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利用SLAF-BSA技术鉴定与普冰3228穗粒数相关的分子标记和候选区域

Identification of molecular markers and candidate regions associated with grain number per spike in Pubing3228 using SLAF-BSA.

作者信息

Wang Jiansheng, Wang Erwei, Cheng Shiping, Ma Aichu

机构信息

College of Chemistry and Environment Engineering, Pingdingshan University, Pingdingshan, Henan, China.

Henan Key Laboratory of Germplasm Innovation and Utilization of Eco-economic Woody Plant, Pingdingshan, Henan, China.

出版信息

Front Plant Sci. 2024 Mar 5;15:1361621. doi: 10.3389/fpls.2024.1361621. eCollection 2024.

Abstract

Grain number per spike, a pivotal agronomic trait dictating wheat yield, lacks a comprehensive understanding of its underlying mechanism in Pubing3228, despite the identification of certain pertinent genes. Thus, our investigation sought to ascertain molecular markers and candidate regions associated with grain number per spike through a high-density genetic mapping approach that amalgamates site-specific amplified fragment sequencing (SLAF-seq) and bulked segregation analysis (BSA). To facilitate this, we conducted a comparative analysis of two wheat germplasms, Pubing3228 and Jing4839, known to exhibit marked discrepancies in spike shape. By leveraging this methodology, we successfully procured 2,810,474 SLAF tags, subsequently resulting in the identification of 187,489 single nucleotide polymorphisms (SNPs) between the parental strains. We subsequently employed the SNP-index association algorithm alongside the extended distribution (ED) association algorithm to detect regions associated with the trait. The former algorithm identified 24 trait-associated regions, whereas the latter yielded 70. Remarkably, the intersection of these two algorithms led to the identification of 25 trait-associated regions. Amongst these regions, we identified 399 annotated genes, including three genes harboring non-synonymous mutant SNP loci. Notably, the APETALA2 (AP2) transcription factor families, which exhibited a strong correlation with spike type, were also annotated. Given these findings, it is plausible to hypothesize that these genes play a critical role in determining spike shape. In summation, our study contributes significant insights into the genetic foundation of grain number per spike. The molecular markers and candidate regions we have identified can be readily employed for marker-assisted breeding endeavors, ultimately leading to the development of novel wheat cultivars possessing enhanced yield potential. Furthermore, conducting further functional analyses on the identified genes will undoubtedly facilitate a comprehensive elucidation of the underlying mechanisms governing spike development in wheat.

摘要

穗粒数是决定小麦产量的关键农艺性状,尽管已鉴定出一些相关基因,但对普冰3228中其潜在机制仍缺乏全面了解。因此,我们的研究旨在通过结合位点特异性扩增片段测序(SLAF-seq)和混合分组分析法(BSA)的高密度遗传图谱方法,确定与穗粒数相关的分子标记和候选区域。为此,我们对两个已知穗形存在显著差异的小麦种质普冰3228和京4839进行了比较分析。通过这种方法,我们成功获得了2810474个SLAF标签,随后在亲本菌株之间鉴定出187489个单核苷酸多态性(SNP)。我们随后采用SNP-index关联算法以及扩展分布(ED)关联算法来检测与该性状相关的区域。前一种算法鉴定出24个与性状相关的区域,而后一种算法得出70个。值得注意的是,这两种算法的交集导致鉴定出25个与性状相关的区域。在这些区域中,我们鉴定出399个注释基因,包括三个含有非同义突变SNP位点的基因。值得注意的是,还注释了与穗型密切相关的APETALA2(AP2)转录因子家族。鉴于这些发现,可以合理推测这些基因在决定穗形方面起着关键作用。总之,我们的研究为穗粒数的遗传基础提供了重要见解。我们鉴定出的分子标记和候选区域可很容易地用于标记辅助育种工作,最终培育出具有更高产量潜力的新型小麦品种。此外,对已鉴定基因进行进一步的功能分析无疑将有助于全面阐明小麦穗发育的潜在机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/16c6/10948542/0b5e824b29c2/fpls-15-1361621-g001.jpg

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