52个马铃薯遗传多样性分析及SSR分子身份证构建

doi:10.13560/j.cnki.biotech.bull.1985.2023-0512

摘要/Abstract

摘要：

马铃薯（Solanum tuberosum L.）种质资源遗传多样性评价是开展遗传育种的前提和基础，为探究马铃薯种质资源的遗传背景，进而为新品种的选育提供育种材料。以52个马铃薯品种（系）为研究材料，采用SSR标记分析亲缘关系，进而评价其遗传多样性，并构建指纹图谱。从25对SSR引物中筛选出多态性丰富、稳定性好的16对引物，并对52份马铃薯品种（系）进行SSR扩增分析，16对SSR引物扩增出147个多态性条带，引物平均多态条带比率为64.71%，平均Nei's遗传多样性（Nei's genetic diversity, H）和平均Shannon's指数（Shannon index, I）分别为0.14和0.24。在遗传相似系数0.23处，52份材料经聚类分析分为3大类，类群I包括49个品种（系），46个是北方品种（系），3个国外引进品种（系），均适宜晋北地区种植；类群II包括2个品种，‘北方016’和‘晋薯16号’，特点是薯皮黄色、干物质含量高；类群III仅包括‘晋薯15号’，在大类的划分上未按照地理来源进行聚类。在遗传相似系数0.36处，类群I又可以分为7个亚类。基于最少引物鉴定最多种质的原则，利用C59、S25、C33、S151共4对引物组合可区分全部供试品种，构建出基于SSR标记的数字化指纹图谱及分子身份证。利用SSR标记技术多态性引物可以评价马铃薯种质资源的遗传多样性，通过分子身份证能够准确鉴别以及溯源不同马铃薯品种（系）。

关键词: 马铃薯, 品种（系）, 遗传多样性, SSR标记, 聚类分析, 指纹图谱, 分子身份证

Abstract:

The evaluation on the genetic diversity of potato germplasm resources（Solanum tuberosum L.）is the prerequisite and basis for genetic breeding, which may provide breeding materials for exploring the genetic background of potato germplasm resources and breeding new varieties. The genetic relationship and diversity of 52 potato varieties（lines）was evaluated by SSR markers, and the fingerprint was constructed. Sixteen primers with abundant polymorphism and good stability were selected from 25 pairs of SSR primers, and 147 polymorphic bands were amplified by 16 pairs of SSR primers from 52 potato varieties（lines）, and the average polymorphic band ratio was 64.71%. The average Nei's genetic diversity（H）and Shannon's index（I）were 0.14 and 0.24, respectively. At the similar genetic coefficient of 0.23, the 52 potato materials were divided into three categories by cluster analysis. Group I included 49 varieties（lines）, 46 varieties（lines）from North China, and 3 imported varieties（lines）from abroad, all of them were suitable for planting in North Shanxi province. Group II included two varieties, ‘Beifang 016’ and ‘Jinshu 16’, characterized by yellow potato skin and high dry matter content. Group III only included ‘Jinshu 15’, and was not clustered according to geographical origin. At the genetic similarity coefficient of 0.36, group I can be divided into 7 subclasses. Based on the principle of identifying the most varieties with the least primers, C59, S25, C33 and S151 were used to distinguish all varieties tested, and the digital fingerprints and molecular identity cards based on SSR markers were constructed. The genetic diversity of potato germplasm resources can be evaluated by using SSR markers, and different potato varieties（lines）can be accurately identified and traced by molecular identification cards.

Key words: potato, varieties（lines）, genetic diversity, SSR markers, clustering analysis, fingerprints, molecular identity card

安苗, 王彤彤, 付逸婷, 夏俊俊, 彭锁堂, 段永红. 52个马铃薯遗传多样性分析及SSR分子身份证构建[J]. 生物技术通报, 2023, 39(12): 136-147.

