‘粤绿肥1号’紫云英转录组测序及生物信息学分析

    Transcriptome Sequencing and Bioinformatic Analysis of Astragalus sinicus ‘Yuelvfei No. 1’

    • 摘要:
      目的 紫云英(Astragalus sinicus L.)能固氮养地和改良土壤,是生态农业的核心物种。广东省推广“冬种紫云英-春耕深翻”模式3万hm2,形成“绿肥-减排-增产”生态经济。紫云英种质资源开发对维持耕地质量提升系统的可持续性具有战略价值。利用高通量测序技术和生物信息学方法分析紫云英的转录组和基因表达特征,为紫云英的资源鉴定与利用提供遗传信息。
      方法 提取‘粤绿肥1号’紫云英的根、茎、叶和种子的总RNA,混合构建文库并进行高通量测序,组装获得紫云英的unigenes。通过序列注释和比对分析‘粤绿肥1号’紫云英转录组特征。
      结果 从‘粤绿肥1号’紫云英转录组中共鉴定出30 792条unigenes,平均长度为1 163 bp,N50值为1 946 bp。紫云英unigenes与鹰嘴豆(Cicer arietinum)具有最高序列相似性。在功能注释分析中,20 930条unigenes获得功能注释,其中KEGG和GO数据库分别注释13 858条和17 439条。GO注释结果显示,这些基因主要参与细胞结构、分子结合及代谢过程等生物学功能。类黄酮是豆科植物与根瘤菌开始信号交流的第一步,共鉴定出紫云英中类黄酮合成通路的关键酶基因41个;R基因在豆科植物与根瘤菌互作中发挥重要作用,鉴定出R基因中最主要的一类NBS基因成员64个。将紫云英unigenes与plantTFDB数据库比对,鉴定出MYB、AP2/ERF、WRKY和bHLH等转录因子367个。通过MISA分析13 047条长度大于1 000 bp的unigenes,鉴定出4 187个SSR位点,并设计相应引物。
      结论 通过转录组测序分析鉴定‘粤绿肥1号’紫云英的根瘤菌互作和转录因子等基因,并设计SSR引物,可为深入探索紫云英资源鉴定和遗传改良提供数据支持。

       

      Abstract:
      Objective Chinese milk vetch (Astragalus sinicus L.), capable of nitrogen fixation to nourish soil, improve soil quality, and support ecological conservation, is a core species in ecological agriculture. Guangdong Province has promoted the "winter cultivation of Chinese milk vetch followed by spring ploughing and deep tillage" model across 30, 000 hm2, establishing a "green manure-emission reduction-yield increase" eco-economic system. The development of Chinese milk vetch germplasm resources holds strategic value for sustaining and enhancing the cropland quality improvement system.To analyze the transcriptome and gene expression characteristics of A.sinicus using high-throughput sequencing technology and bioinformatics methods, providing genetic information for resource identification and utilization of A.sinicus.
      Method Total RNA was extracted from the roots, stems, leaves, and seeds of A.sinicus cultivar 'Yuelvfei No. 1, , pooled to construct a library, and subjected to high-throughput sequencing. The unigenes of A.sinicus were assembled. The transcriptomic characteristics of 'Yuelvfei No. 1, were analyzed through sequence annotation and alignment.
      Result A total of 30 792 unigenes were identified in the transcriptome of 'Yuelvfei No. 1, , with an average length of 1 163 bp and an N50 of 1 946 bp. The unigenes of A.sinicus showed the highest similarity to those of Cicer arietinum (chickpea). Among the unigenes, 20 930 were functionally annotated in databases, including 13 858 annotated in the KEGG database and 17 439 annotated in the GO database, covering cellular anatomical entities, binding, and cellular processes. Flavonoids are the first step in signal exchange between legumes and rhizobia, and this study identified 41 key enzyme genes involved in the flavonoid biosynthesis pathway in A.sinicus. R genes also play a crucial role in the interaction between legumes and rhizobia, and we identified 64 members of the largest class of R genes, NBS genes. Additionally, by aligning A.sinicus unigenes with the plantTFDB database, 367 transcription factors, including MYB, AP2/ERF, WRKY, and bHLH, were identified. MISA analysis of 13 047 unigenes longer than 1 000 bp identified 4 187 SSR loci, for which corresponding primers were designed.
      Conclusion The transcriptomic information of A.sinicus cultivar 'Yuelvfei No. 1, provides valuable data support for further exploration of resource identification and genetic improvement in A.sinicus.

       

    /

    返回文章
    返回