应用分子标记技术改进作物品种保护和监管

doi:10.3724/SP.J.1006.2022.23001

作物学报 ›› 2022, Vol. 48 ›› Issue (8): 1853-1870.doi: 10.3724/SP.J.1006.2022.23001

• 综述 • 下一篇

应用分子标记技术改进作物品种保护和监管

徐云碧¹^,⁶^,^*(), 王冰冰², 张健³, 张嘉楠⁴, 李建生⁵^,^*()

¹中国农业科学院作物科学研究所, 中国北京 100081
²长沙百奥云数据科技有限公司, 中国湖南长沙 410221
³先正达集团中国, 中国北京102206
⁴石家庄博瑞迪生物技术有限公司, 中国河北石家庄 050035
⁵中国农业大学国家玉米改良中心, 中国北京100193
⁶国际玉米小麦改良中心, 墨西哥特斯科科 56130

收稿日期:2022-01-05 接受日期:2022-02-22 出版日期:2022-08-12 网络出版日期:2022-03-01
通讯作者: 徐云碧,李建生
基金资助:
国家重点研发计划项目(2016YFD0101803);石家庄市科技孵化计划项目(191540089A);河北省创新能力提升计划项目新型研发机构建设专项(19962911D);中央级公益性科研院所基本科研业务费专项(Y2020PT20);中国农业科学院农业科技创新计划(CAAS-XTCX2016009);中国农业科学院作物科学研究所中央非公益类基础研究项目, 比尔盖茨基金会和CGIAR MAIZE项目资助

Enhancement of plant variety protection and regulation using molecular marker technology

XU Yunbi¹^,⁶^,^*(), WANG Bing-Bing², ZAHNG Jian³, ZHANG Jia-Nan⁴, LI Jian-Sheng⁵^,^*()

¹Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
²Biobin Data Sciences, Changsha 410221, Hunan, China
³Syngenta Group China, Beijing 102206, China
⁴MolBreeding Biotechnology Co., Ltd., Shijiazhuang 050035, Hebei, China
⁵National Maize Improvement Center of China, China Agricultural University, Beijing 100193, China
⁶International Maize and Wheat Improvement Center (CIMMYT), El Batan Texcoco 56130, Mexico

Received:2022-01-05 Accepted:2022-02-22 Published:2022-08-12 Published online:2022-03-01
Contact: XU Yunbi,LI Jian-Sheng
Supported by:
National Key Research and Development Program of China(2016YFD0101803);Shijiazhuang Science and Technology Incubation Program(191540089A);Hebei Innovation Capability Enhancement Project(19962911D);Central Public-interest Scientific Institution Basal Research Fund(Y2020PT20);Agricultural Science and Technology Innovation Program (ASTIP) of the Chinese Academy of Agricultural Sciences (CAAS)(CAAS-XTCX2016009);Central Non-public-interest Basic Research Fund of the Institute of Crop Science of CAAS, Bill and Melinda Gates Foundation, and CGIAR Research Program MAIZE

摘要/Abstract

摘要：

植物品种保护是植物品种知识产权的重要组成部分。品种特异性、一致性和稳定性(distinctness, uniformity, and stability, DUS)检测和实质性派生品种(essentially derived variety, EDV)评价是植物品种保护中的2个重要概念。DUS-EDV评价经历了从形态性状和系谱为主过渡到了综合利用形态、系谱和分子标记信息的阶段, 并将发展到以分子检测为主的阶段。分子标记的主要类型也从RFLP发展到SSR和SNP。基于靶向测序-液相捕获的芯片技术, 具有分析成本低、标记组配灵活、适合用于不同植物的DUS-EDV评价。利用分子标记检测进行DUS-EDV评价有2个重要策略, 一是在全基因组范围内进行全局性的比较和分析, 二是利用与重要表型有关的功能位点特异性进行局部性、特异性检测。应该针对DUS和EDV分别建立各自的评价标准。DUS可以根据分子标记提供的特殊指纹图谱、单倍型、特有等位基因、特异基因组区段、特异功能标记、最低遗传纯合度、品种内单株间的遗传差异作出判断。EDV的主要评价指标是利用全基因组均匀分布的高密度标记所获得的材料间的遗传相似性, 而不是差异的标记数。所采用的分子标记的数量和基因组覆盖率是决定分子检测效率和可靠性的关键。利用少量标记所获得的品种比较具有较大的抽样误差。区别EDV和非EDV品种的相似性指标可因作物物种和所需要的品种保护水平而异。本文回顾了世界各国主要的作物品种保护实践, 建议在利用分子标记保护品种知识产权和监管品种中, 建立由各方面专家组成的咨询委员会, 根据分子检测技术的发展和功能标记的开发, 不断完善和加强DUS-EDV评价体系, 建立权威的品种保护数据库系统, 有序提供查询和比对服务, 以鼓励种业创新。

