不同来源葡萄种质资源果实叶酸含量及ISSR比较分析

doi:10.6048/j.issn.1001-4330.2023.01.014

新疆农业科学 ›› 2023, Vol. 60 ›› Issue (1): 116-126.DOI: 10.6048/j.issn.1001-4330.2023.01.014

不同来源葡萄种质资源果实叶酸含量及ISSR比较分析

梁雎¹(), 刘国宏¹, 王保民², 何桥³, 阿布来克·尼亚孜¹, 郭红梅¹, 李兴婷¹^,³, 任红松⁴()

1.新疆农业科学院吐鲁番农业科学研究所,新疆吐鲁番 838000
2.中国农业大学,北京 100083
3.西南大学,重庆 400715
4.新疆农业科学院综合试验场,乌鲁木齐,830000

收稿日期:2022-05-19 出版日期:2023-01-20 发布日期:2023-03-07
通信作者: 任红松(1977-),男,新疆尉犁人,研究员,博士,研究方向为果树分子辅助育种,(E-mail)1134037950@qq.com
作者简介:梁雎(1978-),男,新疆吐鲁番人,正高级农艺师,硕士,研究方向为葡萄种质资源、遗传育种及栽培,(E-mail)147302120@qq.com
基金资助:
2021年天山创新团队计划“新疆葡萄种质资源创新利用及栽培技术研究创新团队”(2020D14033);2021年天山青年计划-优秀青年科技人才培养项目“高含量叶酸葡萄品种资源评价和利用”(2020Q023);自治区公益性科研院所基本科研业务经费“葡萄叶酸合成关键基因克隆与分析”(KY2020131)

Comparative Analysis of Fruit Folic Acid Content and ISSR in Different Grape Germplasm Resources

LIANG Ju¹(), LIU Guohong¹, WANG Baomin², HE Qiao³, Abrake Niazi¹, GUO Hongmei¹, LI Xingting¹^,³, REN Hongsong⁴()

1. Turpan Institute of Agricultural Sciences, Xinjiang Academy of Agricultural Sciences, Turpan Xinjiang 838000, China
2. China Agricultural University, Beijing 100083, China
3. Southwest University, Chongqing 400715, China
4. Comprehensive Test Field of Xinjiang Academy of Agricultural Sciences,Urumqi 830000, China

Received:2022-05-19 Online:2023-01-20 Published:2023-03-07
Correspondence author: REN Hongsong(1977-), male, from Yuli County, Xinjiang, researcher, doctor, his research direction is molecular assisted breeding of fruit trees,(E-mail)1134037950@qq.com
Supported by:
Tianshan Innovation Team Project in 2021 "Xinjiang Grape Germplasm Resources Innovation and utilization and Cultivation Technology Research Innovation Team"(2020D14033);2021 Tianshan Youth Plan - Excellent Young Scientific and Technological Talents Training Project "Evaluation And Utilization of High Folic Acid Grape Variety Resources(2020Q023);2021 Basic Scientific R & D Program of Public Welfare Research Institutions of Xinjiang Uygur Autonomous Region of China(KY2020131)

摘要/Abstract

摘要：

【目的】比较不同来源葡萄果实中叶酸积累的差异,并利用分子标记技术对高叶酸种质资源进行评价和筛选。【方法】采用ELISA技术检测22份葡萄种质资源果实叶酸含量;利用ISSR分子标记对22份葡萄种质资源进行基因组多态性分析和聚类分析;分析检测出叶酸含量与聚类结果。【结果】22份葡萄种质资源果实叶酸含量的范围在1.93~46.95 ng/g,在遗传相似系数为0.621处,22份葡萄材料明显分为2大类群。第1大类叶酸含量平均为35.79 ng/g;其余为第2大类,叶酸含量平均为15.71 ng/g。在遗传相似系数为0.622处,21份葡萄材料又明显分为2大类群。第1大类叶酸含量平均为16.53 ng/g;第2大类叶酸含量平均为12.24 ng/g。欧亚种的叶酸含量较欧美杂种稍高3.998 7 ng/g;从30条引物中筛选出9条扩增稳定且多态性丰富的引物用于葡萄的ISSR扩增。共扩增出78条条带,其中多态性条带70条,多态性百分率为 89.7%。22 份葡萄种质资源的遗传相似系数为0.474 4~0.846 2,平均遗传相似系数为0.654 5。在遗传相似系数为0.621处,22份葡萄材料明显分为2大类群。第1大类包含1个欧亚种(红巴拉多),其余为第2大类。在遗传相似系数为0.622处,21份葡萄材料又明显分为2大类群。第1大类包含11个欧亚种(炎兴无核、灰比诺、瑞都翠霞、京蜜、黑奇无核、无核白、1124、京翠、红宝石、无核紫、达米娜),6个欧美杂种(杂1、夏黑、蜜红、红标无核、洛浦早生、宇选1号)。第2大类包含4个欧亚种(美人指、粉红亚都蜜、木纳格、里扎马特)。【结论】群体1的叶酸含量比群体2的叶酸含量高,欧亚种的叶酸含量较欧美杂种稍高。

