Jung Ho Won, Kim Ki Deok, Hwang Byung Kook
Division of Bioscience and Technology, College of Life and Environmental Sciences, Korea University, Anam-dong, Sungbuk-ku, Seoul, 136-713, Korea.
Planta. 2005 Jun;221(3):361-73. doi: 10.1007/s00425-004-1461-9. Epub 2005 Jan 15.
The 5' flanking region of the CALTPI gene, which encodes a basic lipid transfer protein, was isolated and characterized from the genomic DNA of Capsicum annuum. Four different regions of the promoter sequence of the CALTPI gene were fused to the beta-glucuronidase (GUS) coding region. In an Agrobacterium-mediated transient expression assay, the transcriptional activations of the promoter deletions were examined in tobacco leaves after infection with Pseudomonas syringae pv. tabaci, and treatment with ethylene and salicylic acid. The -808 bp region of the CALTPI gene promoter sequence exhibited full promoter activity. The W-box and ERE-box elements, which are essential for induction by all signals, were localized in the region between -555 bp and -391 bp upstream of the translation initiation site. A CALTPI transgene was then introduced under the control of the 35S promoter into the Arabidopsis ecotype Col-0. Transgenic Arabidopsis lines expressing the CALTPI gene developed rapidly compared to the wild-type plants, indicating that CALTPI may be involved in plant development. Overexpression of the CALTPI gene enhanced the resistance against infection by P. syringae pv. tomato and Botrytis cinerea. The transgenic plants expressing the CALTPI gene also showed high levels of tolerance to NaCl and drought stresses at various vegetative growth stages. No transcription of the PR-1, PR-2, PR-5, thionin, and RD29A genes was observed in untreated leaf tissues of the transgenic plants. The enhanced resistance to pathogen and environmental stresses in transgenic Arabidopsis correlated with the enhanced expression of the CALTPI gene.
编码一种碱性脂质转移蛋白的辣椒CALTPI基因的5'侧翼区,是从辣椒的基因组DNA中分离并鉴定出来的。CALTPI基因启动子序列的四个不同区域与β-葡萄糖醛酸酶(GUS)编码区融合。在农杆菌介导的瞬时表达试验中,用丁香假单胞菌番茄致病变种感染烟草叶片,并分别用乙烯和水杨酸处理后,检测启动子缺失片段的转录激活情况。CALTPI基因启动子序列的-808 bp区域表现出完整的启动子活性。对所有信号诱导至关重要的W-box和ERE-box元件,位于翻译起始位点上游-555 bp至-391 bp之间的区域。然后,在35S启动子的控制下,将CALTPI转基因导入拟南芥生态型Col-0。与野生型植物相比,表达CALTPI基因的转基因拟南芥株系生长迅速,这表明CALTPI可能参与植物发育。CALTPI基因的过表达增强了对番茄丁香假单胞菌和灰霉病菌感染的抗性。在各个营养生长阶段,表达CALTPI基因的转基因植物对NaCl和干旱胁迫也表现出高度耐受性。在转基因植物未处理的叶片组织中,未观察到PR-1、PR-2、PR-5、硫堇和RD29A基因的转录。转基因拟南芥对病原体和环境胁迫抗性的增强与CALTPI基因表达的增强相关。