AN Miao, WANG Tong-tong, FU Yi-ting, XIA Jun-jun, PENG Suo-tang, DUAN Yong-hong. Genetic Diversity Analysis and Molecular Identity Card Construction by SSR Markers of 52 Solanum tuberosum L. Varieties（Lines）[J]. Biotechnology Bulletin, 2023, 39(12): 136-147.

图/表 9

参考文献 41

[1]	宋峥, 王崇, 徐颖华, 等. 马铃薯地方种质资源的SSR遗传多样性分析[J]. 分子植物育种, 2022, 20(21): 7143-7153.
	Song Z, Wang C, Xu YH, et al. SSR analysis on genetic diversity for local potato germplasm resources[J]. Mol Plant Breed, 2022, 20(21): 7143-7153.
[2]	王若秋, 赵朋, 王冬冬, 等. 基于SSR标记的彩色马铃薯亲缘关系分析及指纹图谱构建[J]. 西北植物学报, 2018, 38(2): 249-257.
	Wang RQ, Zhao P, Wang DD, et al. Genetic relationship analysis and fingerprint profile construction of pigmented potato based on SSR markers[J]. Acta Bot Boreali Occidentalia Sin, 2018, 38(2): 249-257.
[3]	孙小琼, 王芳, 王崇, 等. 马铃薯品种(系)资源的SSR遗传多样性分析及指纹图谱构建[J/OL]. 分子植物育种, 2022. https://kns.cnki.net/kcms/detail/46.1068.S.20220428.1540.021.html.
	Sun XQ, Wang F, Wang C, et al. SSR genetic diversity analysis and fingerprint construction of potato variety(line)resources[J/OL]. Mol Plant Breed, 2022. https://kns.cnki.net/kcms/detail/46.1068.S.20220428.1540.021.html.
[4]	段艳凤, 刘杰, 卞春松, 等. 中国88个马铃薯审定品种SSR指纹图谱构建与遗传多样性分析[J]. 作物学报, 2009, 35(8): 1451-1457. doi: 10.3724/SP.J.1006.2009.01451
	Duan YF, Liu J, Bian CS, et al. Construction of fingerprinting and analysis of genetic diversity with SSR markers for eighty-eight approved potato cultivars(Solanum tuberosum L.) in China[J]. Acta Agron Sin, 2009, 35(8): 1451-1457. doi: 10.3724/SP.J.1006.2009.01451 URL
[5]	时启冬, 石瑛, 李欣, 等. 东北地区部分马铃薯品种遗传多样性的SSR分析[J]. 中国农学通报, 2014, 30(21): 240-245.
	Shi QD, Shi Y, Li X, et al. Genetic diversity analysis of part of the potato varieties in northeast region by SSR marker[J]. Chin Agric Sci Bull, 2014, 30(21): 240-245. doi: 10.11924/j.issn.1000-6850.2013-3307
[6]	Norero N, Malleville J, Huarte M, et al. Cost efficient potato(Solarium tuberosum L.) cultivar identification by microsatellite amplification[J]. Potato Res, 2002, 45(2/3/4): 131-138. doi: 10.1007/BF02736108 URL
[7]	金兴红, 于卓, 张霞, 等. 基于BSA法开发马铃薯高淀粉相关SSR标记及候选基因筛选[J/OL]. 分子植物育种, 2022. https://kns.cnki.net/kcms/detail/46.1068.S.20220629.1035.010.html.
	Jin XH, Yu Z, Zhang X, et al. Development of potato SSR markers associated with high starch based on BSA method and screening of candidate genes[J/OL]. Mol Plant Breed. https://kns.cnki.net/kcms/detail/46.1068.S.20220629.1035.010.html.
[8]	张霞. 彩色马铃薯花青素含量相关SSR分子标记的开发研究[D]. 呼和浩特: 内蒙古农业大学, 2022.
	Zhang X. Development of SSR molecular markers related to anthocyanin content in colored potato[D]. Hohhot: Inner Mongolia Agricultural University, 2022.
[9]	张海雯, 巩檑, 马斯霜, 等. 基于SSR标记的马铃薯种质遗传多样性及群体结构分析[J]. 分子植物育种, 2020, 18(12): 4144-4152.
	Zhang HW, Gong L, Ma SS, et al. Genetic diversity and population structure analysis of potato(Solanum tuberosum)germplasm based on SSR markers[J]. Mol Plant Breed, 2020, 18(12): 4144-4152.
[10]	Perera PIP, Perera L, Hocher V, et al. Use of SSR markers to determine the anther-derived homozygous lines in coconut[J]. Plant Cell Rep, 2008, 27(11): 1697-1703. doi: 10.1007/s00299-008-0592-z pmid: 18712524
[11]	Diao WP, Jia YY, Song H, et al. Efficient embryo induction in cucumber ovary culture and homozygous identification of the regenetants using SSR markers[J]. Sci Hortic, 2009, 119(3): 246-251. doi: 10.1016/j.scienta.2008.08.016 URL
[12]	解华云, 高崇敏, 叶云峰, 等. 基于TP-M13-SSR分子标记技术的133份薄皮甜瓜种质资源遗传多样性分析[J]. 南方农业学报, 2022, 53(9): 2547-2556.
	Xie HY, Gao CM, Ye YF, et al. Genetic diversity analysis of 133 pellicle melon germplasm resources based on TP-M13-SSR molecular marker technology[J]. J South Agric, 2022, 53(9): 2547-2556.
