Establishment of DNA fingerprint for sunflower by SSR-HRM technique

doi:10.3969/j.issn.1004-1524.2022.04.03

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (4): 678-686.DOI: 10.3969/j.issn.1004-1524.2022.04.03

• Crop Science • Previous Articles Next Articles

Establishment of DNA fingerprint for sunflower by SSR-HRM technique

LIU Yihan¹^,²(), MOU Qingshan¹^,², CHEN Shanyu³, RUAN Guanhai³, HU Jin¹^,², GUAN Yajing¹^,²^,^*()

1. College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China
2. Hainan Institute, Zhejiang University, Sanya 572025,Hainan,China
3. Institute of Crops and Nuclear Technology Utilization, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2021-06-23 Online:2022-04-25 Published:2022-04-28
Contact: GUAN Yajing

Abstract

Abstract:

Combined with high-resolution melting curve analysis technology, 17 pairs of core simple sequence repeats (SSR) primers were screened out for genetic diversity analysis and DNA fingerprint construction of 32 sunflower lines collected from Zhejiang Province.The results of cluster analysis showed that genetic diversity among the 32 materials was relatively low, and genetic relationships among most of 32 materials were consistent with their geographical sources and types. This study innovated the identification method of sunflower germplasm resources, and laid a foundation for further research on identification of sunflower germplasm by microsatellite high-resolution melting technique.

Key words: sunflower, SSR markers, high-resolution melting curve, fingerprint map, genetic diversity

CLC Number:

S565.5

LIU Yihan, MOU Qingshan, CHEN Shanyu, RUAN Guanhai, HU Jin, GUAN Yajing. Establishment of DNA fingerprint for sunflower by SSR-HRM technique[J]. Acta Agriculturae Zhejiangensis, 2022, 34(4): 678-686.

Figures/Tables 7

References 18

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编号 No.	名称 Name	类型 Species	特性 Trait	来源 Origin	编号 No.	名称 Name	类型 Species	特性 Trait	来源 Origin
1	油葵M985 Oil sunflower M985	杂交种 Hybrid	油用 Oil	河北 Hebei	17	红晕向日葵 Red sunflower	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
2	油葵T777 Oil sunflower T777	杂交种 Hybrid	油用 Oil	河北 Hebei	18	多头向日葵 Multipoint sunflower	杂交种 Hybrid	观赏 Ornamental	江苏 Jiangsu
3	美国至尊矮大头 American best dwarf	杂交种 Hybrid	油用 Oil	香港 Hong Kong	19	玩具熊向日葵 Teddy bear sunflower	杂交种 Hybrid	观赏 Ornamental	江苏 Jiangsu
4	YK18-6(油葵杂交组合) YK18-6 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	20	紫色向日葵 Purple sunflower	杂交种 Hybrid	观赏 Ornamental	美国 America
	hybrid combination)
5	YK18-9(油葵杂交组合) YK18-9 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	21	全阳改良 Quanyanggailiang	杂交种 Hybrid	观赏 Ornamental	日本 Japan
	hybrid combination)
6	YK18-10(油葵杂交组合) YK18-10 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	22	富阳酸橙 Fuyangsuancheng	杂交种 Hybrid	观赏 Ornamental	日本 Japan
	hybrid combination)
7	YK18-11(油葵杂交组合) YK18-11 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	23	观赏S5 Ornamental sunflower S5	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
8	YK18-13(油葵杂交组合) YK18-13 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	24	观赏S8 Ornamental sunflower S8	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
9	YK18-15(油葵杂交组合) YK18-15 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	25	观赏S9 Ornamental sunflower S9	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
10	YK18-16(油葵杂交组合) YK18-16 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	26	龙赏葵1号 Longshangkui No.1	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
11	YK18-17(油葵杂交组合) YK18-17 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	27	JZ072	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
12	龙葵杂4号 Longkuiza No.4	杂交种 Hybrid	油用 Oil	黑龙江 Heilongjiang	28	SH361(S粒型) SH361 (S grain shape)	杂交种 Hybrid	食用 Edible	内蒙 Inner Mongolia
13	龙葵杂8号 Longkuiza No.8	杂交种 Hybrid	油用 Oil	黑龙江 Heilongjiang	29	SH363(S粒型) SH363 (S grain shape)	杂交种 Hybrid	食用 Edible	内蒙 Inner Mongolia
14	龙葵杂10号 Longkuiza No.10	杂交种 Hybrid	油用 Oil	黑龙江 Heilongjiang	30	S116(食葵杂交组合) S116 (Edible sunflower hybrid combination)	杂交种 Hybrid	食用 Edible	黑龙江 Heilongjiang
15	龙赏葵5号 Longshangkui No.5	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang	31	S117(食葵杂交组合) S117 (Edible sunflower hybrid combination)	杂交种 Hybrid	食用 Edible	黑龙江 Heilongjiang
16	龙赏葵7号 Longshangkui No.7	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang	32	S144(食葵杂交组合) S144 (Edible sunflower hybrid combination)	杂交种 Hybrid	食用 Edible	黑龙江 Heilongjiang

