Department of Plant Molecular Biology, University of Delhi, South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi, 110 021, India.
Plant Mol Biol. 2011 Jan;75(1-2):35-51. doi: 10.1007/s11103-010-9702-8. Epub 2010 Oct 23.
To elucidate the effect of high temperature, wheat plants (Triticum aestivum cv. CPAN 1676) were given heat shock at 37 and 42°C for 2 h, and responsive genes were identified through PCR-Select Subtraction technology. Four subtractive cDNA libraries, including three forward and one reverse subtraction, were constructed from three different developmental stages. A total of 5,500 ESTs were generated and 3,516 high quality ESTs submitted to Genbank. More than one-third of the ESTs generated fall in unknown/no hit category upon homology search through BLAST analysis. Differential expression was confirmed by cDNA macroarray and by northern/RT-PCR analysis. Expression analysis of wheat plants subjected to high temperature stress, after 1 and 4 days of recovery, showed fast recovery in seedling tissue. However, even after 4 days, recovery was negligible in the developing seed tissue after 2 h of heat stress. Ten selected genes were analyzed in further detail including one unknown protein and a new heat shock factor, by quantitative real-time PCR in an array of 35 different wheat tissues representing major developmental stages as well as different abiotic stresses. Tissue specificity was examined along with cross talk with other abiotic stresses and putative signalling molecules.
为了阐明高温的影响，将小麦植株（Triticum aestivum cv. CPAN 1676）在 37 和 42°C 下热休克 2 小时，并通过 PCR-Select 消减技术鉴定响应基因。从三个不同的发育阶段构建了四个消减 cDNA 文库，包括三个正向和一个反向消减。共产生了 5500 个 EST，其中 3516 个高质量的 EST 提交给 Genbank。通过 BLAST 分析的同源性搜索，生成的 EST 中有超过三分之一属于未知/无命中类别。通过 cDNA 宏阵列和 northern/RT-PCR 分析证实了差异表达。对高温胁迫后 1 天和 4 天的小麦植株进行表达分析，结果表明幼苗组织恢复迅速。然而，即使在 2 小时的热胁迫后 4 天，发育中的种子组织的恢复也可以忽略不计。通过对代表主要发育阶段以及不同非生物胁迫的 35 种不同小麦组织的阵列进行定量实时 PCR，对 10 个选定基因进行了更详细的分析，包括一个未知蛋白和一个新的热休克因子。还研究了组织特异性以及与其他非生物胁迫和假定信号分子的相互作用。
