应用体细胞不亲和性试验和SSR方法综合鉴定闽南地区双孢蘑菇种质资源

袁滨; 柯丽娜; 陈光祥; 连燕萍; 张志鸿; 纪鹏伟; 吴振强

doi:10.19303/j.issn.1008-0384.2020.09.005

应用体细胞不亲和性试验和SSR方法综合鉴定闽南地区双孢蘑菇种质资源

doi: 10.19303/j.issn.1008-0384.2020.09.005

1.
漳州市农业科学研究所，福建　漳州　363005
2.
漳州市角美农业发展有限公司，福建　漳州　363107

基金项目: 福建省科技计划农业引导性（重点）项目（2019N0052）；国家现代农业产业技术体系建设专项（CARS-20）

详细信息

作者简介:
袁滨（1984−），男，副研究员，研究方向：食药用菌育种和栽培（E-mail：yuanbin07031@163.com）

通讯作者:
吴振强（1977−），男，高级农艺师，研究方向：主要从事食用菌栽培研究（E-mail：wuzhenqiang77@sina.com）

中图分类号: S 646
计量
- 文章访问数: 900
- HTML全文浏览量: 248
- PDF下载量: 19
- 被引次数: 0
出版历程
- 收稿日期: 2020-08-15
- 修回日期: 2020-09-09
- 刊出日期: 2020-09-28

Agaricus bisporus Germplasms in Southern Fujian Identified by Somatic Incompatibility Test and SSR

1.
Zhangzhou Institute of Agricultural Science, Zhangzhou, Fujian　363005, China
2.
Zhangzhou Jiaomei Agricultural Development Co., Ltd., Zhangzhou, Fujian　363107, China

摘要

摘要: 目的分析评价引进双孢蘑菇种质资源间的亲缘关系与遗传多样性，为闽南地区双孢蘑菇种质资源的鉴定、保藏与分子育种提供理论基础。方法结合体细胞不亲和性试验和SSR分子标记技术，对供试的9个双孢蘑菇菌株进行鉴定和遗传多样性分析。结果体细胞不亲和性试验结果表明，供试菌株之间拮抗作用较明显，根据拮抗反应的有无，供试菌株分为5组。SSR结果表明，当相似性系数达到0.76水平时，供试菌株聚成4个类群，第1类群包括HK；第2类群包括901和A15；第3类群包括福蘑52；第4类群包括As2796、W192、W2000、福蘑58、福蘑38。结论拮抗反应与SSR分子标记2种方法的结果基本一致。综合这2种方法，推测菌株HK与其他供试菌株亲缘关系较远，尤其与供试的其他国内菌株存在较大差异，可以作为一个育种亲本材料。
- 双孢蘑菇 /
- 亲缘关系 /
- 拮抗 /
- SSR /
- 聚类分析
Abstract: Objective Genetic relationship and diversity of Agaricus bisporus germplasms recently introduced to southern Fujian were studied to facilitate the identification, preservation, and breeding of the mushroom resource. Method Both somatic incompatibility test and SSR molecular marker technology were used to determine the genetic diversity of 9 varieties of A. bisporus. Result Somatic incompatibilities of the germplasms allowed the separation of 5 distinct groups by the presence or absence of antagonistic effect among them. Utilizing a similarity coefficient of 0.77, SSR analysis clustered the germplasms into Group 1 that consisted of HK, Group 2 that included 901 and A15, Group 3 that had Fumo 52, and Group 4 that comprised of As2796, W192, W2000, Fumo 58, and Fumo 38. Conclusion From the results obtained by the somatic incompatibility test and SSR molecular marker technology, HK appeared to be remotely related to the other 8 germplasms, especially the domestic varieties. Hence, it could be a potential candidate for breeding purpose.
- Agaricus bisporus /
- genetic relationship /
- antagonistic effect /
- SSR /
- clustering analysis

HTML全文

图 1 供试菌株拮抗类型

注：І-拮抗线图谱，ІІ- 沟状拮抗图谱，ІІІ-隆起拮抗图谱

Figure 1. Types of antagonistic germplasms

Note: І-Uplift antagonistic map; ІІ-Antagonistic line map; ІІІ-Groove antagonistic map.

