RAPD分子标记技术用于螺旋藻(Spirulina)分类的研究

CLASSIFICATION WITH RAPD MARKER IN SPIRULINA

用 1 0 6条随机引物对Sp 1等 7株钝顶螺旋藻进行RAPD分析。结果表明 :(1 )绝大多数引物的PCR扩增结果均相同 ,与它们同属于钝顶螺旋藻种的事实相符。 (2 )筛选出 2 3条具有明显多态性的随机引物 ,扩增出的总条带数为 1 5 1 ,平均每条引物的扩增条带数为6 5 7;其中多态性条带为 1 2 0 ,占总带数的 79 5 %。 (3 ) 7株材料经聚类分析可聚成 2大类 ,Sp 1和Sp 2与Sp 6聚成第Ⅰ类 ,Sp 3、Sp 4及Sp 5与Sp 7聚成第Ⅱ类。 (4)第Ⅰ类和第Ⅱ类之间的遗传距离为 9 76 3 9,而两大类内部各藻株间的遗传距离仅为 2 6 4 5 8— 4 5 82 8,说明两大类间的遗传背景差异较大 ,而两大类内部各藻株间的遗传背景差异相对较小。上述分类结果与 7株材料的某些生理生化和分子生物学特性相吻合 ,但与依据藻丝体形态特征的经典分类结果有较大出入。本研究首次将RAPD分子标记技术用于螺旋藻的分类 ,为在DNA水平建立螺旋藻分类与新品种 (系 )

Seven strains of Spirulina platensis (Sp-1-Sp-7) were analyzed with 106 random primers using random amplified polymorphic DNA (RAPD) technique.The results were as follows:(1) The amplification bands were mostly same which accorded with that they belonged to the same species.(2) From the 106 arbitrary primers used,23 primers revealed scorable polymorphisms and generated a total of 151 fragments with a mean of 6.57 bands per primer.120(79.5%)of which were polymorphic.(3) The result of cluster analysis showed that 7 strains of Spirulina platensis could be divided into 2 groups,Sp-1,Sp-2 and Sp-6 belonged to group Ⅰ,while Sp-3,Sp-4,Sp-5 and Sp-7 belonged to group Ⅱ.(4) The genetic distances were 9.7639 between group Ⅰ and group Ⅱ and 2.6458-4.5828 within group Ⅰ or group Ⅱ demonstrated that the genetic relationship were remarkable different in the former whereas relatively close among the later.These results were consistent with some of their physiological and biochemical characters as well as some molecular biological characteristics,but dissimilar with the classification by their morphological characteristics.It was the first time that RAPD marker was used in the classification in Spirulina,it provides a new alternative at DNA molecular level for classification and identification of new strains in Spirulina.