Department of Agricultural Biotechnology, Assam Agricultural University, Jorhat, 785013, India.
Office of the ICAR-National Professor (Norman Borlaug Chair), and DBT-AAU Centre, Assam Agricultural University, Jorhat, 785013, India.
Mol Biol Rep. 2019 Dec;46(6):5713-5722. doi: 10.1007/s11033-019-05005-0. Epub 2019 Aug 28.
The differentially expressed genes in the chickpea pod wall have been identified for the first time using a forward suppression subtractive hybridization (SSH) library. In all, 226 clones of SSH library were sequenced and analyzed. A total of 179 high-quality expressed sequence tags (ESTs) were generated and based on the CAP3 assembly of these ESTs, 126 genes (97 singletons and 29 contigs) were computationally annotated. The mapping of 88.26% ESTs by gene ontology (GO) annotation distributed them into 751 GO terms of three categories, cellular location, molecular function, and biological process. The KEGG pathway analysis revealed 45 ESTs are involved in 49 different biological pathways. Also, 67 ESTs encodes four different classes of enzymes such as oxidoreductases (29), transferase (20), hydrolases (16) and isomerase (2). Six genes were selected and subjected to qPCR analysis, of these, two genes (FHG Floral homeotic AGAMOUS-like isoform X2, MADS1 MADS-box transcription factor) showed significant up-regulation in the pod wall compared to leaves. Surprisingly, one of the MADS1 box gene, FHG (CaAGLX2), responsible for flower development expressed in the pod wall. Therefore, understanding its specific role in the pod wall could be interesting. Thus, the transcript dynamics of the chickpea pod wall revealed differentially expressed genes in the pod wall, which may be participating in the metabolic build-up of both pod wall and seeds.
首次使用正向抑制差减杂交 (SSH) 文库鉴定了鹰嘴豆荚壁中的差异表达基因。总共对 226 个 SSH 文库克隆进行了测序和分析。共生成了 179 个高质量表达序列标签 (EST),并基于这些 EST 的 CAP3 组装,计算注释了 126 个基因 (97 个单基因和 29 个基因簇)。GO 注释的基因本体 (GO) 映射将 88.26%的 ESTs 分配到三个类别中的 751 个 GO 术语,分别是细胞位置、分子功能和生物过程。KEGG 途径分析显示 45 个 ESTs 参与了 49 个不同的生物途径。此外,67 个 ESTs 编码了氧化还原酶 (29 个)、转移酶 (20 个)、水解酶 (16 个)和异构酶 (2 个)等四类不同的酶。选择了 6 个基因进行 qPCR 分析,其中两个基因 (FHG Floral homeotic AGAMOUS-like isoform X2, MADS1 MADS-box transcription factor) 在荚壁中与叶片相比表现出明显的上调。令人惊讶的是,MADS1 盒基因之一 FHG (CaAGLX2) 负责花发育,在荚壁中表达。因此,了解其在荚壁中的特定作用可能会很有趣。因此,鹰嘴豆荚壁的转录动态揭示了荚壁中差异表达的基因,这些基因可能参与了荚壁和种子的代谢构建。