苦瓜品种SSR分子标记鉴定技术体系构建与应用

doi:10.3864/j.issn.0578-1752.2024.11.014

Abstract

Abstract:

【Objective】In order to provide a practical support for authenticity identification, purity identification and protection of new variety rights of bitter gourd, a set of SSR core primers were screened, and then, the molecular identification system and SSR fingerprint database were established.【Method】Firstly, by using 8 representative varieties with large phenotypic differences, the preliminary 138 pairs SSR primer screening were carried out through 6% polyacrylamide-gel electrophoresis, and the preliminarily screened primers were synthesized into fluorescent primers. Secondly, the primer re-screening was carried out by fluorescence capillary electrophoresis against 95 bitter gourd varieties with diverse geographical sources. Then, the discrimination rate, PIC value, number of alleles, and other parameters of each re-screened primer were calculated respectively, and a set of SSR primer combinations with high discrimination rate and good polymorphism were selected. Finally, the total of 208 bitter gourd varieties were tested by the re-screened primers and a set of SSR core primers were further selected and used to construct a DNA fingerprint database of bitter gourd varieties.【Result】45 pairs of SSR primers with high specificity, high polymorphism and clear bands were screened by polyacrylamide gel electrophoresis. 20 pairs of SSR core primers were finally screened from small populations to large populations by fluorescence capillary electrophoresis. The core primers were divided into 4 groups for multiple electrophoresis, and DNA fingerprint data of 208 bitter gourd varieties were collected. The total of 65 alleles and 102 genotypes were detected by 20 pairs of SSR core primers. 12 reference varieties were selected, which could cover all alleles. Through phylogenetic tree construction, 208 bitter gourd varieties were divided into 2 categories, indicating that 20 pairs of SSR core primers were suitable for genetic diversity analysis of bitter gourd population. 202 of 208 bitter gourd varieties could be identified by core primers, and the discrimination rate was 97.11%. 11 groups of parents and hybrid materials were used to analyze genetic relationship and then conformed to Mendel’s genetic law. There were only 1 locus of fragment loss in each 2 groups, which provided a reference for the decision threshold value of identification of bitter gourd hybrids.【Conclusion】In this study, the identification system of bitter gourd varieties constructed based on 20 pairs of SSR core primers had superior applicability and excellent identification effect, and could also be used for authenticity identification, purity identification, hybrids identification of bitter gourd varieties, similar varieties selection in DUS test and protection of new variety rights of bitter gourd.

Key words: bitter gourd, SSR marker, DNA fingerprints, capillary electrophoresis

SU GuoZhao, LI AiAi, LIU ZhongHua, CHEN YuHua, ZHANG XiuJie, MA YingXue, YANG XuHong, DENG Chao, XU ZhenJiang. Construction and Application of SSR Marker Identification System for Bitter Gourd Varieties[J].Scientia Agricultura Sinica, 2024, 57(11): 2227-2242.

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Figures/Tables 11

Table 1

Information of 208 bitter gourd varieties"

