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Abstract：
Cotton encounters long-term drought stress problems resulting in major yield losses. Transcription factors (TFs) plays an important role in response to biotic and abiotic stresses. The coexpression patterns of gene networks associated with drought stress tolerance were investigated using transcriptome profiles. Applying a weighted gene coexpression network analysis, we discovered a salmon module with 144 genes strongly linked to drought stress tolerance. Based on coexpression and RT-qPCR analysis GH_D01G0514 was selected as the candidate gene, as it was also identified as a hub gene in both roots and leaves with a consistent expression in response to drought stress in both tissues. For validation of GH_D01G0514, Virus Induced Gene Silencing was performed and VIGS plants showed significantly higher excised leaf water loss and ion leakage, while lower relative water and chlorophyll contents as compared to WT (Wild type) and positive control plants. Furthermore, the WT and positive control seedlings showed higher CAT and SOD activities, and lower activities of hydrogen peroxide and MDA enzymes as compared to the VIGS plants. Gh_D01G0514 (GhNAC072) was localized in the nucleus and cytoplasm. Y2H assay demonstrates that Gh_D01G0514 has a potential of auto activation. It was observed that the Gh_D01G0514 was highly upregulated in both tissues based on RNA Seq and RT-qPCR analysis. Thus, we inferred that, this candidate gene might be responsible for drought stress tolerance in cotton. This finding adds significantly to the existing knowledge of drought stress tolerance in cotton and deep molecular analysis are required to understand the molecular mechanisms underlying drought stress tolerance in cotton.
摘要：
棉花遇到长期干旱胁迫问题，导致严重的产量损失。转录因子（TF）在响应生物和非生物胁迫中起重要作用。使用转录组图谱研究了与干旱胁迫耐受性相关的基因网络的共表达模式。应用加权基因共表达网络分析，我们发现了一个鲑鱼模块，有144个基因与干旱胁迫耐受性密切相关。基于共表达和RT-qPCR分析，选择GH_D01G0514作为候选基因，因为它也被鉴定为根和叶中的hub基因，在两个组织中均响应干旱胁迫而具有一致的表达。为了验证GH_D01G0514，进行了病毒诱导的基因沉默，VIGS植物显示出显着更高的切除叶片水分损失和离子渗漏，与WT（野生型）和阳性对照植物相比，相对水分和叶绿素含量较低。此外，WT和阳性对照幼苗显示出较高的CAT和SOD活性，与VIGS植物相比，过氧化氢和MDA酶的活性较低。Gh_D01G0514(GhNAC072)定位于细胞核和细胞质中。Y2H测定证明Gh_D01G0514具有自激活的潜力。基于RNASeq和RT-qPCR分析,观察到Gh_D01G0514在两种组织中高度上调。因此，我们推断，该候选基因可能与棉花的干旱胁迫耐受性有关。这一发现大大增加了棉花耐旱性的现有知识，需要进行深入的分子分析才能了解棉花耐旱性的分子机制。