关键词: 不同来源; 葡萄种质资源果实; 叶酸含量; 检测; ISSR

Abstract:

【Objective】 To compare the differences of folic acid accumulation in grape fruits from different sources and to evaluate and screen high folic acid germplasm resources by molecular marker technology. 【Methods】 The folic acid content of 22 grape germplasm resources was detected by ELISA; The genomic polymorphism and cluster analysis of 22 grape germplasm resources were carried out by ISSR molecular markers; The detected folic acid content was analyzed with the clustering results. 【Results】 The fruit folic acid content of 22 grape germplasm resources ranged from 1.93 to 46.95 ng/g.Compared with the clustering results, the 22 grape germplasm resources were obviously divided into two groups at the genetic similarity coefficient of 0.621.The average folic acid content of the first category was 35.79 ng/g; The rest belonged to the second category, with an average folate content of 15.71 ng/g.At the genetic similarity coefficient of 0.622, the 21 grape materials were obviously divided into 2 groups.The average folic acid content of the first category was 16.53 ng/g and the average folic acid content of the second category was 12.24 ng/g.At the same time, the analysis of variance showed that the folic acid content of Eurasian species was slightly higher than that of European and American hybrids by 3.998,7 ng/g; Through ISSR molecular marker technology, 9 primers with stable amplification and abundant polymorphism were selected from 30 primers for ISSR amplification of grape.A total of 78 bands were amplified, including 70 polymorphic bands, with a polymorphic percentage of 89.7%.According to the ISSR amplification results, the Jaccard similarity coefficient was analyzed by NTSYSpc2.10e software, and the cluster analysis was carried out by unweighted paired arithmetic mean (UPGMA) to establish the genetic relationship map.The genetic similarity coefficient of 22 grape germplasm resources was between 0.474,4-0.846,2, and the average genetic similarity coefficient was 0.654,5.At the genetic similarity coefficient of 0.621, 22 grape materials were obviously divided into 2 groups.The first category included one Eurasian species (red balado), and the rest was the second category.At the genetic similarity coefficient of 0.622, the 21 grape materials were obviously divided into 2 groups.The first category included 11 Eurasian species (Yanxing seedless, Pinot Grigio, Ruidu Cuixia, Jingmi, Heiqi Seedless, Seedless White, 1124, Jingcui, Ruby, Seedless Purple, Damina), and 6 European and American hybrids (ZA 1, Xiahei, Mihong, Hongbiao Seedless, Luopu Zaosheng and Yuxuan 1).The second category included 4 Eurasian species (Beauty finger, Pink Yadumi, Munag and Rizamat). 【Conclusion】 Through comparing and analyzing the fruit folic acid content and clustering results of different grape germplasm resources, it can be seen that the folic acid content of population 1 is higher than that of population 2.The folic acid content of Eurasian species was slightly higher than that of European and American hybrids.

Key words: different sources; grape germplasm resources fruit; folic acid content; detection; ISSR

中图分类号:

S663.1

梁雎, 刘国宏, 王保民, 何桥, 阿布来克·尼亚孜, 郭红梅, 李兴婷, 任红松. 不同来源葡萄种质资源果实叶酸含量及ISSR比较分析[J]. 新疆农业科学, 2023, 60(1): 116-126.

