基于BSA-seq技术定位绿豆种皮颜色基因
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山西农业大学农学院

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山西省基础研究计划项目(20210302124504);山西农业大学博士科研启动项目(2021BQ43);山西省博士毕业生、博士后研究人员来晋工作奖励资金科研项目(SXBYKY2021050);山西省农业科学院作物科学研究所博士基金计划(ZB1101)


Mapping of A Seed Coat Color Related Gene by BSA-seq in Mung Bean (Vigna radiata L.)
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College of Agronomy, Shanxi Agricultural University

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Shanxi Basic Research Program Project (20210302124504),the Ph.D. of Shanxi Agricultural University Scientific Research Start-up Project (2021BQ43),the Shanxi Province Doctoral Graduates, Postdoctoral Researchers Come to Shanxi Province to Work Incentive Fund Scientific Research Project (SXBYKY2021050),the Shanxi Academy of Agricultural Sciences Institute of Crop Science Doctoral Fund Program (ZB1101)

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    摘要:

    种皮颜色是作物重要的驯化性状和形态标记,绿豆种皮颜色与黄酮类化合物含量有关。挖掘绿豆种皮颜色相关基因有助于开发新品种,提高绿豆种皮的应用价值。本研究以冀绿9号(黑色种皮)和资源330(黄色种皮)为亲本构建F2分离群体,采用BSA-seq方法进行初定位。结果表明,SNPs和InDels关联区域交集位于4号染色体共3.26 Mb的区段,包含324个基因,其中非同义突变基因共49个,移码突变基因15个。进一步开发KASP分子标记进行精细定位,筛选出高质量KASP引物11对。最终将绿豆种皮色位点定位于4号染色体上的KASP330-KASP421之间,物理距离为16,302,330~18,013,421 bp(1.71 Mb)。结合转录组数据分析和qRT-PCR表达分析共筛选出6个候选基因,其中基因LOC106758748基因注释为MYB90,其功能为参与调控黄酮类化合物生物合成,故可能是调控绿豆种皮颜色的关键基因。本研究结果可为绿豆种皮色相关基因的克隆和在育种中的利用奠定基础。

    Abstract:

    Seed coat color is an important agronomic trait that associates with crop domestication and serves as morphological marker. In mung bean, the seed coat color was related to the content of flavonoids. Cloning and application of seed coat color-related genes becomes of interest in development of new mung bean varieties with improved nutritional properties. In this study, the varieties ‘Jilv 9’ (black seed coat) and ‘Ziyuan 330’ (yellow seed coat) were used as parents to generate an F2 segregating population. The BSA-seq approach was applied for preliminary mapping of the genes underlying the seed coat color. The association analysis using integrated SNPs and InDels suggested an interval of 3.26 Mb harboring 324 predicted genes, of which 49 genes were found with non-synonymous mutations and 15 genes were detected with frameshift mutation. By further use of 11 high-quality KASP markers in fine mapping, the candidate interval was finally delimited between KASP330 and KASP421 in the physical interval of 16,302,330–18,013,421 bp (1.71 Mb) on chromosome 4. The transcriptome data analysis and qRT-PCR expression analysis suggested six differently-expressed candidate genes, of which the LOC106758748 was annotated as a transcription factor MYB90 that was reported with a function in the flavonoid biosynthesis and served as key candidate gene regulating the seed coat color in mung bean. This study laid a solid foundation for further cloning of mung bean seed coat color related gene(s) and the utilization in breeding.

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  • 收稿日期:2022-10-25
  • 最后修改日期:2022-11-15
  • 录用日期:2022-11-28
  • 在线发布日期: 2022-12-12
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