scholarly journals Revisiting the Specificity of PCR Primers for Diagnostics of Xanthomonas citri pv. citri by Experimental and In Silico Analyses

Plant Disease ◽  
2013 ◽  
Vol 97 (3) ◽  
pp. 373-378 ◽  
Author(s):  
Suzy Delcourt ◽  
Christian Vernière ◽  
Claudine Boyer ◽  
Olivier Pruvost ◽  
Bruno Hostachy ◽  
...  
Keyword(s):  
Dna Sequences ◽  
In Silico ◽  
Large Strain ◽  
In Silico Analysis ◽  
Citrus Canker ◽  
Pcr Primers ◽  
Xanthomonas Citri ◽  
Canker Disease ◽  
Strain Collection ◽  
New Findings

Asiatic citrus canker disease, caused by Xanthomonas citri pv. citri, seriously impacts citrus production worldwide. Two pathogenic variants, A and A*/Aw, have been described within this pathovar. Two additional pathovars of X. citri with a limited geographic distribution and reduced pathogenicity, namely X. citri pvs. aurantifolii and bilvae, are also pathogenic to citrus and some rutaceous species. Rapid and reliable identification is required for these citrus pathogens, which are classified as a quarantine organism in citrus-producing countries. The specificity of nine polymerase chain reaction primers previously designed for the identification of X. citri pv. citri or citrus bacterial canker strains (both pvs. citri and aurantifolii) was assayed on a large strain collection (n = 87), including the two pathotypes of X. citri pv. citri, other genetic related or unrelated pathogenic xanthomonads, and saprophytic xanthomonads. This study gave congruent results with the original articles when testing the same strains or pathovars but the use of a broad inclusivity and exclusivity panel of strains highlighted new findings. Particularly, primers 2/3, 4/7, and KingF/R failed to provide amplification for three strains from the pathotype A*/Aw. Moreover, all pairs of primers detected at least one non-target strain. These data were supported by in silico analysis of the DNA sequences available from National Center for Biotechnology Information databases.

scholarly journals Modified Monosaccharides Content of Xanthan Gum Impairs Citrus Canker Disease by Affecting the Epiphytic Lifestyle of Xanthomonas citri subsp. citri

2021 ◽  
Vol 9 (6) ◽  
pp. 1176
Author(s):  
Simone Cristina Picchi ◽  
Laís Moreira Granato ◽  
Maria Júlia Festa Franzini ◽  
Maxuel Oliveira Andrade ◽  
Marco Aurélio Takita ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Xanthan Gum ◽  
Wild Type Strain ◽  
Natural Infection ◽  
Citrus Canker ◽  
Wild Type ◽  
Xanthomonas Citri ◽  
Canker Disease ◽  
Key Enzyme ◽  
Citrus Canker Disease

Xanthomonas citri subsp. citri (X. citri) is a plant pathogenic bacterium causing citrus canker disease. The xanA gene encodes a phosphoglucomutase/phosphomannomutase protein that is a key enzyme required for the synthesis of lipopolysaccharides and exopolysaccharides in Xanthomonads. In this work, firstly we isolated a xanA transposon mutant (xanA::Tn5) and analyzed its phenotypes as biofilm formation, xanthan gum production, and pathogenesis on the sweet orange host. Moreover, to confirm the xanA role in the impaired phenotypes we further produced a non-polar deletion mutant (ΔxanA) and performed the complementation of both xanA mutants. In addition, we analyzed the percentages of the xanthan gum monosaccharides produced by X. citri wild-type and xanA mutant. The mutant strain had higher ratios of mannose, galactose, and xylose and lower ratios of rhamnose, glucuronic acid, and glucose than the wild-type strain. Such changes in the saccharide composition led to the reduction of xanthan yield in the xanA deficient strain, affecting also other important features in X. citri, such as biofilm formation and sliding motility. Moreover, we showed that xanA::Tn5 caused no symptoms on host leaves after spraying, a method that mimetics the natural infection condition. These results suggest that xanA plays an important role in the epiphytical stage on the leaves that is essential for the successful interaction with the host, including adaptive advantage for bacterial X. citri survival and host invasion, which culminates in pathogenicity.

