基于ISSR分子标记和农艺性状的大球盖菇种质资源遗传多样性分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-1351

摘要/Abstract

摘要：

目的大球盖菇（Stropharia rugosoannulata）栽培群体存在种质资源混杂、优异品种短缺的问题。解析不同地理来源栽培菌株的遗传多样性与表型变异特征，筛选优良种质，为大球盖菇品种改良提供科学依据。方法以67个不同地理来源的大球盖菇栽培菌株为材料，采用10条ISSR引物进行遗传多样性分析，同时测定10个农艺性状，结合相关性分析、主成分分析及聚类分析开展综合评价。结果 10条ISSR引物共扩增出65条清晰条带，其中多态性条带51条，多态性位点百分率为55.56%-100%，Nei’s基因多样性指数平均值为0.27，香农多样性指数平均值为0.43。UPGMA聚类分析可划分为5个类群，遗传相似系数为0.43-0.96，主成分分析累计解释28%的遗传变异，核心类群菌株分布紧密，遗传背景高度相似。农艺性状中，菌盖硬度变异系数最大（124.03%），产量香农多样性指数最高（1.88）；相关性分析显示产量与单朵重、菌盖直径等7个性状呈极显著正相关（P<0.01），菌丝锁状联合数量与所有农艺性状无显著相关性。主成分分析筛选出菌盖直径和菌柄直径为表型综合评价核心指标，前3个主成分累计贡献率达74.51%；综合得分排名前3的菌株为BIPP21340027、BIPP21340016和BIPP21340020，在产量和商品性状上协同优异。结论供试大球盖菇菌株具有遗传基础较窄、表型性状变异丰富的特征，筛选出菌盖直径、菌柄直径为表型评价核心指标，获得3份优良菌株，可为大球盖菇优异种质筛选、杂交亲本选配及新品种选育提供参考。

关键词: 大球盖菇, ISSR, 农艺性状, 遗传多样性, 综合评价

Abstract:

Objective Commercial cultivated populations of Stropharia rugosoannulata face challenges including genetic admixture and a lack of superior varieties. This study aimed to characterize the genetic diversity and phenotypic variation of cultivated strains from diverse geographic origins, screen for elite germplasm, and provide a scientific foundation for variety improvement. Method 67 S. rugosoannulata strains were collected from different geographical regions. Genetic diversity was analyzed using 10 ISSR primers, and 10 agronomic traits were measured. A comprehensive evaluation was conducted by combining correlation analysis, principal component analysis, and cluster analysis. Result A total of 65 clear bands were amplified by ISSR markers, among which 51 were polymorphic, with the percentage of polymorphic loci per primer ranging from 55.56% to 100%. The average Nei’s gene diversity index and Shannon diversity index were 0.27 and 0.43, respectively. UPGMA cluster analysis identified five clusters, with genetic similarity coefficients ranging from 0.43 to 0.96. Principal component analysis accounted for 28% of the total genetic variation, and strains within the core clusters were closely clustered and shared highly similar genetic backgrounds. For agronomic traits, pileus hardness exhibited the highest coefficient of variation (124.03%), while yield had the highest Shannon diversity index (1.88). Yield was extremely significantly positively correlated with seven traits, including single fruiting body weight and pileus diameter (P<0.01), whereas the number of hyphal clamp connections showed no significant correlation with any agronomic trait. The first three phenotypic principal components contributed 74.51% of the total cumulative variance, and pileus diameter and stipe diameter were identified as the core indicators for phenotypic evaluation. Three strains (BIPP21340027, BIPP21340016, and BIPP21340020) ranked top in the comprehensive evaluation, featuring both high yield and excellent commercial traits. Conclusion This study revealed that the tested Stropharia rugosoannulata strains have a narrow genetic base yet rich phenotypic variation. Pileus diameter and stipe diameter were designated as the core indicators for phenotypic evaluation, and three elite strains were screened out. The results provide a reference for screening superior germplasm, selecting hybrid parents, and breeding new varieties of Stropharia rugosoannulata.

