利用SRAP标记快速检测西瓜杂交种子纯度

doi:10.6048/j.issn.1001-4330.2017.09.007

新疆农业科学 ›› 2017, Vol. 54 ›› Issue (9): 1621-1626.DOI: 10.6048/j.issn.1001-4330.2017.09.007

利用SRAP标记快速检测西瓜杂交种子纯度

韩宏伟, 王浩, 佘建华, 王强, 庄红梅, 刘会芳, 李宁

新疆农业科学院园艺作物研究所,乌鲁木齐 830011

收稿日期:2017-07-05 发布日期:2020-07-22
通信作者: 王浩(1970-),男,山东人,研究员,研究方向为蔬菜栽培与生理,(E-mail)Wanghao183@163.com
作者简介:韩宏伟(1986-),男,河南人,助理研究员,研究方向为特色瓜菜种质资源创新与新品种选育,(E-mail)hhwei2010@sohu.com
基金资助:
新疆维吾尔自治区公益性科研院所基本科研业务费专项“蔬菜品种收集引进及筛选利用研究”(KYGY2016119);新疆农业科学院青年基金项目“基于SRAP标记的西瓜杂交种早佳(84-24)纯度的快速检测体系构建”(xjnkq-2015008)

Using SRAP Markers for Rapid Detection of Zaojia(84-24) Watermelon Hybrid Seed Purity

HAN Hong-wei, WANG Hao, SHE Jian-hua, WANG Qiang, ZHUANG Hong-mei, LIU Hui-fang, LI Ning

Research Institute of Horticultural Crops, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China

Received:2017-07-05 Published:2020-07-22
Correspondence author: WANG Hao(1970-), Professor,Bachelor of Agriculture, Vegetable cultivation and physiology. (E-mail) Wanghao183@163.com
Supported by:
The Basic Research Funding for the Public Welfare Research Institutes of Xinjiang Uygur Autonomous Region "Study on Collection, Introduction and Selection of Vegetable Germplasm" (KYGY2016119); The Youth Fund Project of Xinjiang Academy of Agricultural Sciences "The Construction of Rapid Detection System to Detect the Purity of Zaojia 84-24 Watermelon Seed by SRAP Markers (xjnkq-2015008)

摘要/Abstract

摘要： 【目的】以设施、露地兼用西瓜杂交品种早佳(84-24)及其父本、母本为试验材料,利用SRAP分子标记技术进行DNA指纹图谱构建,筛选出能够快速检测西瓜早佳(84-24)杂交种子纯度的SRAP标记。【方法】选取28对SRAP引物,对供试西瓜品种早佳(84-24)及其父、母本的基因组DNA进行扩增,筛选出扩增多态性好、条带清晰、稳定的SRAP引物作为候选标记,利用100粒杂交种子进行纯度检测准确性验证。【结果】28对供试引物中有22对引物在F₁及其父、母本之间扩增出多态性条带,多态性比率为78.6%,其中有1对为父本显示,母本缺失的显性SRAP引物(Me6F/ Em2R)。用显性引物(Me6F/ Em2R)对100粒西瓜杂交种子进行纯度检测,经聚丙烯酰胺凝胶电泳分析,纯度为99.0%,与田间形态学鉴定结果(99.1%)基本一致。【结论】利用SRAP标记(Me6F/ Em2R),结合聚丙烯酰胺凝胶电泳检测,可以对西瓜杂交种早佳(84-24)的纯度进行快速、准确鉴定。SRAP分子标记技术在西瓜杂交种子纯度室内快速检测中有很大的应用前景。

关键词: 西瓜; SRAP标记; 杂交种子纯度; 快速检测

Abstract: 【Objective】 To screen out one SRAP marker to detect seed purity,the watermelon hybrids of Zaojia (84-24) and its paternal and female lines were used to study the polymorphorrism of amplifiable spectral bands.【Method】Polymorphic primer pairs were selected from 28 SRAP primer pairs for Zaojia (84-24) hybrids and their parents in this experiment. The SRAP primers with good polymorphism, clear bands and stable primers were selected as candidate markers, and the purity of the 100 hybrid seeds was tested and verified.【Result】The results showed that 28 pairs of primers had 22 pairs of primers, and polymorphic bands were amplified between F₁ and its parents. The ratio of polymorphism was 78.6%, among which, there was only one pair of complementary type SRAP primer(Me6F/ Em2R)with male display and lack of female parent, The result showed the purity of hybrid seeds was 99.0%, which was basically the same as the field morphological identification (99.1%).【Conclusion】SRAP primer (Me6F/Em2R), in combination with Polyacrylamide gel electrophoresis can be used to detect the purity of Zaojia (84-24) hybrid seed rapidly and accurately. There is a great prospect of application in using this method.

Key words: watermelon; SRAP molecular marker; hybrid seed purity; rapid detection

中图分类号:

S651

韩宏伟, 王浩, 佘建华, 王强, 庄红梅, 刘会芳, 李宁. 利用SRAP标记快速检测西瓜杂交种子纯度[J]. 新疆农业科学, 2017, 54(9): 1621-1626.