scholarly journals GC-TOF/MS-based metabolomics analysis to investigate the changes driven by N-Acetylcysteine in the plant-pathogen Xanthomonas citri subsp. citri

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Simone Cristina Picchi ◽  
Mariana de Souza e Silva ◽  
Luiz Leonardo Saldanha ◽  
Henrique Ferreira ◽  
Marco Aurélio Takita ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cell Proliferation ◽  
Plant Pathogen ◽  
Model Organism ◽  
Citrus Canker ◽  
Bacterial Cells ◽  
Xanthomonas Citri ◽  
Canker Disease ◽  
Potential Use

AbstractN-Acetylcysteine (NAC) is an antioxidant, anti-adhesive, and antimicrobial compound. Even though there is much information regarding the role of NAC as an antioxidant and anti-adhesive agent, little is known about its antimicrobial activity. In order to assess its mode of action in bacterial cells, we investigated the metabolic responses triggered by NAC at neutral pH. As a model organism, we chose the Gram-negative plant pathogen Xanthomonas citri subsp. citri (X. citri), the causal agent of citrus canker disease, due to the potential use of NAC as a sustainable molecule against phytopathogens dissemination in citrus cultivated areas. In presence of NAC, cell proliferation was affected after 4 h, but damages to the cell membrane were observed only after 24 h. Targeted metabolite profiling analysis using GC–MS/TOF unravelled that NAC seems to be metabolized by the cells affecting cysteine metabolism. Intriguingly, glutamine, a marker for nitrogen status, was not detected among the cells treated with NAC. The absence of glutamine was followed by a decrease in the levels of the majority of the proteinogenic amino acids, suggesting that the reduced availability of amino acids affect protein synthesis and consequently cell proliferation.

scholarly journals Diversity Analysis of Endophytic Bacterial Community in Two Citrus Cultivars With Different Resistance to Citrus Canker

2021 ◽  
Author(s):  
Bing Liu ◽  
Jiahao Lai ◽  
Simeng Wu ◽  
Junxi Jiang ◽  
Weigang Kuang
Keyword(s):  
Bacterial Community ◽  
Endophytic Bacteria ◽  
Citrus Canker ◽  
Navel Orange ◽  
Xanthomonas Citri ◽  
Canker Disease ◽  
Satsuma Mandarin ◽  
Endophytic Bacterial Community ◽  
Dominant Bacteria ◽  
The Relationship

Abstract The selective infection of Xanthomonas citri pv. citri to citrus cultivars is universally known, but it is not clarified whether there is a relationship between endophytic bacteria and the resistance of host variety to canker disease. In order to explore the relationship, Satsuma mandarin and Newhall navel orange were collected respectively as samples of resistant or susceptible cultivars to citrus canker disease, and endophytic bacterial community of two citrus cultivars were analyzed by using a next-generation, Illumina-based sequencing approach. Simultaneously, the seasonal dynamics of endophytic bacterial community and dominant genera were analyzed. The results showed that there were four dominant groups including Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes in all samples at phylum level. Endophytic bacteria were the most abundant in spring samples, then in summer and autumn samples. There were some differences between endophytic bacterial community of resistant citrus and that of susceptible citrus to canker disease, and the endophytic bacteria of Satsuma mandarin are more abundant than that of Newhall navel orange. According to the analysis of dominant bacteria in two citrus cultivars, it was found that some endophytic bacteria with antagonistic characteristics existed universally in all samples, although the dominant bacteria in different seasonal sample were different. However, in Newhall navel orange of susceptible citrus to canker disease, there were not only some bacteria against Xanthomonas citri pv. citri, but also some cooperative bacteria of canker occurrence like Stenotrophomonas.

scholarly journals Biocontrol of Citrus Canker Disease Caused by Xanthomonas citri subsp. citri Using an Endophytic Bacillus thuringiensis

