Department of Plant Molecular Biology, University of Delhi, New Delhi, India.
Plant Cell Rep. 2012 Jan;31(1):237-51. doi: 10.1007/s00299-011-1160-5. Epub 2011 Oct 5.
Molecular dissection and a deeper analysis of the heat stress response mechanism in wheat have been poorly understood so far. This study delves into the molecular basis of action of TaMIPS, a heat stress-inducible enzyme that was identified through PCR-select subtraction technology, which is named here as TaMIPS2. MIPS (L-Myo-inositol-phosphate synthase) is important for the normal growth and development in plants. Expression profiling showed that TaMIPS2 is expressed during different developing seed stages upon heat stress. Also, the transcript levels increase in unfertilized ovaries and significant amounts are present during the recovery period providing evidence that MIPS is crucial for its role in heat stress recovery and flower development. Alternatively spliced forms from rice and Arabidopsis were also identified and their expression analysis revealed that apart from heat stress, some of the spliced variants were also inducible by drought, NaCl, Cold, ABA, BR, SA and mannitol. In silico promoter analysis revealed various cis-elements that could contribute for the differential regulation of MIPS in different plant systems. Phylogenetic analysis indicated that MIPS are highly conserved among monocots and dicots and TaMIPS2 grouped specifically with monocots. Comparative analyses was undertaken by different experimental approaches, i.e., semi-quantitative RT-PCR, quantitative RT-PCR, Genevestigator as a reference expression tool and motif analysis to predict the possible function of TaMIPS2 in regulating the different aspects of plant development under abiotic stress in wheat.
迄今为止,小麦耐热反应机制的分子解析和深入分析仍知之甚少。本研究通过 PCR-select 差减技术鉴定了一种热诱导酶 TaMIPS 的作用分子基础,将其命名为 TaMIPS2。MIPS(L-肌醇磷酸合酶)对植物的正常生长发育很重要。表达谱分析表明,TaMIPS2 在受到热胁迫时不同发育种子阶段表达。此外,在未受精的子房中转录水平增加,在恢复期有大量表达,这表明 MIPS 在耐热恢复和花发育中起关键作用。还鉴定了来自水稻和拟南芥的选择性剪接形式,其表达分析表明,除了热胁迫外,一些剪接变体还可被干旱、NaCl、冷、ABA、BR、SA 和甘露醇诱导。启动子的计算机分析揭示了各种顺式元件,这些元件可能有助于 MIPS 在不同植物系统中的差异调控。系统发育分析表明 MIPS 在单子叶植物和双子叶植物中高度保守,TaMIPS2 与单子叶植物特异性聚集。通过不同的实验方法进行了比较分析,即半定量 RT-PCR、定量 RT-PCR、Genevestigator 作为参考表达工具和基序分析,以预测 TaMIPS2 在调节小麦非生物胁迫下植物发育的不同方面的可能功能。