利用SSR标记鉴定香菇单核体及杂交后代

Identification of Xianggu (Lentinula edodes) monokaryons and hybrid progenies using SSR markers

摘要:

【目的】研究简单重复序列(Simple sequence repeat，SSR)分子标记方法用于香菇原生质体单核体、孢子单核体及其杂交后代的分离和鉴定。【方法】利用基于香菇全基因组序列信息开发的SSR标记，分析由香菇品种“L808”双核菌丝制备的原生质体单核体、孢子单核体及其杂交后代的SSR指纹。【结果】对制备的原生质体单核体的鉴定中，在不经过杂交配对的情况下，鉴定出“L808”的两种不同极性的原生质体单核体，其分离比例为191:1，该鉴定结果得到SSR标记、随机扩增多态性DNA (Random amplified polymorphismic DNA，RAPD)标记及传统方法的验证。另外，开发的香菇SSR标记还能以多位点组合的方式，用于对孢子单核体及其杂交后代的鉴定。【结论】应用SSR标记可加快香菇单核体的制备进程，并提高鉴定单核体及相关杂交菌株的准确性，促进香菇遗传育种研究。

Abstract:

[Objective] Simple sequence repeat (SSR) molecule markers were used to separate and identify protoplast monokaryons, sporulated monokaryons and their hybrid progenies of two Xianggu (Lentinula edodes) strains. [Methods] SSR primers developed from the whole genome sequence of Xianggu were used to identify the genotypes of different monokaryons and hybrids, and then to separate them according different genotype information. [Results] Employing Lefp-55 SSR markers, we obtained both protoplast monokaryons of strain “L808” without matching crossing of monokaryons, and we found the segregation ratio of both protoplast monokaryons was high to 191:1. This result was confirmed by other SSR markers, random amplified polymorphismic DNA (RAPD) markers and tranditional method. In the identification of sporulated monokaryons and their hybrid progenies, cooperation of several SSR markers was able to identify numerous hybrids, and it was useful in genetic and breeding research. [Conclusion] Protoplast monokaryons separation would be fast and accurate by using SSR markers, which also can be used in identification of sporulated monokaryons and their hybrid progenies of Xianggu in relevant genetic and breeding research.