编号 No.	品种名称 Variety name	地区 Region	编号 No.	品种名称 Variety name	地区 Region	编号 No.	品种名称 Variety name	地区 Region	编号 No.	品种名称 Variety name	地区 Region
1	广良2号 Guangliang 2 hao^A	广东 Guangdong	53	M3吉强 M3 Jiqiang^A	广东 Guangdong	105	莆田苦瓜 Putiankugua^B	福建 Fujian	157	桂农科8号 Guinongke 8 hao^A	广西 Guangxi
2	齐胜 Qisheng^A	广东 Guangdong	54	利农52号 Linong 52 hao^A	广东 Guangdong	106	良苦1402 Liangku 1402^A	浙江 Zhejiang	158	桂农科12号 Guinongke 12 hao^A	广西 Guangxi
3	碧俊 Bijun^A	广东 Guangdong	55	利农G4 Linong G4^A	广东 Guangdong	107	桂农科育1号 Guinongkeyu 1 hao^B	广西 Guangxi	159	桂农科17号 Guinongke 17 hao^A	广西 Guangxi
4	青俊 Qingjun^A	广东 Guangdong	56	阿宝7号 Abao 7 hao^A	广东 Guangdong	108	桂农科育2号 Guinongkeyu 2 hao^B	广西 Guangxi	160	桂农科1号 Guinongke 1 hao^A	广西 Guangxi
5	领俊 Lingjun^A	广东 Guangdong	57	E39吉早 E39 Jizao^A	广东 Guangdong	109	丰绿苦瓜 Fenglükugua^A	广东 Guangdong	161	桂农科18号 Guinongke 18 hao^A	广西 Guangxi
6	雅俊 Yajun^A	广东 Guangdong	58	利农9号 Linong 9 hao^A	广东 Guangdong	110	广良3号 Guangliang 3 hao^A	广东 Guangdong	162	桂农科5号 Guinongke 5 hao^A	广西 Guangxi
7	赏玉 Shangyu^A	广东 Guangdong	59	宝玛88 Baoma 88^A	广东 Guangdong	111	MC1-6-12^B	广西 Guangxi	163	桂农科20号 Guinongke 20 hao^A	广西 Guangxi
8	广良108 Guangliang 108^A	广东 Guangdong	60	利农3号 Linong 3 hao^A	广东 Guangdong	112	真功夫13号 Zhengongfu 13 hao^A	广东 Guangdong	164	春晓4号 Chunxiao 4 hao^A	福建 Fujian
9	广良177 Guangliang 117^A	广东 Guangdong	61	汕江大顶 Shanjiangdading^A	广东 Guangdong	113	奇胜105 Qisheng 105^A	福建 Fujian	165	闽绿1号 Minlü 1 hao^A	福建 Fujian
10	广良315 Guangliang 315^A	广东 Guangdong	62	汕海明珠 Shanhaimingzhu^A	广东 Guangdong	114	奇胜201 Qisheng 201^A	福建 Fujian	166	闽研8号 Minyan 8 hao^A	福建 Fujian
11	绿健 Lüjian^A	广东 Guangdong	63	利农22号 Linong 22 hao^A	广东 Guangdong	115	奇胜202 Qisheng 202^A	福建 Fujian	167	C19^B	广东 Guangdong
12	碧珠2号 Bizhu 2 hao^A	广东 Guangdong	64	阿宝1号 Abao 1 hao^A	广东 Guangdong	116	奇胜419 Qisheng 419^A	福建 Fujian	168	C129^B	广东 Guangdong
13	秀珠 Xiuzhu^A	广东 Guangdong	65	翡丽2号 Feili 2 hao^A	广东 Guangdong	117	奇胜122 Qisheng 122^A	福建 Fujian	169	如玉178 Ruyu 178^A	福建 Fujian
14	广良934 Guangliang 934^A	广东 Guangdong	66	E37吉早 E37 Jizao^A	广东 Guangdong	118	田美9号 Tianmei 9 hao^A	福建 Fujian	170	亮丽三号 Liangli 3 hao^A	福建 Fujian
15	广良995 Guangliang 995^A	广东 Guangdong	67	阿宝12号 Abao 12 hao^A	广东 Guangdong	119	田美5号 Tianmei 5 hao^A	福建 Fujian	171	大白苦瓜 Dabaikugua^C	四川 Sichuan
16	碧珠3号 Bizhu 3 hao^A	广东 Guangdong	68	龙头2号 Longtou 2 hao^A	广东 Guangdong	120	奇胜302 Qisheng 302^A	福建 Fujian	172	长汀苦瓜 Changtingkugua^C	福建 Fujian
17	碧珠5号 Bizhu 5 hao^A	广东 Guangdong	69	早翠 Zaocui^A	广东 Guangdong	121	田美31号 Tianmei 31 hao^A	福建 Fujian	173	光泽米黄苦瓜 Guangzemihuangkugua^C	福建 Fujian
18	碧珠8号 Bizhu 8 hao^A	广东 Guangdong	70	利农11号 Linong 11 hao^A	广东 Guangdong	122	新翠 Xincui^A	福建 Fujian	174	南平青苦瓜 Nanpingqingkugua^C	福建 Fujian
19	广良900 Guangliang 900^A	广东 Guangdong	71	阿宝10号 Abao 10 hao^A	广东 Guangdong	123	如玉5号 Ruyu 5 hao^A	福建 Fujian	175	永定大顶苦瓜 Yongdingdadingkugua^C	福建 Fujian
20	广良106 Guangliang 106^A	广东 Guangdong	72	奇胜389 Qisheng 389^A	福建 Fujian	124	丰绿3号 Fenglü 3 hao^A	广东 Guangdong	176	文县苦瓜 Wenxiankugua^C	甘肃 Gansu
21	绿健30 Lüjian 30^A	广东 Guangdong	73	田美油亮一号 Tianmeiyouliang 1 hao^A	福建 Fujian	125	吉美1号 Jimei 1 hao^A	广东 Guangdong	177	潮安大瘤苦瓜 Chaoandaliukugua^C	广东 Guangdong
22	珍珠 Zhenzhu^A	广东 Guangdong	74	田美黑珍珠 Tianmeiheizhenzhu^A	福建 Fujian	126	槟城苦瓜 Penangkugua^B	引自马来西亚 Malaysia	178	长滑苦瓜 Changhuakugua^C	广东 Guangdong
23	阿宝11号 Abao11hao^A	广东 Guangdong	75	亮丽五号 Liangli 5 hao^A	福建 Fujian	127	热科2号 Reke 2 hao^A	海南 Hainan	179	苦瓜 Kugua^C	河北 Hebei
24	利农E36 Linong E36^A	广东 Guangdong	76	亮丽六号 Liangli 6 hao^A	福建 Fujian	128	热研黑玉 Reyanheiyu^B	海南 Hainan	180	白苦瓜 Baikugua^C	黑龙江 Heilongjiang
25	翠竹 Cuizhu^A	广东 Guangdong	77	巴奈特 Banaite^A	福建 Fujian	129	利农珍珠 Linongzhenzhu^A	广东 Guangdong	181	邵阳长白苦瓜 Shaoyangchangbaikugua^C	湖南 Hunan
