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

Abstract

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

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

Enhancement of plant variety protection and regulation using molecular marker technology

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