The ColRS-Regulated Membrane Protein Gene XAC1347 Is Involved in Copper Homeostasis and hrp Gene Expression in Xanthomonas citri subsp. citri

Fan, Xuetong; Guo, Jing; Zhou, Yinghui; Zhuo, Tao; Hu, Xun; Zou, Huasong

doi:10.3389/fmicb.2018.01171

Cited by 11 publications

(14 citation statements)

References 40 publications

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“…In contrast, no transference of the copper tolerance phenotype to Cu S strains was observed. These results reinforce the hypothesis that the tolerance phenotype is related to conserved genes located on the chromosome, such as cohLAB or other genes (Fan et al ., ), rather than on plasmids. Horizontal transfer of genes located on plasmids, for example those that are responsible for resistance to metals or antibiotics, is a common process among bacteria of the same species or even between different species.…”

Section: Discussionmentioning

confidence: 99%

Description of copper tolerantXanthomonas citrisubsp.citriand genotypic comparison with sensitive and resistant strains

et al. 2019

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The copper-based products widely used for control of citrus canker may lead to the development of Xanthomonas citri subsp. citri (X. citri) resistant to copper (Cu R ). However, the study of copper sensitivity of X. citri strains from Paran a state, Brazil, did not reveal the existence of Cu R , but copper tolerant (Cu T ) strains. The aim of this study was to describe for the first time the existence of Cu T X. citri and compare the genetic determinants that differentiate the Cu T strains from the sensitive (Cu S ) and Cu R strains. Cu T strains supported intermediate concentrations of copper in comparison to Cu S and Cu R . Cu T strains lack the gene clusters copLAB or copABCD responsible for copper resistance in Cu R strains and the large plasmids (c. ≥200 kb) that normally carry these genes. The nucleotide sequences of chromosomal homologous genes cohLAB, involved in copper homeostasis, were 100% similar in strains of all phenotypes. Cu T strains differed from Cu S strains by the higher expression of the homologous chromosomal genes cohA and cohB in the presence of copper. Cu T X. citri strains are not precursors of Cu R strains and do not pose a threat to the efficient use of copper-based bactericides for management of citrus canker in citrus orchards. Copper resistance and tolerance are distinct phenotypes and should not be used as synonyms. The proper characterization of the sensitivity to copper leads to a more confident monitoring of the distribution of copper resistant populations of X. citri and adoption of containment measures only when necessary.

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Section: Discussionmentioning

confidence: 99%

Description of copper tolerantXanthomonas citrisubsp.citriand genotypic comparison with sensitive and resistant strains

et al. 2019

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“…R. solanacearum was isolated from a diseased tobacco plant in Sanmin, Fujian province, China. Nutrient-rich (NB) medium was used to cultivate R. solanacearum at 28°C (21). N. tobacum NC89 was grown in a greenhouse at 28°C with a photoperiod of 16 h of light (7.5 μmol/m 2 ·s) and 8 h of darkness.…”

Section: Methodsmentioning

confidence: 99%

“…The resulting construct contained the insertion sequence of the hypothetical gene interrupted by the ripAA gene. After the introduction of the construct into wild-type P. mosselii A1 by electroporation, a standard two-step homologous recombination procedure was used to isolate the marker-free ripAA insertion mutant (21).…”

Section: Methodsmentioning

confidence: 99%

An improved control efficacy against tobacco bacterial wilt by an engineered Pseudomonas mosselii expressing the ripAA gene from phytopathogenic Ralstonia solanacearum

Zhuo

Chen

Fan

et al. 2019

Preprint

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The environmental bacterium Pseudomonas mosselii produces antagonistic secondary metabolites with inhibitory effects on multiple plant pathogens, including Ralstonia solanacearum, the causal agent of bacterial wilt. In this study, an engineered P. mosselii strain was generated to express R. solanacearum ripAA, which determines incompatible interactions with tobacco plants. The ripAA gene together with its native promoter was integrated into the P. mosselii chromosome. The resulting strain showed no difference in antimicrobial activity against R. solanacearum. Promoter-LacZ fusion and RT-PCR experiments demonstrated that the ripAA gene was transcribed in culture media. Compared with that of the wild type, the engineered strain reduced the disease index by 9.1% for bacterial wilt on tobacco plants. A transcriptome analysis was performed to identify differentially expressed genes in tobacco plants, and the results revealed that ethylene- and jasmonate-dependent defense signaling pathways were induced. These data demonstrated that the engineered P. mosselii expressing ripAA enables improved biological control against tobacco bacterial wilt by the activation of host defense responses.