[13]	Tiwari JK, Ali N, Devi S, et al. Analysis of allelic variation in wild potato(Solanum)species by simple sequence repeat(SSR)markers[J]. 3 Biotech, 2019, 9(7): 262. doi: 10.1007/s13205-019-1785-6
[14]	de Haan S, Núñez J, Bonierbale M, et al. A simple sequence repeat(SSR)marker comparison of a large in- and ex-situ potato Landrace cultivar collection from Peru reaffirms the complementary nature of both conservation strategies[J]. Diversity, 2013, 5(3): 505-521. doi: 10.3390/d5030505 URL
[15]	Sharma V, Nandineni MR. Assessment of genetic diversity among Indian potato(Solanum tuberosum L.) collection using microsatellite and retrotransposon based marker systems[J]. Mol Phylogenet Evol, 2014, 73: 10-17. doi: 10.1016/j.ympev.2014.01.003 URL
[16]	段艳凤. 马铃薯种质资源遗传多样性评价及抗晚疫病相关基因分析[D]. 北京: 中国农业科学院, 2019.
	Duan YF. Genetic diversity assessment of germplasm resources and analysis of resistance-related genes to Phytophthora infestans in potato[D]. Beijing: Chinese Academy of Agricultural Sciences, 2019.
[17]	王鹏, 李芳弟, 郭天顺, 等. 马铃薯种质资源遗传关系分析及指纹图谱构建[J]. 农业生物技术学报, 2020, 28(5): 794-810.
	Wang P, Li FD, Guo TS, et al. Genetic relationship analysis and fingerprint construction of potato(Solanum tuberosum)germplasm resources[J]. J Agric Biotechnol, 2020, 28(5): 794-810.
[18]	高玉坤, 崔江慧, 项晓冬, 等. 65个马铃薯品种(系)指纹图谱构建和遗传多样性分析[J]. 农业生物技术学报, 2020, 28(8): 1363-1378.
	Gao YK, Cui JH, Xiang XD, et al. Fingerprint construction and genetic diversity analysis for 65 potato(Solanum tuberosum)varieties(lines)[J]. J Agric Biotechnol, 2020, 28(8): 1363-1378.
[19]	刘春雷. 马铃薯EST-SSR标记开发及育种资源的指纹图谱构建[D]. 雅安: 四川农业大学, 2016.
	Liu CL. Development and utility of EST-SSR marker and construction of potato(Solanum tuberosum L.)germplasm fingerprinting[D]. Ya'an: Sichuan Agricultural University, 2016.
[20]	Jian YQ, Yan WY, Xu JF, et al. Genome-wide simple sequence repeat markers in potato: abundance, distribution, composition, and polymorphism[J]. DNA Res, 2021, 28(6): dsab020. doi: 10.1093/dnares/dsab020 URL
[21]	刘宇飞. 5个马铃薯新品系的主要性状、染色体构型及SSR遗传差异分析[D]. 呼和浩特: 内蒙古农业大学, 2021.
	Liu YF. Major traits, chromosomal configurations and SSR analysis of 5 new potato strains[D]. Hohhot: Inner Mongolia Agricultural University, 2021.
[22]	于佳乐. 18个马铃薯品种的抗旱性评价及遗传多样性分析[D]. 呼和浩特: 内蒙古农业大学, 2022.
	Yu JL. Drought resistance evaluation and genetic diversity analysis of 18 potato varieties[D]. Hohhot: Inner Mongolia Agricultural University, 2022.
[23]	李靓. 6个彩色马铃薯新品系的细胞学、重要性状及SSR遗传差异分析[D]. 呼和浩特: 内蒙古农业大学, 2021.
	Li L. Analysis of cytology, important characters and SSR genetic differences of 6 new colored potato strains[D]. Hohhot: Inner Mongolia Agricultural University, 2021.
[24]	王绍鹏, 邱彩玲, 李勇, 等. 马铃薯SSR标记聚丙烯酰胺凝胶电泳分析[J]. 中国马铃薯, 2009, 23(3): 143-147.
	Wang SP, Qiu CL, Li Y, et al. Analysis of potato with polyacrylamide gel electrophoresis based on SSR markers[J]. Chin Potato J, 2009, 23(3): 143-147.
[25]	黄先群, 冉江, 黄团, 等. 马铃薯引进品种及变异材料的SSR遗传多样性分析[J]. 西南农业学报, 2014, 27(4): 1399-1403.
	Huang XQ, Ran J, Huang T, et al. Genetic diversity analysis of introduced varieties and mutation materials by SSR markers in potato[J]. Southwest China J Agric Sci, 2014, 27(4): 1399-1403.
[26]	张海龙, 于卓, 祁娜, 等. 10个马铃薯新育成品种的SSR分析[J]. 种子, 2022, 41(6): 34-38.
	Zhang HL, Yu Z, Qi N, et al. SSR analysis on ten newly bred potato varieties[J]. Seed, 2022, 41(6): 34-38.
[27]	刘毅强, 田再民, 祁利潘, 等. 20份马铃薯品种(系)指纹图谱构建和遗传多样性分析[J]. 江苏农业科学, 2022, 50(21): 41-46.
	Liu YQ, Tian ZM, Qi LP, et al. Fingerprint construction and genetic diversity analysis of 20 potato varieties(lines)[J]. Jiangsu Agric Sci, 2022, 50(21): 41-46.
[28]	姜超. 彩色马铃薯优良新品系培育及花青素组分的HPLC-MS分析[D]. 呼和浩特: 内蒙古农业大学, 2017.