HAN Hong-wei, WANG Hao, SHE Jian-hua, WANG Qiang, ZHUANG Hong-mei, LIU Hui-fang, LI Ning. Using SRAP Markers for Rapid Detection of Zaojia(84-24) Watermelon Hybrid Seed Purity[J]. Xinjiang Agricultural Sciences, 2017, 54(9): 1621-1626.

参考文献

[1] 文方德,李卓杰,傅家瑞.种子纯度鉴定进展及评论[J].种子,1995,(5),36-38.
   WEN Fang-de, LI Zuo-jie, FU Jia-rui. (1995). The Advances and Comments of Hybrid purity test [J]. Seed, (5):36-38.
[2] 李晓慧,王从彦,常高正.分子标记技术在西瓜遗传育种上的应用 [J].分子植物育种,2008,2(6):329-334.
   LI Xiao-hui, WANG Cong-yan, CHANG Gao-zheng. (2008). Applications of Molecular Marker on Watermelon Genetics and Breeding Programs [J]. Molecular Plant Breeding, 2(6):329-334.
[3] Li, G., & Quiros, C. F. (2001). Sequence-related amplified polymorphism (srap), a new marker system based on a simple pcr reaction: its application to mapping and gene tagging in brassica. Theoretical & Applied Genetics, 103(2-3): 455-461.
[4] Li, G., Gao, M., Yang, B., & Quiros, C. F. (2003). Gene for gene alignment between the brassica and arabidopsis genomes by direct transcriptome mapping. Theoretical & Applied Genetics, 107(1): 168-180.
[5] Ferriol, M., Picó, B., & Nuez, F. (2003). Genetic diversity of a germplasm collection of cucurbita pepo using srap and aflp markers. Theoretical & Applied Genetics, 107(2): 271-282.
[6] Lin, Z., Zhang, X., Nie, Y., He, D., & Wu, M. (2003). Construction of a genetic linkage map for cotton based on srap. Chinese Science Bulletin, 48(19): 2,064-2,068.
[7] Ferriol, M., Picó, B., & Nuez, F. (2003). Genetic diversity of a germplasm collection of cucurbita pepo using srap and aflp markers. Theoretical & Applied Genetics, 107(2):271-282.
[8]刘泽发,孙小武,董亚静. SRAP标记鉴定西瓜种子纯度方法研究[J].中国瓜菜,2009,(1):5-8.
   LIU Ze-fa, SUN Xiao-wu, DONG Ya-jing. (2009). Testing of watermelon hybrid purity using sequence-related amplified polymorphism [J]. China Cucurbits and Vegetables, (1):5-8. (in Chinese)
[9] 刘成平.瓤瓜EST-SSR、SRAP、ISSR和RAPD反应体系的建立及其纯度鉴定应用的研究[D].武汉:华中农业大学硕士学位论文,2008.
   LIU Cheng-ping. (2008). Establishment of EST-SSR, SRAP, ISSR and RAPD Reaction System and Studies on the Application of purity Identification in bottle Gourd[D].Master Thesis. Huazhong Agricultural University, Wuhan. (in Chinese)
[10] 张建成,王传堂,焦坤,等.SRAP标记技术在花生种子纯度鉴定中的应用[J].中国农学通报,2005, 21(12):35-39.
   ZHAN Jian-cheng, WANG Chuan-tang, JIAO Kun, et al. (2005). Study on the Seed Test Using Sequence-related Amplification Polymorphisms (SRAP) in Cultivated Peanut (Arachis hypogaea L.) [J]. Chinese Agricultural Science Bulletin, 21(12):35-39. (in Chinese)
[11]郭庆华,郭美丽.与红花苞叶刺性状紧密连锁的 SRAP 分子标记[J].药学学报,2007,42(7): 794-797.
   GUO Qing-hua, GUO Mei-li. (2007). SRAP fragments linked to spines ofCarthamus tinctorius[J]. Acta Pharmaceutica Sinica, 42(7): 794-797. (in Chinese)
[12] Murray, M. G., & Thompson, W. F. (1980). Rapid isolation of high weight plant dna. Nucleic Acids Research, 8(19):4,321-4,325.
[13]姜陆,卢浩,李冠,等.快速检测甜瓜西州蜜17号种子纯度的 SSR标记[J].新疆农业科学,2016,53(3):407-411.
   JIANG Lu, LU Hao, LI Guan, et al. (2016). SSR Marker for Rapid Detection of Xizhoumi 17 Muskmelon's Seed Purity [J]. Xinjiang Agricultural Sciences, 53(3): 407-411. (in Chinese)

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利用SRAP标记快速检测西瓜杂交种子纯度

Using SRAP Markers for Rapid Detection of Zaojia(84-24) Watermelon Hybrid Seed Purity

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