Towards transcript profiling of desiccation tolerance in Xerophyta humilis:: Construction of a normalized 11 k X. humilis cDNA set and microarray expression analysis of 424 cDNAs in response to dehydration
Towards transcript profiling of desiccation tolerance in Xerophyta humilis:: Construction of a normalized 11 k X. humilis cDNA set and microarray expression analysis of 424 cDNAs in response to dehydration
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DOI：
10.1111/j.1399-3054.2004.00381.x
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发表时间：
2004-09-01
影响因子：
6.4
通讯作者：
Illing, N
中科院分区：
文献类型：
--
作者：
Collett, H;Shen, A;Illing, N
Xerophyta humilis (Bak.) Dur and Schinz 是南非本土的复活植物，能够保护自己免受与极度脱水相关的压力。为了分析响应干燥和补水恢复的基因表达的整体模式，我们生成了一个标准化的 10 900 个文库，代表来自根和叶组织的在脱水-补水循环期间表达的基因。对于此处描述的小规模微阵列分析，对 424 个 cDNA 进行了测序、注释、排列并与水合和脱水的叶特异性 RNA 杂交。使用反向 Northern 印迹作为比较表达结果的替代方法。结合两种方法的结果，总共鉴定了 55 个脱水诱导 cDNA。对 55 个脱水上调 cDNA 中的 14 个进行 Northern 印迹分析，验证了所有 14 个基因的表达状态。脱水上调的cDNA包括那些与编码金属硫蛋白、半乳糖醇合酶、醛糖还原酶和乙二醛酶的已知脱水应激反应基因同源的cDNA。还鉴定出大量编码晚期胚胎丰富蛋白（LEA）、脱水蛋白和干燥相关蛋白的基因，这表明提供机械和抗氧化保护以防止水分流失的蛋白质在干燥的矮叶草叶组织中的 mRNA 群体中占主导地位。本研究中鉴定的脱水上调基因（之前未涉及脱水反应）包括编码假定叶绿体 RNA 结合蛋白和含有 SNF2/解旋酶结构域的蛋白质的 cDNA。与描述干燥敏感植物脱水反应的微阵列数据进行比较，揭示了矮矮芥菜、拟南芥和水稻之间表达模式的差异，这可以为干燥耐受和敏感表型的潜在机制提供线索。 X. humilis 文库有望成为耐干燥植物脱水和复水反应转录谱分析的有用资源。
Xerophyta humilis (Bak.) Dur and Schinz is an indigenous Southern African resurrection plant which is able to protect itself from the stresses associated with extreme dehydration. For the purpose of analysing global patterns of gene expression in response to desiccation and recovery on rehydration, we have generated a normalized 10 900 library, representing genes from root and leaf tissue that are expressed during the dehydration-rehydration cycle. For the small-scale microarray analysis described here, 424 cDNAs were sequenced, annotated, arrayed and hybridized with hydrated and dehydrated, leaf-specific RNA. Reverse Northern blots were used as an alternative method to compare the expression results. A total of 55 dehydration-inducible cDNAs were identified combining the results of both methods. Northern blot analysis of 14 of the 55 the dehydration-upregulated cDNAs verified the expression status of all 14 genes. Dehydration-upregulated cDNAs included those homologous to known dehydration stress-responsive genes encoding metallothioneins, galactinol synthases, an aldose reductase and a glyoxalase. A large number of genes encoding late embryonic abundant proteins (LEAs), dehydrins and desiccation-related proteins were also identified, suggesting that proteins that provide mechanical and antioxidant protection against water loss dominate the mRNA population in desiccated X. humilis leaf tissue. Dehydration-upregulated genes identified in this study, and not previously implicated in the dehydration response, include cDNAs encoding a putative chloroplast RNA-binding protein and a protein containing SNF2/helicase domains. Comparisons with microarray data, which profile the dehydration response in desiccation-sensitive plants, reveal differences in expression patterns between X. humilis and Arabidopsis and rice that could provide clues as to the mechanisms underlying desiccation-tolerant and -sensitive phenotypes. The X. humilis library promises to be a useful resource for transcript profiling the dehydration and rehydration response in a desiccation-tolerant plant.