26	点翠 Diancui^A	广东 Guangdong	78	奇胜543 Qisheng 543^A	福建 Fujian	130	阿宝1号 Abao 1 hao^A	广东 Guangdong	182	沅江青苦瓜 Yuanjiangqingkugua^C	湖南 Hunan
27	宝马99 Baoma 99^A	广东 Guangdong	79	奇胜553 Qisheng 553^A	福建 Fujian	131	青脆1号 Qingcui 1 hao^A	上海 Shanghai	183	三江口苦瓜 Sanjiangkoukugua^C	江西 Jiangxi
28	阿宝3号 Abao 3 hao^A	广东 Guangdong	80	奇胜555 Qisheng 555^A	福建 Fujian	132	热科1号 Reke 1 hao^A	海南 Hainan	184	青皮苦瓜 Qingpikugua^C	新疆引 Xinjiang
29	泰来石头 Tailaishitou^A	广东 Guangdong	81	田美油亮二号 Tianmeiyouliang 2 hao^A	福建 Fujian	133	闽研6号 Minyan 6 hao^A	福建 Fujian	185	白皮苦瓜 Baipikugua^C	四川 Sichuan
30	玉帅绿白 Yushuailübai^A	广东 Guangdong	82	奇胜572 Qisheng 572^A	福建 Fujian	134	奇胜370 Qisheng 370^A	福建 Fujian	186	白苦瓜 Baikugua^C	四川 Sichuan
31	翡丽 Feili^A	广东 Guangdong	83	奇胜573 Qisheng 573^A	福建 Fujian	135	安其罗 Anqiluo^A	福建 Fujian	187	长白苦瓜 Changbaikugua^C	四川 Sichuan
32	强野 Qiangye^A	广东 Guangdong	84	田美明珠 Tianmeimingzhu^A	福建 Fujian	136	亮丽一号 Liangli 1 hao^A	福建 Fujian	188	二白苦瓜 Erbaikugua^C	四川 Sichuan
33	海宝 Haibao^A	广东 Guangdong	85	奇胜245 Qisheng 245^A	福建 Fujian	137	奇胜383 Qisheng 383^A	福建 Fujian	189	绵阳苦瓜 Mianyangkugua^C	四川 Sichuan
34	吉美1号 Jimei 1 hao^A	广东 Guangdong	86	奇胜279 Qisheng 279^A	福建 Fujian	138	田美115 Tianmei 115^A	福建 Fujian	190	苦瓜 Kugua^C	四川 Sichuan
35	澄选珍珠 Chengxuanzhenzhu^A	广东 Guangdong	87	奇胜219 Qisheng 219^A	福建 Fujian	139	奇胜346 Qisheng 346^A	福建 Fujian	191	长白苦瓜 Changbaikugua^C	四川 Sichuan
36	阿宝19号 Abao 19 hao^A	广东 Guangdong	88	金晓601 Jinxiao 601^A	福建 Fujian	140	亮丽二号 Liangli 2 hao^A	福建 Fujian	192	汉源苦瓜 Hanyuankugua^C	四川 Sichuan
37	真功夫11号 Zhengongfu11hao^A	广东 Guangdong	89	田美127 Tianmei127^A	福建 Fujian	141	金晓301 Jinxiao 301^A	福建 Fujian	193	南平长苦瓜 Nanpingchangkugua^C	福建 Fujian
38	汕海2号 Shanhai 2 hao^A	广东 Guangdong	90	农福759 Nongfu 759^A	福建 Fujian	142	KTCQ36^A	福建 Fujian	194	樟木苦瓜 Zhangmukugua^C	广西 Guangxi
39	吉早2号 Jizao 2 hao^A	广东 Guangdong	91	农运来1897 Nongyunlai 1897^A	福建 Fujian	143	春宝 Chunbao^A	福建 Fujian	195	青皮苦瓜 Qingpikugua^C	广西 Guangxi
40	36号油身 36 Haoyoushen^A	广东 Guangdong	92	绿脆1号 Lücui 1 hao^A	上海 Shanghai	144	金晓402 Jinxiao 402^A	福建 Fujian	196	白苦瓜 Baikugua^C	广西 Guangxi
41	利农12号 Linong 12 hao^A	广东 Guangdong	93	青脆2号 Qingcui 2 hao^A	上海 Shanghai	145	奇胜148 Qisheng 148^A	福建 Fujian	197	长苦瓜 Changkugua^C	湖北 Hubei
42	利农17号 Linong 17 hao^A	广东 Guangdong	94	明绿1号 Minglü 1 hao^A	福建 Fujian	146	白富美 Baifumei^A	福建 Fujian	198	白苦瓜 Baikugua^C	四川 Sichuan
43	利农21号 Linong 21 hao^A	广东 Guangdong	95	百利1437 Baili 1437^A	福建 Fujian	147	田美21号 Tianmei 21 hao^A	福建 Fujian	199	长白苦瓜 Changbaikugua^C	宁夏 Ningxia
44	春雅大顶 Chunyadading^A	广东 Guangdong	96	秀玉7号 Xiuyu 7 hao^A	江苏 Jiangsu	148	奇胜127 Qisheng 127^A	福建 Fujian	200	白苦瓜 Baikugua^C	北京 Beijing
45	玉珠1号 Yuzhu 1 hao^A	广东 Guangdong	97	碧绿3号 Bilü 3 hao^A	广东 Guangdong	149	奇胜小玲珑 Qishengxiaolinglong^A	福建 Fujian	201	白苦瓜 Baikugua^C	北京 Beijing
46	明珠3号 Mingzhu 3 hao^A	广东 Guangdong	98	寿光疙瘩绿 Shouguanggedalü^B	山东 Shandong	150	白珍珠二号 Baizhenzhu 2 hao^A	福建 Fujian	202	大白苦瓜 Dabaikugua^C	北京 Beijing
47	超油3号 Chaoyou 3 hao^A	广东 Guangdong	99	良苦1403 Liangku 1403^A	浙江 Zhejiang	151	越高56号 Yuegao 56 hao^A	福建 Fujian	203	绿苦瓜 Lükugua^C	北京 Beijing
48	碧丽 Bili^A	广东 Guangdong	100	良苦1405 Liangku 1405^A	浙江 Zhejiang	152	越高94号 Yuegao 94 hao^A	福建 Fujian	204	Layiz^C	引自泰国 Thailand
49	假珍珠13号 Jiazhenzhu 13 hao^A	广东 Guangdong	101	良苦1406 Liangku 1406^A	浙江 Zhejiang	153	金晓405 Jinxiao 405^A	福建 Fujian	205	金晓401 Jinxiao 401^A	福建 Fujian
50	107#2#假珍珠 107#2#Jiazhenzhu^A	广东 Guangdong	102	云霄苦瓜 Yunxiaokugua^B	福建 Fujian	154	碧绿9号 Bilü 9 hao^A	广东 Guangdong	206	秀玉5号 Xiuyu 5 hao^A	江苏 Jiangsu
51	14#23#假珍珠 14#23#Jiazhenzhu^A	广东 Guangdong	103	良苦1401 Liangku 1401^A	浙江 Zhejiang	155	绿冰清 Lübingqing^A	广东 Guangdong	207	超群3号 Chaoqun 3 hao^A	福建 Fujian
52	M1吉强 M1 Jiqiang^A	广东 Guangdong	104	疙瘩绿 Gedalü^B	浙江 Zhejiang	156	桂农科10号 Guinongke 10 hao^A	广西 Guangxi	208	金晓403 Jinxiao 403^A	福建 Fujian