下载: 全尺寸图片幻灯片

图 2 供试双孢蘑菇菌株SSR分子标记聚类图

Figure 2. SSR-based dendrogram of A. bisporus germplasms

下载: 全尺寸图片幻灯片

表 1 供试的双孢蘑菇菌株编号

Table 1. Sample codes for A. bisporus germplasms

编号 No.	菌株名称 Strain	来源 Origin
1	As2796	漳州市农科所
2	W2000	福建省农科院
3	W192	福建省农科院
4	福蘑38	福建省农科院
5	福蘑52	福建省农科院
6	福蘑58	福建省农科院
7	901	制种户
8	A15	制种户
9	HK	香港

下载: 导出CSV

表 2 供试菌株鉴定所用SSR引物

Table 2. SSR primers for variety identification

引物名 Primer	序列（5′ to 3′）　　 Sequence（5′ to 3′）	5′端修饰 5′end modification	重复基序 Repeat motifs	片段大小/bp Fragment size	退火温度/℃ Annealing temperature
AbSSR005-F	CTCTGGGATATGGACGAGGA	5′6-FAM	（GATGAG）6	118	56
AbSSR013-F	GACTGCCTGATTGACGGATT	5′HEX	（TA）6	162	57
AbSSR015-F	CTCGAGTCGACGAAGGAAAC	5′HEX	（GA）7	238	58
AbSSR016-F	TGTCTGGTTTTGCTCACGTC	5′HEX	（TC）12	242	55
AbSSR018-F	TGGCTCTTTACAGCCTTGGT	5′6-FAM	（CAT）6	122	55
AbSSR084-F	CGACCCATCATCAACTTCCT	5′HEX	（GAA）6	234	59
AbSSR6-F	ACCACATTCTGGAAAACGAA	5′HEX	（GCT）8	181	55
AbSSR36-F	CGTTGATGGAGTTCACTGAG	5′HEX	（GAAG）4（GAAAAG）（GAAG）	148	58
L11-F	ATAAAAAAGCATAATCACAAATG	5′6-FAM	（TC）13	228	50
L15-F	GCAGGTCCAGTGTGAACGG	5′6-FAM	（TCC）8	192	57
L17-F	ATCCAATTCACCAACCAGC	5′6-FAM	（A）11gagaataagaaattgaaaattg（A）16	186	52
L23-F	CTTTTCAGGGGAAGACAACG	5′6-FAM	（GTG）7	174	55

下载: 导出CSV

表 3 供试菌株间的拮抗反应情况

Table 3. Antagonistic reactions among tested germplasms

菌株 Strain	As2796 As2796	W2000 W2000	W19 2W192	福蘑38 Fomo 38	福蘑52 Fomo 52	福蘑58 Fomo 58	901	A15	HK
As2796 As2796	−
W2000 W2000	−	−
W192 W192	−	−	−
福蘑38 Fomo 38	−	−	−	−
福蘑52 Fomo 52	+	+	+	+	−
福蘑58 Fomo 58	−	−	+	−	+	−
901	+	+	+	+	+	+	−
A15	+	+	+	+	+	+	−	−
HK	+	+	+	+	+	+	+	+	−
注：‘+’表示有拮抗性；‘−’表示无拮抗 Note: + presence of antagonism; − absence of antagonism.

下载: 导出CSV

表 4 相似性系数

Table 4. List of similarity coefficients

菌株 Strain	福蘑38 Fomo 38	福蘑52 Fomo 52	福蘑58 Fomo 58	901	A15	As2796	HK	W192	W2000
福蘑38 Fomo 38	1
福蘑52 Fomo 52	0.615	1
福蘑58 Fomo 58	0.9	0.595	1
901	0.541	0.588	0.629	1
A15	0.462	0.722	0.486	0.765	1
As2796	0.821	0.667	0.865	0.529	0.556	1
HK	0.526	0.686	0.5	0.667	0.743	0.571	1
W192	0.905	0.718	0.8	0.432	0.564	0.821	0.632	1
W2000	0.81	0.769	0.8	0.486	0.615	0.821	0.684	0.905	1

下载: 导出CSV

参考文献(32)