Key words: Stropharia rugosoannulata, ISSR, agronomic traits, genetic diversity, comprehensive evaluation

李诗琪, 武月园, 张殿朋, 郭远, 刘宇, 宋忠娟, 王守现, 宋爽. 基于ISSR分子标记和农艺性状的大球盖菇种质资源遗传多样性分析[J]. 生物技术通报, 2026, 42(11): 252-262.

LI Shi-qi, WU Yue-yuan, ZHANG Dian-peng, GUO Yuan, LIU Yu, SONG Zhong-juan, WANG Shou-xian, SONG Shuang. Genetic Diversity Analysis of Stropharia rugosoannulata Germplasm Resources Based on ISSR Markers and Agronomic Traits[J]. Biotechnology Bulletin, 2026, 42(11): 252-262.

图/表 11

参考文献 32

[32]	Liang XD, Wu J, Liu JL, et al. Correlation and regression analysis of Lentinula edodes yield and its constituent factors [J]. China Cucurbits Veg, 2024, 37(10): 56-62.
[33]	曹雪莲, 陈影, 彭卫红, 等. 毛头鬼伞种质资源农艺性状遗传多样性分析及综合评价 [J]. 食用菌学报, 2022, 29(5): 53-64.
	Cao XL, Chen Y, Peng WH, et al. Genetic diversity analysis and comprehensive evaluation of agronomic traits of Coprinus comatus germplasm resources [J]. Acta Edulis Fungi, 2022, 29(5): 53-64.
[34]	王瑞娟, 尚晓冬, 杨慧, 等. 不同保藏方法对金针菇“上研1号”栽培特性的影响 [J]. 食用菌学报, 2019, 26(1): 11-17.
	Wang RJ, Shang XD, Yang H, et al. Effect of different preservation methods on the main cultivation characteristics of Flammulina velutipes “shangyan 1” [J]. Acta Edulis Fungi, 2019, 26(1): 11-17.
[35]	董锡文, 朱德旋, 杜春梅, 等. 4种维生素对黑木耳锁状联合菌丝形成的影响 [J]. 安徽农业科学, 2019, 47(5): 56-57, 61.
	Dong XW, Zhu DX, Du CM, et al. The effects of four kinds of vitamin on the forming of hyphal clamp connection of aurieuzaria auricular [J]. J Anhui Agric Sci, 2019, 47(5): 56-57, 61.
[1]	罗信昌, 陈士瑜. 中国菇业大典 [M]. 北京: 清华大学出版社, 2010: 986-1003.
	Luo XC, Chen SY. Collection of mushroom industry in China [M]. Beijing: Tsinghua University Press, 2010: 986-1003.
[2]	张颖. 大球盖菇北方棚内反季栽培技术 [J]. 中国林副特产, 2014(6): 54-55.
	Zhang Y. Off-season cultivation techniques of Pleurotus ostreatus in northern greenhouse [J]. For Prod Speciality China, 2014(6): 54-55.
[3]	Brodzinska Z., Lasota W. Chemical composition of cultivated mushrooms Part I. Stropharia rugoso-annulata Farlow ex Murr [J]. Bromatol. I Chem. Toksykol. 1981, 14, 229-238.
[4]	中国预防医学科学院营养与食品卫生研究所. 食物成分表[M]. 北京: 人民卫生出版社, 2002: 52-53.
	Institute of Nutrition and Food Safety, Chinese Academy of Preventive Medicine. Food Composition Table[M]. Beijing: People’s Medical Publishing House, 2002: 52-53.
[5]	王晓炜, 詹巍, 陶明煊, 等. 大球盖菇营养成分、抗氧化活性物质分析 [J]. 食用菌, 2007, 29(6): 62-63.
	Wang XW, Zhan W, Tao MX, et al. Analysis of nutritional components and antioxidant active substances of Pleurotus ostreatus [J]. Edible Fungi, 2007, 29(6): 62-63.
[6]	Liu YT, Sun J, Luo ZY, et al. Chemical composition of five wild edible mushrooms collected from Southwest China and their antihyperglycemic and antioxidant activity [J]. Food Chem Toxicol, 2012, 50(5): 1238-1244.
[7]	李淑荣, 王丽, 倪淑君, 等. 大球盖菇不同部位氨基酸含量测定及营养评价 [J]. 食品研究与开发, 2017, 38(8): 95-99.
	Li SR, Wang L, Ni SJ, et al. The amino acids content of different part of Stropharia rugoso-annulata and their nutrition evaluation [J]. Food Res Dev, 2017, 38(8): 95-99.
[8]	Wei L, Jing BN, Li X, et al. Evaluation of nutritional ingredients, biologically active materials, and pharmacological activities of Stropharia rugosoannulata grown under the bamboo forest and in the greenhouse [J]. J Food Qual, 2021, 2021: 5478227.
[9]	Wu J, Fushimi K, Tokuyama S, et al. Functional-food constituents in the fruiting bodies of Stropharia rugosoannulata [J]. Biosci Biotechnol Biochem, 2011, 75(8): 1631-1634.
[10]	Yan QX, Huang MX, Sun P, et al. Steroids, fatty acids and ceramide from the mushroom Stropharia rugosoannulata Farlow apud Murrill [J]. Biochem Syst Ecol, 2020, 88: 103963.
[11]	颜淑婉, 苏诗垂. 大球盖菇人工栽培研究简报 [J]. 江苏食用菌, 1995(3): 2.
	Yan SW, Su SC. Brief report on artificial cultivation of Pleurotus ostreatus [J]. Jiangsu Edible Fungi, 1995(3): 2.
[12]	Zhang Y, Ni JP, Yang J, et al. Citrus stand ages regulate the fraction alteration of soil organic carbon under a Citrus/Stropharua rugodo-annulata intercropping system in the Three Gorges Reservoir area, China [J]. Environ Sci Pollut Res, 2017, 24(22): 18363-18371.
[13]	任纪帆, 朱静娴, 王庆佶, 等. 大球盖菇“山农球盖3号”品种的选育 [J]. 菌物学报, 2020, 39(6): 977-982.
	Ren JF, Zhu JX, Wang QJ, et al. Breeding of Stropharia rugosoannulata “shannongqiugai No.3” [J]. Mycosystema, 2020, 39(6): 977-982.
[14]	宋爽, 刘宇, 武月园, 等. 野生白灵侧耳种质资源遗传多样性分析和评价 [J]. 食用菌学报, 2025, 32(3): 120-130.