编号 No.	名称 Name	类型 Species	特性 Trait	来源 Origin	编号 No.	名称 Name	类型 Species	特性 Trait	来源 Origin
1	油葵M985 Oil sunflower M985	杂交种 Hybrid	油用 Oil	河北 Hebei	17	红晕向日葵 Red sunflower	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
2	油葵T777 Oil sunflower T777	杂交种 Hybrid	油用 Oil	河北 Hebei	18	多头向日葵 Multipoint sunflower	杂交种 Hybrid	观赏 Ornamental	江苏 Jiangsu
3	美国至尊矮大头 American best dwarf	杂交种 Hybrid	油用 Oil	香港 Hong Kong	19	玩具熊向日葵 Teddy bear sunflower	杂交种 Hybrid	观赏 Ornamental	江苏 Jiangsu
4	YK18-6(油葵杂交组合) YK18-6 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	20	紫色向日葵 Purple sunflower	杂交种 Hybrid	观赏 Ornamental	美国 America
	hybrid combination)
5	YK18-9(油葵杂交组合) YK18-9 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	21	全阳改良 Quanyanggailiang	杂交种 Hybrid	观赏 Ornamental	日本 Japan
	hybrid combination)
6	YK18-10(油葵杂交组合) YK18-10 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	22	富阳酸橙 Fuyangsuancheng	杂交种 Hybrid	观赏 Ornamental	日本 Japan
	hybrid combination)
7	YK18-11(油葵杂交组合) YK18-11 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	23	观赏S5 Ornamental sunflower S5	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
8	YK18-13(油葵杂交组合) YK18-13 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	24	观赏S8 Ornamental sunflower S8	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
9	YK18-15(油葵杂交组合) YK18-15 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	25	观赏S9 Ornamental sunflower S9	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
10	YK18-16(油葵杂交组合) YK18-16 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	26	龙赏葵1号 Longshangkui No.1	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
11	YK18-17(油葵杂交组合) YK18-17 (Oil sunflower	杂交种 Hybrid	油用 Oil	吉林 Jilin	27	JZ072	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang
	hybrid combination)
12	龙葵杂4号 Longkuiza No.4	杂交种 Hybrid	油用 Oil	黑龙江 Heilongjiang	28	SH361(S粒型) SH361 (S grain shape)	杂交种 Hybrid	食用 Edible	内蒙 Inner Mongolia
13	龙葵杂8号 Longkuiza No.8	杂交种 Hybrid	油用 Oil	黑龙江 Heilongjiang	29	SH363(S粒型) SH363 (S grain shape)	杂交种 Hybrid	食用 Edible	内蒙 Inner Mongolia
14	龙葵杂10号 Longkuiza No.10	杂交种 Hybrid	油用 Oil	黑龙江 Heilongjiang	30	S116(食葵杂交组合) S116 (Edible sunflower hybrid combination)	杂交种 Hybrid	食用 Edible	黑龙江 Heilongjiang
15	龙赏葵5号 Longshangkui No.5	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang	31	S117(食葵杂交组合) S117 (Edible sunflower hybrid combination)	杂交种 Hybrid	食用 Edible	黑龙江 Heilongjiang
16	龙赏葵7号 Longshangkui No.7	杂交种 Hybrid	观赏 Ornamental	黑龙江 Heilongjiang	32	S144(食葵杂交组合) S144 (Edible sunflower hybrid combination)	杂交种 Hybrid	食用 Edible	黑龙江 Heilongjiang

