Linthorst H J, Melchers L S, Mayer A, van Roekel J S, Cornelissen B J, Bol J F
Department of Biochemistry, Leiden University, The Netherlands.
Proc Natl Acad Sci U S A. 1990 Nov;87(22):8756-60. doi: 10.1073/pnas.87.22.8756.
Healthy tobacco plants accumulate beta-1,3-glucanases (glucan endo-1,3-beta-glucosidase; EC 3.2.1.39) in their roots and in specific parts of the flowers. After infection with tobacco mosaic virus, acidic and basic beta-1,3-glucanases are induced in the inoculated and virus-free leaves of the plant. An analysis of cDNA clones demonstrated that at least five genes for acidic beta-1,3-glucanases are induced after tobacco mosaic virus infection. Southern blot analysis indicated that the tobacco genome contains approximately eight genes for acidic beta-1,3-glucanases and a smaller number of genes encoding basic beta-1,3-glucanases. Genes from both gene families were cloned and sequenced. The basic isozymes contain a C-terminal extension that is cleaved off during their targeting to the vacuoles. This extension is absent in the acidic isozymes, which accumulate extracellularly. Northern blot hybridization showed that genes encoding acidic and basic beta-1,3-glucanases are strongly induced after tobacco mosaic virus infection or salicylate treatment of tobacco. The cloning of these genes is a first step toward the identification of regulatory elements involved in their coordinate induction.
健康的烟草植株在其根部和花朵的特定部位积累β-1,3-葡聚糖酶(葡聚糖内切-1,3-β-葡糖苷酶;EC 3.2.1.39)。感染烟草花叶病毒后,在植株接种病毒的叶片和未感染病毒的叶片中会诱导产生酸性和碱性β-1,3-葡聚糖酶。对cDNA克隆的分析表明,烟草花叶病毒感染后至少有五个酸性β-1,3-葡聚糖酶基因被诱导。Southern印迹分析表明,烟草基因组中大约有八个酸性β-1,3-葡聚糖酶基因和较少数量的编码碱性β-1,3-葡聚糖酶的基因。这两个基因家族的基因都被克隆并测序。碱性同工酶含有一个C末端延伸区,在其靶向液泡的过程中会被切除。酸性同工酶中不存在这种延伸区,它们在细胞外积累。Northern印迹杂交表明,编码酸性和碱性β-1,3-葡聚糖酶的基因在烟草花叶病毒感染或水杨酸处理烟草后被强烈诱导。这些基因的克隆是鉴定参与其协同诱导的调控元件的第一步。