	Song S, Liu Y, Wu YY, et al. Genetic diversity analysis and evaluation of wild Pleurotus tuoliensis germplasm resources [J]. Acta Edulis Fungi, 2025, 32(3): 120-130.
[15]	陆欢, 尚晓冬, 宋春艳, 等. 基于ISSR标记和农艺性状的金顶侧耳种质资源遗传多样性分析 [J]. 食用菌学报, 2022, 29(6): 14-24.
	Lu H, Shang XD, Song CY, et al. Genetic diversity analysis of Pleurotus citrinopilestu germplasm resources based on ISSR markers and agronomic traits [J]. Acta Edulis Fungi, 2022, 29(6): 14-24.
[16]	Yan PS, Jiang JH, Li GF, et al. Mating system and DNA polymorphism of monokaryons with different mating type of Stropharia rugoso-annulata [J]. World J Microbiol Biotechnol, 2003, 19(7): 737-740.
[17]	朱静娴, 李丽君, 赵淑芳, 等. 基于SRAP分子标记的23株大球盖菇遗传多样性和亲缘关系分析 [J]. 山东农业科学, 2018, 50(3): 22-28.
	Zhu JX, Li LJ, Zhao SF, et al. Genetic diversity and phylogenetic relationship analysis of 23 Stropharia rugosoannulata strains based on SRAP molecular marker [J]. Shandong Agric Sci, 2018, 50(3): 22-28.
[18]	陈绪涛, 章炉军, 孙鹏, 等. 我国主栽皱环球盖菇菌株遗传多样性及栽培特性 [J]. 菌物学报, 2021, 40(12): 3081-3095.
	Chen XT, Zhang LJ, Sun P, et al. Genetic diversity and cultivation characteristics of the main cultivars of Stropharia rugosoannulata in China [J]. Mycosystema, 2021, 40(12): 3081-3095.
[19]	曾宇宇. 大球盖菇核心种质库构建及其基质降解能力分析 [D]. 武汉: 华中农业大学, 2023.
	Zeng YY. Establishment of core collections and analysis of substrate degradation ability of Stropharia rugosoannulata [D]. Wuhan: Huazhong Agricultural University, 2023.
[20]	鲍大鹏. 我国食用菌遗传学的发展及展望 [J]. 菌物学报, 2021, 40(4): 806-821.
	Bao DP. Development and prospect of genetics of edible mushrooms in China [J]. Mycosystema, 2021, 40(4): 806-821.
[21]	仇律雯, 杨扬, 范亚明, 等. 国家东南区鲜食糯玉米品质及农艺性状与SSR标记遗传多样性分析 [J]. 江苏农业科学, 2022, 50(18): 130-135.
	Qiu L, Yang Y, Fan YM, et al. Genetic diversity analysis of quality, agronomic traits and SSR markers of fresh waxy maize in Southeastern China [J]. Jiangsu Agric Sci, 2022, 50(18): 130-135.
[22]	赫卫, 张慧. 基于表型性状和SRAP标记的观赏用辣椒种质资源遗传多样性分析 [J]. 中国瓜菜, 2022, 35(1): 16-23.
	He W, Zhang H. Analysis of genetic diversity of pepper germplasm resources for ornamental based on phenotypic traits and SRAP markers [J]. China Cucurbits Veg, 2022, 35(1): 16-23.
[23]	李辉玲, 张建文, 李守明. 基于SSR标记的新疆大蒜及其野生蒜种遗传多样性及聚类分析 [J]. 江苏农业科学, 2022, 50(18): 158-163.
	Li HL, Zhang JW, Li SM. Genetic diversity and clustering analysis of Xinjiang garlic and wild garlic resources based on SSR markers [J]. Jiangsu Agric Sci, 2022, 50(18): 158-163.
[24]	陈兆贵, 张荃锋, 卢晓琪, 等. 基于ISSR分子标记的冬种马铃薯品种遗传多样性分析 [J]. 湖南农业科学, 2022(1): 8-11.
	Chen ZG, Zhang QF, Lu XQ, et al. Genetic diversity analysis of winter potato varieties based on ISSR molecular markers [J]. Hunan Agric Sci, 2022(1): 8-11.
[25]	Gong WB, Zeng YY, Li XR, et al. Molecular profiling of rice straw degradability discrepancy in Stropharia rugosoannulata core germplasm [J]. J Agric Food Chem, 2024, 72(45): 25379-25390.
[26]	杨瑞恒, 吴莹莹, 宋春艳, 等. 基于公共数据库信息对中国野生香菇种质资源地理分布的再分析 [J]. 菌物学报, 2018, 37(3): 335-351.
	Yang RH, Wu YY, Song CY, et al. Reanalysis of geographical distribution of wild Lentinula edodes in China based on the information from public databases [J]. Mycosystema, 2018, 37(3): 335-351.
[27]	刘天宇. 马尾松种子园亲本GBS-SNP开发及群体遗传多样性研究 [D]. 长沙: 中南林业科技大学, 2023.
	Liu TY. Genetic diversity of parent in seed orchard of Pinus massoniana using SNP calling from genotyping by sequencing [D]. Changsha: Central South University of Forestry & Technology, 2023.
[28]	Xiao Y, Cheng XJ, Liu J, et al. Population genomic analysis uncovers environmental stress-driven selection and adaptation of Lentinula edodes population in China [J]. Sci Rep, 2016, 6: 36789.
[29]	宋莹, 刘俊杰, 刘岩岩, 等. 辽宁省主栽香菇菌株ISSR遗传差异性分析 [J]. 北方园艺, 2017(5): 82-85.
	Song Y, Liu JJ, Liu YY, et al. ISSR analysis of genetic diversity of main cultivated strains of Lentinula edodes in Liaoning [J]. North Hortic, 2017(5): 82-85.
[30]	王洪秀, 章炉军, 孙鹏, 等. 对92个茶树菇菌株的遗传多样性分析和农艺性状的评价 [J]. 微生物学杂志, 2024, 44(2): 19-32.
	Wang HX, Zhang LJ, Sun P, et al. Genetic diversity and agronomic traits evaluation of 92 tea tree mushroom (Agrocybe aegerita) strains [J]. J Microbiol, 2024, 44(2): 19-32.
[31]	刘芹, 孔维丽, 崔筱, 等. 基于模糊数学和聚类分析的香菇品种综合品质评价 [J]. 中国瓜菜, 2021, 34(11): 37-46.
	Liu Q, Kong WL, Cui X, et al. Comprehensive quality evaluation of Lentinula edodes using fuzzy mathematics and cluster analysis [J]. China Cucurbits Veg, 2021, 34(11): 37-46.
[32]	梁雪迪, 吴杰, 刘金玲, 等. 香菇产量及其构成因素的相关性和回归分析 [J]. 中国瓜菜, 2024, 37(10): 56-62.