2019 ◽  
Vol 35 (5) ◽  
pp. 486-497 ◽  
Author(s):  
Md. Nurul Islam ◽  
Md. Sarafat Ali ◽  
Seong-Jin Choi ◽  
Jae-Wook Hyun ◽  
Kwang-Hyun Baek
Keyword(s):  
Bacillus Thuringiensis ◽  
Citrus Canker ◽  
Xanthomonas Citri ◽  
Canker Disease ◽  
Citrus Canker Disease

scholarly journals From Local Surveys to Global Surveillance: Three High-Throughput Genotyping Methods for Epidemiological Monitoring of Xanthomonas citri pv. citri Pathotypes

2008 ◽  
Vol 75 (4) ◽  
pp. 1173-1184 ◽  
Author(s):  
Lan Bui Thi Ngoc ◽  
Christian Verni�re ◽  
Philippe Jarne ◽  
Sylvain Brisse ◽  
Fabien Gu�rin ◽  
...  
Keyword(s):  
Host Range ◽  
Phylogenetic Signal ◽  
Variable Number Tandem Repeat ◽  
Citrus Canker ◽  
Variable Number ◽  
Aflp Analysis ◽  
Xanthomonas Citri ◽  
Epidemiological Monitoring ◽  
Or Groups ◽  
Strain Collection

ABSTRACT Asiatic citrus canker is a major disease worldwide, and its causal agent, Xanthomonas citri pv. citri, is listed as a quarantine organism in many countries. Analysis of the molecular epidemiology of this bacterium is hindered by a lack of molecular typing techniques suitable for surveillance and outbreak investigation. We report a comparative evaluation of three typing techniques, amplified fragment length polymorphism (AFLP) analysis, insertion sequence ligation-mediated PCR (IS-LM-PCR) typing, and multilocus variable-number tandem-repeat analysis (MLVA), with 234 strains originating from Asia, the likely center of origin of the pathogen, and reference strains of pathotypes A, A*, and Aw, which differ in host range. The typing techniques were congruent in describing the diversity of this strain collection, suggesting that the evolution pattern of the bacterium may be clonal. Based on a hierarchical analysis of molecular variance, the AFLP method best described the genetic variation found among pathotypes whereas MLVA best described the variation found among individual strains from the same countries or groups of neighboring countries. IS-LM-PCR data suggested that the transposition of insertion sequences in the genome of X. citri pv. citri occurs rarely enough not to disturb the phylogenetic signal. This technique may be useful for the global surveillance of non-epidemiologically related strains. Although pathological characteristics of strains could be most often predicted from genotyping data, we report the occurrence in the Indian peninsula of strains genetically related to pathotype A* strains but with a host range similar to that of pathotype A, which makes the classification of this bacterium even more complicated.

scholarly journals The ecnA Antitoxin Is Important Not Only for Human Pathogens: Evidence of Its Role in the Plant Pathogen Xanthomonas citri subsp. citri

2019 ◽  
Vol 201 (20) ◽  
Author(s):  
Laís Moreira Granato ◽  
Simone Cristina Picchi ◽  
Maxuel de Oliveira Andrade ◽  
Paula Maria Moreira Martins ◽  
Marco Aurélio Takita ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Host Plants ◽  
Citrus Canker ◽  
Human Pathogens ◽  
Xanthomonas Citri ◽  
Content Type ◽  
Canker Disease ◽  
Bacterial Pathogenicity ◽  
Wide Range ◽  
Host Infection

ABSTRACT Xanthomonas citri subsp. citri causes citrus canker disease worldwide in most commercial varieties of citrus. Its transmission occurs mainly by wind-driven rain. Once X. citri reaches a leaf, it can epiphytically survive by forming a biofilm, which enhances the persistence of the bacteria under different environmental stresses and plays an important role in the early stages of host infection. Therefore, the study of genes involved in biofilm formation has been an important step toward understanding the bacterial strategy for survival in and infection of host plants. In this work, we show that the ecnAB toxin-antitoxin (TA) system, which was previously identified only in human bacterial pathogens, is conserved in many Xanthomonas spp. We further show that in X. citri, ecnA is involved in important processes, such as biofilm formation, exopolysaccharide (EPS) production, and motility. In addition, we show that ecnA plays a role in X. citri survival and virulence in host plants. Thus, this mechanism represents an important bacterial strategy for survival under stress conditions. IMPORTANCE Very little is known about TA systems in phytopathogenic bacteria. ecnAB, in particular, has only been studied in bacterial human pathogens. Here, we showed that it is present in a wide range of Xanthomonas sp. phytopathogens; moreover, this is the first work to investigate the functional role of this TA system in Xanthomonas citri biology, suggesting an important new role in adaptation and survival with implications for bacterial pathogenicity.

