以香菇保藏菌种庆元135以及庆元出菇的135新鲜子实体组织分离得到的菌丝为材料制备原生质体,分别获得58和83个原生质体单核体;经交配型鉴定后,用已获得的135特异SCAR引物对单核进行PCR扩增,统计SCAR条带的分布。结果显示:原生质体单核体分为A1B1和A2B2两种交配型,而特异条带仅存在于后者中,证明135特异SCAR标记是稳定遗传的,也为鉴定以带标记核为亲本之一的杂交后代提供了依据,这就进一步拓宽了分子标记应用于菌株鉴定的适用范围。
Refrigerated mycelium and fresh fruitbody tissue-isolated mycelium of Lentinula edodes strain 135were used to prepare protoplast monokaryons. Fifty-eight and 83 monokaryons were isolated respectively and their mating types were determined. PCR amplification employing specific primers was used to establish the distribution among the protoplast monokaryons of a SCAR marker specific for strain 135. Protoplast monokaryons segregated into either A1B1 or A2B2 mating type, and the SCAR marker was detected only in the latter. The outcome is a regular distribution pattern that demonstrates the stable inheritance of the SCAR marker and is fundamental to identification of offspring arising from crosses between protoplast monokaryons of this particular mating type and other karyons.