Citri Strains Research Articles

Analysis of the Genetic Variation and Geographic Distribution Patterns of Xanthomonas citri pv. citri Strains in Citrus Production in Burkina Faso.

It is essential to have a thorough knowledge of the genetic variation among different strains of Xanthomonas citri pv. citri, which is responsible for causing citrus bacterial canker. This understanding is important for studying disease characteristics, population structure, and evolution and ultimately for developing sustainable methods of control. A total of 48 strains obtained from citrus production areas in Burkina Faso in 2012, 2020, and 2021 were subjected to Polymerase Chain reaction (PCR) tests using specific primers. The aim was to examine the distribution of type 3 effectors and determine the geographical origins of the strains. The examination of the distribution of type 3 non-transcription-activator-like effectors (TALEs) revealed a broader range of strains obtained in 2020 and 2021 than in 2012. However, all the strains possessed a shared set of three genes, specifically, XopE2, XopN, and AvrBs2. Furthermore, all examined effectors were observed in the Bobo-Dioulasso region. Regarding the characterization of TALEs, two profiles containing two to three TALEs were discovered. Profile 1, consisting of two TALEs, was found in 37 X. citri pv. citri strains, whereas Profile 2, comprising three TALEs, was detected in 11 strains. Among the three TALEs (A, B, and C) that were identified, TALEs B and C were present in all the strains. The correlation matrix indicated a positive association between the type 3 effector content of strains and the duration of their isolation. Principal component analysis revealed a limited organization of the strains under investigation. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

A UDP-glycosyltransferase gene PcUGT202A9 was associated with abamectin resistance in Panonychus citri (McGregor)

Panonychus citri (McGregor) strains have developed a high level of resistance to abamectin, but the underlying molecular mechanism is unknown. Uridine diphosphate (UDP)-glycosyltransferases (UGTs) are critical for the removal of a variety of exogenous and endogenous substances. In this study, an enzyme activity assay revealed that UGTs potentially contribute to P. citri abamectin resistance. Spatiotemporal expression profiles showed that only PcUGT202A9 was significantly overexpressed in the abamectin-resistant strain (AbR) at all developmental stages. Moreover, UGT activity decreased significantly, whereas abamectin susceptibility increased significantly, in AbR after PcUGT202A9 was silenced. Three-dimensional modeling and molecular docking analyses revealed that PcUGT202A9 can bind stably to abamectin. Recombinant PcUGT202A9 activity was detected when α-naphthol was used, but the enzymatic activity was inhibited by abamectin (50 % inhibitory concentration: 803.3 ± 14.20 μmol/L). High-performance liquid chromatography and mass spectrometry analyses indicated that recombinant PcUGT202A9 can effectively degrade abamectin and catalyze the conjugation of UDP-glucose to abamectin. These results imply PcUGT202A9 contributes to P. citri abamectin resistance.

A class of photosensitizer highly effective to control bacterial infection in plants even on rainy days with dim light

Bacterial infections of plants are widespread and highly virulent, causing serious economic losses in agricultural production worldwide. These infections become particularly prevalent during rainy weather, characterized by high humidity and low light intensity (about 2–10% of a sunny day). Here we identified a class of photosensitizers that were effective in inhibiting plant bacterial infections under rainy weather with dim light. One of these compounds, compound 1, demonstrated exceptional efficacy by eliminating more than 4 logs (99.99%) of either Xanthomonas perforans or Xanthomonas citri subsp. citri (Xcc) strain at a concentration of 6.25 μM under rainy weather conditions with a light exposure of 28.51 J/cm2. For Clavibacter michiganensis (Cm), compound 1 demonstrated a comparable bactericidal effect at a lower concentration of 3.13 μM. Furthermore, compound 1 was effective in controlling plant leaf infections during rainy weather. Moreover, compound 1 exhibited a potent inhibitory effect on the biofilms pertinent to tomato spot disease and citrus canker under natural illumination on rainy days. Additionally, compound 1 displayed remarkable resilience to rain-wash on plant leaves, retaining 41.20% of its initial concentration following exposure to three simulated rain events. These findings provide a new strategy for plant pathogen disease management highlight its feasibility even during rainy weather.