Table 1

Fig. 1

Table 2

Table 3

Table 4

Fig. 2

Fig. 3

Table 5

Fig. 4

Table 6

Fig. 5

References 42

[1]	GB/T 39917-2021. 主要农作物品种真实性和纯度SSR分子标记检测稻. 北京: 中华人民共和国农业农村部, 2021.
	GB/T 39917-2021. Protocol for identification of rice varieties-SSR marker method. Beijing: Ministry of Agriculture and Rural Affairs of the People’s Republic of China, 2021. (in Chinese)
[2]	王凤格, 杨扬, 易红梅, 赵久然, 任洁, 王璐, 葛建镕, 江彬, 张宪晨, 田红丽, 侯振华. 中国玉米审定品种标准SSR指纹库的构建. 中国农业科学, 2017, 50(1): 1-14. doi: 10.3864/j.issn.0578-1752.2017.01.001.
	WANG F G, YANG Y, YI H M, ZHAO J R, REN J, WANG L, GE J R, JIANG B, ZHANG X C, TIAN H L, HOU Z H. Construction of an SSR-based standard fingerprint database for corn variety authorized in China. Scientia Agricultura Sinica, 2017, 50(1): 1-14. doi: 10.3864/j.issn.0578-1752.2017.01.001. (in Chinese)
[3]	郑永胜, 张晗, 王东建, 孙加梅, 王雪梅, 段丽丽, 李华, 王玮, 李汝玉. 基于荧光检测技术的小麦品种SSR鉴定体系的建立. 中国农业科学, 2014, 47(19): 3725-3735. doi: 10.3864/j.issn.0578-1752.2014.19.001.
	ZHENG Y S, ZHANG H, WANG D J, SUN J M, WANG X M, DUAN L L, LI H, WANG W, LI R Y. Development of a wheat variety identification system based on fluorescently labeled SSR Markers. Scientia Agricultura Sinica, 2014, 47(19): 3725-3735. doi: 10.3864/j.issn.0578-1752.2014.19.001. (in Chinese)
[4]	李益, 马先锋, 唐浩, 李娜, 江东, 龙桂友, 李大志, 牛英, 韩瑞玺, 邓子牛. 柑橘品种鉴定的SSR标记开发和指纹图谱库构建. 中国农业科学, 2018, 51(15): 149-159. doi: 10.3864/j.issn.0578-1752.2018.15.012.
	LI Y, MA X F, TANG H, LI N, JIANG D, LONG G Y, LI D Z, NIU Y, HAN R X, DENG Z N. SSR markers screening for identification of citrus cultivar and construction of DNA fingerprinting library. Scientia Agricultura Sinica, 2018, 51(15): 149-159. doi: 10.3864/j.issn.0578-1752.2018.15.012. (in Chinese)
[5]	NY/T 2474-2013. 黄瓜品种鉴定技术规程 SSR分子标记法. 北京: 中华人民共和国农业农村部, 2013.
	NY/T 2474-2013. Protocol for the identification of cucumber varieties-SSR marker method. Beijing: Ministry of Agriculture and Rural Affairs of the People’s Republic of China, 2013. (in Chinese)
[6]	NY/T 2472-2013. 西瓜品种鉴定技术规程 SSR分子标记法. 北京: 中华人民共和国农业农村部, 2013.
	NY/T 2472-2013. Identification of watermelon varieties-SSR marker method. Beijing: Ministry of Agriculture and Rural Affairs of the People’s Republic of China, 2013. (in Chinese)
[7]	王国莉, 徐毓璇, 黄梅花. 基于SSR和SRAP标记的苦瓜品种鉴定及亲缘关系分析. 分子植物育种, 2016, 14(2): 501-510.
	WANG G L, XU Y X, HUANG M H. Identification and genetic relationship analysis of bitter gourd varieties based on SSR and SPAP. Molecular Plant Breeding, 2016, 14(2): 501-510. (in Chinese)
[8]	姚春鹏, 张晓爱, 吴廷全, 邓洁, 金庆敏, 李海达, 王瑞娟, 张长远. 用于长绿2号苦瓜种子纯度鉴定的SSR分子标记的开发. 分子植物育种, 2019, 17(11): 3660-3664.
	YAO C P, ZHANG X A, WU T Q, DENG J, JIN Q M, LI H D, WANG R J, ZHANG C Y. Development of SSR molecular markers for seed purity identification of bitter gourd hybrid Changlü No.2. Molecular Plant Breeding, 2019, 17(11): 3660-3664. (in Chinese)
[9]	DHILLON N P S, SANGUANSIL S, SCHAFLEITNER R, WANG Y W, MCCREIGHT J D. Diversity among a wide Asian collection of bitter gourd landraces and their genetic relationships with commercial hybrid cultivars. Journal of the American Society for Horticultural Science, 2016, 141(5): 475-484.
[10]	CUI J J, CHENG J W, NONG D G, PENG J Z, HU Y F, HE W M, ZHOU Q J, DHILLON N P S, HU K L. Genome-wide analysis of simple sequence repeats in bitter gourd (Momordica charantia). Frontiers in Plant Science, 2017, 8: 1103.
[11]	POWELL W, MORGANTE M, ANDRE C, HANAFEY M, VOGEL J, TINGEY S, RAFALSKI A. The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis. Molecular Breeding, 1996, 2(3): 225-238.
[12]	DAR A A, MAHAJAN R, SHARMA S. Molecular markers for characterization and conservation of plant genetic resources. The Indian Journal of Agricultural Sciences, 2019, 89(11): 1755-1763.
[13]	WANG S Z, PAN L, HU K, CHEN C Y, DING Y. Development and characterization of polymorphic microsatellite markers in Momordica charantia (Cucurbitaceae). American Journal of Botany, 2010, 97(8): e75-e78.
[14]	GUO D L, ZHANG J P, XUE Y M, HOU X G. Isolation and characterization of 10 SSR markers of Momordica charantia (Cucurbitaceae). American Journal of Botany, 2012, 99(5): e182-e183.
[15]	SAXENA S, SINGH A, ARCHAK S, BEHERA T K, JOHN J K, MESHRAM S U, GAIKWAD A B. Development of novel simple sequence repeat markers in bitter gourd (Momordica charantia L.) through enriched genomic libraries and their utilization in analysis of genetic diversity and cross-species transferability. Applied Biochemistry and Biotechnology, 2015, 175(1): 93-118.
[16]	JI Y, LUO Y M, HOU B W, WANG W Z, ZHAO J F, YANG L M, XUE Q Y, DING X Y. Development of polymorphic microsatellite loci in Momordica charantia (Cucurbitaceae) and their transferability to other cucurbit species. Scientia Horticulturae, 2012, 140: 115-118.
[17]	温常龙, 罗江, 韩向阳, 张建, 杨静静. 一种鉴定苦瓜品种真实性的方法及其专用SSR引物组合: CN202010209970.8. 北京: 北京市农林科学院, 2021.
	WEN C L, LUO J, HAN X Y, ZHANG J, YANG J J. A method for identifying the authenticity of bitter gourd varieties and its special SSR primer combination: CN202010209970.8. Beijing: Beijing Academy of Agriculture and Forestry Sciences, 2021. (in Chinese)
[18]	邓俭英, 王绪, 方锋学, 张曼, 孙德利. 不同性别类型苦瓜基因组DNA提取及RAPD标记初步研究. 广西农业科学, 2007, 38(3): 223-226.
	DENG J Y, WANG X, FANG F X, ZHANG M, SUN D L. Effective extraction of genomic DNA and RAPD analysis on different sex types of Momordica charantia L. Guangxi Agricultural Sciences, 2007, 38(3): 223-226. (in Chinese)
[19]	LIU K J, MUSE S V. PowerMarker: An integrated analysis environment for genetic marker analysis. Bioinformatics, 2005, 21(9): 2128-2129. doi: 10.1093/bioinformatics/bti282 pmid: 15705655
[20]	PEAKALL R, SMOUSE P E. GenAlEx 6.5: Genetic analysis in Excel. Population genetic software for teaching and research: an update. Bioinformatics, 2012, 28(19): 2537-2539.