	Jiang C. Breeding of new strains of colored potato and HPLC-MS analysis of anthocyanin components[D]. Hohhot: Inner Mongolia Agricultural University, 2017.
[29]	Dean Goldring JP. The roles of acetic acid and methanol during fixing and staining proteins in an SDS-polyacrylamide electrophoresis gel[J]. Methods Mol Biol, 2018, 1853: 15-18. doi: 10.1007/978-1-4939-8745-0_2 pmid: 30097924
[30]	岳东, 于卓, 于肖夏, 等. 彩色马铃薯新品系细胞遗传学观测及SSR分析[J]. 华北农学报, 2022, 37(6): 108-114. doi: 10.7668/hbnxb.20193007
	Yue D, Yu Z, Yu XX, et al. Cytological observation and SSR analysis of new colored potato strains[J]. Acta Agric Boreali Sin, 2022, 37(6): 108-114.
[31]	Pérez-Figueroa A, Rodríguez-Ramilo ST, Caballero A. Analysis and management of gene and allelic diversity in subdivided populations using the software program METAPOP[J]. Methods Mol Biol, 2012, 888: 261-275. doi: 10.1007/978-1-61779-870-2_15 pmid: 22665286
[32]	张招娟, 邓跃, 黄泷健, 等. 福建省马铃薯主栽品种SSR遗传多样性分析[J]. 分子植物育种, 2019, 17(22): 7420-7427.
	Zhang ZJ, Deng Y, Huang LJ, et al. Genetic diversity analysis of main potato varieties in Fujian Province based on SSR[J]. Mol Plant Breed, 2019, 17(22): 7420-7427.
[33]	马庆国, 宋晓波, 贺君星, 等. 基于SSR分子标记的核桃种质资源分子身份证构建[J]. 植物资源与环境学报, 2023, 32(2): 1-9.
	Ma QG, Song XB, He JX, et al. Establishment of molecular identity cards of walnut(Juglans spp.) germplasm resources based on SSR molecular marker[J]. J Plant Resour Environ, 2023, 32(2): 1-9.
[34]	王宇晴, 李乔乔, 阚文亮, 等. 利用SSR分子标记构建甜菜登记品种的分子身份证[J]. 江苏农业科学, 2022, 50(18): 289-294.
	Wang YQ, Li QQ, Kan WL, et al. Construction of molecular identity cards of sugar beet registration varieties using SSR molecular markers[J]. Jiangsu Agric Sci, 2022, 50(18): 289-294.
[35]	刘冠群, 吴祠平, 谭礼强, 等. 利用SSR分子标记构建名山茶树基因身份证[J]. 四川农业大学学报, 2019, 37(4): 469-474, 503.
	Liu GQ, Wu CP, Tan LQ, et al. Construction of SSR-based molecular IDs for tea planted in Mingshan[J]. J Sichuan Agric Univ, 2019, 37(4): 469-474, 503.
[36]	侯丽媛, 董艳辉, 邓舒, 等. 部分苹果属种质遗传多样性分析及分子身份证构建[J]. 山西农业科学, 2020, 48(8): 1171-1179.
	Hou LY, Dong YH, Deng S, et al. Study on genetic diversity and construction of molecular identity card for some Malus mill. germplasm resource[J]. J Shanxi Agric Sci, 2020, 48(8): 1171-1179.
[37]	李清, 罗永坚, 吴柔贤, 等. 广东省大豆种质资源遗传多样性分析及DNA分子身份证构建[J]. 广东农业科学, 2020, 47(12): 221-228.
	Li Q, Luo YJ, Wu RX, et al. Analysis on genetic diversity and construction of DNA molecular identity card of soybean germplasm resources in Guangdong Province[J]. Guangdong Agric Sci, 2020, 47(12): 221-228.
[38]	秦瑞英, 许学, 张立平, 等. 小麦SSR指纹图谱及品种身份证的构建——基于毛细管电泳分析[J]. 中国农学通报, 2017, 33(34): 46-55. doi: 10.11924/j.issn.1000-6850.casb17010102
	Qin RY, Xu X, Zhang LP, et al. Construction of wheat variety SSR fingerprint and ID: based on capillary electrophoresis[J]. Chin Agric Sci Bull, 2017, 33(34): 46-55. doi: 10.11924/j.issn.1000-6850.casb17010102
[39]	管俊娇, 杨龙, 木万福, 等. 利用SSR标记构建青花菜的分子身份证[J]. 中国蔬菜, 2021(12): 39-45.
	Guan JJ, Yang L, Mu WF, et al. Molecular identity card establishment for broccoli(Brassica oleracea L. var. italica plenck)accessions using SSR markers[J]. China Veg, 2021(12): 39-45.
[40]	刘文林, 张举梅, 盛万民, 等. 52份俄罗斯引进马铃薯种质资源的遗传多样性与分子身份证构建[J]. 分子植物育种, 2016, 14(1): 251-258.
	Liu WL, Zhang JM, Sheng WM, et al. Genetic diversity analysis and molecular identities establishment of 52 Russian potato varieties[J]. Mol Plant Breed, 2016, 14(1): 251-258.
[41]	程林涛, 孙晓阳, 谢福春, 等. 基于表型性状和SSR分子标记的草地早熟禾遗传多样性分析和分子身份证构建[J]. 中国草地学报, 2022, 44(9): 1-10.
	Cheng LT, Sun XY, Xie FC, et al. Genetic diversity analysis and molecular ID construction of Kentucky bluegrass based on phenotypic traits and SSR molecular markers[J]. Chin J Grassland, 2022, 44(9): 1-10.

