棉花MNP标记位点开发及其在DNA指纹图谱构建中的应用
Development and application of cotton MNP marker for fingerprint cons-truction
艾莎 1李莎 2方治伟 3李论 3李甜甜 3高利芬 3陈利红 3肖华锋 3万人静 3闫多子 4武星廷 4彭海 3韩瑞玺 4周俊飞3
作者信息
- 1. 江汉大学生命科学学院,湖北武汉 430056
- 2. 三峡工程鱼类资源保护湖北省重点实验室,湖北宜昌 443100;中国长江三峡集团有限公司中华鲟研究所,湖北宜昌 443100
- 3. 江汉大学生命科学学院,湖北武汉 430056;农业农村部华中生物DNA指纹鉴定公共研发中心,湖北武汉 430056
- 4. 农业农村部科技发展中心,北京 100176
- 折叠
摘要
棉花是重要的经济作物,棉属植物基因组复杂,分子标记开发困难且滞后于其他二倍体农作物.本研究利用多态性好、准确性高的MNP(multiple nucleotide polymorphisms,MNP)标记法,构建我国棉花主栽品种指纹图谱,为品种鉴定、遗传多样性分析、分子育种等提供技术指导和数据支撑.本研究根据现有棉花基因组数据开发了 940 个MNP位点,对 98 份棉花样品进行多重PCR扩增,构建文库并测序,提取每个样品中 940 个MNP标记的基因型,构建棉花品种指纹图谱.对 98 个样品进行两两比对,获得每 2 个样品之间的遗传差异.利用遗传差异比例对 98 个棉花样品绘制系统发育树.940个MNP位点在98份棉花样品中均获得有效扩增产物,扩增产物长度分布在163～274 bp,平均长度270 bp.在98份样品中平均区分度(discriminative power,DP)为0.46,任意2个品种间两两比较遗传差异,平均有 410.26 个MNP位点存在差异,平均差异为 44.84%.从 98 份棉花品种的系统发育树可以看出,利用MNP标记可以依据地理分布将其分为 3 个类群,大致分布于黄河流域、长江流域和新疆地区.本研究开发的 940 个MNP标记可以对棉花品种进行精准分型,利用 940 个棉花MNP标记构建的 98 份棉花品种DNA指纹图谱,可为品种鉴定、品种权保护及种质创新提供技术支撑.
Abstract
Cotton is an important economic crop with complex genomes,yet the development of cotton molecular markers lags behind that of other diploid crops,posing challenges.This study utilized the highly polymorphic and accurate Multiple Dispersed Nucleotide Polymorphisms(MNP)marker approach to construct fingerprints for the main cotton varieties in China.This achievement provides a technical foundation for variety identification,genetic diversity analysis,and molecular breeding.Using public cotton genome information,a panel of 940 MNP markers was developed.Subsequently,98 cotton samples underwent mul-tiplex PCR amplification,library construction,sequencing,and genotyping.As a result,a fingerprint database for each cotton variety was established.Pairwise comparisons based on MNP genotypes were conducted to determine genetic differences,fol-lowed by a sequential phylogenetic analysis.Effective amplification products were obtained from all 940 MNP loci in the 98 cot-ton samples,with amplification product lengths ranging from 163 to 274 bp and an average of 270 bp.A total of 940 polymorphic loci were detected in the 98 samples,with an average discriminative power(DP)of 0.46.When comparing genetic differences between any two varieties,an average of 410.26 MNP loci showed differences,accounting for an average difference of 44.84%.The NJ phylogenetic tree revealed that the 98 cotton samples could be divided into three groups based on MNP markers,roughly corresponding to their distribution in the Yellow River basin,the Yangtze River basin,and the Xinjiang region.The 940 MNP markers developed in this study can accurately classify cotton varieties,and the DNA fingerprints of the 98 cotton varieties con-structed using these markers provide technical support for variety identification,variety rights protection,and germplasm innova-tion.
关键词
棉花/分子标记/MNP/指纹图谱/品种鉴定Key words
cotton/marker/MNP/fingerprint/variety identification引用本文复制引用
基金项目
湖北省科技重大项目(2022ABA001)
江汉大学一流学科建设重大专项资助计划(2023XKZ024)
三峡工程鱼类资源保护湖北省重点实验室开放课题项目(2021041-ZHX)
出版年
2024
国家科技期刊平台
NSTL
万方数据