紫芝体细胞不亲和性与遗传差异分析

蒋晓涵; 杨春艳; 王车昭; 刘新锐

紫芝体细胞不亲和性与遗传差异分析

福建农林大学生命科学学院/福建农林大学菌物研究中心，福建　福州　350002

基金项目: 福建省科技重大专项专题项目（2022NZ029015）

详细信息

作者简介:
蒋晓涵（2000 —），女，在读研究生，主要从事食用菌遗传育种研究，E-mail：978234858@qq.com

通讯作者:
刘新锐（1980 —），男，博士，副研究员，主要从事食用菌遗传育种研究，E-mail： 121376381@qq.com

中图分类号: Q36
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出版历程
- 收稿日期: 2024-03-18
- 修回日期: 2024-04-10
- 网络出版日期: 2024-06-26

Somatic incompatibility and genetic difference of Ganoderma sinense

College of Life Sciences, Fujian Agriculture and Forestry University/Mycological Research Center of Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China

摘要

摘要: 目的评价紫芝菌株间体细胞不亲和反应与遗传差异的关系，为应用体细胞不亲和性评价紫芝菌株间的遗传差异提供依据。方法以7个不同交配基因型的紫芝单核体为亲本，应用单向或者双向核迁移技术构建双核体菌株；通过在PDA培养基配对检测双核体菌株间的体细胞不亲和性，并采用ISSR、RAPD和SRAP 3种分子标记综合分析双核体菌株间的遗传差异。结果构建了5类遗传背景清晰的不同双核体菌株11个，它们之间的体细胞不亲性反应分为亲和、不亲和，其中不亲和反应出现隔离区、隔离区带线状和类似墙式结构的3种类型；3种分子标记综合分析显示，11个菌株间的遗传相似系数0.29～0.97，UPMGA聚类树状图能很好的展示11个双核体菌株间的遗传差异，并与亲本来源相一致。结论紫芝菌株间的体细胞不亲性反应主要受细胞核的影响，而细胞质的影响极小，并且紫芝菌株间的体细胞不亲性反应类型与菌株间的遗传差异相对应，在以后的紫芝种质资源遗传差异评价中，可应用操作简便的体细胞不亲和性进行初步分析。
- 拮抗 /
- 遗传关系 /
- 分子标记
Abstract: Objective The relation between somatic incompatibility (SI) and genetic difference of Ganoderma sinense was investigated. This will provide the basis for evaluating genetic differences of G.sinense strains by SI test. Method Construction of dikaryon strains using unidirectional or bidirectional nuclear migration test with seven monokaryon of G.sinense as parents. Then SI reaction was tested on the PDA medium, and the genetic differences between them were analyzed by ISSR, RAPD and SRAP molecular markers. Result Eleven dikaron strains of five types with clear genetic relationship were obtained. The SI reactions were result in compatibility and incompatibility. And the incompatibility emerged three types of gap, gap with line and wall-like structure. The combined analysis of ISSR, RAPD and SRAP molecular markers genetic similarity coefficients ranging from 0.29 to 0.97 among the eleven strains.The UPMGA clustering diagram was able to demonstrate the genetic relationship of eleven trains, and these genetic relationships correspond to those of the parental sources. Conclusion The nucleus has a large effect on the SI response of G.sinense, whereas the cytoplasm has a minimal effect on it. The type of SI reaction of G. sinense strains corresponds to their genetic differences. So in the future evaluation of genetic differences in G. sinense germplasm resources, a simple and convenient method of SI could be applied for preliminary analysis.
- antagonism /
- genetic relationship /
- molecular marker

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图 1 紫芝双核体在PDA培养基上的SI反应

正面(F)/背面(B)。

Figure 1. Somatic incompatibility gap reactions between heterokaryotic colonies of G.sinense paired on PDA medium

Front (F)/back (B).

