基于ISSR-PCR体系鉴别樟芝单核体交配型  

Identification of Monokaryon Mating Type in Taiwanofungus camphoratus Based on ISSR-PCR System

作　　者：李晓晖[1] 盖舒萍 汪雯翰[2] 琚建伟 张守兵 丁保安 李燕[2] 贾薇[2,4] LI Xiaohui;GAI Shuping;WANG Wenhan;JU Jianwei;ZHANG Shoubing;DING Baoan;LI Yan;JIA Wei(College of Food Sctences&Technology,Shanghai Ocean University,Shanghai 201306,China;Institute of Edible Fungi,Shanghai Academy of Agricultural Sciences/Key Laboratory of Edible Fungi Resources and Utilization(South),Ministry of Agriculture and Rural Affairs,P.R.China/National Engineering Research Center of Edible Fungi/National R&D Center for Edtble Fungal Processing,Shanghai 201403,China;Shanghai Tongyan Industrial Co.,Ltd.,Shanghai 201417,China;Shanghai Guosen Bio-tech Co.,Ltd.,Shanghai 201403,China)

机构地区：[1]上海海洋大学食品学院,上海201306 [2]上海市农业科学院食用菌研究所/农业农村部南方食用菌资源利用重点实验室/国家食用菌工程技术研究中心/国家食用菌加工技术研发分中心,上海201403 [3]上海彤颜实业有限公司,上海201417 [4]上海国森生物科技有限公司,上海201403

出　　处：《中国食用菌》2024年第2期68-75,共8页Edible Fungi of China

基　　金：上海市自然科学基金(20ZR1422900)。

摘　　要：通过原生质体单核化技术获得樟芝单核体,基于内转录间隔区(internal transcribed spacer,ITS)序列进行鉴定,采用14个引物对简单重复序列区间(inter-simple sequence repeats,ISSR)进行多态性扩增,筛选条带清晰、重复性好的引物用于樟芝单核体的交配型鉴定。结果表明,通过原生质体单核化技术获得31个樟芝单核体,经ITS序列分析确定获得的单核体为樟芝。以单核体S2、S10、S14、S27的DNA为模板,对14条ISSR引物进行初步筛选,得到4个条带清晰、重复性好的引物P7、P9、P21、P25用于交配型鉴定。单核体S9和S25为同一交配型,两者与S2为不同交配型,通过镜检进一步验证S9、S25可以与S2形成具有锁状联合的双核菌株。该方法可明显缩短樟芝单核体交配型的鉴定时间。The monokaryon of Taiwanofungus camphoratus was obtained by protoplast monokaryon technique,and the sequence was identified based on internal transcribed interval(ITS),14 inter-simple sequence repeats(ISSR)markers were used to select clear and reproducible primers for mating type identification of monokaryon of T.camphoratus.The results showed that 31 monokaryons were obtained by protoplast monokaryon technique,and the monokaryons were identified as T.camphoratus by ITS sequence identification,14 ISSR primers were screened by using the DNA of monokaryons S2,S10,S14,S27 as templates,and 4 primers P7,P9,P21 and P25 with clear bands and good repeatability were obtained for mating type identification.Monokaryon S9 and S25 are of the same mating type and of different mating type from S2,and it was further verified by microscopic examination that monokaryon S9 and S25 could form binucleate strains with lock association with S2.This method can shorten the identification time of monokaryon mating type of T.camphoratus.

关 键 词：樟芝 单核体 交配型 ISSR ITS 

分 类 号：S646.2[农业科学—蔬菜学]