LIANG Ju, LIU Guohong, WANG Baomin, HE Qiao, Abrake Niazi, GUO Hongmei, LI Xingting, REN Hongsong. Comparative Analysis of Fruit Folic Acid Content and ISSR in Different Grape Germplasm Resources[J]. Xinjiang Agricultural Sciences, 2023, 60(1): 116-126.

图/表 6

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品种名称 Variety name	来源 Source	父母本 Male female parent	产地 Place of origin
炎兴无核Yanxing seedness	欧亚种	不详	中国
灰比诺Pinot Grigio	欧亚种	Pinot Noir芽变	法国
瑞都翠霞Ruiducuixia	欧亚种	京秀×香妃	中国
京蜜Jingmi	欧亚种	京秀×香妃	中国
黑奇无核Heiqiseedness	欧亚种	不详	美国
无核白Thompson seedless	欧亚种	不详	小亚细亚
1124	欧亚种	葡萄园皇后×康耐诺	中国
京翠Jingcui	欧亚种	京秀×香妃	中国
红宝石Rubyseedness	欧亚种	皇帝× Pirovano75	美国
无核紫Blackmonukka	欧亚种	不详	印度
达米娜Damina	欧亚种	比坎× 玫瑰香	罗马尼亚
美人指Manicure finger	欧亚种	优尼坤 × 巴拉蒂	日本
粉红亚都蜜Yatomi Rosa	欧亚种	ペソノニアキェ ×(ヮーベローザ×ローザワスット)	日本
木纳格Munage	欧亚种	不详	中国
里扎马特Lizhamate	欧亚种	可口甘 × 帕尔肯特	前苏联
红巴拉多Hongbaladuo	欧亚种	巴拉多×京秀杂	日本
杂1 Za1	欧美杂种	红地球×克瑞森无核	中国
夏黑Summer black	欧美杂种	巨峰 × 无核白	日本
蜜红Mihong	欧美杂种	井川682芽变	日本
红标无核Hongbiaoseedness	欧美杂种	郑州早红×巨峰	中国
洛浦早生Luopuzaosheng	欧美杂种	京亚芽变	中国
宇选1号Yuxuan1hao	欧美杂种	巨峰芽变选育	中国

品种名称 Variety name	来源 Source	父母本 Male female parent	产地 Place of origin
炎兴无核Yanxing seedness	欧亚种	不详	中国
灰比诺Pinot Grigio	欧亚种	Pinot Noir芽变	法国
瑞都翠霞Ruiducuixia	欧亚种	京秀×香妃	中国
京蜜Jingmi	欧亚种	京秀×香妃	中国
黑奇无核Heiqiseedness	欧亚种	不详	美国
无核白Thompson seedless	欧亚种	不详	小亚细亚
1124	欧亚种	葡萄园皇后×康耐诺	中国
京翠Jingcui	欧亚种	京秀×香妃	中国
红宝石Rubyseedness	欧亚种	皇帝× Pirovano75	美国
无核紫Blackmonukka	欧亚种	不详	印度
达米娜Damina	欧亚种	比坎× 玫瑰香	罗马尼亚
美人指Manicure finger	欧亚种	优尼坤 × 巴拉蒂	日本
粉红亚都蜜Yatomi Rosa	欧亚种	ペソノニアキェ ×(ヮーベローザ×ローザワスット)	日本
木纳格Munage	欧亚种	不详	中国
里扎马特Lizhamate	欧亚种	可口甘 × 帕尔肯特	前苏联
红巴拉多Hongbaladuo	欧亚种	巴拉多×京秀杂	日本
杂1 Za1	欧美杂种	红地球×克瑞森无核	中国
夏黑Summer black	欧美杂种	巨峰 × 无核白	日本
蜜红Mihong	欧美杂种	井川682芽变	日本
红标无核Hongbiaoseedness	欧美杂种	郑州早红×巨峰	中国
洛浦早生Luopuzaosheng	欧美杂种	京亚芽变	中国
宇选1号Yuxuan1hao	欧美杂种	巨峰芽变选育	中国