编号 No.	标记名 Marker name	上游引物 Forward primer (5'-3')	下游引物 Reverse primer (5'-3')	位置 Position	等位基因数量 Number of alleles
1	ORS543	CCAAGTTTCAGTTACAATCCATGA	GGTCATTAGGAGTTTGGGATCA	Linkage group 1	5
2	ORS456	CCAAGGAATTCTAACAAGAGTTTAAG	GATTTCTCACTTCACTCCTCTATGC	Linkage group 8	3
3	ORS342	CACCAGCATAGCCATTCAAA	TGTTCATCAGGTTTGTCTCCA	Linkage group 2	3
4	ORS656	ACGGACGTAGAGTGGTGGAG	TCGTGGTAAGGGAAGACAACA	Linkage group 16	6
5	ORS844	ACGATGCAAAGAATATACTGCAC	CATGTTTAATAGGTTTTAATTCTAGGG	Linkage group 9	4
6	ORS505	AGTGATGGCATTCCCAATTT	GTGCGTTGGCTCTTATGGAT	Linkage group5	5
7	ORS665	GCACATGAGGTATGGATCTCCT	TGCAAATACAACTCGGGAAA	Linkage group 3	5
8	ORS899	CGAATACAGACTCGATAAACGACA	GCCACGTATAACTGACTATGACCA	Linkage group 16	5
9	ORS229	GACCCGAATGAGACCCAAACTG	TCCGACCCGAATCTTATGAACC	Linkage group 2	5
10	ORS1265	ACCCTTGGAGTTTAGGGATCA	GGGTTTAGCAAATAATAGGCACA	Linkage group 9	7
11	ORS925	ATGATTCTAAGTTGCGGTAGTGC	GTTGGGTTTAAGTTGTTGCTTCC	Linkage group 2	7
12	ORS894	TGCATTACCTAATTTCTAGTGGGTTT	TTTCCTCATGATCCCGATTCTAT	Linkage group 8	4
13	ORS938	ACCAACTCCCATGCAACCTAA	GCGTTCTCACCGTTCTAACACTT	Linkage group 9	5
14	ORS1161	CAACTACGTCACGATACTCGCC	GGAGCTGAAGCTGAAGACAAATC	Linkage group 8	5
15	ORS691	ACCGTCCTTAGCTCTTGTGAG	GCATCTGAGCAACTGCGTTA	Linkage group 10	3
16	ORS423	AAGCAACCATAATGCATCAGAA	TCATATGGAGGGATCTGTTGG	Linkage group 2	4
17	ORS1114	AGATGGTGGCAGGAGAGTTAAAG	GCAGAAACAGATCAGGAGGGTAT	Linkage group 3	3

编号 No.	标记名 Marker name	上游引物 Forward primer (5'-3')	下游引物 Reverse primer (5'-3')	位置 Position	等位基因数量 Number of alleles
1	ORS543	CCAAGTTTCAGTTACAATCCATGA	GGTCATTAGGAGTTTGGGATCA	Linkage group 1	5
2	ORS456	CCAAGGAATTCTAACAAGAGTTTAAG	GATTTCTCACTTCACTCCTCTATGC	Linkage group 8	3
3	ORS342	CACCAGCATAGCCATTCAAA	TGTTCATCAGGTTTGTCTCCA	Linkage group 2	3
4	ORS656	ACGGACGTAGAGTGGTGGAG	TCGTGGTAAGGGAAGACAACA	Linkage group 16	6
5	ORS844	ACGATGCAAAGAATATACTGCAC	CATGTTTAATAGGTTTTAATTCTAGGG	Linkage group 9	4
6	ORS505	AGTGATGGCATTCCCAATTT	GTGCGTTGGCTCTTATGGAT	Linkage group5	5
7	ORS665	GCACATGAGGTATGGATCTCCT	TGCAAATACAACTCGGGAAA	Linkage group 3	5
8	ORS899	CGAATACAGACTCGATAAACGACA	GCCACGTATAACTGACTATGACCA	Linkage group 16	5
9	ORS229	GACCCGAATGAGACCCAAACTG	TCCGACCCGAATCTTATGAACC	Linkage group 2	5
10	ORS1265	ACCCTTGGAGTTTAGGGATCA	GGGTTTAGCAAATAATAGGCACA	Linkage group 9	7
11	ORS925	ATGATTCTAAGTTGCGGTAGTGC	GTTGGGTTTAAGTTGTTGCTTCC	Linkage group 2	7
12	ORS894	TGCATTACCTAATTTCTAGTGGGTTT	TTTCCTCATGATCCCGATTCTAT	Linkage group 8	4
13	ORS938	ACCAACTCCCATGCAACCTAA	GCGTTCTCACCGTTCTAACACTT	Linkage group 9	5
14	ORS1161	CAACTACGTCACGATACTCGCC	GGAGCTGAAGCTGAAGACAAATC	Linkage group 8	5
15	ORS691	ACCGTCCTTAGCTCTTGTGAG	GCATCTGAGCAACTGCGTTA	Linkage group 10	3
16	ORS423	AAGCAACCATAATGCATCAGAA	TCATATGGAGGGATCTGTTGG	Linkage group 2	4
17	ORS1114	AGATGGTGGCAGGAGAGTTAAAG	GCAGAAACAGATCAGGAGGGTAT	Linkage group 3	3

