Jun Sung-Hoon, Han Min-Jung, Lee Shinyoung, Seo Young Sam, Kim Woo Taek, An Gynheung
National Research Laboratory of Plant Functional Genomics, Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang, 790-784 Korea.
Plant Cell Physiol. 2004 Mar;45(3):281-9. doi: 10.1093/pcp/pch033.
EIN2 is a central signal transducer in the ethylene-signaling pathway, and a unique membrane-anchored protein. By screening a cDNA library, we have isolated a cDNA clone (OsEIN2) that encodes the rice EIN2 homolog. The full-length ORF clone was obtained by reverse transcriptase-polymerase chain reaction. OsEIN2 shares significant amino acid sequence similarity with Arabidopsis EIN2 (57% similarity and 42% identity). Both the numbers and positions of introns and exons in the OsEIN2 and AtEIN2 coding regions are also conserved. To address whether this structural similarity is indicative of functional conservation of the corresponding proteins, we also generated transgenic lines expressing the antisense construct of OsEIN2. Those plants were stunted and shoot elongation was severely inhibited. Their phenotypes were similar to that found with wild-type rice seedlings that were treated with AgNO3, an ethylene signal inhibitor. In the OsEIN2 antisense plants, the expression levels of two ethylene-responsive genes, SC129 and SC255, were decreased compared with the wild types. These results suggest that OsEIN2 is a positive component of the ethylene-signaling pathway in rice, just as AtEIN2 is in Arabidopsis: Our antisense transgenic plants produced approximately 3.5 times more ethylene than the wild-type plants. Expression analysis of rice ACS and ACO genes showed that the transcript levels of OsACS1 and OsACO1 were elevated in the transgenic plants.
EIN2是乙烯信号通路中的核心信号转导因子,是一种独特的膜锚定蛋白。通过筛选cDNA文库,我们分离出一个编码水稻EIN2同源物的cDNA克隆(OsEIN2)。通过逆转录聚合酶链反应获得了全长开放阅读框克隆。OsEIN2与拟南芥EIN2具有显著的氨基酸序列相似性(相似性为57%,同一性为42%)。OsEIN2和AtEIN2编码区的内含子和外显子的数量和位置也保守。为了研究这种结构相似性是否表明相应蛋白质的功能保守,我们还构建了表达OsEIN2反义构建体的转基因株系。这些植株发育不良,茎伸长受到严重抑制。它们的表型与用乙烯信号抑制剂硝酸银处理的野生型水稻幼苗相似。在OsEIN2反义植株中,与野生型相比,两个乙烯响应基因SC129和SC255的表达水平降低。这些结果表明,OsEIN2是水稻乙烯信号通路中的一个正向组分,就像AtEIN2在拟南芥中一样:我们的反义转基因植株产生的乙烯比野生型植株多约3.5倍。水稻ACS和ACO基因的表达分析表明,转基因植株中OsACS1和OsACO1的转录水平升高。