菌株编号Strain number	来源Origin	菌株编号Strain number	来源Origin	菌株编号Strain number	来源Origin	菌株编号Strain number	来源Origin
BIPP21340001	北京市海淀区	BIPP21340023	北京市密云区	BIPP21340040	北京市房山区	BIPP21340061	山东省
BIPP21340002	北京市通州区	BIPP21340024	北京市密云区	BIPP21340041	河北邢台临县	BIPP21340062	山东省
BIPP21340004	北京市房山区	BIPP21340025	北京市房山区	BIPP21340042	河北邢台临县	BIPP21340063	山东省
BIPP21340005	北京市房山区	BIPP21340026	北京市房山区	BIPP21340043	北京市房山区	BIPP21340064	山东省
BIPP21340008	北京市密云区	BIPP21340027	北京市密云区	BIPP21340044	陕西省安康市	BIPP21340065	山东省
BIPP21340010	北京市大兴区	BIPP21340028	北京市房山区	BIPP21340045	福建省	BIPP21340066	新疆维吾尔自治区
BIPP21340011	北京市大兴区	BIPP21340029	北京市顺义区	BIPP21340050	福建省	BIPP21340067	新疆维吾尔自治区
BIPP21340012	北京市房山区	BIPP21340030	北京市平谷区	BIPP21340051	山东省聊城市	BIPP21340068	新疆维吾尔自治区
BIPP21340013	北京市丰台区	BIPP21340031	北京市平谷区	BIPP21340052	北京市海淀区	BIPP21340069	山东省
BIPP21340015	北京市顺义区	BIPP21340032	北京市丰台区	BIPP21340053	四川省	BIPP21340070	北京市海淀区
BIPP21340016	北京市顺义区	BIPP21340033	北京市大兴区	BIPP21340054	四川省	BIPP21340072	北京市海淀区
BIPP21340017	北京市顺义区	BIPP21340034	北京市海淀区	BIPP21340055	山东省	BIPP21340074	福建省
BIPP21340018	北京市顺义区	BIPP21340035	北京市海淀区	BIPP21340056	山东省	BIPP21340081	云南省
BIPP21340019	北京市顺义区	BIPP21340036	北京市房山区	BIPP21340057	黑龙江省	BIPP21340082	云南省
BIPP21340020	北京市顺义区	BIPP21340037	北京市房山区	BIPP21340058	黑龙江省	BIPP21340085	山东省
BIPP21340021	北京市平谷区	BIPP21340038	北京市房山区	BIPP21340059	山东省	BIPP21340090	黑龙江省
BIPP21340022	北京市延庆区	BIPP21340039	北京市海淀区	BIPP21340060	山东省

