Phytopathology. 1997 May;87(5):494-9. doi: 10.1094/PHYTO.1997.87.5.494.
ABSTRACT A chimeric gene fusion cassette, consisting of a secretory sequence from barley alpha-amylase joined to a modified cecropin (MB39) coding sequence and placed under control of the promoter and terminator from the potato proteinase inhibitor II (PiII) gene, was introduced into tobacco by Agrobacterium-mediated transformation. Transgenic and control plants reacted differently when inoculated with tobacco wildfire pathogen Pseudomonas syringae pv. tabaci at various cell concentrations. With control plants (transformed with a PiII-GUS [beta-D-glucuronidase] gene fusion), necrosis was clearly visible in leaf tissue infiltrated with bacterial inoculum levels of 10(2), 10(3), 10(4), 10(5), and 10(6) CFU/ml. With MB39-transgenic plants, however, necrosis was observed only in the areas infiltrated with the two highest levels (10(5) and 10(6) CFU/ml). No necrosis was evident in areas infiltrated with bacterial concentrations of 10(4) CFU/ml or less. Bacterial multiplication in leaves of MB39-transgenic plants was suppressed more than 10-fold compared to control plants, and absence of disease symptom development was associated with this growth suppression. We conclude that the pathogen-induced promoter and the secretory sequence were competent elements for transforming a cecropin gene into an effective disease-control gene for plants.
摘要 一个嵌合基因融合盒,由大麦α-淀粉酶的分泌序列与修饰的抗菌肽(MB39)编码序列组成,并置于马铃薯蛋白酶抑制剂 II(PiII)基因的启动子和终止子的控制之下,通过根癌农杆菌介导的转化被引入烟草中。当用不同细胞浓度的烟草野火病原体丁香假单胞菌 pv。tabaci 接种时,转基因和对照植物的反应不同。用对照植物(转化为 PiII-GUS[β-D-葡萄糖醛酸酶]基因融合体),用 10(2)、10(3)、10(4)、10(5)和 10(6)CFU/ml 的细菌接种物水平渗透的叶片组织中可明显观察到坏死。然而,在用 MB39-转基因植物处理时,仅在渗透两个最高水平(10(5)和 10(6)CFU/ml)的区域观察到坏死。在渗透细菌浓度为 10(4)CFU/ml 或更低的区域,没有明显的坏死。与对照植物相比,MB39-转基因植物叶片中的细菌繁殖被抑制了 10 倍以上,并且没有疾病症状的发展与这种生长抑制有关。我们得出结论,病原体诱导的启动子和分泌序列是将抗菌肽基因转化为植物有效疾病控制基因的功能元件。