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首页> 外文期刊>Molecular genetics and genomics: MGG >A systematic exploration of high-temperature stress-responsive genes in potato using large-scale yeast functional screening
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A systematic exploration of high-temperature stress-responsive genes in potato using large-scale yeast functional screening

机译:大规模酵母功能筛选系统研究马铃薯高温胁迫响应基因

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摘要

Potato (S. tuberosum) is a highly heat-sensitive crop; a slight rise from optimal temperature can lead to drastic decline in tuber yield. Despite several advancements made in breeding for thermo-tolerant potato, molecular mechanisms governing thermo-tolerance is poorly understood. The first step towards understanding the thermo-tolerance mechanism is to identify the key genes involved in it. Here we used a yeast-based functional screening method to identify, characterize and classify potato genes with potentials to impart heat tolerance. We constructed two cDNA expression libraries from heat-stressed potato plants (35 A degrees C) after 2 and 48 h of treatment. 95 potential candidate genes were identified based on enhanced ability of yeast cells over-expressing heterologous potato cDNA sequences to tolerate heat stress. Cross-resistance analysis of these heat-tolerant yeast clones to other abiotic stresses indicated that 20 genes were responsive to drought, 14 to salt and 11 to heat/drought/salt stresses. Comparison of 95 genes with reported whole potato transcriptome data showed that majority of them have varying expression patterns under heat, drought and salt stresses. The expression pattern was validated by analyzing the expression of 22 randomly selected genes under various stresses using qPCR. Gene ontology (GO) enrichment analysis of these 95 genes indicated that most of them are involved in various cellular metabolism, signal transduction, response to stress and protein folding, suggesting possible role of these genes in heat tolerance of potato. Genes identified from this study can be potential candidates for engineering heat tolerance as well as broad-spectrum abiotic stress tolerance of potato.
机译:马铃薯(S. tuberosum)是高度热敏感的作物;最佳温度略有升高会导致块茎产量大幅下降。尽管在耐热马铃薯的育种方面取得了一些进展,但对耐热性的分子机制了解甚少。理解耐热机制的第一步是确定其中涉及的关键基因。在这里,我们使用基于酵母的功能筛选方法来鉴定,表征和分类具有潜在耐热性的马铃薯基因。在处理2和48小时后,我们从热胁迫的马铃薯植株(35 A摄氏度)构建了两个cDNA表达文库。基于酵母细胞过表达异源马铃薯cDNA序列耐受热胁迫的增强能力,鉴定了95个潜在候选基因。这些耐热酵母克隆对其他非生物胁迫的交叉抗性分析表明,有20个基因对干旱有响应,有14个基因对盐有响应,有11个基因对热/干旱/盐胁迫有响应。将95个基因与已报道的整个马铃薯转录组数据进行比较,结果表明它们中的大多数在高温,干旱和盐胁迫下具有不同的表达模式。通过使用qPCR分析22种随机选择的基因在各种胁迫下的表达来验证表达模式。对这95个基因的基因本体论(GO)富集分析表明,它们中的大多数都参与了各种细胞代谢,信号转导,对压力的反应和蛋白质折叠,这暗示了这些基因在马铃薯耐热性中的可能作用。这项研究确定的基因可能是马铃薯工程耐热性和广谱非生物胁迫耐受性的潜在候选者。

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