Lee G J, Shin R, Park C J, Yoo T H, Paek K H
Graduate School of Biotechnology, Korea University, Seoul.
Mol Cells. 2001 Oct 31;12(2):250-6.
A cDNA library was constructed with mRNA extracted from TMV resistant hot pepper plants 24 and 48 h after inoculation by TMV. The library was screened differentially with radio-labeled cDNA synthesized with mRNA from the leaves of either TMV-inoculated or mock-inoculated hot pepper plants. CaSAR8.2 clone was one of the clones isolated by this differential screening. The predicted amino acid sequence of CaSAR8.2 has a homology of 52% similarity to that of tobacco SAR8.2 genes. Southern blot analysis showed that a multigene family of CaSAR8.2 was present in the hot pepper genome. Transcripts homologous to CaSAR8.2 accumulated abundantly in the leaves and the flowers, but little in other tissues. CaSAR8.2 gene expression was induced by avirulent pathotype TMV-P0 inoculation but not by virulent TMV-P1.2 inoculation. Effects of exogenously applied abiotic elicitors on CaSAR8.2 expression were also examined. Salicylic acid and ethephon treatments caused a rapid accumulation of CaSAR8.2 transcripts in pepper leaves and methyl jasmonate treatment slightly induced the expression of CaSAR8.2. A strain of Xanthomonas campestris pv. vesicatoria (Xcv) that contains an avirulence gene avrBs2, was infiltrated into the leaves of a pepper cultivar containing the Bs2 resistance gene. A marked induction of CaSAR8.2 gene expression was observed in Xcv-infiltrated leaves. These results suggest possible roles of CaSAR8.2 as pathogenesis-related protein against varieties of pathogens including virus and bacteria.
用从接种烟草花叶病毒(TMV)24小时和48小时后的抗TMV辣椒植株中提取的mRNA构建了一个cDNA文库。用从接种TMV或模拟接种的辣椒植株叶片中合成的放射性标记cDNA对该文库进行差异筛选。CaSAR8.2克隆是通过这种差异筛选分离出的克隆之一。CaSAR8.2预测的氨基酸序列与烟草SAR8.2基因的氨基酸序列有52%的相似性。Southern杂交分析表明,CaSAR8.2在辣椒基因组中存在一个多基因家族。与CaSAR8.2同源的转录本在叶片和花中大量积累,但在其他组织中很少。无毒型TMV-P0接种可诱导CaSAR8.2基因表达,而有毒型TMV-P1.2接种则不能。还检测了外源施加的非生物激发子对CaSAR8.2表达的影响。水杨酸和乙烯利处理导致辣椒叶片中CaSAR8.2转录本迅速积累,茉莉酸甲酯处理轻微诱导了CaSAR8.2的表达。将含有无毒基因avrBs2的野油菜黄单胞菌辣椒斑点病致病型菌株(Xcv)渗入含有Bs2抗性基因的辣椒品种叶片中。在Xcv渗入的叶片中观察到CaSAR8.2基因表达的显著诱导。这些结果表明CaSAR8.2作为一种病程相关蛋白可能对包括病毒和细菌在内的多种病原体发挥作用。