.squamigera,L.haywardii,L.houdyshelii and L.straminea,but also matched well with the results of chromosomal,karyotypical,and morphological classification.Except of L.albiflora,these species whose basic chromosome number similar or equal to x =11 and karyotypes including 11t or multiples were clustered together respectively,while other species whose basic chromosome number was not x =11 but with 6M＋10T karyotypes were clustered together.The same species L.anhuiensis at different localities or L.1ongituba which has different flower color can be distinguished easily by SSR-PCR technique,and can also the morphological very similar species such as L.radiata and L.radiata var.pumila,L.aurea and L.chinensis.According to the results,we could conclude that L.anhuiensis as an independent species was not supported,it should be recognized as a variety or hybrid of L.longituba due to their very close relationships,and could provide reference for classification,especially germplasm identification,and the cross breeding of Lycoris.