Wang Ren, Shen Wenbiao, Liu Linglong, Jiang Ling, Liu Yuqiang, Su Ning, Wan Jianmin
State Key Laboratory for Crop Genetics & Germplasm Enhancement, Nanjing Agricultural University; Research Center of Plant Gene Engineering, Nanjing, Jiangsu Province 210095, PR China.
Plant Mol Biol. 2008 Mar;66(4):401-14. doi: 10.1007/s11103-007-9278-0. Epub 2008 Jan 5.
OsLOX1 is a novel full-length cDNA isolated from developing rice seeds. We have examined its biochemical properties and expression patterns. The protein has dual positional specificity, as it releases both C-9 and C-13 oxidized products in a 4:3 ratio. OsLOX1 transcripts were detected at low abundance in immature seeds and newly germinated seedlings, but accumulate rapidly and transiently in response to wounding or brown planthopper (BPH) attack, reaching a peak 3 h after wounding and 6 h after insect feeding. We produced transgenic rice lines carrying either sense or antisense constructs under the control of a cauliflower mosaic virus 35S promoter, and these rice lines showed altered OsLOX1 activity. In all of the antisense lines and more than half of the sense lines the expression levels of OsLOX1, the levels of enzyme activity, and the levels of the endogenous OsLOX1 products (jasmonic acid, (Z)-3-hexenal and colneleic acid) at 6, 48, and 48 h after BPH feeding respectively, were below the levels found in non-transgenic control plants; yet, the levels in the remaining sense transformants were enhanced relative to controls. Transformants with a lower level of OsLOX1 expression were less able to tolerate BPH attack, while those with enhanced OsLOX1 expression were more resistant. Our data suggest that the OsLOX1 product is involved in tolerance of the rice plant to wounding and BPH attack.
OsLOX1是从发育中的水稻种子中分离出的一种新型全长cDNA。我们已经研究了它的生化特性和表达模式。该蛋白质具有双重位置特异性,因为它以4:3的比例释放C-9和C-13氧化产物。在未成熟种子和新发芽的幼苗中检测到低丰度的OsLOX1转录本,但在受到创伤或褐飞虱(BPH)攻击后会迅速且短暂地积累,在受伤后3小时和昆虫取食后6小时达到峰值。我们构建了在花椰菜花叶病毒35S启动子控制下携带正义或反义构建体的转基因水稻品系,这些水稻品系显示出OsLOX1活性的改变。在所有反义品系和超过一半的正义品系中,分别在褐飞虱取食后6、48和48小时,OsLOX1的表达水平、酶活性水平以及内源性OsLOX1产物(茉莉酸、(Z)-3-己烯醛和科尼酸)的水平均低于非转基因对照植物中的水平;然而,其余正义转化体中的水平相对于对照有所提高。OsLOX1表达水平较低的转化体对褐飞虱攻击的耐受性较差,而OsLOX1表达增强的转化体则更具抗性。我们的数据表明,OsLOX1产物参与了水稻植株对创伤和褐飞虱攻击的耐受性。