《植物生理学报》 2018, 54(2): 265-272

利用分子标记对‘淮稻5号’进行品种真实性鉴定

顾伟航¹, 田佳琪¹, 朱明超^2,*, 文正怀², 严卫古², 王兴龙³, 张大兵^1,3, 袁政^1,3,*

¹上海交通大学生命科学技术学院, 上海200240; ² 江苏徐淮地区淮阴农业科学研究所, 淮安市农业科学研究院, 江苏淮安223001; ³淮安市作物分子辅助育种重点实验室, 江苏省区域现代农业与环境保护协同创新中心, 江苏天丰种业有限公司, 江苏淮安223001

通信作者：朱明超；E-mail: 574937902@qq.com; zyuan@sjtu.edu.cn

摘　要：

‘淮稻5号’具有抗倒性强、高产稳产、米质优良等优点, 自推广以来, 以其独有的优越性在粳稻市场中占有重要竞争份额。因此, 开发可用于该品种真实性鉴定的分子标记具有重要的经济和应用价值。本研究利用前期筛选获得的99对水稻InDel分子标记对‘淮稻5号’标准品(H1)和4个生产中与‘淮稻5号’无明显表型差异的供测品种进行基因型分析, 筛选获得4对‘淮稻5号’品种特异性InDel分子标记: chr1_3386-1、chr5_751、chr8_6491-1和 chr11_5641-0。为进一步验证这4对分子标记鉴定结果的可靠性, 对供测品种基因组进行了2b-RAD测序分析, 测序结果与InDel分子标记检测结果一致, 供测品种H2、H4虽然表型与其他供测品种几乎一致, 但与标准品(H1)及其他供测品种(H3、H5)在基因型上存在明显差异。在此基础上, 利用这4对InDel分子标记对来自不同地区以及举报的‘淮稻5号’问题材料进行了基因型检测, 结果进一步验证本研究获得的4对分子标记可用于 ‘淮稻5号’品种真实性的快速检测鉴定。因此, 本研究结果为加强‘淮稻5号’品种权的保护及种子质量监测工作提供了理论依据和技术支持。

关键词：‘淮稻5号’; 分子标记; InDel; 2b-RAD; 真实性

收稿：2017-11-20   修定：2017-12-29

资助：国家重点研发计划(2016YFD0101107)和上海市水稻产业技术体系建设项目(沪农科产字(2017)第3号)。

Development and application of variety-specific molecular markers in authenticity identification of japonica rice ‘Huaidao 5’

GU Wei-Hang¹, TIAN Jia-Qi¹, ZHU Ming-Chao^2,*, WEN Zheng-Huai², YAN Wei-Gu², WANG Xing-Long³, ZHANG Da-Bing^1,3, YUAN Zheng^1,3,*

¹School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China; ²Huaiyin Institute of Agricultural Science of Xuhuai Region in Jiangsu, Huaian Academy of Agricultural Sciences, Huaian, Jiangsu 223001, China; ³Key Laboratory of Crop Marker-Assisted Breeding of Huaian Municipality, Jiangsu Collaborative Innovation Center of Regional Modern Agriculture and Environmental Protection, Jiangsu Tianfeng Seed Company, Huaian, Jiangsu 223001, China

Corresponding author: ZHU Ming-Chao; E-mail: 574937902@qq.com; zyuan@sjtu.edu.cn

Abstract:

‘Huaidao 5’ is one of the best-welcomed rice varieties with advantage of strong lodging resistance, high and stable yield, and high grain quality etc. In Jiangsu province, the planting area of ‘Huaidao 5’ is always higher than other varieties since it had been authorized by the local government. Therefore, it is important to develop and apply variety-specific molecular markers in authenticity identification of ‘Huaidao 5’ due to economic and intellectual property value. In this study, we screened and obtained 4 pairs of InDel markers, chr1_3386-1, chr5_751, chr8_6491-1 and chr11_5641-0 from our 99 pairs InDel markers pool to perform genotyping analysis on standard ‘Huaidao 5’ (H1) and other four test samples (H2–H5), whose growth phenotypes seem identical to H1 under natural planting condition. To further validate our InDel genotyping results, we employed 2b-RAD sequencing technique to detect the genomic-level DNA polymorphisms in H1–H5 samples. It’s not surprising to find that consistent to the InDel markers analysis data, the H2 and H4 varieties have different DNA polymorphisms compared to that of H1, H3 and H4 varieties although they have similar growth phenotypes. Furthermore, we applied these four InDel markers to perform blind samples detection, including different ‘Huaidao 5’ samples collected from different planting areas of Jiangsu province and wild type varieties such as ‘Wuyungene 7’ etc. And the results further confirmed that the four InDel markers we developed in this study could be used for authenticity identification of the ‘Huaidao 5’. Therefore, our results provide theoretical evidences and technical supports for seed quality monitoring and variety rights protection of the ‘Huaidao 5’.

Key words: ‘Huaidao 5’; molecular markers; InDel; 2b-RAD; authenticity