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图 2 11株紫芝ISSR、RAPD和SRAP分析UPMGA聚类树状图

Figure 2. UPMGA dendrogram based on ISSR, RAPD and SRAP analysis of 11 G.sinense strains

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表 1 引物序列

Table 1. sequences of primers used for molecular markers

分子标记 Molecular marker	引物 primer	序列（5′-3′） sequences	分子标记 Molecular marker	引物 primer	序列（5′-3′） sequences
ISSR	ISSR6	DHB (CGA)₅	SRAP	ME4	TGAGTCCAAACCGGACC
ISSR	ISSR17	(TG)₈RC	SRAP	ME5	TGAGTCCAAACCGGAAG
ISSR	ISSR18	VHV (GT)₈	SRAP	ME6	TGAGTCCAAACCGGTAG
RAPD	S17	AGGGAACGAG	SRAP	EM8	GACTGCGTACGAATTAGC
RAPD	S18	CCACAGCAGT	SRAP	EM13	GACTGCGTACGAATTGGT
RAPD	S367	AGCGAGCAAG	SRAP	EM14	GACTGCGTACGAATTCAG
SRAP	ME3	TGAGTCCAAACCGGAAT	SRAP	EM17	GACTGCGTACGAATTCCA
ISSR引物中的单字母简写代表多碱基混合位点。位置: D= (A, G, T), H= (A, C, T), B= (C, G, T), R= (A, G), V= (A, C, G). Single letter abbreviations of ISSR primers for mixed base. Positions: D= (A, G, T), H= (A, C, T), B= (C, G, T), R= (A, G), V= (A, C, G).

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表 2 测试菌株信息

Table 2. Information of tested strains

菌株 strain	亲本和交配型 Parents and mating types	菌株 strain	亲本和交配型 Parent and mating type
Sh2	G.s0007-31^FP A₁B₇、G.s0011-16 A₃B₃	Sh13	G.s0007-31 A₁B₇、G.s0011-3^FP A₂B₂
Sh3	G.s0011-16^FP A₃B₃、G.s0012-28 A₅B₅	Sh14	G.s0007-31^FP A₁B₇、G.s0012-26 A₄B₄
Sh4	G.s0011-16 A₃B₃、G.s0012-28^FP A₅B₅	Sh15	G.s0007-31 A₁B₇、G.s0012-26^FP A₄B₄
Sh9	G.s0004-11 A₁B₁、G.s0011-16^FP A₃B₃	Sh21	G.s0011-3 A₂B₂、G.s0012-28^FP A₅B₅
Sh10	G.s0004-11^FP A₁B₁、G.s0011-16 A₃B₃	Sh24	G.s0004-11 A₁B₁、G.s0014-36^FP A₇B₇
Sh12	G.s0007-31^FP A₁B₇、G.s0011-3 A₂B₂
FP代表母本。 Female parent (FP).

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表 3 测试菌株的体细胞不和性与遗传关系

Table 3. Somatic incompatibility and genetic relationship between strains

异核体类别 Type of heterokaryon	配对 pairing	拮抗信息 Information of antagonism	遗传相似系数 Genetic similarity coefficient
Ⅰ 同核异质体 Same dikaryon in different FP	Sh3×Sh4	-	0.97
	Sh12×Sh13	-	0.97
	Sh14×Sh15	-	0.95
Ⅱ 细胞质和一个核相同，另一个核不同 one same monokaryon with one different monokaryon in the same FP	Sh2×Sh14	G+	0.66
	Sh2×Sh12	G+	0.74
	Sh3×Sh9	GL++	0.66
Ⅲ 细胞质相同，细胞核不同 Different dikaryon in the same FP	Sh4×Sh15	G+++	0.46
	Sh3×Sh13	G+++	0.63
	Sh9×Sh13	G++	0.57
Ⅳ 一个核相同，另一个核与细胞质都不同 one same monokaryon with one different monokaryon in different FP	Sh3×Sh10	GL++	0.69
	Sh2×Sh15	G+	0.66
	Sh3×Sh2	W-l S	0.54
Ⅴ 细胞质和细胞核均不同 Different dikaryon in different FP	Sh2×Sh21	G++	0.66
	Sh3×Sh14	G+++	0.49
	Sh14×Sh24	G++++	0.29
FP代表母本；SI反应分为（1）无拮抗-，（2）隔离型G，和（3）隔离型带线状GL和（4）菌丝墙式结构W-l S；“+”表示弱拮抗，“++”中等拮抗，“+++”强拮抗，“++++”非常强拮抗。 Female parent (FP), Three types of SI reaction, (1) no antagonism -, (2) [G] gap, (3) [GL] gap with line (4) hyphal wall-like structure (W-l S). + slight reaction, ++ moderate reaction, +++ strong reaction, ++++ more strong reaction.

