USDA-ARS Plant Protection Research Unit, Department of Plant Pathology, Cornell University, Ithaca, NY 14853, USA.
Mol Plant Pathol. 2007 Jul;8(4):423-36. doi: 10.1111/j.1364-3703.2007.00403.x.
Emerging evidence suggests that plant cell-wall-modifying enzymes induced by root-parasitic nematodes play important roles in feeding cell formation. We previously identified a tobacco endo-beta-1,4-glucanase (cellulase) gene, NtCel7, that was strongly induced in both root-knot and cyst nematode feeding cells. To characterize further the developmental and nematode-responsive regulation of NtCel7, we isolated the NtCel7 promoter and analysed its expression over a time course of nematode infection and in response to auxin, gibberellin, ethylene and sucrose in soybean and tomato hairy roots and in Arabidopsis containing the NtCel7 promoter fused to the beta-glucuronidase (GUS) reporter gene. Histochemical analyses of transgenic plant materials revealed that the NtCel7 promoter exhibited a unique organ-specific expression pattern during plant development suggestive of important roles for NtCel7 in both vegetative and reproductive growth. In all plant species tested, strong GUS expression was observed in root tips and lateral root primordia of uninfected roots with weaker expression in the root vasculature. Further analyses of transgenic Arabidopsis plants revealed expression in shoot and root meristems and the vasculature of most organs during plant development. We also determined that the NtCel7 promoter was induced by auxin, but not gibberellin, ethylene or sucrose. Moreover, strong GUS activity was observed in both cyst and root-knot nematode-induced feeding sites in transgenic roots of soybean, tomato and Arabidopsis. The conserved developmental and nematode-responsive expression of the NtCel7 promoter in heterologous plants indicates that motifs of this regulatory element play a fundamental role in regulating NtCel7 gene expression within nematode feeding sites and that this regulation may be mediated by auxin.
新出现的证据表明,根寄生线虫诱导的植物细胞壁修饰酶在形成取食细胞中发挥重要作用。我们之前鉴定了一种烟草内切-β-1,4-葡聚糖酶(纤维素酶)基因 NtCel7,该基因在根结线虫和孢囊线虫的取食细胞中均被强烈诱导。为了进一步研究 NtCel7 的发育和线虫响应调节,我们分离了 NtCel7 启动子,并分析了其在线虫感染过程中的表达,以及在大豆和番茄毛状根和含有 NtCel7 启动子与β-葡萄糖醛酸酶(GUS)报告基因融合的拟南芥中对生长素、赤霉素、乙烯和蔗糖的响应。对转基因植物材料的组织化学分析表明,NtCel7 启动子在植物发育过程中表现出独特的器官特异性表达模式,提示 NtCel7 在营养和生殖生长中均具有重要作用。在所测试的所有植物物种中,未感染根的根尖和侧根原基中观察到强烈的 GUS 表达,根系脉管系统中的表达较弱。对转基因拟南芥植物的进一步分析表明,在植物发育过程中,茎和根分生组织以及大多数器官的脉管系统中均有表达。我们还确定 NtCel7 启动子被生长素诱导,但不被赤霉素、乙烯或蔗糖诱导。此外,在大豆、番茄和拟南芥的转基因根中,在孢囊线虫和根结线虫诱导的取食部位也观察到强烈的 GUS 活性。NtCel7 启动子在异源植物中的保守发育和线虫响应表达表明,该调节元件的基序在调节线虫取食部位的 NtCel7 基因表达中发挥着重要作用,并且这种调节可能是由生长素介导的。