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利用 RIL 群体和兵豆(Lens culinaris Medik.)基因型进行全基因组 InDel 发现和基于 PCR 的 InDel 标记的早熟性验证。

Genome-wide discovery of InDels and validation of PCR-Based InDel markers for earliness in a RIL population and genotypes of lentil (Lens culinaris Medik.).

机构信息

Division of Genetics, Indian Agricultural Research Institute, New Delhi, India.

Indian Council of Forestry Research and Education-Institute of Forest Biodiversity, Hyderabad, India.

出版信息

PLoS One. 2024 May 22;19(5):e0302870. doi: 10.1371/journal.pone.0302870. eCollection 2024.

Abstract

The systematic identification of insertion/deletion (InDel) length polymorphisms from the entire lentil genome can be used to map the quantitative trait loci (QTL) and also for the marker-assisted selection (MAS) for various linked traits. The InDels were identified by comparing the whole-genome resequencing (WGRS) data of two extreme bulks (early- and late-flowering bulk) and a parental genotype (Globe Mutant) of lentil. The bulks were made by pooling 20 extreme recombinant inbred lines (RILs) each, derived by crossing Globe Mutant (late flowering parent) with L4775 (early flowering parent). Finally, 734,716 novel InDels were identified, which is nearly one InDel per 5,096 bp of lentil genome. Furthermore, 74.94% of InDels were within the intergenic region and 99.45% displayed modifier effects. Of these, 15,732 had insertions or deletions of 20 bp or more, making them amenable to the development of PCR-based markers. An InDel marker I-SP-356.6 (chr. 3; position 356,687,623; positioned 174.5 Kb from the LcFRI gene) was identified as having a phenotypic variance explained (PVE) value of 47.7% for earliness when validated in a RIL population. Thus, I-SP-356.6 marker can be deployed in MAS to facilitate the transfer of the earliness trait to other elite late-maturing cultivars. Two InDel markers viz., I-SP-356.6 and I-SP-383.9 (chr. 3; linked to LcELF3a gene) when tested in 9 lentil genotypes differing for maturity duration, clearly distinguished three early (L4775, ILL7663, Precoz) and four late genotypes (Globe Mutant, MFX, L4602, L830). However, these InDels could not be validated in two genotypes (L4717, L4727), suggesting either absence of polymorphism and/or presence of other loci causing earliness. The identified InDel markers can act as valuable tools for MAS for the development of early maturing lentil varieties.

摘要

从整个兵豆基因组中系统地鉴定插入/缺失(InDel)长度多态性可用于定位数量性状基因座(QTL),并进行各种连锁性状的标记辅助选择(MAS)。通过比较两个极端群体(早开花群体和晚开花群体)和兵豆亲本基因型(Globe Mutant)的全基因组重测序(WGRS)数据,鉴定了 InDels。这些群体是通过将 Globe Mutant(晚开花亲本)与 L4775(早开花亲本)杂交,然后将每个亲本的 20 个极端重组自交系(RIL)进行混合而制成的。最后,鉴定出了 734,716 个新的 InDels,这几乎是兵豆基因组每 5,096 bp 就有一个 InDel。此外,74.94%的 InDels位于基因间区,99.45%显示修饰效应。其中,有 15,732 个 InDels 的插入或缺失长度为 20 bp 或更长,这使得它们适合开发基于 PCR 的标记。在一个 RIL 群体中验证时,发现一个 InDel 标记 I-SP-356.6(chr. 3;位置 356,687,623;距离 LcFRI 基因 174.5 Kb)的早发性表型方差解释(PVE)值为 47.7%。因此,I-SP-356.6 标记可以用于 MAS,以促进将早发性性状转移到其他晚熟优良品种中。当在 9 个成熟时间不同的兵豆基因型中测试两个 InDel 标记 I-SP-356.6 和 I-SP-383.9(chr. 3;与 LcELF3a 基因相连)时,它们清楚地区分了三个早熟(L4775、ILL7663、Precoz)和四个晚熟基因型(Globe Mutant、MFX、L4602、L830)。然而,这两个 InDels 在两个基因型(L4717、L4727)中无法得到验证,这表明要么不存在多态性,要么存在导致早发性的其他基因座。鉴定出的 InDel 标记可以作为兵豆早熟品种 MAS 的有用工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04a2/11111061/2983b7f2e0f7/pone.0302870.g001.jpg

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