菌株编号Strain number	来源Origin	菌株编号Strain number	来源Origin	菌株编号Strain number	来源Origin	菌株编号Strain number	来源Origin
BIPP21340001	北京市海淀区	BIPP21340023	北京市密云区	BIPP21340040	北京市房山区	BIPP21340061	山东省
BIPP21340002	北京市通州区	BIPP21340024	北京市密云区	BIPP21340041	河北邢台临县	BIPP21340062	山东省
BIPP21340004	北京市房山区	BIPP21340025	北京市房山区	BIPP21340042	河北邢台临县	BIPP21340063	山东省
BIPP21340005	北京市房山区	BIPP21340026	北京市房山区	BIPP21340043	北京市房山区	BIPP21340064	山东省
BIPP21340008	北京市密云区	BIPP21340027	北京市密云区	BIPP21340044	陕西省安康市	BIPP21340065	山东省
BIPP21340010	北京市大兴区	BIPP21340028	北京市房山区	BIPP21340045	福建省	BIPP21340066	新疆维吾尔自治区
BIPP21340011	北京市大兴区	BIPP21340029	北京市顺义区	BIPP21340050	福建省	BIPP21340067	新疆维吾尔自治区
BIPP21340012	北京市房山区	BIPP21340030	北京市平谷区	BIPP21340051	山东省聊城市	BIPP21340068	新疆维吾尔自治区
BIPP21340013	北京市丰台区	BIPP21340031	北京市平谷区	BIPP21340052	北京市海淀区	BIPP21340069	山东省
BIPP21340015	北京市顺义区	BIPP21340032	北京市丰台区	BIPP21340053	四川省	BIPP21340070	北京市海淀区
BIPP21340016	北京市顺义区	BIPP21340033	北京市大兴区	BIPP21340054	四川省	BIPP21340072	北京市海淀区
BIPP21340017	北京市顺义区	BIPP21340034	北京市海淀区	BIPP21340055	山东省	BIPP21340074	福建省
BIPP21340018	北京市顺义区	BIPP21340035	北京市海淀区	BIPP21340056	山东省	BIPP21340081	云南省
BIPP21340019	北京市顺义区	BIPP21340036	北京市房山区	BIPP21340057	黑龙江省	BIPP21340082	云南省
BIPP21340020	北京市顺义区	BIPP21340037	北京市房山区	BIPP21340058	黑龙江省	BIPP21340085	山东省
BIPP21340021	北京市平谷区	BIPP21340038	北京市房山区	BIPP21340059	山东省	BIPP21340090	黑龙江省
BIPP21340022	北京市延庆区	BIPP21340039	北京市海淀区	BIPP21340060	山东省

