Effective delivery of a nematode-repellent peptide using a root-cap-specific promoter
Effective delivery of a nematode-repellent peptide using a root-cap-specific promoter
复制标题
DOI：
10.1111/j.1467-7652.2010.00542.x
复制
发表时间：
2011-02-01
影响因子：
13.8
通讯作者：
Urwin, Peter E.
中科院分区：
文献类型：
--
作者：
Lilley, Catherine J.;Wang, Dong;Urwin, Peter E.
研究了拟南芥MDK4-20启动子对分泌型线虫驱避肽直接有效转基因表达的潜力。研究了其在转基因拟南芥和薯类植物中的表达模式。在两个物种中，它指导主要定位于根冠细胞的根特异性β -葡糖苷酶的表达。使用荧光计时器蛋白dsRED-E5确定，在分离的马铃薯根缘细胞中，MDK4-20启动子比常用的组成启动子CaMV35S保持活性的时间更长。在AtMDK4-20或CaMV35S控制下，表达线虫驱避肽的转基因拟南芥株系减少了甜菜囊线虫的建立。使用AtMDK4-20启动子的最佳品系显示出80%的抗性水平，与使用CaMV35S启动子的品系相当。在转基因马铃薯植株中，使用AtMDK4-20启动子对马铃薯包囊线虫（Globodera pallida）的抗性达到94.9 +/- 0.8%，而表达CaMV35S启动子的驱避肽株系的抗性为34.4 +/- 8.4%。这些结果确定了AtMDK4-20启动子的潜力，以限制驱避肽的表达，同时维持甚至提高囊线虫防御的功效。
P>The potential of the MDK4-20 promoter of Arabidopsis thaliana to direct effective transgenic expression of a secreted nematode-repellent peptide was investigated. Its expression pattern was studied in both transgenic Arabidopsis and Solanum tuberosum (potato) plants. It directed root-specific beta-glucuronidase expression in both species that was chiefly localized to cells of the root cap. Use of the fluorescent timer protein dsRED-E5 established that the MDK4-20 promoter remains active for longer than the commonly used constitutive promoter CaMV35S in separated potato root border cells. Transgenic Arabidopsis lines that expressed the nematode-repellent peptide under the control of either AtMDK4-20 or CaMV35S reduced the establishment of the beet cyst nematode Heterodera schachtii. The best line using the AtMDK4-20 promoter displayed a level of resistance > 80%, comparable to that of lines using the CaMV35S promoter. In transgenic potato plants, 94.9 +/- 0.8% resistance to the potato cyst nematode Globodera pallida was achieved using the AtMDK4-20 promoter, compared with 34.4 +/- 8.4% resistance displayed by a line expressing the repellent peptide from the CaMV35S promoter. These results establish the potential of the AtMDK4-20 promoter to limit expression of a repellent peptide whilst maintaining or even improving the efficacy of the cyst-nematode defence.