编号 Code	材料名称 Accession	编号 Code	材料名称 Accession
1	L1150-22	27	蓬勃2号 Pengbo 2
2	陇薯7号 Longshu 7	28	晋薯16号 Jinshu 16
3	陇薯10号 Longshu 10	29	228
4	陇薯14号 Longshu 14	30	冀张薯8号 Jizhangshu 8
5	陇薯20号 Longshu 20	31	北方013 Beifang 013
6	陇薯19号 Longshu 19	32	北方016 Beifang 016
7	石薯6号 Shishu 6	33	青薯10号 Qingshu 10
8	青薯9号Qingshu 9	34	石薯1号 Shishu 1
9	东农310 Dongnong 310	35	石薯3号 Shishu 3
10	同薯20号 Tongshu 20	36	荷兰806 Holland 806
11	中薯306 Zhongshu 306	37	EV
12	克新25 Kexin 25	38	云薯6号 Yunshu 6
13	中薯9号 Zhongshu 9	39	实验1号 Shiyan 1
14	476	40	希森6号 Xisen 6
15	晋薯15号 Jinshu 15	41	中加2号 Zhongjia 2
16	新大坪 Xindaping	42	118
17	大同里外黄 Datongiwaihuang	43	V7
18	民丰红 Minfenghong	44	V9
19	鄂薯3号 Eshu 3	45	雪川红 Xuechuanhong
20	克新23号 Kexin 23	46	黑金刚 Heijingang
21	同薯29号 Tongshu 29	47	紫玫瑰 Zimeigui
22	川引2号 Chuanyin 2	48	丽薯6号 Lishu 6
23	京张薯1号 Jingzhangshu 1	49	冀张薯20号 Jizhangshu 20
24	京张薯3号 Jingzhangshu 3	50	克新13 Kexin 13
25	实验2号 Shiyan 2	51	荷兰14 Holland 14
26	蓬勃1号 Pengbo 1	52	冀张薯12号 Jizhangshu 12

