摘要
茯苓Wolfiporia hoelen是一种食药兼用的大型真菌,在我国栽培历史悠久,但由于长期无性繁殖,导致菌种退化,栽培产量下降,严重影响产业发展。为解决茯苓育种的难题,我们前期建立了茯苓同核体鉴定方法,明确了其交配系统,建立了单孢同核体杂交育种体系,但是部分茯苓菌株子实体形成困难或子实体贴生,导致担孢子收集困难,因此原生质体单核化的研究具有重要意义。本研究以两株存在较大遗传差异且同核体类型不同的菌株776和775为材料,核荧光染色表明超过60%的原生质体具有细胞核,原生质体再生率分别为11.0%和7.6%,不易生长的再生菌株中既有同核也有异核菌株。原生质体单核化获得了两个菌株的同核体菌株,比例可达到10%以上,菌株776获得了两种交配型的同核体菌株(5:3),不存在偏分离(χ~2=0.5),但菌株775原生质体单核化仅获得了一种交配型的同核体菌株,表现出严重的交配型偏分离。通过杂交菌株与两亲本对峙试验及基于rpb2的杂合位点验证,证实获得了原生质体同核体杂交菌株25株,原生质体同核体与单孢同核体杂交菌株50株,表明茯苓原生质体同核体杂交育种的可行性。
Wolfiporia hoelen is an edible and medicinal fungus that has been cultivated in China for a long time and is cultivated currently on a large scale.However strain degeneration and decrease in yield caused by long-term asexual propagation affected the development of the industry.In order to solve the difficult breeding problem,distinguishing methods of homokaryotic strains and single-spore homokaryotic strain crossbreeding systems were constructed,and the mating system was disclosed in our previous studies.However,the difficulty of fruiting in some strains or attachment of some fruiting bodies to the media resulted in the difficulty of collecting basidiospores still remains unsettled.Therefore,the research on protoplast monokaryogenesis is important and meaningful.In the present study,the two strains,775 and 776,with distant genetic relationship and different homokaryotic types,were used as experimental strains.Nuclear fluorescence staining revealed that more than 60% of protoplasts had nuclei,and the protoplast regeneration ratio was 7.6% and 11.0% respectively.The regenerative strains with growth stagnancy included both homokaryotic and heterokaryotic strains.The homokaryotic strains were collected with a ratio over 10%,and the homokaryotic strain 776 with different mating type was collected with a ratio of 5:3 with no partial separation(χ~2=0.5),and strain 775 with only one mating type was collected with a significant partial separation.By crossing of protoplast homokaryotic strains derived from protoplast and single spore,25 and 50 hybrids were produced separately,which were confirmed by antagonism tests and rpb2 specific loci.This protoplast homokaryotic strain crossbreeding method built in W.hoelen was proved to be practicable.
作者
李寿建
董彩虹
LI Shoujian;DONG Caihong(State Key Laboratory of Mycology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China;University of Chinese Academy of Sciences,Beijing 100049,China)
出处
《菌物学报》
CAS
CSCD
北大核心
2023年第6期1258-1272,共15页
Mycosystema
基金
贵州省科技计划项目(黔科合支撑[2021]一般249)。
关键词
原生质体单核化
交配型因子偏分离
原生质体核荧光染色
rpb2
protoplast monokaryogenesis
segregation distortion of mating-type factors
nuclear fluorescence staining of protoplasts
rpb2
