RAPD分子标记在酱油生产菌系统发育分析中的应用

Application of RAPD Molecular Marker in the Phylogenetic Analysis of Soy Sauce Strains

摘要:

以米曲霉沪酿3.042（AS3.951）、酱油曲霉AS3.495为参照，对从商品酱油曲精中分离到的8株酱油生产菌进行RAPD分析。实验筛选到6个扩增产物谱带多、特征好、覆盖面广的引物：Primer1、Primer5、Primer6、Primer7、Primer8、Primer9，重复实验证明其RAPD-PCR扩增图谱具有较好的稳定性，扩增产物谱带一般4～8条，各实验菌株主带1～4条,次带丰富。根据RAPD-PCR扩增图谱构建的系统进化树较好地吻合了传统的形态分类学，证实了RAPD分子标记在酱油生产菌系统发育分析中应用的可行性。

Abstract:

RAPD analysis of eight soy sauce strains isolated from commercial soy sauce koji was done with random primers,using Aspergillus oryzae AS3.951 and Aspergillus sojae AS3.495 as controls.Six appropriate primers (Primer1,Primer5,Primer6,Primer7,Primer8,Primer9) for RAPD-PCR were screened from nine random primers,and repetitive experiments demonstrated that their RAPD-PCR polymorphic patterns were stable.There were usually 4～8 bands in their RADP-PCR patterns,where the number of the main bands was 1～4 and the secondary bands were abundant.The phylogenetic tree of these strains was reconstructed according to their RAPD-PCR patterns,and it basically corresponded to traditional morphological taxology,demonstrating that the application of RAPD molecular marker in the phylogenetic analysis of soy sauce strains was feasible.