基于转录组测序解析甜玉米耐低温发芽的相关基因

    Analysis of Genes Related to Low-temperature Germination Ability in Sweet Corn Based on Transcriptome Sequencing

    • 摘要:
      目的 通过转录组测序分析低温下萌发能力不同的甜玉米自交系,鉴定甜玉米耐低温萌发的相关基因。
      方法 利用低温萌发能力强的甜玉米自交系Scil-H7th为供体构建近等基因系,获得不同低温萌发能力的材料,通过生理指标测定、转录组分析和qRT-PCR,筛选分析与低温萌发相关的基因。
      结果 对自交系在不同萌发时期和同一萌发时期自交系之间的基因表达量进行比较分析,获得不同材料在相同萌发时期4个比较组中共有的差异表达基因410个,其中,65个基因只在高种子活力材料Scil-H7th和近等基因系L18中表达,在低种子活力的轮回亲本Scil-C4th中不表达;这些基因中有32个基因的功能已知,结合生物信息学分析和表达量分析,筛选出有交集的20个DEGs,推测这些基因与低温萌发密切相关。利用qRT-PCR对其中最有可能参与低温萌发的7个上调表达基因进行表达量验证,其中编码UDP-葡萄糖基转移酶的基因Zm00001d037383(UGT1)在DP胚部表达明显高于其他组织,推测该基因在甜玉米耐低温萌发过程中起关键作用。
      结论 通过转录组测序分析,鉴定了甜玉米耐低温萌发的相关基因,为后续创制低温萌发能力更强的甜玉米种质资源奠定了基础。

       

      Abstract:
      Objective The study was carried out to identify genes related to low-temperature germination ability in sweet corn through transcriptome sequencing of inbred lines with different germination ability at low-temperature.
      Method Using the sweet corn inbred line Scil-H7th with strong low-temperature germination ability as a donor, NILs (near-isogenic lines) were constructed and NILs with different low-temperature germination ability were obtained, then genes related to low-temperature germination were selected through physiological determination, transcriptomic analysis and qRT-PCR.
      Result The differentially expressed genes (DEGs) within lines in different germination stages and DEGs among lines in the same germination stages were comparatively analyzed. Among different materials at the same germination stage, a total of 410 DEGs in all the four comparisons were obtained, among which 65 genes were only expressed in the high seed vigor material Scil-H7th and the near-isogenic line L18 but not expressed in the low seed vigor rotation parent (Scil-C4th). 32 of these genes were functionally annotated. Combined with bioinformatics analysis and expression analysis, 20 DEGs were identified to be highly associated with low-temperature germination. From the 20 candidates, expression profile of 7 genes with upper-regulated transcriptome sequencing were analyzed with qRT-PCR; among them, the Zm00001d037383, encoding UDP-glucosyltransferase, was highly expressed in embryos of DP and hypothesized to play a key role in low-temperature tolerant germination in sweet corn.
      Conclusion The transcriptome sequencing analysis identified genes related to low-temperature germination ability in sweet corn, which laid a foundation for the development of sweet corn with better germination ability at low temperature.

       

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