Akiyama Takashi, Jin Shigeki, Yoshida Midori, Hoshino Tamotsu, Opassiri Rodjana, Ketudat Cairns James R
National Agricultural Research Center for Hokkaido Region, 1 Hitsujigaoka, Toyohira-ku, Sapporo 062-8555, Japan.
J Plant Physiol. 2009 Nov 1;166(16):1814-25. doi: 10.1016/j.jplph.2009.06.002. Epub 2009 Jun 30.
We isolated two rice endo-(1,3;1,4)-beta-glucanase genes, denoted OsEGL1 and OsEGL2, which encoded proteins that shared 64% amino acid sequence identity. Both the OsEGL1 and OsEGL2 genes were successfully expressed in Escherichia coli to produce functional proteins. Purified OsEGL1 and OsEGL2 proteins hydrolyzed (1,3;1,4)-beta-glucans, but not (1,3;1,6)-beta-linked or (1,3)-beta-linked glucopolysaccharides nor carboxymethyl cellulose, similar to previously characterized grass endo-(1,3;1,4)-beta-glucanases. RNA blot analysis revealed that the OsEGL1 gene is expressed constitutively not only in young roots of rice seedlings, but also in mature roots of adult rice plants. Little or no expression of the OsEGL2 gene was observed in all tissues or treatments tested, but database and RT-PCR analysis indicated it is expressed in ripening panicle. In rice seedling leaves, OsEGL1 gene expression significantly increased in response to methyl jasmonate, abscisic acid, ethephon and mechanical wounding. Mechanical wounding also increased the leaf elongation rate in rice seedlings by 16% relative to that of control seedlings at day 4 after treatment. The increase in the leaf elongation rate of rice seedlings treated under mechanical wounding was concomitant with an increase in OsEGL1 expression levels in seedling leaves.
我们分离出了两个水稻内切 -(1,3;1,4)-β - 葡聚糖酶基因,分别命名为OsEGL1和OsEGL2,它们编码的蛋白质氨基酸序列同一性为64%。OsEGL1和OsEGL2基因均在大肠杆菌中成功表达,产生了具有功能的蛋白质。纯化后的OsEGL1和OsEGL2蛋白能够水解(1,3;1,4)-β - 葡聚糖,但不能水解(1,3;1,6)-β - 连接或(1,3)-β - 连接的葡聚糖多糖以及羧甲基纤维素,这与之前鉴定的禾本科内切 -(1,3;1,4)-β - 葡聚糖酶相似。RNA印迹分析表明,OsEGL1基因不仅在水稻幼苗的幼根中组成型表达,在成年水稻植株的成熟根中也有表达。在所有测试的组织或处理中,几乎未观察到OsEGL2基因的表达,但数据库和逆转录 - 聚合酶链反应分析表明它在成熟穗中表达。在水稻幼苗叶片中,茉莉酸甲酯、脱落酸、乙烯利和机械损伤均能显著提高OsEGL1基因的表达。与处理后第4天的对照幼苗相比,机械损伤还使水稻幼苗的叶片伸长率提高了16%。水稻幼苗在机械损伤处理下叶片伸长率的增加与幼苗叶片中OsEGL1表达水平的增加是同步的。