引物名称 Primer name	引物序列 Primer sequence （5′-3′）
ISSR1	CTCCTCCTCCTCSC
ISSR2	BDBACAACAACAACAACA
ISSR4	CACCACCACCACSC
ISSR5	VDHTCGTCGTCGTCGTCG
ISSR6	DHBCGACGACGACGACGA
ISSR12	AGAGAGAGAGAGAGAGT
ISSR13	GAGAGAGAGAGAGAGAC
ISSR15	CACACACACACACACAG
ISSR17	TGTGTGTGTGTGTGTGRC
ISSR18	VHVGTGTGTGTGTGTGTGT

引物名称 Primer name	引物序列 Primer sequence （5′-3′）
ISSR1	CTCCTCCTCCTCSC
ISSR2	BDBACAACAACAACAACA
ISSR4	CACCACCACCACSC
ISSR5	VDHTCGTCGTCGTCGTCG
ISSR6	DHBCGACGACGACGACGA
ISSR12	AGAGAGAGAGAGAGAGT
ISSR13	GAGAGAGAGAGAGAGAC
ISSR15	CACACACACACACACAG
ISSR17	TGTGTGTGTGTGTGTGRC
ISSR18	VHVGTGTGTGTGTGTGTGT

引物Primer	扩增出的条带数 Number of amplified bands	多态性条带数 Number of polymorphic bands	多态位点百分率Percentage of polymorphic loci	观测等位基因数 Observed number of alleles	有效等位基因数 Effective number of alleles	Nei’s基因多样性指数 Nei’s gene diversity index	香农多样性指数 Shannon diversity index（I）
ISSR1	9	8	88.89%	1.89	1.59	0.34	0.67
ISSR2	7	6	85.71%	1.86	1.51	0.31	0.47
ISSR4	6	4	66.67%	1.67	1.29	0.18	0.29
ISSR5	7	5	71.43%	1.71	1.43	0.27	0.40
ISSR6	9	5	55.56%	1.56	1.22	0.15	0.23
ISSR12	5	3	60.00%	1.60	1.33	0.21	0.32
ISSR13	7	7	100%	2.00	1.83	0.45	0.64
ISSR15	4	4	100%	2.00	1.59	0.34	0.51
ISSR17	6	6	100%	2.00	1.59	0.36	0.55
ISSR18	5	3	60.00%	1.60	1.16	0.12	0.19