园艺学报2025，Vol.52Issue(12)：3202-3218,17.DOI:10.16420/j.issn.0513-353x.2024-0881
番茄重要性状HRM分子标记开发及不同反应体系的分析效果评价
Development of HRM Molecular Markers for Important Traits in Tomato and Assessment of Analytical Effects Across Various Reaction Systems
摘要
Abstract
High resolution melting(HRM)genotyping technology can identify the genotypes of different samples by forming different melting curves,be characterized by its simplicity,rapid processing,high throughput,and superior resolution.This study aims to develop HRM molecular markers for identifying superior alleles associated with important traits in tomato and to assess the impact of various factors on HRM analysis outcomes.A total of 23 HRM molecular markers were successfully developed based on the sequence characteristics of 23 superior alleles of specific genes or their adjacent regions.These genes include those associated with disease resistance,such as Bwr-12,Frl,I-7,Pto,Rx4,Sm,Ty-1;genes conferring abiotic stress tolerance,including SlACR11A,SlBBX31,SlHAK20,SlSOS2;genes related to fruit traits,such as Aft,LC,GF,OVATE,PT,R,SlALMT9,STP1,Y;and genes associated with plant type,including SD1,SP,SP5G.Furthermore,an assessment of the impact of varying reaction volumes,template DNA concentrations,and DNA extraction methods on HRM analysis revealed that reaction volumes of 5,7.5,10,12.5 and 15 µL demonstrated comparable analytical performance to the recommended reaction volume of 20 µL specified in the kit.Notably,within the 5 µL reaction volume,DNA concentrations ranging from 12.5 ng·µL-1 to 1 600 ng·µL-1 exhibited robust analytical performance.Additionally,DNA extracted using the NaOH method was found to be suitable for HRM analysis.The findings of this study will facilitate the identification of superior alleles and marker-assisted breeding in tomato.关键词
番茄/重要性状/优异等位基因/HRM/分子标记Key words
tomato/important trait/superior allele/HRM/molecular marker分类
农业科技引用本文复制引用
刘畅,李君明,刘磊,李鑫,朱璨,黄泽军,魏凯,章力,路菲菲,余庆同,王梦雅,王孝宣,杜永臣,国艳梅..番茄重要性状HRM分子标记开发及不同反应体系的分析效果评价[J].园艺学报,2025,52(12):3202-3218,17.基金项目
国家重点研发计划项目(2022YFD1200500) （2022YFD1200500）
国家自然科学基金项目(31872949) （31872949）
现代农业产业技术体系建设专项资助(CARS-23-A06) （CARS-23-A06）
