Rancé I, Fournier J, Esquerré-Tugayé M T
Unite Mixte de Recherche 5546, Centre National de la Recherche Scientifique-Universite Paul Sabatier, Centre de Physiologie Vegetale, 118 Route de Narbonne, 31062 Toulouse Cedex 4, France.
Proc Natl Acad Sci U S A. 1998 May 26;95(11):6554-9. doi: 10.1073/pnas.95.11.6554.
Nicotiana tabacum 46-8 cultivar displays an incompatible interaction with race 0 of Phytophthora parasitica var. nicotianae (Ppn), a fungal pathogen of most tobacco cultivars. At the plant level, incompatibility is characterized by the induction of lipoxygenase (LOX, EC = 1.13.11.12) activity and localized hypersensitive cell death before defense gene activation. To evaluate the involvement of LOX in the onset of plant defense, tobacco 46-8 plants were genetically engineered using full-length or partial-length antisense (AS) tobacco LOX cDNA constructs. AS expression strongly reduced elicitor- and pathogen-induced LOX activity. Eight independent AS-LOX lines were selected and assayed for their response to Ppn. After root or stem inoculation with race 0, all AS-LOX lines but one displayed a compatible phenotype whereas control transformed plants, not containing the AS-LOX cassette, showed the typical incompatible reaction. The presence of the fungus in transgenic lines was demonstrated by PCR amplification of a Ppn-specific genomic sequence. A linear relationship was found between the extent of LOX suppression and the size of the lesion caused by the fungus. The AS-LOX plants also showed enhanced susceptibility toward the compatible fungus Rhizoctonia solani. The results demonstrate the strong involvement of LOX in the establishment of incompatibility in plant-microorganism interactions, consistent with its role in the defense of host plants.
烟草品种46 - 8与寄生疫霉烟草变种(Ppn)的0号生理小种表现出不亲和互作,Ppn是大多数烟草品种的一种真菌病原体。在植物水平上,不亲和表现为在防御基因激活之前脂氧合酶(LOX,EC = 1.13.11.12)活性的诱导和局部过敏细胞死亡。为了评估LOX在植物防御起始中的作用,利用全长或部分长度的反义(AS)烟草LOX cDNA构建体对烟草46 - 8植株进行了基因工程改造。AS表达强烈降低了激发子和病原体诱导的LOX活性。选择了8个独立的AS - LOX株系,并检测它们对Ppn的反应。用0号生理小种对根或茎进行接种后,除一个株系外,所有AS - LOX株系均表现出亲和表型,而不含AS - LOX盒的对照转化植株则表现出典型的不亲和反应。通过对Ppn特异性基因组序列的PCR扩增证明了转基因株系中真菌的存在。发现LOX抑制程度与真菌引起的病斑大小之间存在线性关系。AS - LOX植株对亲和真菌立枯丝核菌也表现出增强的敏感性。结果表明LOX在植物 - 微生物互作的不亲和建立中起重要作用,这与其在宿主植物防御中的作用一致。