基于等位基因InDel快速检测黑木耳原生质体单核体

以黑木耳 Auricularia auricula-judae栽培菌株Au916三磷酸甘油醛脱氢酶（GAPDH）两个等位基因中的InDel为基础，以原生质体再生菌株为研究材料，建立了基于InDel的黑木耳原生质体单核体检测技术，并采用荧光显微镜检和配对试验对此方法进行了验证。应用InDel能清晰地区分黑木耳双核体和两种原生质体单核体，且不同的单核体之间可以进行交配，其杂交双核体能正常出耳，结果表明基于等位基因InDel标记可以快速准确地检测黑木耳原生质体单核体。在大规模测序获得真菌基因组信息的基础上，InDel标记在原生质体单核体鉴定中的应用将更加普遍。

Rapid identification of protoplast-regenerated monokaryotic isolates of Auricularia auricula-judae based on an allele InDel marker

An insertion-deletion length polymorphism (InDel) marker was designed based on the two alleles of glyceraldehyde-3- phosphate dehydrogenase gene isolated from Auricularia auricula-judae strain Au916. The protoplast regenerated strains from Au916 were examined to identify the monokaryons based on the InDel marker. Fluorescence microscopic examination and paired test were also conducted to verify the results of InDel detection. The results showed that the dikaryons and two different types of monokaryons from parent strain were separated from each other by the PCR patterns due to presence or absence of the InDel. Different genotype monokaryons could mate with each other to form clamp connection for producing fruiting body. In conclusion, the method based on the allele’s InDel could quickly and accurately identify the monokaryon from protoplast regenerated strains of A. auricula-judae. Based on large-scale sequencing of the fungal genomes, identification and application of InDel markers in monokaryon identification will become more universal.