品种名称 Variety name	叶酸含量均值 Folic acid average content (ng/g)	种 Species
黑奇无核Heiqiseedness	46.95^aA	欧亚种
红巴拉多Hongbaladuo	35.96^abAB	欧亚种
炎兴无核Yanxing seedness	35.69^abAB	欧亚种
红宝石Rubyseedness	27.06^bcBC	欧亚种
夏黑Summer black	21.56^bcdBCD	欧美杂种
达米拉Damina	19.92^cdeBCD	欧亚种
本纳格Munage	17.83^cdefBCD	欧亚种
蜜红Mihong	18.55^cdefBCD	欧美杂种
无核紫Blackmonukka	17.37^cdefBCD	欧亚种
瑞都翠霞Ruiducuixia	16.68^cdefBCD	欧亚种
宇选一号Yuxuan1hao	14.27^cdefCD	欧美杂种
京密Jingmi	13.5^cdefCD	欧亚种
里扎马特Lizhamate	12.76^cdefCD	欧亚种
洛浦早生Luopuzaosheng	11.40^cdefCD	欧美杂种
粉红亚都蜜Yatomi Rosa	10.11^defCD	欧亚种
杂1 Za1	8.79^defCD	欧美杂种
美人指Manicure	8.27^defCD	欧亚种
无核白Thompson seedless	7.98^defCD	欧亚种
红标无核Hongbiaoseedness	7.65^defCD	欧美杂种
1124	7.16^defCD	欧亚种
京翠Jingcui	5.03^efD	欧亚种
灰比诺Pinot Grigio	1.93^fD	欧亚种

品种名称 Variety name	叶酸含量均值 Folic acid average content (ng/g)	种 Species
黑奇无核Heiqiseedness	46.95^aA	欧亚种
红巴拉多Hongbaladuo	35.96^abAB	欧亚种
炎兴无核Yanxing seedness	35.69^abAB	欧亚种
红宝石Rubyseedness	27.06^bcBC	欧亚种
夏黑Summer black	21.56^bcdBCD	欧美杂种
达米拉Damina	19.92^cdeBCD	欧亚种
本纳格Munage	17.83^cdefBCD	欧亚种
蜜红Mihong	18.55^cdefBCD	欧美杂种
无核紫Blackmonukka	17.37^cdefBCD	欧亚种
瑞都翠霞Ruiducuixia	16.68^cdefBCD	欧亚种
宇选一号Yuxuan1hao	14.27^cdefCD	欧美杂种
京密Jingmi	13.5^cdefCD	欧亚种
里扎马特Lizhamate	12.76^cdefCD	欧亚种
洛浦早生Luopuzaosheng	11.40^cdefCD	欧美杂种
粉红亚都蜜Yatomi Rosa	10.11^defCD	欧亚种
杂1 Za1	8.79^defCD	欧美杂种
美人指Manicure	8.27^defCD	欧亚种
无核白Thompson seedless	7.98^defCD	欧亚种
红标无核Hongbiaoseedness	7.65^defCD	欧美杂种
1124	7.16^defCD	欧亚种
京翠Jingcui	5.03^efD	欧亚种
灰比诺Pinot Grigio	1.93^fD	欧亚种

引物编号 Primr numbr	引物碱基序列 Primer base sequence	扩增总带数 Total number of amplified bands	多态性带数 Number of polymor phic bands	多态位点百分率 Polymor phic locus percenta (%)
ubc811	GAGAGAGAGAGAGAGAC	4	1	25
ubc815	CTCTCTCTCTCTCTCTG	9	9	100
ubc825	ACACACACACACACACT	10	9	90
ubc834	AGAGAGAGAGAGAGAGYT	10	10	100
ubc842	GAGAGAGAGAGAGAGAYG	10	9	90
ubc855	ACACACACACACACACYT	7	6	85
ubc856	ACACACACACACACACYA	4	3	75
ubc881	GGGTGGGGTGGGGTG	13	13	100
ubc807	AGAGAGAGAGAGAGAGT	11	10	90
总计Totel		78	70	-
平均Average		8.6	7.7	90

不同来源葡萄种质资源果实叶酸含量及ISSR比较分析

Comparative Analysis of Fruit Folic Acid Content and ISSR in Different Grape Germplasm Resources

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