[1]	黄毅. 食用菌工厂化栽培实践[M]. 福州: 福建科学技术出版社, 2014.
[2]	WANG Z S, LIAO J H, CHEN M Y, et al. Breeding and Industrial Development of Agaricus bisporus [J]. Acta Edulis Fungi, 2012, 19(3): 1−14.
[3]	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.
[4]	郑丹丹, 胡扬扬, 王琦. 双孢蘑菇活性成分研究进展 [J]. 食用菌学报, 2016, 23(2):94−103. ZHENG D D, HU Y Y, WANG Q. Recent research on bioactive components from Agaricus bisporus [J]. Acta Edulis Fungi, 2016, 23(2): 94−103.(in Chinese
[5]	袁滨, 柯丽娜, 洪丽明, 等. 双孢蘑菇新品种筛选试验与示范研究 [J]. 热带农业科学, 2018, 38(5):62−65, 70. YUAN B, KE L N, HONG L M, et al. Screening trial and demonstration of new strains of Agaricus bisporus [J]. Chinese Journal of Tropical Agriculture, 2018, 38(5): 62−65, 70.(in Chinese
[6]	陈世通, 李荣春. 食用菌育种方法的研究现状·存在的问题及展望 [J]. 安徽农业科学, 2012, 40(10):5850−5852. doi: 10.3969/j.issn.0517-6611.2012.10.049 CHEN S T, LI R C. Present research on breeding methods of edible fungi and the problems in it as well as the prospects [J]. Journal of Anhui Agricultural Sciences, 2012, 40(10): 5850−5852.(in Chinese doi: 10.3969/j.issn.0517-6611.2012.10.049
[7]	张天镇. 作物育种学总论[M]. 北京: 中国农业出版社, 2005: 60—63.
[8]	杨明霞, 杨萍, 任瑞, 等. 山楂的生殖生物学和杂交育种研究进展 [J]. 中国农学通报, 2018, 34(36):70−74. doi: 10.11924/j.issn.1000-6850.casb17110009 YANG M X, YANG P, REN R, et al. Reproductive Biology and Crossbreeding of Hawthorn Research Progress [J]. Chinese Agricultural Science Bulletin, 2018, 34(36): 70−74.(in Chinese doi: 10.11924/j.issn.1000-6850.casb17110009
[9]	翟阳阳, 宋金俤, 吴成龙, 等. 阿魏蘑杂交育种研究 [J]. 食药用菌, 2018, 26(6):367−374. ZHAI Y Y, SONG J D, WU C L, et al. Study on cross breeding of Pleurotus nebrodensis [J]. Edible and Medicinal Mushrooms, 2018, 26(6): 367−374.(in Chinese
[10]	江玉姬, 赵书光, 谢宝贵, 等. 金针菇杂交育种中亲本菌株的选择模式 [J]. 福建农林大学学报(自然科学版), 2010, 39(4):403−408. JIANG Y J, ZHAO S G, XIE B G, et al. Model of selecting parent strains for Flammulina velutipes hybridization breeding [J]. Journal of Fujian Agricultural and Forestry University, 2010, 39(4): 403−408.(in Chinese
[11]	RAJARAM S. Prospects and promise of wheat breeding in the 21st century [J]. Euphytica, 2001, 119(1): 3−15.
[12]	李娜娜, 张德平, 戴思兰. 遗传距离及聚类分析在切花菊亲本选配中的应用 [J]. 东北林业大学学报, 2011, 39(11):46−49. doi: 10.3969/j.issn.1000-5382.2011.11.013 LI N N, ZHANG D P, DAI S L. Application of Genetic Distance and Cluster Analysis to Parent Selection for Hybridization in Cut Chrysanthemum [J]. Journal of northeast forestry university, 2011, 39(11): 46−49.(in Chinese doi: 10.3969/j.issn.1000-5382.2011.11.013
[13]	纪永民, 王书平, 赵晋铭, 等. 优质高产濉科系列大豆品种选育的经验和体会 [J]. 大豆科技, 2018, 12(4):22−29. doi: 10.3969/j.issn.1674-3547.2018.04.006 JI Y M, WANG S P, ZHAO J M, et al. Experience and Lessons of Breeding High Quality and High Yield Suike Soybean Series Varieties [J]. Soybean Bulletin, 2018, 12(4): 22−29.(in Chinese doi: 10.3969/j.issn.1674-3547.2018.04.006
