Department of Phytopathology, University of Passo Fundo (UPF), BR 285, PO BOX 611, Passo Fundo, RS, 99052-900, Brazil.
World J Microbiol Biotechnol. 2012 Apr;28(4):1475-81. doi: 10.1007/s11274-011-0949-2. Epub 2011 Nov 19.
Based on the premise of symbiotic control, we genetically modified the citrus endophytic bacterium Methylobacterium extorquens, strain AR1.6/2, and evaluated its capacity to colonize a model plant and its interaction with Xylella fastidiosa, the causative agent of Citrus Variegated Chlorosis (CVC). AR1.6/2 was genetically transformed to express heterologous GFP (Green Fluorescent Protein) and an endoglucanase A (EglA), generating the strains ARGFP and AREglA, respectively. By fluorescence microscopy, it was shown that ARGFP was able to colonize xylem vessels of the Catharanthus roseus seedlings. Using scanning electron microscopy, it was observed that AREglA and X. fastidiosa may co-inhabit the C. roseus vessels. M. extorquens was observed in the xylem with the phytopathogen X. fastidiosa, and appeared to cause a decrease in biofilm formation. AREglA stimulated the production of resistance protein, catalase, in the inoculated plants. This paper reports the successful transformation of AR1.6/2 to generate two different strains with a different gene each, and also indicates that AREglA and X. fastidiosa could interact inside the host plant, suggesting a possible strategy for the symbiotic control of CVC disease. Our results provide an enhanced understanding of the M. extorquens-X. fastidiosa interaction, suggesting the application of AR1.6/2 as an agent of symbiotic control.
基于共生控制的前提,我们对柑橘内生菌甲基杆菌(Methylobacterium extorquens)菌株 AR1.6/2 进行了基因改造,并评估了其定植模式植物的能力及其与黄单胞菌(Xylella fastidiosa)的相互作用,后者是柑橘斑驳黄化病(Citrus Variegated Chlorosis,CVC)的病原体。AR1.6/2 被基因改造以表达异源 GFP(绿色荧光蛋白)和内切葡聚糖酶 A(EglA),分别生成菌株 ARGFP 和 AREglA。通过荧光显微镜观察到,ARGFP 能够定植长春花幼苗的木质部导管。通过扫描电子显微镜观察到,AREglA 和 X. fastidiosa 可能共同栖息在长春花的导管中。在含有植物病原菌 X. fastidiosa 的木质部中观察到甲基杆菌,并且似乎导致生物膜形成减少。AREglA 刺激接种植物中抗性蛋白、过氧化氢酶的产生。本文报道了成功地对 AR1.6/2 进行了基因改造,以产生带有不同基因的两种不同菌株,并且还表明 AREglA 和 X. fastidiosa 可以在宿主植物内相互作用,提示了一种用于柑橘斑驳黄化病共生控制的可能策略。我们的结果提供了对甲基杆菌 - X. fastidiosa 相互作用的增强理解,表明了 AR1.6/2 作为共生控制剂的应用。