Castresana C, de Carvalho F, Gheysen G, Habets M, Inzé D, Van Montagu M
Laboratorium voor Genetica, Rijksuniversiteit Gent, Belgium.
Plant Cell. 1990 Dec;2(12):1131-43. doi: 10.1105/tpc.2.12.1131.
The Nicotiana plumbaginifolia gn1 gene encoding a beta-1,3-glucanase isoform has been characterized. The gn1 product represents an isoform distinct from the previously identified tobacco beta-1,3-glucanases. By expressing gn1 in Escherichia coli, we have determined directly that the encoded protein does, indeed, correspond to a beta-1,3-glucanase. In N. plumbaginifolia, gn1 was found to be expressed in roots and older leaves. Transgenic tobacco plants containing the 5'-noncoding region of gn1 fused to the beta-glucuronidase (GUS) reporter gene also showed maximum levels of GUS activity in roots and older leaves. No detectable activity was present in the upper part of the transgenic plants with the exception of stem cells at the bases of emerging shoots. The expression conferred by the gn1 promoter was differentially induced in response to specific plant stress treatments. Studies of three plant-bacteria interactions showed high levels of GUS activity when infection resulted in a hypersensitive reaction. Increased gene expression was confined to cells surrounding the necrotic lesions. The observed expression pattern suggests that the characterized beta-1,3-glucanase plays a role both in plant development and in the defense response against pathogen infection.
已对编码β-1,3-葡聚糖酶同工型的烟草垂花烟草gn1基因进行了表征。gn1产物代表一种与先前鉴定的烟草β-1,3-葡聚糖酶不同的同工型。通过在大肠杆菌中表达gn1,我们直接确定了编码的蛋白质确实对应于一种β-1,3-葡聚糖酶。在烟草垂花烟草中,发现gn1在根和老叶中表达。含有与β-葡萄糖醛酸酶(GUS)报告基因融合的gn1 5'-非编码区的转基因烟草植株在根和老叶中也显示出最高水平的GUS活性。除了新出现芽基部的干细胞外,转基因植物上部未检测到活性。gn1启动子赋予的表达在响应特定的植物胁迫处理时受到差异诱导。对三种植物-细菌相互作用的研究表明,当感染导致过敏反应时,GUS活性水平很高。基因表达的增加局限于坏死病变周围的细胞。观察到的表达模式表明,所表征的β-1,3-葡聚糖酶在植物发育和对病原体感染的防御反应中均起作用。