[21]	TAMURA K, STECHER G, KUMAR S. MEGA11: Molecular evolutionary genetics analysis version 11. Molecular Biology and Evolution, 2021, 38(7): 3022-3027. doi: 10.1093/molbev/msab120 pmid: 33892491
[22]	LETUNIC I, BORK P. Interactive Tree Of Life (iTOL): An online tool for phylogenetic tree display and annotation. Bioinformatics, 2007, 23(1): 127-128. doi: 10.1093/bioinformatics/btl529 pmid: 17050570
[23]	LYTTLE T W. Segregation distorters. Annual Review of Genetics, 1991, 25: 511-557. pmid: 1812815
[24]	BOTSTEIN D, WHITE R L, SKOLNICK M, DAVIS R W. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. American Journal of Human Genetics, 1980, 32(3): 314-331. pmid: 6247908
[25]	ALHARIRI A, BEHERA T K, JAT G S, DEVI M B, BOOPALAKRISHNAN G, HEMEDA N F, TELEB A A, ISMAIL E, ELKORDY A. Analysis of genetic diversity and population structure in bitter gourd (Momordica charantia L.) using morphological and SSR markers. Plants, 2021, 10(9): 1860.
[26]	聂兴华, 李伊然, 田寿乐, 王雪峰, 苏淑钗, 曹庆芹, 邢宇, 秦岭. 中国板栗品种(系)DNA指纹图谱构建及其遗传多样性分析. 园艺学报, 2022, 49(11): 2313-2324. doi: 10.16420/j.issn.0513-353x.2021-0863
	NIE X H, LI Y R, TIAN S L, WANG X F, SU S C, CAO Q Q, XING Y, QIN L. Construction of DNA fingerprint map and analysis of genetic diversity for Chinese chestnut cultivars(lines). Acta Horticulturae Sinica, 2022, 49(11): 2313-2324. (in Chinese)
[27]	李晓娟, 赵文菊, 赵孟良, 郭怡婷, 马一栋, 马雪杰, 任延靖. 球茎甘蓝SSR分子标记开发及指纹图谱构建. 南方农业学报, 2023, 54(1): 22-33.
	LI X J, ZHAO W J, ZHAO M L, GUO Y T, MA Y D, MA X J, REN Y J. SSR molecular markers development and fingerprints construction in kohlrabi(Brassica oleracea var. gongylodes L.). Journal of Southern Agriculture, 2023, 54(1): 22-33. (in Chinese)
[28]	郭栋梁, 黄石连, 王静, 韩冬梅, 李建光. 基于SSR分子标记的龙眼种质资源遗传多样性分析及其指纹图谱构建. 中国农学通报, 2022, 38(36): 67-73. doi: 10.11924/j.issn.1000-6850.casb2022-0009
	GUO D L, HUANG S L, WANG J, HAN D M, LI J G. Longan germplasms resources: Genetic diversity analysis and fingerprint construction based on SSR markers. Chinese Agricultural Science Bulletin, 2022, 38(36): 67-73. (in Chinese) doi: 10.11924/j.issn.1000-6850.casb2022-0009
[29]	王瑞, 洪文娟, 罗华, 赵丽娜, 陈颖, 王君. 石榴品种SSR指纹图谱构建及杂种父本鉴定. 园艺学报, 2023, 50(2): 265-278. doi: 10.16420/j.issn.0513-353x.2021-1008
	WANG R, HONG W J, LUO H, ZHAO L N, CHEN Y, WANG J. Construction of SSR fingerprints of pomegranate cultivars and male parent identification of hybrids. Acta Horticulturae Sinica, 2023, 50(2): 265-278. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-1008
[30]	孙泽硕, 蒋冬月, 柳新红, 沈鑫, 李因刚, 屈雨飞, 李永华. 基于SSR标记的42份樱花品种的聚类分析及DNA指纹图谱构建. 园艺学报, 2023, 50(3): 657-668. doi: 10.16420/j.issn.0513-353x.2021-1057
	SUN Z S, JIANG D Y, LIU X H, SHEN X, LI Y G, QU Y F, LI Y H. Cluster analysis and construction of DNA fingerprinting of 42 oriental cultivars of flowering cherry based on SSR markers. Acta Horticulturae Sinica, 2023, 50(3): 657-668. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-1057
[31]	崔竣杰, 程蛟文, 曹毅, 胡开林. 基于SSR标记和表型性状构建苦瓜核心种质的研究. 中国蔬菜, 2022(2): 25-32.
	CUI J J, CHENG J W, CAO Y, HU K L. Studies on construction of bitter gourd core collection based on SSR markers and phenotypic traits. China Vegetables, 2022(2): 25-32. (in Chinese)
[32]	MANGELSDORF P C, JONES D F. The expression of Mendelian factors in the gametophyte of maize. Genetics, 1926, 11(5): 423-455. doi: 10.1093/genetics/11.5.423 pmid: 17246465
[33]	MATSUSHITA S, ISEKI T, FUKUTA Y, ARAKI E, KOBAYASHI S, OSAKI M, YAMAGISHI M. Characterization of segregation distortion on chromosome 3 induced in wide hybridization between indica and japonica type rice varieties. Euphytica, 2003, 134(1): 27-32.
[34]	WANG G, HE Q Q, XU Z K, SONG R T. High segregation distortion in maize B73 x teosinte crosses. Genetics and Molecular Research, 2012, 11(1): 693-706.
[35]	MANABE M, INO T, KASAYA M, TAKUMI S, MORI N, OHTSUKA I, NAKAMURA C. Segregation distortion through female gametophytes in interspecific hybrids of tetraploid wheat as revealed by RAPD analysis. Hereditas, 2004, 131(1): 47-53.
[36]	ZAMIR D, TANKSLEY S D, JONES R A. Haploid selection for low temperature tolerance of tomato pollen. Genetics, 1982, 101(1): 129-137. doi: 10.1093/genetics/101.1.129 pmid: 17246078
[37]	LAMBRIDES C J, GODWIN I D, LAWN R J, IMRIE B C. Segregation distortion for seed testa color in mungbean (Vigna radiata L. wilcek). Journal of Heredity, 2004, 95(6): 532-535.
[38]	谷艳鹏, 张泽人, 孙涛, 韩庆军, 栗宁宁, 鲁仪增, 窦德泉, 郑健. 基于EST-SSR标记的花楸树和少叶花楸杂交F₁代群体的杂种鉴定及遗传关系分析. 植物资源与环境学报, 2022, 31(4): 65-73.
	GU Y P, ZHANG Z R, SUN T, HAN Q J, LI N N, LU Y Z, DOU D Q, ZHENG J. Hybrid identification and genetic relationship analysis of F₁ hybrid population of Sorbus pohuashanensis and Sorbus hupehensis var. paucijuga based on EST-SSR marker. Journal of Plant Resources and Environment, 2022, 31(4): 65-73. (in Chinese)
[39]	周宁宁, 李淑斌, 李远波, 蹇洪英, 晏慧君, 王其刚, 陈敏, 邱显钦, 张颢, 唐开学. 二倍体月季F₁群体的SSR鉴定与遗传分析. 园艺学报, 2017, 44(1): 151-160. doi: 10.16420/j.issn.0513-353x.2016-0413
	ZHOU N N, LI S B, LI Y B, JIAN H Y, YAN H J, WANG Q G, CHEN M, QIU X Q, ZHANG H, TANG K X. Hybrids identification and genetic analysis in diploid roses population (F₁) using SSR markers. Acta Horticulturae Sinica, 2017, 44(1): 151-160. (in Chinese)
[40]	汪国云, 沈禹彤, 贾慧敏, 焦云, 柴春燕, 包劲松, 孙德利, 贾惠娟, 高中山. ‘荸荠’×‘东魁’杨梅杂交群体构建与SSR杂种鉴定. 果树学报, 2015, 32(4): 555-560, 737.
	WANG G Y, SHEN Y T, JIA H M, JIAO Y, CHAI C Y, BAO J S, SUN D L, JIA H J, GAO Z S. Construction of a crossing population between two Chinese bayberry cultivars ‘Biqi’ and ‘Dongkui’ and hybrid identification by polymorphic SSRs. Journal of Fruit Science, 2015, 32(4): 555-560, 737. (in Chinese)
[41]	王哲. 植物杂交后代中基因偏分离的产生原因及其进化意义. 遗传, 2016, 38(9): 801-810.
	WANG Z. Distorted segregation in plant hybrids and its implication for evolution. Hereditas, 2016, 38(9): 801-810. (in Chinese)
[42]	TAYLOR D R, INGVARSSON P K. Common features of segregation distortion in plants and animals. Genetica, 2003, 117(1): 27-35. doi: 10.1023/a:1022308414864 pmid: 12656570