[14]	刘盛荣, 张维瑞, 柯斌榕, 等. 基于拮抗试验的杏鲍菇菌株分类研究 [J]. 食药用菌, 2015, 23(1):33−36. LIU S R, ZHANG W R, KE B R. Study on classification of Pleurotus eryngii strains based on antagonism test [J]. Edible and Medicinal Mushrooms, 2015, 23(1): 33−36.(in Chinese
[15]	杨和川, 谭一罗, 苏文英, 等. 金针菇菌株农艺性状评价及遗传多样性分析 [J]. 南方农业学报, 2018, 49(12):2371−2378. doi: 10.3969/j.issn.2095-1191.2018.12.04 YANG H C, TAN Y L, SU W Y, et al. Agronomic traits evaluation and genetic diversity analysis of Flammulina filiformis [J]. Journal of Southern Agriculture, 2018, 49(12): 2371−2378.(in Chinese doi: 10.3969/j.issn.2095-1191.2018.12.04
[16]	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. 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, 2005, 32(5): 72−76.(in Chinese doi: 10.3969/j.issn.0253-2654.2005.05.015
[17]	刘昆, 宋小亚, 蒋俊路, 等. 食用菌拮抗反应的定量分析 [J]. 食药用菌, 2015, 23(6):364−366. LIU K, SONG X Y, JIANG J L, et al. Quantitative analysis of antagonistic reaction of edible fungi [J]. Edible and medicina1 mushroom, 2015, 23(6): 364−366.(in Chinese
[18]	王翠, 郭仲杰, 尤洁, 等. 双孢蘑菇产、质量性状相关分子标记的初步研究 [J]. 福建农业科技, 2018(7):1−5. WANG C, GUO Z J, YOU J, et al. A preliminary study on molecular markers related to yield and quality traits of Agaricus bisporus [J]. Fujian Agricultural Science and Technology, 2018(7): 1−5.(in Chinese
[19]	林媛, 陈文炳, 邵碧英, 等. 双孢蘑菇RAPD标记的遗传多样性分析 [J]. 食品科学, 2009, 30(20):272−276. LIN Y, CHEN W B, SHAO B Y, et al. Detection of genetic diversity of Agaricus bisporus using RAPD markers [J]. Food Science, 2009, 30(20): 272−276.(in Chinese
[20]	王金斌, 刘华, 郭陈莉, 等. 中国工厂化主栽双孢蘑菇品种的SSR标记遗传多样性分析 [J]. 分子植物育种, 2019, 17(5):1589−1596. WANG J B, LIU H, GUO C L, et al. Genetic Diversity Analysis of Industrial Leading Cultivated Agaricus bisporus Varieties in China by SSR Markers [J]. Molecular Plant Breeding, 2019, 17(5): 1589−1596.(in Chinese
[21]	王新新, 李丹, 宋冰, 等. 双孢蘑菇种质SSR分子身份证的构建 [J]. 食用菌学报, 2016, 23(2):6−11. WANG X X, LI D, SONG B, et al. Development of a Single Sequence Repeat-based Molecular ID System for Differentiating Agaricus bisporus Strains [J]. Acta Edulis Fungi, 2016, 23(2): 6−11.(in Chinese
[22]	顾敏, 沈颖越, 金群力, 等. 双孢蘑菇SSR分子标记开发及其在遗传多样性分析中的应用 [J]. 浙江农业学报, 2013, 25(5):987−993. doi: 10.3969/j.issn.1004-1524.2013.05.14 GU M, SHEN Y Y, JIN Q L, et al. Development and application of SSR markers in Agaricus bisporus [J]. Acta Agricuhurae Zhejiangensis, 2013, 25(5): 987−993.(in Chinese doi: 10.3969/j.issn.1004-1524.2013.05.14
[23]	FOULONGNE-ORIOL M, SPATARO C, SAVOIE J M. Novel microsatellite markers suitable for genetic studies in the white button mushroom Agaricus bisporus [J]. Applied Microbiology and Biotechnology, 2009, 84(6): 1125−1135. doi: 10.1007/s00253-009-2030-8
