Barthels N, van der Lee F M, Klap J, Goddijn O J, Karimi M, Puzio P, Grundler F M, Ohl S A, Lindsey K, Robertson L, Robertson W M, Van Montagu M, Gheysen G, Sijmons P C
Department of Genetics, Flanders Interuniversity Institute for Biotechnology (VIB), Universiteit Gent, Belgium.
Plant Cell. 1997 Dec;9(12):2119-34. doi: 10.1105/tpc.9.12.2119.
In the quest for plant regulatory sequences capable of driving nematode-triggered effector gene expression in feeding structures, we show that promoter tagging is a valuable tool. A large collection of transgenic Arabidopsis plants was generated. They were transformed with a beta-glucuronidase gene functioning as a promoter tag. Three T-DNA constructs, pGV1047, p delta gusBin19, and pMOG553, were used. Early responses to nematode invasion were of primary interest. Six lines exhibiting beta-glucuronidase activity in syncytia induced by the beet cyst nematode were studied. Reporter gene activation was also identified in galls induced by root knot and ectoparasitic nematodes. Time-course studies revealed that all six tags were differentially activated during the development of the feeding structure. T-DNA-flanking regions responsible for the observed responses after nematode infection were isolated and characterized for promoter activity.
在寻找能够驱动线虫触发的效应基因在取食结构中表达的植物调控序列的过程中，我们发现启动子标签是一种有价值的工具。我们构建了大量转基因拟南芥植株。这些植株用作为启动子标签的β-葡萄糖醛酸酶基因进行转化。使用了三种T-DNA构建体，即pGV1047、pΔgusBin19和pMOG553。我们主要关注对线虫入侵的早期反应。研究了六个在甜菜孢囊线虫诱导的合胞体中表现出β-葡萄糖醛酸酶活性的株系。在根结线虫和外寄生线虫诱导的虫瘿中也鉴定到了报告基因的激活。时间进程研究表明，在取食结构发育过程中，所有六个标签均被差异激活。分离出负责线虫感染后观察到的反应的T-DNA侧翼区域，并对其启动子活性进行了表征。