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参考文献(21)

[1]	黄年来, 林志彬, 陈国梁. 中国食药用菌学[M]. 上海: 上海科技文献出版社, 2010: 1623-1673. [1] 黄年来, 林志彬, 陈国良, 等. 中国食药用菌学[M]. 上海: 上海科学技术文献出版社, 2010: 1623-1673.
[2]	刘新锐, 王圣铕, 谢宝贵, 等. 紫芝不亲和性因子分析 [J]. 菌物学报, 2014, 33(2):464−468. [2] 刘新锐, 王圣铕, 谢宝贵, 等. 紫芝不亲和性因子分析[J]. 菌物学报, 2014, 33(2): 464-468. Liu X R, Wang S Y, Xie B G, , et al. Incompatibility factors of Ganoderma sinense[J]. Mycosystema, 2014, 33(2): 464-468. (in Chinese)[知网中文][知网英文] Liu X R, Wang S Y, Xie B G, et al. Incompatibility factors of Ganoderma sinense [J]. Mycosystema, 2014, 33(2): 464−468. (in Chinese)
[3]	Liming T, Chan W , Baokai C , et al. Lanostane triterpenoids from mycelia-associated Ganoderma sinense and their anti-inflammatory activity. [J]. Phytochemistry, 2023, 215: 113870-113870. [3] TENG L M, WANG C, CUI B K, et al. Lanostane triterpenoids from mycelia-associated Ganoderma sinense and their anti-inflammatory activity[J]. Phytochemistry, 2023, 215: 113870. [PubMed]
[4]	Gao S Y, Zhang P, Zhang C Y, et al. Meroterpenoids from Ganoderma sinense protect hepatocytes and cardiomyocytes from oxidative stress induced injuries[J]. Fitoterapia, 2018, 131: 73-79. [4] GAO S Y, ZHANG P, ZHANG C Y, et al. Meroterpenoids from Ganoderma sinense protect hepatocytes and cardiomyocytes from oxidative stress induced injuries[J]. Fitoterapia, 2018, 131: 73-79. [PubMed]
[5]	WU N, PENG B, LI T, et al. Rapid Simultaneous Determination of Four Ganoderic Acids in Ganoderma (Chinese Name: Lingzhi) by Direct Infusion–Multiple Reaction Monitoring Cubed[J]. Journal of Analysis and Testing, 2024, 8(1): 52-62. [5] WU N, PENG B, LI T, et al. Rapid simultaneous determination of four ganoderic acids in Ganoderma (chinese Name: Lingzhi) by direct infusion–multiple reaction monitoring cubed[J]. Journal of Analysis and Testing, 2024, 8(1): 52-62. [LinkOut]
[6]	Mei R Q, Zuo F J, Duan X Y, et al. Ergosterols from Ganoderma sinense and their anti-inflammatory activities by inhibiting NO production[J]. Phytochemistry Letters, 2019, 32: 177-180. [6] MEI R Q, ZUO F J, DUAN X Y, et al. Ergosterols from Ganoderma sinense and their anti-inflammatory activities by inhibiting NO production[J]. Phytochemistry Letters, 2019, 32: 177-180. [LinkOut]
[7]	Jiang Y F, Chang Y J, Liu Y, et al. Overview of Ganoderma sinense polysaccharide-an adjunctive drug used during concurrent Chemo/Radiation therapy for cancer treatment in China[J]. Biomedicine & Pharmacotherapy, 2017, 96: 865-870. [7] JIANG Y F, CHANG Y J, LIU Y, et al. Overview of Ganoderma sinense polysaccharide-an adjunctive drug used during concurrent Chemo/Radiation therapy for cancer treatment in China[J]. Biomedicine & Pharmacotherapy = Biomedecine & Pharmacotherapie, 2017, 96: 865-870. [PubMed]