编号 Code	材料名称 Accession	编号 Code	材料名称 Accession
1	L1150-22	27	蓬勃2号 Pengbo 2
2	陇薯7号 Longshu 7	28	晋薯16号 Jinshu 16
3	陇薯10号 Longshu 10	29	228
4	陇薯14号 Longshu 14	30	冀张薯8号 Jizhangshu 8
5	陇薯20号 Longshu 20	31	北方013 Beifang 013
6	陇薯19号 Longshu 19	32	北方016 Beifang 016
7	石薯6号 Shishu 6	33	青薯10号 Qingshu 10
8	青薯9号Qingshu 9	34	石薯1号 Shishu 1
9	东农310 Dongnong 310	35	石薯3号 Shishu 3
10	同薯20号 Tongshu 20	36	荷兰806 Holland 806
11	中薯306 Zhongshu 306	37	EV
12	克新25 Kexin 25	38	云薯6号 Yunshu 6
13	中薯9号 Zhongshu 9	39	实验1号 Shiyan 1
14	476	40	希森6号 Xisen 6
15	晋薯15号 Jinshu 15	41	中加2号 Zhongjia 2
16	新大坪 Xindaping	42	118
17	大同里外黄 Datongiwaihuang	43	V7
18	民丰红 Minfenghong	44	V9
19	鄂薯3号 Eshu 3	45	雪川红 Xuechuanhong
20	克新23号 Kexin 23	46	黑金刚 Heijingang
21	同薯29号 Tongshu 29	47	紫玫瑰 Zimeigui
22	川引2号 Chuanyin 2	48	丽薯6号 Lishu 6
23	京张薯1号 Jingzhangshu 1	49	冀张薯20号 Jizhangshu 20
24	京张薯3号 Jingzhangshu 3	50	克新13 Kexin 13
25	实验2号 Shiyan 2	51	荷兰14 Holland 14
26	蓬勃1号 Pengbo 1	52	冀张薯12号 Jizhangshu 12