Establishment of DNA fingerprint for sunflower by SSR-HRM technique

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品种编号 Accession number	17对SSR核心标记 17 pairs of SSR core markers
品种编号 Accession number	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17
1	1	1	1	2	1	1	4	1	1	1	1	1	2	1	1	1	1
2	1	1	0	2	1	1	4	1	1	1	1	1	2	1	1	1	1
3	1	1	1	2	1	2	1	1	1	1	1	1	2	1	1	1	1
4	1	4	1	1	1	5	1	4	1	1	2	2	2	1	1	3	5
5	1	2	4	1	2	3	4	1	1	1	2	1	2	2	1	5	2
6	1	1	2	1	2	1	1	2	1	1	1	2	3	1	2	2	3
7	1	2	2	1	2	3	1	3	1	1	1	6	3	1	2	1	2
8	1	6	1	1	1	7	2	1	1	1	1	1	3	1	2	1	2
9	1	1	1	5	2	1	2	1	2	1	3	2	2	4	4	1	3
10	1	3	1	1	1	1	3	1	1	1	2	1	2	2	4	1	3
11	1	4	2	1	1	1	4	3	1	1	1	2	3	1	5	1	2
12	1	1	1	1	2	8	1	1	2	1	1	1	2	2	2	1	3
13	1	1	1	1	4	3	3	2	2	1	1	1	1	1	1	2	1
14	1	1	1	1	3	4	3	2	1	1	1	1	1	1	2	2	1
15	1	2	2	0	1	2	3	1	1	1	2	1	1	2	3	3	1
16	1	1	1	2	1	1	1	1	1	1	1	1	2	1	1	1	1
17	2	3	2	2	3	6	1	2	3	1	1	3	1	2	2	2	1
18	1	5	1	8	5	6	6	1	1	3	1	5	1	2	3	8	1
19	1	4	2	1	3	1	5	3	2	1	2	8	1	3	4	9	6
20	1	1	1	6	6	0	7	5	2	2	2	4	4	2	1	10	3
21	1	1	1	3	2	3	1	3	2	1	1	1	2	2	1	6	1
22	1	1	2	3	2	1	1	1	1	1	1	1	1	1	6	4	4
23	1	2	1	4	1	1	2	2	2	1	2	1	1	1	5	11	6
24	1	2	2	1	2	1	2	1	2	1	2	1	1	3	5	3	1
25	1	7	1	7	1	3	3	1	4	2	2	3	1	1	2	4	1
26	1	2	1	1	3	2	2	2	1	1	2	1	1	1	5	1	0
27	1	1	1	1	2	4	3	4	3	1	1	3	1	1	2	6	2
28	1	2	2	2	1	2	2	1	2	1	2	2	1	2	4	1	1
29	2	2	3	2	2	1	3	1	3	1	2	2	2	2	3	1	1
30	1	3	2	3	2	1	1	2	2	1	1	1	1	1	2	7	1
31	1	3	3	4	2	1	1	1	2	1	2	7	2	2	3	1	1
32	1	1	3	1	1	2	2	2	1	1	2	1	1	1	3	0	1

