Yau Chi Ping, Wang Lingjian, Yu Manda, Zee Sze Yong, Yip Wing Kin
Department of Botany, University of Hong Kong, Pokfulam Road, Hong Kong.
J Exp Bot. 2004 Mar;55(397):547-56. doi: 10.1093/jxb/erh055. Epub 2004 Jan 30.
Five ethylene receptor genes, OS-ERS1, OS-ERS2, OS-ETR2, OS-ETR3, and OS-ETR4 were isolated and characterized from rice. The genomic structure of OS-ERS1 and OS-ERS2 revealed that the introns within the coding sequences occurred in conserved positions to those of At-ETR1 and At-ERS1, whereas each of the OS-ETR2, OS-ETR3, and OS-ETR4 genes contained 1 intron within its coding region located at a position equivalent to those of At-ERS2, At-ETR2, and At-EIN4. Deduced amino acid sequences of OS-ERS1, OS-ERS2, OS-ETR2, OS-ETR3, and OS-ETR4 showed that they exhibited significant homology to the prokaryotic two-component signal transducer and a wide range of ethylene receptors in a variety of plant species. Northern analysis revealed that the level of OS-ETR2 mRNA was markedly elevated either by the exogenous application of IAA or by ethylene treatment in young etiolated rice seedlings, whereas the OS-ERS1 transcript level was only slightly induced under the same experimental conditions. Pretreatment with silver prevented IAA-induced and ethylene-induced accumulation of both mRNAs (OS-ERS1 and OS-ETR2). However, the abundance of OS-ERS2 mRNA was shown to be down-regulated by both IAA and ethylene treatments, indicating that it was not positively regulated by ethylene. Analysis of the expression of the three ethylene receptor genes in different tissues of rice has unravelled their corresponding tissue-specificity in which OS-ERS1 was constitutively expressed in considerable amounts in all tissues studied, while OS-ERS2 and OS-ETR2 exhibited differential expression patterns in different tissues of rice. Moreover, higher levels of these three mRNAs were commonly observed in anthers when compared with their corresponding levels in other tissues, suggesting the important role played by ethylene involved in the regulation of pollen development in rice. Among the five ethylene receptor genes, the expression levels of both OS-ETR3 and OS-ETR4 were too low to be detected by the northern blot analysis. Results from RT-PCR illustrated that both mRNAs were present in young green rice seedlings and anthers.
从水稻中分离并鉴定了五个乙烯受体基因,即OS-ERS1、OS-ERS2、OS-ETR2、OS-ETR3和OS-ETR4。OS-ERS1和OS-ERS2的基因组结构显示,编码序列中的内含子出现在与At-ETR1和At-ERS1的内含子保守位置相同的位置,而OS-ETR2、OS-ETR3和OS-ETR4基因中的每一个在其编码区域内都含有1个内含子,其位置与At-ERS2、At-ETR2和At-EIN4的内含子位置相当。OS-ERS1、OS-ERS2、OS-ETR2、OS-ETR3和OS-ETR4的推导氨基酸序列表明,它们与原核双组分信号转导器以及多种植物物种中的多种乙烯受体具有显著同源性。Northern分析表明,在幼嫩黄化水稻幼苗中,外源施加IAA或乙烯处理均可显著提高OS-ETR2 mRNA的水平,而在相同实验条件下,OS-ERS1转录本水平仅略有诱导。用银预处理可阻止IAA诱导和乙烯诱导的两种mRNA(OS-ERS1和OS-ETR2)的积累。然而,OS-ERS2 mRNA的丰度显示在IAA和乙烯处理下均下调,表明它不受乙烯的正向调控。对水稻不同组织中三个乙烯受体基因表达的分析揭示了它们相应的组织特异性,其中OS-ERS1在所有研究组织中均大量组成型表达,而OS-ERS2和OS-ETR2在水稻不同组织中表现出差异表达模式。此外,与其他组织中的相应水平相比,在花药中通常观察到这三种mRNA的水平更高,这表明乙烯在水稻花粉发育调控中发挥重要作用。在这五个乙烯受体基因中,OS-ETR3和OS-ETR4的表达水平太低,无法通过Northern印迹分析检测到。RT-PCR结果表明,两种mRNA都存在于幼嫩绿色水稻幼苗和花药中。