序号No.	引物Primer	正向引物Forword primer（5'-3'）	反向引物Reverse primer（5'-3'）
1	C33	TCCCCTTACCCCTCTTCAAAT	AACCCGTGTCAACTTCTGCT
2	C59	TCATCACAACGTGACCCCA	GGGCTTGAATGATGTGAAGCTC
3	S25	AGAGATCGATGTAAAACACGT	GTGGCATTTTGATGGATT
4	C42	AGGCAAACCCCGTAAATCTT	TCTTCCTCACTCGTCCCACT
5	S151	GCTGCTAAACACTCAAGCAGAA	CAACTACAAGATTCCATCCACAC
6	STM-2022	GCGTCAGCGATTTCAGTACTA	TTCAGTCAACTCCTGTTGCG
7	C54	TGGCTGCAGAAATCCAAGATG	TGAAACCACTGGGAGGACTT
8	C58	GCTGCTAAACACTCAAGCAGAA	CAACTACAAGATTCCATCCACAC
9	C3	CCGGAGAATTAGACGATGA	TTTATGGGGAGCAGTTGAGG
10	S153	CCCTTACCCCTCTTCAATC	TCTCTTCTCAGCTGCCGTCT
11	UGP	TGGGGTTCCATCAAAC	GAAACTGCTGCCGGTGC
12	STM1016	TTCTGATTTCATGCATGTTTCC	ATGCTTGCCATGTGATGTGT
13	Patain	TGGTCTGGTGCATTAGAAAAAA	CAACCAACAAGGTAAATGGTACC
14	S170	CGCAAATCTTCATCCGATTC	TCCGGCGGATAATACTTGTT
15	S118	AGAGATCGATGTAAAACACGT	GTGGCATTTTGATGCATT
16	STM0030	AGAGATCGATGTAAAACACGT	GTGGCATTTTGATGGATT
17	C57	AGCTGGCTGACCCTGAACTC	GACAAAATTACAGGAACTGCAAA
18	S148	CAGCAAAATCAGAACCCGAT	GGATCATCAAATTCACCGCT
19	STG0016	AGCTGCTCAGCATCAAGAGA	ACCACCTCAGGCACTTCATC
20	S174	TGAGGGTTTTCAGAAAGGGA	CATCCTTGCAACAACCTCCT
21	StI0003	ACCATCCACCATGTCAATGC	CTCATGGATGGTGTCATTGG
22	SSI	TCTCTTGACACGTGTCACTGAAAC	TCACCGATTACAGTAGGCAAGAGA
23	S189	CCTTGTAGTACAGCAGTGGTC	TCCGCCAAGACTGATGCA
24	STM0014	CAGTCTTCAGCCCATAGG	TAAACAATGGTAGACAAGACAAA
25	STM2020	GCGTCAGCGATTTCAGTACTA	TTCAGTCAACTCCTGTTGCG

