Abstract:
Drought and high temperature stress may occur concomitantly or individually in succession causing cellular dysfunctions. Abscisic acid (ABA) is a key stress regulator, and its responsive genes are controlled by ABRE (Abscisic acid Responsive Element)-binding factors (ABFs)and G-Box Regulatory factors (GRFs). Here, we identify ABFs, GRFs and targeting miRNAs in desi and kabuli chickpea. To validate their role after drought priming and subsequent high temperature stress, two contrasting chickpea varieties (PBG1 and PBG5) were primed and exposed to 32 °C, 35 °C and 38 °C for 12, 6 and 2 h respectively and analyzed for Physio-biochemical, expression of ABFs, GRFs and MiRNAs, and GC-MS based metabolite analysis. To ascertain the ABF-GRF protein-protein interactions, docking studies were carried out between the ABF3 and GRF14. Genome-wide analysis identified total 9 & 11 ABFs, and 11 GRFsin desi and kabuli respectively. Their gene structure, and motif composition were conserved in all subfamilies and only 10 and 12 genes have undergone duplication in both desi and kabuli chickpea respectively. These genes were differentially expressed in-silico. MiR172 and miR396 were identified to target ABFs and GRFs respectively. Protein-protein interaction (ABF3 and GRF14) might be successful only when the ABF3 was phosphorylated. Drought priming downregulated miR172 and miR396 and eventually upregulated targeting ABFs, and GRFs. Metabolite profiling (GC-MS) revealed the accumulation of 87 metabolites in Primed (P) and Non-Primed (NP) Chickpea plants. Tolerant cultivar (PBG5) responded better in all respects however both severity of stress and exposure are important factors and can produce broadly similar cellular response.
摘要:
干旱和高温胁迫可能同时或单独连续发生,导致细胞功能障碍。脱落酸(ABA)是一种关键的应激调节剂,其响应基因受ABRE(脱落酸响应元件)结合因子(ABFs)和G-Box调节因子(GRFs)控制。这里,我们识别ABF,desi和kabuli鹰嘴豆中的GRFs和靶向miRNA。为了验证它们在干旱引发和随后的高温胁迫后的作用,两个对比鹰嘴豆品种(PBG1和PBG5)被引发并暴露于32°C,35°C和38°C分别持续12、6和2小时,并进行生理生化分析,ABFs的表达,GRFs和miRNA,和基于GC-MS的代谢物分析。为了确定ABF-GRF蛋白质-蛋白质相互作用,在ABF3和GRF14之间进行了对接研究。全基因组分析确定总共9和11ABF,和11GRFsindesi和kabuli。它们的基因结构,和基序组成在所有亚家族中都是保守的,在desi和kabuli鹰嘴豆中分别只有10和12个基因发生了重复。这些基因在计算机上差异表达。MiR172和miR396被鉴定为分别靶向ABFs和GRFs。蛋白质-蛋白质相互作用(ABF3和GRF14)可能只有在ABF3被磷酸化时才成功。干旱引发下调miR172和miR396,并最终上调靶向ABF,和GRF。代谢物谱分析(GC-MS)揭示了87种代谢物在引发(P)和非引发(NP)鹰嘴豆植物中的积累。耐受品种(PBG5)在所有方面的反应都更好,但是胁迫的严重程度和暴露都是重要因素,并且可以产生广泛相似的细胞反应。