Wang Ningbo, Liu Mengjie, Guo Lihua, Yang Xiufen, Qiu Dewen
The State Key Laboratory for Biology of Plant Disease and Insect Pests, Institute of Plant protection, Chinese Academy of Agricultural sciences, No. 12 Zhongguancun South Street, Beijing 100081, China.
Int J Biol Sci. 2016 Apr 28;12(6):757-67. doi: 10.7150/ijbs.14333. eCollection 2016.
Here we reported a novel protein elicitor from Bacillus amyloliquefaciens NC6 induced systemic resistance (ISR) in tobacco. The purification was executed by ion-exchange chromatography, native-page extraction and HPLC, and the amino acid sequence was identified by mass spectrometry. This recombinant elicitor protein, expressed in Escherichia coli by an E1 expression vector, had good thermal stability, and the elicitor caused a clearly defined hypersensitive response (HR) necrosis in tobacco leaves. It could also trigger early defence events, including generation of reactive oxygen species (H2O2 and O2 (-)) and phenolic-compound accumulation. Quantitative real-time PCR (Q-RT-PCR) results indicated that several plant defence genes, including the salicylic acid (SA)-responsive PR1a, PR1b, PR5, and phenylalanine ammonia lyase (PAL), as well as the jasmonic acid (JA)-responsive PDF1.2 and CORONATINE INSENSITIVE 1 (COI1), were all up-regulated. Moreover, infiltration conferred systemic resistance against a broad spectrum of pathogens, including Tobacco mosaic virus (TMV) and the fungal pathogen Botrytis cinerea.
在此，我们报道了一种来自解淀粉芽孢杆菌NC6的新型蛋白激发子，其可诱导烟草产生系统抗性（ISR）。通过离子交换色谱、非变性聚丙烯酰胺凝胶电泳提取和高效液相色谱进行纯化，并通过质谱鉴定氨基酸序列。这种重组激发子蛋白由E1表达载体在大肠杆菌中表达，具有良好的热稳定性，且该激发子在烟草叶片中引起了明确的过敏反应（HR）坏死。它还能引发早期防御事件，包括活性氧（H2O2和O2(-)）的产生以及酚类化合物的积累。实时定量聚合酶链反应（Q-RT-PCR）结果表明，包括水杨酸（SA）响应的PR1a、PR1b、PR5和苯丙氨酸解氨酶（PAL），以及茉莉酸（JA）响应的PDF1.2和冠菌素不敏感蛋白1（COI1）在内的多个植物防御基因均上调。此外，浸润赋予了对多种病原体的系统抗性，包括烟草花叶病毒（TMV）和真菌病原体灰葡萄孢。
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