Volatile compounds of hexane extract from Pterodon pubescens Benth seeds and its significant in vitro potential against different bacterial strains

Pterodon pubescens Benth is a Brazilian medicinal plant (sucupira, in Brazilian Portuguese). This paper aims to determine the volatile composition and antibacterial activities of hexane extract from P. pubescens seeds (HE-PP). Antibacterial activities were screened by the microdilution broth method in 96-well culture plates and MIC values were expressed as µg/mL. HE-PP was active against several oral bacteria whose MIC values ranged between 12.5 µg/mL and 50 µg/mL and against three mycobacterial strains (MIC = 125 µg/mL and 500 µg/mL). In addition, HE-PP was active against Xanthomonas citri strain (MIC = 100 µg/mL). Cytotoxic activity of the extract was evaluated in human tumour and non-tumour cell lines. HE-PP showed selective cytotoxicity to cervical adenocarcinoma (HeLa cells – IC50 = 53.47 µg/mL). Its major constituents were identified by GC-MS and GC-FID: E-caryophyllene, vouacapane, E-geranylgeraniol and dehydroabietol. Results reinforce the biological potential of HE-PP against a broad spectrum of pathogenic and phytopathogenic bacteria.

CYP4CL2 Confers Metabolic Resistance to Pyridaben in the Citrus Pest Mite Panonychus citri.

The citrus red mite Panonychus citri has developed strong resistance to acaricides. Cytochrome P450 monooxygenases (P450s) can detoxify pesticides and are involved in pesticide resistance in many insects. Here, a pyridaben-resistant P. citri strain showed cross-resistance to cyenopyrafen, bifenazate, fenpyroximate, and tolfenpyrad. Piperonyl butoxide, a P450 inhibitor, significantly increased the toxicity of pyridaben to resistant (Pyr_Rs) and susceptible (Pyr_Control) P. citri strains. P450 activity was significantly higher in Pyr_Rs than in Pyr_Control. Analyses of RNA-Seq data identified a P450 gene (CYP4CL2) that is potentially involved in pyridaben resistance. Consistently, it was up-regulated in two field-derived resistant populations (CQ_WZ and CQ_TN). RNA interference-mediated knockdown of CYP4CL2 significantly decreased the pyridaben resistance in P. citri. Transgenic Drosophila melanogaster expressing CYP4CL2 showed increased pyridaben resistance. Molecular docking analysis showed that pyridaben could bind to several amino acids at substrate recognition sites in CYP4CL2. These findings shed light on P450-mediated pyridaben resistance in pest mites.

Diversity of copper resistant Xanthomonas citri subsp. citri strains, the causal agent of Asiatic citrus canker, in Iran

Diversity of copper resistant Xanthomonas citri subsp. citri strains, the causal agent of Asiatic citrus canker, in Iran

Chemical composition and anti-Xanthomonas citri activities of essential oils from Schinus molle L. fresh and dry leaves and of its major constituent spathulenol

Citrus canker, which is caused by Xanthomonas citri, is a severe disease that affects citrus plants worldwide. This paper aimed to compare, for the first time, the chemical composition and anti-Xanthomonas citri activities of essential oils from Schinus molle fresh and dry leaves (EO-FL and EO-DL, respectively). Anti-X. citri activity of spathulenol, the major constituent of oils, was also evaluated. Activities were screened by the broth microdilution method on 96‐well culture plates. Three major constituents were identified in EO-FL and EO-DL by GC-MS and GC-FID: spathulenol, β-caryophyllene and caryophyllene oxide. EO-DL (MIC = 31.25 µg/mL), EO-FL (MIC = 62.5 µg/mL) and spathulenol (MIC = 100 µg/mL) were active against X. citri strains (resistant, tolerant and sensitive to copper). Even though results showed that in vitro potential of EO-FL, EO-DL and spathulenol against X. citri, further in vivo studies are needed to prove their applicability to the biocontrol of citrus canker.