关键词: 植物品种保护, 特异性-一致性-稳定性(DUS), 实质性派生品种(EDV), 分子标记, 分子检测, 遗传相似性

Abstract:

Plant variety protection is one of the important approaches for plant intellectual property protection. The distinctness, uniformity and stability (DUS) and essentially derived variety (EDV) are two major concepts in plant variety protection. DUS-EDV has been evaluated largely through morphological traits and pedigrees at the very beginning, to an integrated approach using morphological traits, pedigrees and molecular marker information and now to a stage largely driven by molecular diagnostics. Molecular diagnostic technology has been evolved from RFLP to SSR and SNP marker systems. The liquid SNP chip, represented by genotyping by target sequencing through capture in solution, has advantages of low cost, high flexibility in marker combinations and wide suitability for DUS-EDV evaluation across plant species. There are two important strategies in DUS-EDV evaluation, one being examined based on the analysis and comparison at the whole genome level and the other being examined at specific genomic regions for target functional loci associated with important phenotypes. Evaluation criteria should be established separately for DUS and EDV. The former can be evaluated based on the criteria constructed for specific fingerprint maps, haplotypes, unique alleles, genomic regions, target functional markers, minimum genetic homozygosity, and within-variety variation, whereas the latter can be examined by the genetic similarity between the potential EDV and check variety estimated using a large number of molecular markers evenly distributed across the genome, rather than by the number of markers. The number and the genomic coverage of molecular markers are two key factors affecting the efficiency and reliability in DUS and EDV assessment. Using only a small number of markers in such assessment will likely result in a large sampling error for the estimates. The threshold of genetic similarity required for distinguishing EDV and non-EDV can vary greatly across plant species and with the levels of plant variety protection. After reviewed the current status of plant variety protection across countries, the authors proposed that a national consultant expert committee should be established for consistent support to implement and improve DUS-EDV system, and an official database system should be constructed for public service and comparison of variety DNA fingerprint data to facilitate innovative activities in plant breeding.

Key words: plant variety protection, distinctness-uniformity-stability (DUS), essentially derived variety (EDV), molecular marker, molecular diagnostics, genetic similarity

徐云碧, 王冰冰, 张健, 张嘉楠, 李建生. 应用分子标记技术改进作物品种保护和监管[J]. 作物学报, 2022, 48(8): 1853-1870.

XU Yunbi, WANG Bing-Bing, ZAHNG Jian, ZHANG Jia-Nan, LI Jian-Sheng. Enhancement of plant variety protection and regulation using molecular marker technology[J]. Acta Agronomica Sinica, 2022, 48(8): 1853-1870.

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世代 Generation (years)	支撑技术 Support technique	代表分子标记 Representative molecular markers	检测方式 Diagnostics	特征 Characteristics
G1 (1980s)	凝胶电泳 Gel electrophoresis	RFLP, RAPD	琼脂糖和聚丙烯酰胺凝胶电泳, EB和银染 Agarose and polyacrylamide gel electrophoresis with EB and silver staining	手动, 实验室, 极高成本, 不灵活 Manual, exp. laboratory, very high cost, not flexible, 100 DP/D
G2 (1990s)	荧光检测 Florescence electrophoresis	SSR, AFLP, KASP, SNP	聚丙烯酰胺凝胶电泳加荧光检测 Polyacrylamide gel electrophoresis and florescence detection	半自动, 实验台, 高成本, 不太灵活 Semi-automatic, exp. station, high cost, little flexible, 1000 DP/D
G3 (2000s)	固相芯片 Solid chips	SNP, 包括InDel SNP, including InDels	探针杂交和荧光检测 Probe hybridization and florescence detection	自动, 工作站, 低成本, 比较灵活 Automatic, workstation, low cost, less flexible, 1M⁺ DP/D
G4 (2010s)	液相芯片 Liquid chips	SNP, 包括InDel SNP, including InDels	靶向测序基因型检测和液相捕获 Genotyping by target sequencing (GBTS) and captured in solution	自动, 工作站, 极低成本, 非常灵活 Automatic, workstation, very low cost, very flexible, 1M⁺ DP/D
G5 (2020s)	全测序 Fully sequencing	SNP和所有序列变异 SNP and all sequence variations	序列阅读和比较 Sequence reading and comparison	高度自动, 便携式, 几乎零成本, 超灵活 Highly automatic, carry-on, almost no cost, extremely flexible, 1B⁺ DP/D

应用分子标记技术改进作物品种保护和监管

Enhancement of plant variety protection and regulation using molecular marker technology

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