茶树菇B交配型位点结构分析

doi:10.11924/j.issn.1000-6850.casb20200300292

中国农学通报 ›› 2021, Vol. 37 ›› Issue (2): 14-19.doi: 10.11924/j.issn.1000-6850.casb20200300292

所属专题：食用菌；食用菌；园艺

茶树菇B交配型位点结构分析

钱蓉¹^,²(), 陈卫民²^,³^,⁴, 柴红梅²^,³^,⁴, 陶南²^,³^,⁴, 赵永昌²^,³^,⁴, 陈玉惠¹()

¹西南林业大学生命科学学院,昆明 650224
²云南省农业科学院生物技术与种质资源研究所,昆明 650221
³云南省农业生物技术重点实验室,昆明 650223
⁴农业部西南作物基因资源与种质创制重点实验室,昆明 650223

收稿日期:2020-03-31 修回日期:2020-05-20 出版日期:2021-01-15 发布日期:2021-01-14
通讯作者: 陈玉惠
作者简介:钱蓉,女,1993年出生,云南曲靖人,硕士研究生,主要从事大型食用真菌遗传学方面的研究。通信地址：650205 昆明市盘龙区北京路延长线2238号云南省农业科学院生物技术与种质资源研究所,Tel：0871-65174681,E-mail：1433549562@qq.com。
基金资助:
国家自然科学基金“信息素受体多等位基因在田头菇属真菌生物种识别中的作用研究”(31360024);现代农业(食用菌)产业技术体系(CARS20)

B Mating Locus from Agrocybe aegerita: Structural Analysis

Qian Rong¹^,²(), Chen Weimin²^,³^,⁴, Chai Hongmei²^,³^,⁴, Tao Nan²^,³^,⁴, Zhao Yongchang²^,³^,⁴, Chen Yuhui¹()

¹College of Life Science Southwest Forestry University, Kunming 650224
²Biotechnology and Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming 650221
³Yunnan Provincial Key Lab of Agricultural Biotechnology, Kunming 650223
⁴Key Lab of Southwestern Crop Gene Resources and Germplasm Innovation, Ministry of Agriculture, Kunming 650223

Received:2020-03-31 Revised:2020-05-20 Online:2021-01-15 Published:2021-01-14
Contact: Chen Yuhui

摘要/Abstract

摘要：

为了探明茶树菇有性生殖内在分子机制,包括交配极性转换产生的同核体出菇问题。以茶树菇YSG同核体子代为实验菌株,依据基因组注释信息,利用嵌套引物验证B交配位点,设计引物扩增同核体群体,获得的扩增谱与菌株交配型进行比对。结果表明：（1）同核体培养后出现3种类型,菌丝型、无盖型和菌柄型,后两种为畸形子实体,不进行有性生殖;（2）依据交配位点B设计的引物扩增谱显示,B交配位点完整遗传到子代菌株中,没有观察到重组事件发生;（3）交配型分析显示,该B交配位点决定菌株的交配型,没有新的交配极性出现。本研究推断供试菌株茶树菇YSG同核体只包含一个B交配位点,结果与已报道的3个亚位点不同。这些结果差异是否由于类群差异造成,还需要更多的出菇实验及交配位点鉴定予以确认。

关键词: 茶树菇, 同核体出菇, B交配基因, 交配型转换, 有性生殖

Abstract:

The aim is to investigate the molecular mechanism of sexual reproduction including mating type switching in Agrocybe aegerita. Homokaryon offspring isolated from Agrocybe aegerita YSG strain was employed in this study. Based on the genome annotation, B mating locus was identified using overlapping primers. Primer pairs for B mating genes were used for amplifying the monokaryotic population, and the amplification spectrum was compared with mating type. The results showed: (1) three types including mycelia, capless and stripe-like fruiting bodies occurred after the cultivation of homokaryon, and the latter two were the abnormal fruiting bodies without going through the sexual reproduction; (2) the amplified spectrum using the primer pairs based on the mating gene sequences showed that the complete B mating locus was inherited into the offspring strains without recombination event occurrence; (3) mating type analysis showed that B mating locus governed the mating behavior, and new mating type was not found. It is inferred that Agrocybe aegerita YSG only contained one B mating locus, which is also different from the three reported sublocus. More fruiting tests and mating loci identification are needed for clarifying whether the differences are caused by the group varieties.

Key words: Agrocybe aegerita, homokaryon fruiting, B mating gene, mating type switching, sexual reproduction

中图分类号:

S646.9

钱蓉, 陈卫民, 柴红梅, 陶南, 赵永昌, 陈玉惠. 茶树菇B交配型位点结构分析[J]. 中国农学通报, 2021, 37(2): 14-19.

Qian Rong, Chen Weimin, Chai Hongmei, Tao Nan, Zhao Yongchang, Chen Yuhui. B Mating Locus from Agrocybe aegerita: Structural Analysis[J]. Chinese Agricultural Science Bulletin, 2021, 37(2): 14-19.

图/表 4

图1. 同核体菌株出菇特征 A为菌丝型同核体;B为无盖型同核体;C为菌柄型同核体

引物名称	序列(5’-3’)	Tm/℃	长度/bp
YS8RCB1f	GCTGTGCTCCTCCAACTCTATT	57.7	5162
YS8RCB1r	ACTTCTCGTCTGCCTTCTTCTC	57.7	5162
YS8RCB2f	TTGTCGTTGGCGTCCACCTT	63.7	2151
YS8RCB2r	TCGCCTTTCGTTAGCCCTCC	63.7	2151
YS8RCB3f	ATCCGTGTTCGCACTCGTT	58.2	3567
YS8RCB3r	CGTCGCTCGTATTCGTTGA	58.2	3567
YS8RCB4f	GCCATATCGTGCCTTACCG	57.7	2331
YS8RCB4r	GTGAATAGCGAGCGTCCAG	57.7	2331
YS8RCB5f	ACGGTTCGTGGTTCGTTGAT	59.4	2980
YS8RCB5r	CAGCCGATGAGTTTGTGGAG	59.4	2980
P3af	TGGCTCTTCGTTGCGTTCTTC	62.9	459
P3ar	TAGCGGTGTCTGGTGGTCGTC	62.9	459
P3bf	TGTCGTTGCGATGGAGTTT	56.3	463
P3br	GTGGCGTGTGGGAGTATTC	56.3	463

表1. YS8菌株B交配位点验证及交配型鉴定引物特征

图2. B交配位点结构特征 A为YS8菌株B交配位点结构;B为B2-rcb3a下游和B2-rcb3b上游区域序列互为反向互补;C为B2-rcb3a下游和B2-rcb3b拼接为完整B2-rcb3

图3. 同核体子代B交配位点基因型及交配型 A为分别使用引物对P3af/r和P3bf/r获得的扩增谱;B为各同核体所对应的B交配型

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茶树菇B交配型位点结构分析

B Mating Locus from Agrocybe aegerita: Structural Analysis

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