利用ISSR-SCAR标记鉴定金针菇双-单杂交子  

Identification of Dikaryon-monokaryon Hybrid of Flammulina filiformis by ISSR-SCAR Marker

作　　者：李慧[1] 夏伟伟 胡子钰 王朝江[1] 高春燕[1] LI Hui;XIA Weiwei;HU Ziyu;WANG Chaojiang;GAO Chunyan(Institute of Cash Crops,Hebei Academy of Agriculture and Forestry Sciences,Shijazhuang,Hebei 050051)

机构地区：[1]河北省农林科学院经济作物研究所,河北石家庄050051

出　　处：《北方园艺》2022年第16期121-127,共7页Northern Horticulture

基　　金：河北省重点研发计划资助项目(19226368D);河北省重点研发计划资助项目(21326315D);河北省农林科学院创新工程资助项目(2022KJCXZX-JZS-9);国家食用菌产业技术体系资助项目(CARS-24)。

摘　　要：以2个金针菇菌株为试材,采用原生质体制备与再生技术获得单核体,鉴定其交配型并构建双-单杂交组合,进一步利用ISSR-PCR技术研究亲本、杂交子及其原生质体单核体的遗传差异,以期为金针菇双-单杂交子的准确鉴定提供参考依据。结果表明:2个金针菇菌株分别获得了2种交配型的原生质体单核体,但所占比例差异较大。将FC6的原生质体单核菌株FC6P23与F93配对构建完全亲和双-单杂交组合,获得杂交子FC693。ISSR-PCR扩增图谱结果显示FC693分别含有单核亲本受体核和双核亲本供体核的特异性条带T3和T9,表现为共显性,证明其为真正的杂交子,进一步将特异性条带转化为SCAR标记,利用多重PCR技术可对双-单杂交子进行快速、准确地鉴定。Two strains of Flammulina filiformis were used to create protoplast monokaryons which mating types were determined by mating test.The full compatiable di-mon mating was carried out, and the parents, hybrids and their protoplast monokaryons were used to ISSR-PCR spectrum analysis in order to provide reference for the identification of di-mon hybrid of Flammulina filiformis.The results showed that two types of protoplast monokaryons were obtained for both strains of FC6 and F93 while the ratio of two mating types for monokaryons were different.The hybrid strain of FC693 was obtained by the full compatiable di-mon mating of parent strains of FC6 P23 and F93.ISSR spectrum analysis showed that the specific fragment named T3 was detected in both FC693 and FC6 P23 while the specific fragment named T9 was detected in both FC693 and F93.These specific fragments were designed by SCAR primers and used to multiplex PCR for hybrid and parent strains.These ISSR-SCAR markers could distinguish the di-mon hybrid accurately and rapidly.

关 键 词：金针菇 双单杂交 ISSR标记 SCAR标记 

分 类 号：S646.15[农业科学—蔬菜学]