ITS-RFLP及SRAP标记在灰树花种质评价中的应用

Evaluation of Grifola frondosa Germplasm Using ITS-RFLP and SRAP Markers

以25个灰树花(Grifola frondosa)菌株的子实体为材料提取基因组DNA,ITS扩增后对产物进行酶切,25个菌株的ITS条带处于同一位置,酶切后部分菌株呈现不一样的切点,可将这些菌株大致分为三类。应用扩增清晰、多态性高的47对SRAP引物对这25个菌株进行SRAP分析,共得到138条多态性条带,树状图显示,这些菌株在相似度77%水平上可分为三类,与ITS-RFLP的分析结果类似。SRAP分析结果显示,有10个菌株可以被有效的区分,15个菌株未能找到其相互之间的差异带,说明ITS-RFLP和SRAP标记可以应用于区分菌株,进行种质评价。

Genomic DNA samples prepared from fruit bodies of 25 Grifola frondosa strains were amplified with ITS（internal transcribed spacer）primers.Although the PCR reaction products exhibited identical electrophoresis patterns,hydrolysis using a range of restriction endonucleases revealed that the ITS bands contained different restriction enzyme cutting sites and could be separated into three broad groups based on the sequences of the hydrolytic products.SRAP amplification employing 47 SRAP（sequence related amplified polymorphism） primer pairs generated 138 polymorphic bands.A phylogram constructed using the SRAP data effectively differentiated ten strains whereas no obvious differences were detected between the remaining 15 strains.Our data implied that ITS-RFLP and SRAP markers were effective in G.frondosa strain identification and germplasm evaluation.