scholarly journals Degenerate PCR primers for detecting putative priming glycosyltransferase genes in Bifidobacterium strains

2015 ◽  
Vol 6 (4) ◽  
pp. 553-562 ◽  
Author(s):  
C. Hidalgo-Cantabrana ◽  
I. Ordoñez ◽  
P. Ruas-Madiedo ◽  
A. Margolles
Keyword(s):  
Dna Sequence ◽  
Sequence Homology ◽  
In Silico ◽  
In Silico Analysis ◽  
Pcr Primers ◽  
Degenerate Pcr ◽  
Negative Results ◽  
Galactosyl Transferase ◽  
Undecaprenyl Phosphate ◽  
Silico Analysis

A new PCR-based method to detect putative exopolysaccharide (EPS) producers from the genus Bifidobacterium was developed based on the detection of two priming glycosyltransferase genes: rfbP (undecaprenyl-phosphate sugar phospho-transferase) and cpsD (galactosyl-transferase). An in silico analysis of the genomes of 28 bifidobacterial strains, belonging to 8 different species, allowed us to detect rfbP, cpsD, or both, in the large majority of the genomes. Based on DNA sequence homology studies, 24 degenerated primers were synthesised in order to select the primer pairs with the broadest capacity to detect the presence of these genes. Four primer pairs targeting internal regions of rfbP and cpsD were selected, allowing the detection of at least one of the two genes in 63 out of 99 bifidobacterial strains analysed, whereas control strains from other genera yielded negative results, suggesting that these genes are widely spread in this genus. The use of these primers is recommended to screen for the potential of Bifidobacterium strains to produce EPS.

scholarly journals Comparative in silico analysis of PCR primers suited for diagnostics and cloning of ammonia monooxygenase genes from ammonia-oxidizing bacteria

2008 ◽  
Vol 64 (1) ◽  
pp. 141-152 ◽  
Author(s):  
Pilar Junier ◽  
Ok-Sun Kim ◽  
Verónica Molina ◽  
Petra Limburg ◽  
Thomas Junier ◽  
...  
Keyword(s):  
In Silico ◽  
In Silico Analysis ◽  
Pcr Primers ◽  
Ammonia Oxidizing Bacteria ◽  
Ammonia Monooxygenase ◽  
Silico Analysis

scholarly journals In Vitro and In Vivo Antibacterial Activity of Some Organic and Inorganic Salts Against Asiatic Citrus Canker Agent Xanthomonas Citri Subsp. Citri

2014 ◽  
Vol 2 (6) ◽  
pp. 296
Author(s):  
Vahideh Hasabi ◽  
Hossein Askari ◽  
Seyed Mehdi Alavi ◽  
Masood Soltani Najafabadi
Keyword(s):  
Antibacterial Activity ◽  
Citrus Canker ◽  
Inhibitory Effect ◽  
Inorganic Salts ◽  
Xanthomonas Citri ◽  
Canker Disease ◽  
Growth Inhibitory ◽  
Iron And Zinc

Asiatic citrus canker caused by Xanthomonas citri subsp. citri is becoming a disease of high economic impact, affecting all types of important citrus crops. In this study, the potential antibacterial activity of ten organic and inorganic salts on X. citri subsp. citri and on citrus canker disease development was evaluated. Among the salt compounds, copper, iron and zinc inorganic salts particularly zinc (with the highest diameter of inhibition, the lowest MIC and MBC values and the highest bacterial growth inhibitory effect) had direct antibacterial activity and strongly reduced the development of canker disease and bacterial population of lime plants.

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