[1]	LI Wenlue, LUO Xiahong, LIU Tingting, JIN Guanrong, GE Yaying, CHEN Changli, AN Xia. Determination and comparative analysis of physicochemical properties of different sunflower seeds [J]. Acta Agriculturae Zhejiangensis, 2022, 34(4): 671-677.
[2]	PEI Yun, XU Xiuhong, LU Jinbiao, CHEN Amin, ZHANG Wanping. Genetic diversity analysis of 151 cherry tomato resources in Guizhou Province [J]. Acta Agriculturae Zhejiangensis, 2022, 34(2): 310-316.
[3]	WANG Zhiqi, SUN Jian, LIANG Junchao, ZHAO Yunyan, YAN Tingxian, YAN Xiaowen, WEI Wenliang, LE Meiwang. Study on genetic diversity of sesame germplasm in Jiangxi Province based on molecular markers [J]. Acta Agriculturae Zhejiangensis, 2021, 33(9): 1565-1580.
[4]	MA Jie, QU Wen, CHEN Chunyan, WANG Lei, MA Jun, LIU Zhenshan, MA Wei, ZHOU Ping, HE Yuankuan, SUN Bo. Development of SSR markers based on transcriptome sequencing and genetic diversity analysis of Nainaiqingcai leaf mustard [J]. Acta Agriculturae Zhejiangensis, 2021, 33(9): 1640-1649.
[5]	LIU Shili, BIAN Yuling, JIA Yongyi, CHI Meili, LI Fei, ZHENG Jianbo, CHENG Shun, GU Zhimin. Genetics analysis based on mitochondrial COⅠ sequences in five cultured populations of red-claw crayfish (Cherax quadricarinatus) [J]. Acta Agriculturae Zhejiangensis, 2021, 33(8): 1385-1392.
[6]	ZHANG Jingzhen, WANG Lianjun, LEI Jian, CHAI Shasha, YANG Xinsun, ZHANG Wenying. Genetic diversity analysis and construction of DNA fingerprint of yam (Dioscorea oppositeac Thunb.) germplasm by cpSSR marker [J]. Acta Agriculturae Zhejiangensis, 2021, 33(7): 1222-1233.
[7]	AN Hongwei, SONG Qinfei, NIU Suzhen. Study on genetic diversity,population structure and genetic differentiation of tea germplasm in Guizhou [J]. Acta Agriculturae Zhejiangensis, 2021, 33(7): 1234-1243.
[8]	LIU Xiuhui, ZOU Guihua, ZHAI Guowei, LIU Heqin, ZHENG Xueqiang, CHEN Heyun. Association analysis between SSR markers and germination-related traits in sorghum germplasm resources [J]. Acta Agriculturae Zhejiangensis, 2021, 33(6): 965-973.
[9]	HONG Xia, ZHAO Yongbin, QU Weidong, CHEN Yinlong, QIU Liping, WANG Jiaoyang. Comparative analysis on genetic diversity of Colocasia esculenta germplasm in Zhejiang Province based on phenotype and simple sequence repeats markers [J]. , 2020, 32(9): 1544-1554.
[10]	YAN Fulin, WANG Bo, WEN Di, XU Wenfen, SUN Qingwen, WEI Shenghua. Genetic diversity analysis of Sabia parviflora based on chloroplast gene psbA-trnH sequence [J]. , 2020, 32(5): 810-815.
[11]	GUO Qinwei, ZHANG Ting, LIU Huiqin, ZHANG Xinhui, LI Chaosen, XIANG Xiaomin, ZHAO Dongfeng, WAN Hongjian. Evaluation of genetic diversity in Luffa germplasm resources in China based on ISSR molecular marker [J]. , 2020, 32(4): 616-623.
[12]	ZHAO Xingkai, SHI Haichun, YU Xuejie, YANG Shu, ZHAO Changyun, XIA Wei, KE Yongpei. Breeding potential analyses of 13 new inbred lines in maize [J]. Acta Agriculturae Zhejiangensis, 2020, 32(12): 2119-2127.
[13]	HU Qianwen, XU Yanhao, WANG Rong, ZHANG Wenying, HUA Wei, LYU Chao. Association analysis of four spike traits in barley [J]. , 2020, 32(11): 1941-1953.
[14]	YIN Xiaoxiao, LI Yanfang, GU Jiang, LIAO Yan, XIE Yue, YANG Guangyou, GU Xiaobin. Genetic diversity analysis of Psoroptes ovis var. cuniculi by full-length of mitochondrial ATP6 gene in China [J]. , 2019, 31(8): 1231-1238.
[15]	TANG Shoujie, BI Xiang, ZHANG Feiming, ZHANG Youliang. Genetic diversity analysis and RAPD identification of three successive generations of meio-gynogenetic population in Megalobrama amblycephala [J]. , 2019, 31(8): 1257-1271.