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“…The resultant constructs were transformed into A. tumefaciens GV3101 using electroporation. A pGDG-XAC1347 construct was used for co-transformation with pGDR-SDE1mp to provide additional con rmation that the SDE1 protein is localized to cell membrane [9]. To isolate the interacting protein from N. benthamiana, the coding sequence of SDE1 gene was cloned into pGBKT7 at EcoR I and Pst I sites to generate pGBKT7-SDE1.…”

Section: Dna Manipulation and Plasmid Constructionmentioning

confidence: 99%

“…RNA preparation and cDNA synthesis were performed as previously described [9]. qRT-PCR experiments were performed on a CFX Connect real-time system (Bio-Rad, Hercules, CA, USA) using iTaq Universal SYBR Green Supermix (Bio-Rad, Shanghai, China).…”

Section: Qrt-pcrmentioning

confidence: 99%

Transient expression of 'Candidatus Liberibacter asiaticus' effector SDE1 induces chlorosis by suppressing NbDDX3 gene in Nicotiana benthamiana

Zhou¹,

Li²,

Liu³

et al. 2020

Preprint

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Background: 'Candidatus Liberibacter asiaticus' (Las) is the pathogenic bacterium that causes Huanglongbing in citrus plants, as well as in several types of experimental plants. Las releases a set of effectors to modulate host responses. One of these critical effectors is Sec-delivered effector 1 (SDE1), which induces chlorosis in Nicotiana benthamiana. Results: Four SDE1-interacting proteins were identified from N. benthamiana, including DEAD-box RNA helicase DDX3, 26S proteasome non-ATPase regulatory subunit PSMD14, an ARM repeat protein, and a hypothetical protein. Gene silencing revealed that knockdown of the NbDDX3 gene led to chlorosis in N. benthamiana leaves. Fluorescent signal detection revealed that SDE1 was localized to the cell membrane, cytoplasm, and nucleus. Simultaneously, NbDD3 was expressed in cytoplasmic vesicles, as well as in the cell membrane. The interactions between SDE1 and NbDDX3 were shown to be localized on cell membrane using co-localization and bimolecular fluorescence complementation analysis. Moreover, the transcription of NbDDX3 gene was substantially suppressed in N. benthamiana plants that expressed SDE1. Conclusion: Las effector SDE1 interacts with NbDDX3 at the cell membrane. Most importantly, the transient expression of SDE1 exerts a suppression effect on the transcription of NbDDX3 gene. The silencing of NbDDX3 leads to leaf chlorosis in N. benthamiana. This provided evidence to understand the molecular events in association with chlorosis induced by SDE1.

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The ColRS-Regulated Membrane Protein Gene XAC1347 Is Involved in Copper Homeostasis and hrp Gene Expression in Xanthomonas citri subsp. citri

Cited by 11 publications

References 40 publications

Description of copper tolerantXanthomonas citrisubsp.citriand genotypic comparison with sensitive and resistant strains

Description of copper tolerantXanthomonas citrisubsp.citriand genotypic comparison with sensitive and resistant strains

An improved control efficacy against tobacco bacterial wilt by an engineered Pseudomonas mosselii expressing the ripAA gene from phytopathogenic Ralstonia solanacearum

Transient expression of 'Candidatus Liberibacter asiaticus' effector SDE1 induces chlorosis by suppressing NbDDX3 gene in Nicotiana benthamiana

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