[8]	HAN W, CHEN H, ZHOU L, et al. Polysaccharides form Ganoderma Sinense-rice bran fermentation products and their anti-tumor activities on non-small-cell lung cancer [J]. BMC Complement Medicine Therap1es, 2021, 21(1): 169. [8] HAN W, CHEN H J, ZHOU L, et al. Polysaccharides from Ganoderma Sinense - rice bran fermentation products and their anti-tumor activities on non-small-cell lung cancer[J]. BMC Complementary Medicine and Therapies, 2021, 21(1): 169. [PubMed]
[9]	Lind M, Stenlid J, Olson A. Genetics and QTL mapping of somatic incompatibility and intraspecific interactions in the basidiomycete Heterobasidion annosum s. l[J]. Fungal Genetics and Biology, 2007, 44 (12): 1242-1251. [9] LIND M, STENLID J, OLSON A. Genetics and QTL mapping of somatic incompatibility and intraspecific interactions in the basidiomycete Heterobasidion annosum s. l[J]. Fungal Genetics and Biology: FG & B, 2007, 44(12): 1242-1251. [PubMed]
[10]	Marcais B, Caël O, Delatour C. Genetics of somatic incompatibility in Collybia fusipes[J]. Mycological Research, 2000, 104 (3): 304-310. [10] MARCAIS B, CAËL O, DELATOUR C. Genetics of somatic incompatibility in Collybia fusipes[J]. Mycological Research, 2000, 104(3): 304-310. [LinkOut]
[11]	唐传红, 张劲松, 陈明杰, 等. 利用拮抗试验和RAPD对灵芝属菌株进行分类研究 [J]. 微生物学通报, 2005, 32(5):72−76. [11] 唐传红, 张劲松, 陈明杰, 等. 利用拮抗试验和RAPD对灵芝属菌株进行分类研究[J]. 微生物学通报, 2005, 32(5): 72-76. TANG C H, ZHANG J S, CHEN M J, et al. Study on classification of strains of Ganoderma by anatagonistic effect and rapd[J]. Microbiology, 2005, 32(5): 72-76. (in Chinese)[知网中文][知网英文] doi: 10.3969/j.issn.0253-2654.2005.05.015 Tang C H, Zhang J S, Chen M J, et al. Study on classification of strains of Ganoderma by anatagonistic effect and Rapd [J]. Microbiology China, 2005, 32(5): 72−76. (in Chinese) doi: 10.3969/j.issn.0253-2654.2005.05.015
[12]	李黎. 中国木耳栽培种质资源的遗传多样性研究[D]. 武汉: 华中农业大学, 2016[12] 李黎. 中国木耳栽培种质资源的遗传多样性研究[D]. 武汉: 华中农业大学, 2011. LI L. Studies on Genetic Diversity of Auricularia Auricula-judae Cultivated Germplasm Resources in China[D]. Wuhan: Huazhong Agricultural University, 2011. (in Chinese)[知网博士中文][知网博士英文]
[13]	张瑞颖. 香菇菌株多相鉴定鉴别技术研究[D]. 北京: 中国农业大学, 2004[13] 张瑞颖. 香菇菌株多相鉴定鉴别技术研究[D]. 北京: 中国农业大学, 2004. ZHANG R Y. Study on Polyphasic Strain-typing Technique of Lentinula Edodes[D]. Beijing: China Agricultural University, 2004. (in Chinese)[知网硕士中文][知网硕士英文]
[14]	刘靖宇, 宋秀高, 叶夏, 等. 香菇菌株遗传多样性ISSR、RAPD和SRAP综合分析 [J]. 食用菌学报, 2011, 18(3):1−8. [14] 刘靖宇, 宋秀高, 叶夏, 等. 香菇菌株遗传多样性ISSR、RAPD和SRAP综合分析[J]. 食用菌学报, 2011, 18(3): 1-8. LIU J Y, SONG X G, YE X, et al. Differentiation of Lentinula edodes Strains Using ISSR, RAPD and SRAP markers[J]. Acta Edulis Fungi, 2011, 18(3): 1-8. (in Chinese)[知网中文][知网英文] doi: 10.3969/j.issn.1005-9873.2011.03.001 Liu J Y, Song X G, Ye X, et al. Differentiation of Lentinula edodes strains using ISSR, RAPD, SRAP markers [J]. Acta Edulis Fungi, 2011, 18(3): 1−8. (in Chinese) doi: 10.3969/j.issn.1005-9873.2011.03.001