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序号 No.	引物编号 Primer No.	引物名称 Primer name	染色体定位 Chromosome location	重复基序 SSR motifs	正向引物序列 Forward primer sequences (5′-3′)	反向引物序列 Reverse primer sequences (5′-3′)
1	KG4	N9	1	(TC) 17	ATCCATCCCCACAAGTTGAA	CCATAAGGATATGTTTGCATGG
2	KG51	Bdcs_SSr-0097	2	(TA) 8	AATGCCAGATGTAATTGCTATGGTG	TACACATCCATACCGCTAGAAGAAG
3	KG2	N5	3	(CT) 13	CGTCGCTCTCACAAGAGATAAG	TTTGGTGGAAATCCCCTATT
4	KG41	Bdcs_SSr-0064	4	(AT) 7	GACTTTTGTCCGATCGAACATAACT	TGGCATCCATTTTGTTTTTCTTTCA
5	KG20	AVRDC-BG48	4	(GA) 8	GCAAAAACACTGTCACCCAC	TTCGCTTCTTCCCTCTTCAT
6	KG16	McSSR20	4	(TCT) 9	GGAATTCAGGTGAACCTGACG	CCAGGAGGAAGAGGAACTGC
7	KG49	Bdcs_SSr-0080	5	(TAT) 6	TGCTACTTGAACCAATGAGAAGAAAC	TGATTTGTTCTTTCTATGAAAATGTTGT
8	KG23	AVRDC-BG136	6	(TTC) 7	TCGCAGTCTCATTTCTCAAG	AGTGGCAGAGCGTTTTACCT
9	KG33	Bdcs_SSr-0038	6	(TA) 6	TCTCGAGGTGGGAGATTTAATCTTC	TTATTTGGTTTCTGATGCGTCTGTC
10	KG30	MC07_92319	7	(TC) 15	GCAAATCAAAGAAGCCAAGC	GTAGGGGTTGGGTTGATCCT
11	KG26	MC07_95456	7	(CT) 16	TTCTTGAGAGACGGTTGGCT	GATACAAAGAAACGGTGGCG
12	KG18	AVRDC-BG2	8	(AC) 8	GAGCACACAGAAAATTGGGT	TGATCCACTCCCAATCTTAGC
13	KG43	Bdcs_SSr-0098	8	(AAT) 6	TATTTGTATTTGTCCGTCGGAGGG	CGCCATTTCCTGTTTGATTCAATAT
14	KG42	Bdcs_SSr-0085	8	(TC) 6	ATGGACTCAGAATGAACTGAAAAGG	ACTTTGTTCTCATAAGAGGGGAGTT
15	KG8	S15	8	(AG) 6 (AG) 12	GGGTAGTGGAATGATGGGTT	TAGTGTTTTCGTGAGGGAGG
16	KG22	AVRDC-BG85	8	(GAA) 8	TGCAACCACTTGGGTTCTAA	CACGCCAGTAGCTTCAACAT
17	KG50	Bdcs_SSr-0088	9	(TTA) 7	AAGAAGGTGTACTAGTAGCTCCTCT	CCAAGTGGCATCTTATCAACACATT
18	KG24	MC10_147313	10	(CT) 21	CGGCATGAAGAATGGCTAAT	GGGGTTTTCCCCTAATTGAA
19	KG39	Bdcs_SSr-0094	11	(ATA) 6	ACATATTACTTGGTAATTGCTAGACAA	TGAACATTGTTGAGGCACCTAAAAA
20	KG45	Bdcs_SSr-0025	11	(GA) 5	CCAACAATTATCCACTTGTGCTTCT	GACTTTATTGTGTGGGGACATGAAC

