Development of Novel Polymorphic EST-SSR Markers in Bailinggu (Pleurotus tuoliensis) for Crossbreeding.
Development of Novel Polymorphic EST-SSR Markers in Bailinggu (Pleurotus tuoliensis) for Crossbreeding.
复制标题
白灵菇杂交育种新型多态性EST-SSR标记的开发
作者：
Dai Y;Su W;Yang C;Song B;Li Y;Fu Y
单核体及其交配型的鉴定和杂交后代的鉴别是白灵菇杂交育种的关键环节。然而，传统的杂交育种方法麻烦且耗时。利用RNA-seq技术，开发了白灵菇单核体及其交配型、遗传多样性和杂交后代的EST-SSR标记。我们从一个新测序的白灵菇转录组中鉴定了1110个潜在的EST-SSR位点，然后随机选择100个EST-SSR进行进一步验证。结果表明，分别有39、43和34个新的EST-SSR标记成功区分了单核体和双亲双核体，区分了两种不同的交配型，区分了F1和F2杂种后代。利用18个高信息量的EST-SSR标记在37个单核体中共检测到86个等位基因。每个位点观察到的等位基因数范围从3到7。聚类分析表明，这些单核体具有较高的遗传多样性水平。近缘种Pleuerynaevar.单核体中EST-SSR的转移率阿魏和糙皮侧耳的分离率分别为72%和64%。本研究为白灵菇与近缘种的杂交育种提供了新的SSR标记和有效的方法。
Identification of monokaryons and their mating types and discrimination of hybrid offspring are key steps for the crossbreeding of Pleurotus tuoliensis (Bailinggu). However, conventional crossbreeding methods are troublesome and time consuming. Using RNA-seq technology, we developed new expressed sequence tag-simple sequence repeat (EST-SSR) markers for Bailinggu to easily and rapidly identify monokaryons and their mating types, genetic diversity and hybrid offspring. We identified 1110 potential EST-based SSR loci from a newly-sequenced Bailinggu transcriptome and then randomly selected 100 EST-SSRs for further validation. Results showed that 39, 43 and 34 novel EST-SSR markers successfully identified monokaryons from their parent dikaryons, differentiated two different mating types and discriminated F1 and F2 hybrid offspring, respectively. Furthermore, a total of 86 alleles were detected in 37 monokaryons using 18 highly informative EST-SSRs. The observed number of alleles per locus ranged from three to seven. Cluster analysis revealed that these monokaryons have a relatively high level of genetic diversity. Transfer rates of the EST-SSRs in the monokaryons of closely-related species Pleurotus eryngii var. ferulae and Pleurotus ostreatus were 72% and 64%, respectively. Therefore, our study provides new SSR markers and an efficient method to enhance the crossbreeding of Bailinggu and closely-related species.
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