花生全基因组变异的鉴定和InDel标记的开发及应用
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作者单位:

1.广东省省级现代农业(耕地保育与节水农业)产业技术研发中心/中国热带农业科学院湛江实验站,湛江 524013;2.云南农业大学农学与生物技术学院,昆明 650201;3.湛江市农业科学研究院,广东湛江 524094

作者简介:

研究方向为作物遗传育种,E-mail: zhangxj@catas.cn

通讯作者:

徐志军,研究方向为花生种质资源学,E-mail: zhijunxu1990@163.com

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基金项目:

海南省自然科学基金项目(321QN348);中央级公益性科研院所基本科研业务费专项(1630102024002,1630102023001)


Genome-wide Variation Identification in Peanut and Development of InDel Markers for Genetic Research
Author:
Affiliation:

1.Guangdong Modern Agriculture (Cultivated Land Conservation and Water-saving Agriculture) Industrial Technology Research and Development Center/Zhanjiang Experiment Station, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang 524013;2.College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201;3.Zhanjiang Academy of Agricultural Sciences, Zhanjiang 524094,Guangdong

Fund Project:

Foundation projects: Hainan Provincial Natural Science Foundation of China (321QN348); Central Public-interest Scientific Institution Basal Research Fund(1630102024002,1630102023001)

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    摘要:

    利用13 个花生品种的重测序数据进行花生全基因组变异鉴定,分析InDel在花生基因组上的分布特点,开发、验证并评估InDel标记在花生亲缘关系和品种鉴定中的效率。结果表明,从13 个花生品种中共检测到313432 个高质量SNP和38777 个高质量InDel,平均分布密度分别为123 个/Mb和15.23 个/Mb;InDel和SNP主要分布于基因间,分别占检测SNP和InDel总数的52.347%和60.081%。利用插入或缺失长度≥10 bp的InDel位点进行标记开发,共有3675 个位点可以进行引物设计,这些位点在花生20 条染色体上分布不均匀,平均分布密度为1.48 个/Mb。e-PCR检测表明,InDel引物扩增位点以1个位点为主,有效引物数为2561 对(69.69%),可检测出3133 个位点,根据引物扩增位点在基因组上的位置信息绘制了扩增位点的物理图谱。在100 对随机引物中,共有31 对在4 个亲缘关系较远的花生品种中存在差异条带,31对引物在47 份花生品种(育种系)中共检测到62 个等位变异,主基因频率范围为0.51~0.98,平均为0.77;多态性信息量范围为0.04~0.37,平均为0.24。聚类分析和群体结构分析均可将47 份花生品种(育种系)分为2 个类群,且结果基本一致;47 份材料最少可用7 个标记进行区分,表明开发的InDel标记可以有效地用于花生遗传多样性的评估和品种鉴定。研究结果丰富了花生的分子标记,为InDel标记在花生资源遗传多样性、品种鉴定、指纹图谱构建等遗传研究中的利用提供了帮助。

    Abstract:

    This study aimed to identify genetic variations on the whole genome level using genome resequencing data of 13 peanut cultivars, clarify their distribution characteristic, develop and verify InDel markers, and evaluate the efficiency of InDel markers in peanut cultivar identification. A total of 313432 high-quality SNPs and 38777 high-quality InDel were detected, with an average distribution density of 123/Mb and 15.23/Mb, respectively. The InDels and SNPs were mainly distributed in intergenic regions, with a frequency of 52.35% and 60.08% of the total SNP and InDel, respectively. Primers were designed using InDel with insertion or deletion length ≥10 bp, and 3675 InDel could be used to develop InDel markers. These InDel locus were unevenly distributed on the 20 chromosomes of peanut with an average density of 1.48 /Mb. Using electronic PCR, the InDel primers amplified mainly with 1 locus and 2561 effective primers (69.69%) could amplified 3133 effective loci in the peanut reference genome. The physical map of amplification loci was drawn according to the loci position in cultivated peanut genome. Among 100 pairs of random primers, 31 pairs amplified different bands in the 4 varieties with distant relatives. The 31 InDel primer pairs amplified 62 alleles in 47 peanut cultivars (or breeding lines), the frequency of major genes ranged from 0.51 to 0.98 with an average of 0.77, and the polymorphic information content ranged from 0.04 to 0.37, with an average of 0.24. Both cluster analysis and population structure analysis could divide the 47 peanut cultivars (breeding lines) into two groups, and the 47 materials could be distinguished by at least 7 markers, indicating that the developed InDel markers could be effectively used for the assessment of genetic diversity and variety identification of peanut. The research results enriched the molecular markers of peanut, and was beneficial for the use of InDel markers in genetic studies of peanut resource genetic diversity, variety identification, fingerprint construction.

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引用本文

张雪姣,季木灯,王丽媛,等.花生全基因组变异的鉴定和InDel标记的开发及应用[J].植物遗传资源学报,2024,25(12):2136-2148.

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  • 收稿日期:2024-02-06
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  • 在线发布日期: 2024-12-06
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