Gene Expression and Yeast Two-Hybrid Studies of 1R-MYB Transcription Factor Mediating Drought Stress Response in Chickpea (Cicer arietinum L.).
Gene Expression and Yeast Two-Hybrid Studies of 1R-MYB Transcription Factor Mediating Drought Stress Response in Chickpea (Cicer arietinum L.).
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DOI：
10.3389/fpls.2015.01117
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发表时间：
2015
影响因子：
5.6
通讯作者：
Varshney RK
中科院分区：
文献类型：
--
作者：
Ramalingam A;Kudapa H;Pazhamala LT;Garg V;Varshney RK
干旱胁迫是影响鹰嘴豆产量的主要因素之一。基因组学辅助育种具有精确和高效地加速育种的潜力。为了做到这一点，了解耐旱性的分子机制和候选基因的鉴定是至关重要的。转录因子在植物逆境胁迫相关基因的调控中起着重要作用。在这种情况下，定量真实的时间-PCR（qRT-PCR）被用来研究差异基因表达的选择TF，确定从大规模表达序列标签（EST）分析，在对比干旱响应基因型。将ICC 4958（耐受）、ICC 1882（敏感）、JG 11（优良）和JG 11+（渐渗系）的根组织用于研究。随后，克隆在干旱胁迫下在耐旱基因型中显著诱导的候选单重复MYB（1 R-MYB）转录物（1 R-MYB蛋白质的编码序列区）并进行酵母双杂交（Y2 H）分析。使用1 R-MYB诱饵蛋白用Y2 H筛选根cDNA文库，鉴定了三种CDS编码肽，即半乳糖苷醇-蔗糖半乳糖基转移酶2、CBL（钙调磷酸酶B样）相互作用丝氨酸/苏氨酸蛋白激酶25和阿坝响应性17样，其通过在酵母中共转化来证实。这些研究结果提供了初步的见解，这种1 R-MYB转录因子的能力，共同调节鹰嘴豆耐旱机制。
Drought stress has been one of the serious constraints affecting chickpea productivity to a great extent. Genomics-assisted breeding has a potential to accelerate breeding precisely and efficiently. In order to do so, understanding the molecular mechanisms for drought tolerance and identification of candidate genes are crucial. Transcription factors (TFs) have important roles in the regulation of plant stress related genes. In this context, quantitative real time-PCR (qRT-PCR) was used to study the differential gene expression of selected TFs, identified from large-scale expressed sequence tags (ESTs) analysis, in contrasting drought responsive genotypes. Root tissues of ICC 4958 (tolerant), ICC 1882 (sensitive), JG 11 (elite), and JG 11+ (introgression line) were used for the study. Subsequently, a candidate single repeat MYB (1R-MYB) transcript that was remarkably induced in the drought tolerant genotypes under drought stress was cloned (coding sequence region for the 1R-MYB protein) and subjected to yeast two-hybrid (Y2H) analysis. The screening of a root cDNA library with Y2H using the 1R-MYB bait protein, identified three CDS encoding peptides namely, galactinol-sucrose galactosyltransferase 2, CBL (Calcineurin B-like)-interacting serine/threonine-protein kinase 25, and ABA responsive 17-like, which were confirmed by co-transformation in yeast. These findings provide preliminary insights into the ability of this 1R-MYB transcription factor to co-regulate drought tolerance mechanism in chickpea.