序号No.	引物Primer	正向引物Forword primer（5'-3'）	反向引物Reverse primer（5'-3'）
1	C33	TCCCCTTACCCCTCTTCAAAT	AACCCGTGTCAACTTCTGCT
2	C59	TCATCACAACGTGACCCCA	GGGCTTGAATGATGTGAAGCTC
3	S25	AGAGATCGATGTAAAACACGT	GTGGCATTTTGATGGATT
4	C42	AGGCAAACCCCGTAAATCTT	TCTTCCTCACTCGTCCCACT
5	S151	GCTGCTAAACACTCAAGCAGAA	CAACTACAAGATTCCATCCACAC
6	STM-2022	GCGTCAGCGATTTCAGTACTA	TTCAGTCAACTCCTGTTGCG
7	C54	TGGCTGCAGAAATCCAAGATG	TGAAACCACTGGGAGGACTT
8	C58	GCTGCTAAACACTCAAGCAGAA	CAACTACAAGATTCCATCCACAC
9	C3	CCGGAGAATTAGACGATGA	TTTATGGGGAGCAGTTGAGG
10	S153	CCCTTACCCCTCTTCAATC	TCTCTTCTCAGCTGCCGTCT
11	UGP	TGGGGTTCCATCAAAC	GAAACTGCTGCCGGTGC
12	STM1016	TTCTGATTTCATGCATGTTTCC	ATGCTTGCCATGTGATGTGT
13	Patain	TGGTCTGGTGCATTAGAAAAAA	CAACCAACAAGGTAAATGGTACC
14	S170	CGCAAATCTTCATCCGATTC	TCCGGCGGATAATACTTGTT
15	S118	AGAGATCGATGTAAAACACGT	GTGGCATTTTGATGCATT
16	STM0030	AGAGATCGATGTAAAACACGT	GTGGCATTTTGATGGATT
17	C57	AGCTGGCTGACCCTGAACTC	GACAAAATTACAGGAACTGCAAA
18	S148	CAGCAAAATCAGAACCCGAT	GGATCATCAAATTCACCGCT
19	STG0016	AGCTGCTCAGCATCAAGAGA	ACCACCTCAGGCACTTCATC
20	S174	TGAGGGTTTTCAGAAAGGGA	CATCCTTGCAACAACCTCCT
21	StI0003	ACCATCCACCATGTCAATGC	CTCATGGATGGTGTCATTGG
22	SSI	TCTCTTGACACGTGTCACTGAAAC	TCACCGATTACAGTAGGCAAGAGA
23	S189	CCTTGTAGTACAGCAGTGGTC	TCCGCCAAGACTGATGCA
24	STM0014	CAGTCTTCAGCCCATAGG	TAAACAATGGTAGACAAGACAAA
25	STM2020	GCGTCAGCGATTTCAGTACTA	TTCAGTCAACTCCTGTTGCG

序号 No.	引物名称 Primer name	等位位点 Alleles	多态性位点 Polymorphic alleles	多态位点百分数 Percentage of polymorphic bands/%	PIC	Na	Ne	H	I
1	C33	18	14	77.78	0.08	2.00	1.12	0.08	0.16
2	C59	26	26	100.00	0.17	2.00	1.24	0.17	0.29
3	S25	20	14	70.00	0.11	2.00	1.15	0.11	0.21
4	C42	20	10	50.00	0.11	2.00	1.14	0.11	0.20
5	S151	8	5	62.50	0.02	2.00	1.18	0.11	0.19
6	STM-2022	13	11	84.62	0.24	2.00	1.38	0.24	0.39
7	C54	9	7	77.78	0.12	2.00	1.16	0.12	0.21
8	C58	7	5	71.43	0.14	1.85	1.20	0.12	0.20
9	C3	4	2	50.00	0.30	1.50	1.22	0.15	0.24
10	S153	21	14	66.67	0.18	1.95	1.25	0.17	0.29
11	UGP	13	5	38.46	0.13	2.00	1.19	0.13	0.23
12	STM1016	7	2	28.57	0.20	2.00	1.34	0.20	0.32
13	Patain	5	3	60.00	0.06	2.00	1.07	0.06	0.13
14	S170	18	6	33.33	0.21	2.00	1.21	0.15	0.26
15	S118	14	10	71.43	0.08	2.00	1.31	0.20	0.32
16	STM0030	14	13	92.86	0.17	2.00	1.24	0.12	0.17
合计 Total		217	147	67.74	-	-	-	-	-
平均值Mean		13.56	9.19	64.71	0.15	1.96	1.21	0.14	0.24