引物 Primer	主要等位基因频率 Major allele frequency	基因型数量Number of genotypes	多态性信息含量 Polymorphism information content	等位基因数 Number of observed alleles	有效等位基因数Number of effective alleles	观察杂合度 Observed heterozygosity	期望杂合度 Expected heterozygosity	香农-多样性指数 Shannon- Wiener index
KG16	0.60	3	0.37	2	1.93	0.47	0.48	0.67
KG18	0.60	7	0.41	4	2.04	0.44	0.51	0.81
KG2	0.67	5	0.35	4	1.80	0.43	0.44	0.66
KG20	0.53	4	0.38	3	2.00	0.48	0.50	0.71
KG22	0.71	8	0.34	5	1.72	0.37	0.42	0.67
KG23	0.66	3	0.35	2	1.82	0.39	0.45	0.64
KG24	0.58	12	0.48	8	2.24	0.46	0.55	0.99
KG26	0.68	12	0.45	6	1.97	0.43	0.49	0.96
KG30	0.75	8	0.38	5	1.70	0.36	0.41	0.77
KG33	0.69	3	0.34	2	1.74	0.44	0.43	0.62
KG39	0.65	3	0.35	2	1.84	0.34	0.46	0.65
KG4	0.75	5	0.31	3	1.60	0.36	0.38	0.59
KG41	0.63	3	0.36	2	1.88	0.40	0.47	0.66
KG42	0.55	3	0.37	2	1.98	0.58	0.49	0.69
KG43	0.53	5	0.43	4	2.16	0.45	0.54	0.86
KG45	0.65	3	0.35	2	1.84	0.42	0.46	0.65
KG49	0.64	3	0.36	2	1.86	0.47	0.46	0.66
KG50	0.58	3	0.37	2	1.95	0.37	0.49	0.68
KG51	0.62	3	0.36	2	1.89	0.35	0.47	0.66
KG8	0.74	6	0.34	3	1.67	0.35	0.40	0.67
合计Total	\	102	\	65	37.61	\	\	\
均值Average	0.64	\	0.37	3.25	1.88	0.42	0.46	0.71

