Green tissue-specific co-expression of chitinase and oxalate oxidase 4 genes in rice for enhanced resistance against sheath blight
Green tissue-specific co-expression of chitinase and oxalate oxidase 4 genes in rice for enhanced resistance against sheath blight
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DOI：
10.1007/s00425-015-2398-x
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发表时间：
2016-01-01
期刊：
影响因子：
4.3
通讯作者：
Datta, Karabi
中科院分区：
文献类型：
--
作者：
Karmakar, Subhasis;Molla, Kutubuddin Ali;Datta, Karabi
水稻绿色组织特异性的两种防御相关基因（OsCHI11和OsOXO4）同时过表达，导致水稻对白叶枯病（R. solani）具有显著的抗性，而不会影响任何重要的农艺性状。从水稻中克隆的两个防御相关基因（OsOXO4和OsCHI11）过表达可有效增强水稻对枯丝核菌的抗性。这些基因在两种不同的绿色组织特异性启动子的控制下表达，即玉米磷酸烯醇丙酮酸羧化酶基因启动子PEPC和水稻顺式作用544 bp DNA元件，位于D54O翻译起始位点P （D54O-544）的上游。通过PCR筛选T-0转基因水稻植株，并通过T-1后代植株的Southern杂交证实基因整合。OsOXO4和OsCHI11在所有被试植物中均成功表达。过表达转基因植株中PR基因的表达在病原菌侵染后显著增加。感染后，转基因植株与野生型植株相比，过氧化氢水平升高，活性氧清除酶活性显著改变，膜损伤减少。在枯丝核菌毒素试验、离体叶片接种试验和植物体内生物试验中，与非转基因对照植物相比，转基因植物的疾病症状显著减少。这是首次报道在转基因水稻中，由两种绿色组织特异性启动子驱动的两种不同PR基因过表达，从而增强了水稻的抗纹枯病能力。
Green tissue-specific simultaneous overexpression of two defense-related genes ( OsCHI11 & OsOXO4 ) in rice leads to significant resistance against sheath blight pathogen ( R. solani ) without distressing any agronomically important traits.Overexpressing two defense-related genes (OsOXO4 and OsCHI11) cloned from rice is effective at enhancing resistance against sheath blight caused by Rhizoctonia solani. These genes were expressed under the control of two different green tissue-specific promoters, viz. maize phosphoenolpyruvate carboxylase gene promoter, PEPC, and rice cis-acting 544-bp DNA element, immediately upstream of the D54O translational start site, P (D54O-544) . Putative T-0 transgenic rice plants were screened by PCR and integration of genes was confirmed by Southern hybridization of progeny (T-1) rice plants. Successful expression of OsOXO4 and OsCHI11 in all tested plants was confirmed. Expression of PR genes increased significantly following pathogen infection in overexpressing transgenic plants. Following infection, transgenic plants exhibited elevated hydrogen peroxide levels, significant changes in activity of ROS scavenging enzymes and reduced membrane damage when compared to their wild-type counterpart. In a Rhizoctonia solani toxin assay, a detached leaf inoculation test and an in vivo plant bioassay, transgenic plants showed a significant reduction in disease symptoms in comparison to non-transgenic control plants. This is the first report of overexpression of two different PR genes driven by two green tissue-specific promoters providing enhanced sheath blight resistance in transgenic rice.