[24]	吴明基, 向海涛, 刘华清, 等. 利用形态学性状及SSR分子标记鉴定水稻优良恢复系闽恢3301的亲代来源 [J]. 福建农业学报, 2012, 27(7):673−678. doi: 10.3969/j.issn.1008-0384.2012.07.003 WU M J, XIANG H T, LIU H Q, et a1. Genetic Original Identification of Minhui 3301 by Morphological Traits and SSR Markers [J]. Fujian Journal ofAgricultural Sciences, 2012, 27(7): 673−678.(in Chinese doi: 10.3969/j.issn.1008-0384.2012.07.003
[25]	王让剑, 杨军, 孔祥瑞, 等. 福建15个茶树品种SSR遗传差异分析与指纹图谱建立 [J]. 福建农业学报, 2014, 29(10):970−975. doi: 10.3969/j.issn.1008-0384.2014.10.008 WANG R J, YANG J, KONG X R, et a1. Genetic Variances and DNA Fingerprints of 15 Teas in Fuiian [J]. Fian Journal of Agricultural Sciences, 2014, 29(10): 970−975.(in Chinese doi: 10.3969/j.issn.1008-0384.2014.10.008
[26]	付涛, 严春风, 林乐静, 等. 我国南方野生樱属植物的SSR亲缘关系分析 [J]. 核农学报, 2018, 32(10):1949−1959. doi: 10.11869/j.issn.100-8551.2018.10.1949 FU T, YAN C F, LIN L J, et a1. Analysis of Genetic Relationship of Wild Cerasus in South China With SSR Markers [J]. Journal of Nuclear Agricultural Science, 2018, 32(10): 1949−1959.(in Chinese doi: 10.11869/j.issn.100-8551.2018.10.1949
[27]	陈坚, 郑益平, 郭文杰, 等. 福建省草莓主栽与引进品种的SSR分子标记鉴定 [J]. 福建农业学报, 2018, 33(2):150−153. CHEN J, ZHANG Y P, GUO W J, et a1. Identification of Major Introduced Strawberry Cultivars in Fujian Using SSR Molecular Marker [J]. Fujian Journal of Agricultural Sciences, 2018, 33(2): 150−153.(in Chinese
[28]	周平, 郭瑞, 张小丹, 等. SSR分析50份桃种质资源遗传多样性 [J]. 福建农业学报, 2017, 32(1):47−50. ZHOU P, GUO R, ZHANG X D, et al. Genetic Diversity of 50 Prunus persica Germplasms Aanlyzed by SSR Markers [J]. Fujian Journal of Agricultural Sciences, 2017, 32(1): 47−50.(in Chinese
[29]	王小艳, 王春晖, 姜性坚, 等. 杏鲍菇多孢分离菌株ISSR、RAPD分子标记及体细胞不亲和性分析 [J]. 中国食用菌, 2015, 34(3):57−60. WANG X Y, WANG C H, JIANG X J, et al. Analysis on Multi-spore Isolated Pleurotus eryngii Using ISSR and RAPD Molecular [J]. Edible Fungi of China, 2015, 34(3): 57−60.(in Chinese
[30]	史灵燕, 刘保卫, 顾新颖, 等. 生化和ISSR分子标记在黑木耳品种鉴定中的应用 [J]. 北方园艺, 2019, 24(9):136−141. SHI L Y, LIU B W, GU X Y, et al. Application of Biochemical and ISSR Molecular Markers in Identification of Auricularia auricular [J]. Northern Horticulture, 2019, 24(9): 136−141.(in Chinese
[31]	路新彦, 刘昆, 蒋俊, 等. 7个黑木耳菌株遗传差异性分析 [J]. 中国食用菌, 2017, 31(1):52−55. LU X Y, LIU K, JIANG J, et al. Study on Genetic Polymorphism of 7 Auricularia auricula Strains [J]. Edible Fungi of China, 2017, 31(1): 52−55.(in Chinese
[32]	杨军, 徐吉, 宋一鸣, 等. 耐高温型灰树花菌株筛选及供试灰树花亲缘关系分析 [J]. 基因组学与应用生物学, 2016, 35(2):396−405. YANG J, XU J, SONG Y M, et al. Screen of Grifola frondosa thermotolerant strains and analysis of the genetic relationship of candidate strains [J]. Genomics and Applied Biology, 2016, 35(2): 396−405.(in Chinese