组数 Group	FAM	TAMRA	ROX	HEX
1	KG20 (97-101) KG8 (229-233)	KG51 (276-278)	KG26 (244-258)	KG18 (139-158)
2	KG45 (97-99)	KG43 (297-300) KG22 (245-273)	KG49 (211-214)	KG23 (107-118)
3	KG24 (230-248)	KG2 (158-166)	KG42 (255-257)	KG33 (103-105) KG16 (206-215)
4	KG30 (228-250)	KG50 (259-262)	KG39 (273-276)	KG41 (187-196) KG4 (91-103)

编号 No.	品种名称 Variety name	品种来源 Variety origin	保藏编号 Preservation No.	编号 No.	品种名称 Variety name	品种来源 Variety origin	保藏编号 Preservation No.
1	广良995 Guangliang 995	广东省良种引进服务公司 Guangdong Provincial Improved Variety Introduce Service Corp.	\	7	奇胜小玲珑Qishengxiaolinglong	福建省农业科学院 Fujian Academy of Agricultural Sciences	\
2	碧珠3号 Bizhu 3 hao	广东省良种引进服务公司 Guangdong Provincial Improved Variety Introduce Service Corp.	\	8	桂农科5号 Guinongke 5 hao	福建省农业科学院 Fujian Academy of Agricultural Sciences	\
3	碧珠5号 Bizhu 5 hao	广东省良种引进服务公司 Guangdong Provincial Improved Variety Introduce Service Corp.	\	9	槟城苦瓜Penangkugua	农业农村部植物新品种保藏中心 New Plant Variety Preservation Center，Ministry of Agriculture and Rural Affairs	XIN41090
4	碧珠8号 Bizhu 8 hao	广东省良种引进服务公司 Guangdong Provincial Improved Variety Introduce Service Corp.	\	10	田美115 Tianmei 115	农业农村部植物新品种保藏中心 New Plant Variety Preservation Center，Ministry of Agriculture and Rural Affairs	XIN43633
5	广良900 Guangliang 900	广东省良种引进服务公司 Guangdong Provincial Improved Variety Introduce Service Corp.	\	11	长滑苦瓜Changhuakugua	国家园艺种质资源库 National Horticultural Germplasm Resource Library	V05G0020
6	利农9号 Linong 9 hao	广东和利农生物种业股份有限公司 Guangdong Helinong Biological Seed Industry Co.，Ltd.	\	12	长白苦瓜Changbaikugua	国家园艺种质资源库 National Horticultural Germplasm Resource Library	V05G0159

品种名称 Variety name	位点Locus				品种名称 Variety name	位点Locus
品种名称 Variety name	KG2	KG8	KG33	KG39	品种名称 Variety name	KG2	KG8	KG33	KG39
广良2号父本 Male parent of Guangliang 2 hao	162/162	229/229	103/103	273/273	田美115父本 Male parent of Tianmei 115	162/162	229/229	103/103	273/273
广良2号母本 Female parent of Guangliang 2 hao	164/164	229/229	105/105	273/273	田美115母本 Female parent of Tianmei 115	164/164	231/231	103/105	273/276
广良2号Guangliang 2 hao	162/164	229/229	103/105	273/273	田美115 Tianmei 115	164/164	229/231	103/105	273/273

Construction and Application of SSR Marker Identification System for Bitter Gourd Varieties

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