[15]	徐珍, 章炉军, 尚晓冬, 等. 金针菇品种 DUS 测试性状的分级与评价[J]. 菌物学报, 2019, 38(5): 658-668[15] 徐珍, 章炉军, 尚晓冬, 等. 金针菇品种DUS测试性状的分级与评价[J]. 菌物学报, 2019, 38(5): 658-668. XU Z, ZHANG L J, SHANG X D, et al. Gradation and evaluation for Flammulina filiformis DUS testing traits[J]. Mycosystema, 2019, 38(5): 658-668. (in Chinese)[知网中文][知网英文] Xu Z, Zhang L J, Shang X D, et al. Gradation and evaluation for Flammulina filiformis DUS testing traits[J]. Mycosystema, 2019, 38 (5): 658-668. (in Chinese)
[16]	刘靖宇, 刘新锐, 邓优锦, 等. 双向核迁移在香菇遗传和育种中的应用研究[J]. 菌物学报, 2011, 30(5): 774-781. [16] 刘靖宇, 刘新锐, 邓优锦, 等. 双向核迁移在香菇遗传和育种中的应用研究[J]. 菌物学报, 2011, 30(5): 774-781. LIU J Y, LIU X R, DENG Y J, et al. The application of the ‘bidirectional haploid nuclei migration’ in breeding and genetics of Lentinula edodes[J]. Mycosystema, 2011, 30(5): 774-781. (in Chinese)[知网中文][知网英文] Liu J Y, Liu X R, Deng Y J, et al. The application of the ‘bidirectional haploid nuclei migration’ in breeding and genetics of Lentinula edodes. [J] Mycosystema, , 2011, 30(5): 774-781. (in Chinese)
[17]	Caten C. Vegetative incompatibility and cytoplasmic infection in fungi[J]. Microbiology, 1972, 72(2): 221-229. [17] CATEN C E. Vegetative incompatibility and cytoplasmic infection in fungi[J]. Journal of General Microbiology, 1972, 72(2): 221-229. [PubMed]
[18]	Worrall JJ. Somatic incompatibility in basidionycetes[J]. Mycologia, 1997, 89 (1): 24-36. [18] WORRALL J J. Somatic incompatibility in basidiomycetes[J]. Mycologia, 1997, 89(1): 24-36. [LinkOut]
[19]	Giovannetti M, Sbrana C, Strani P, et al. Genetic diversity of isolates of Glomus mosseae from different geographic areas detected by vegetative compatibility testing and biochemical and molecular analysis[J]. Apply Environment Microbiology, 2003, 69: 616-624. [19] GIOVANNETTI M, SBRANA C, STRANI P, et al. Genetic diversity of isolates of Glomus mosseae from different geographic areas detected by vegetative compatibility testing and biochemical and molecular analysis[J]. Applied and Environmental Microbiology, 2003, 69(1): 616-624. [PubMed]
[20]	May G. Somatic incompatibility and individualism in the coprophilous basidiomycete[J], Coprinus cinereus. Transactions of the British Mycological Society, 1988, 91 (3): 443-451. [20] MAY G. Somatic incompatibility and individualism in the coprophilous Basidiomycete, Coprinus cinereus[J]. Transactions of the British Mycological Society, 1988, 91(3): 443-451. [LinkOut]
[21]	Hansen EM, Stenlid J, Johansson M. Genetic control of somatic incompatibility in the root-rotting basidiomycete Heterobasidion annosum[J]. Mycological Research , 1993, 97 (10): 1229-1233. [21] HANSEN E M, STENLID J, JOHANSSON M. Genetic control of somatic incompatibility in the root-rotting basidiomycete Heterobasidion annosum[J]. Mycological Research, 1993, 97(10): 1229-1233. [LinkOut]