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内生甲基杆菌表达了一种异源的β-1,4-内切葡聚糖酶 A(EglA)在长春花幼苗中,该植物是韧皮部杆菌的模式宿主植物。

Endophytic Methylobacterium extorquens expresses a heterologous β-1,4-endoglucanase A (EglA) in Catharanthus roseus seedlings, a model host plant for Xylella fastidiosa.

机构信息

Department of Phytopathology, University of Passo Fundo (UPF), BR 285, PO BOX 611, Passo Fundo, RS, 99052-900, Brazil.

出版信息

World J Microbiol Biotechnol. 2012 Apr;28(4):1475-81. doi: 10.1007/s11274-011-0949-2. Epub 2011 Nov 19.

DOI:10.1007/s11274-011-0949-2
PMID:22805929
Abstract

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.

摘要

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。

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Endophytic Methylobacterium extorquens expresses a heterologous β-1,4-endoglucanase A (EglA) in Catharanthus roseus seedlings, a model host plant for Xylella fastidiosa.内生甲基杆菌表达了一种异源的β-1,4-内切葡聚糖酶 A(EglA)在长春花幼苗中,该植物是韧皮部杆菌的模式宿主植物。
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本文引用的文献

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Culture and serological detection of the xylem-limited bacterium causing citrus variegated chlorosis and its identification as a strain ofXylella fastidiosa.木质部限制作物黄化脉明细菌的培养和血清学检测及其鉴定为木质部难养细菌的一个菌株。
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Transmission of Methylobacterium mesophilicum by Bucephalogonia xanthophis for paratransgenic control strategy of citrus variegated chlorosis.嗜中甲基杆菌通过黄斑布氏吸虫传播用于柑橘杂色黄化病的共生转基因控制策略
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Nodulation and plant-growth promotion by methylotrophic bacteria isolated from tropical legumes.
未脱皮猪屎豆种子内生菌微生物组:植物有益甲基杆菌的鉴定。
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An endophytic microbe from an unusual volcanic swamp corn seeks and inhabits root hair cells to extract rock phosphate.一种内生微生物从一种不寻常的火山沼泽玉米中寻找并栖息在根毛细胞中以提取磷矿粉。
Sci Rep. 2017 Oct 18;7(1):13479. doi: 10.1038/s41598-017-14080-x.
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The diversity of citrus endophytic bacteria and their interactions with Xylella fastidiosa and host plants.柑橘内生细菌的多样性及其与木质部难养菌和寄主植物的相互作用。
Genet Mol Biol. 2016 Oct-Dec;39(4):476-491. doi: 10.1590/1678-4685-GMB-2016-0056. Epub 2016 Oct 10.
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Fungal endophytes of Catharanthus roseus enhance vindoline content by modulating structural and regulatory genes related to terpenoid indole alkaloid biosynthesis.长春花的真菌内生菌通过调节与萜类吲哚生物碱生物合成相关的结构和调控基因来提高长春质碱含量。
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从热带豆科植物中分离出的甲基营养菌的结瘤作用及促进植物生长的特性
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Can J Microbiol. 2006 May;52(5):419-26. doi: 10.1139/w05-142.
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Methylotrophic metabolism is advantageous for Methylobacterium extorquens during colonization of Medicago truncatula under competitive conditions.在竞争条件下,甲基营养型代谢对豌豆根瘤菌在蒺藜苜蓿定殖过程中具有优势。
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Molecular characterization of a beta-1,4-endoglucanase from an endophytic Bacillus pumilus strain.一株内生短小芽孢杆菌β-1,4-内切葡聚糖酶的分子特征分析
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Diversity of endophytic bacterial populations and their interaction with Xylella fastidiosa in citrus plants.柑橘类植物内生细菌种群的多样性及其与木质部难养菌的相互作用。
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