Complete Genome Sequence of Xanthomonas citri subsp. citri CQ13, an Alternative Model Strain to Study Citrus Bacterial Canker in China

Xanthomonas citri subsp. citri is a quarantine organism causing citrus bacterial canker in many countries. Sharing research materials worldwide became difficult due to local administrative regulations, which hindered the continuity of studies on this disease. Here, we report a complete genome sequence of X. citri subsp. citri strain CQ13 achieved using a combination of Illumina and Nanopore platforms. The CQ13 genome comprises a circular chromosome (5,142,501 bp; G + C content 64.83%) and two circular plasmids, pXAC33 and pXAC64 (34,102 bp and 64,942 bp; G + C content 61.96 and 61.41%, respectively). Annotation revealed 4,243 protein-coding sequences, six rRNA genes, 53 tRNA genes, 40 ncRNA genes, and 168 pseudogenes. Notably, the analyses of average nucleotide identity, host range, pathogenicity, and type III effector repertoire suggest that CQ13 is particularly closely related to the current model strain 306. Therefore, strain CQ13 could serve as an alternative model strain, and its genome sequence could provide a valuable resource for citrus canker research in China. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Occurrence of bean-associated cytorhabdovirus and cowpea mild mottle virus infecting cultivated and wild Passiflora spp. in Brazil

Bean-associated cytorhabdovirus (BaCV) is a strain of Cytorhabdovirus caricae (genus Cytorhabdovirus, family Rhabdoviridae) identified in Brazil, in bean and soybean plants, in mixed infection with cowpea mild mottle virus (CPMMV, genus Carlavirus, family Betaflexiviridae). Ensuing the report of citrus-associated rhabdovirus (CiaRV — a citrus strain of Cytorhabdovirus caricae) infecting passion fruit in China, we speculated that BaCV, and possibly CPMMV, could be present in passion fruit crops in Brazil. This was tested by screening cultivated and wild Passiflora spp., and hybrid by RT-PCR and amplicon sequencing, which confirmed the presence of BaCV and CPMMV in several Passiflora spp. A total of 114 surveyed plants were collected in multiple locations from 2016 to 2021 and BaCV was identified in 4 of 19 cultivated P. edulis plants from a commercial field in Distrito Federal, Central Brazil. Moreover, while BaCV was detected in 4 of the 55 Passiflora accessions sampled at the Germplasm Bank “Flor da Paixão” (BAG-FP), CPMMV was identified in 14 of these 55 Passiflora spp. and hybrid accessions. This is the first record of BaCV in P. edulis in Brazil, and CPMMV and BaCV in other wild Passiflora species in the world. To investigate the potential mixed infection of BaCV and CPMMV-infected plants and other viruses previously identified infecting Passiflora spp. in Brazil, RT-PCR was conducted with specific primers for lettuce chlorosis virus (LCV, genus Crinivirus, family Closteroviridae) and cowpea aphid-borne mosaic virus (CABMV, genus Potyvirus, family Potyviridae). Most of the inspected plants had a mixed infection with these viruses. Our results warrant further studies to evaluate these viruses’ epidemiology, impacts, and interactions in the passion fruit crop.

Orchid fleck dichorhavirus movement protein shows RNA silencing suppressor activity.

To counteract RNA interference-mediated antiviral defence, virus genomes evolved to express proteins that inhibit this plant defence mechanism. Using six independent biological approaches, we show that orchid fleck dichorhavirus citrus strain (OFV-citrus) movement protein (MP) may act as a viral suppressor of RNA silencing (VSR). By using the alfalfa mosaic virus (AMV) RNA 3 expression vector, it was observed that the MP triggered necrosis response in transgenic tobacco leaves and increased the viral RNA (vRNA) accumulation. The use of the potato virus X (PVX) expression system revealed that the cis expression of MP increased both the severity of the PVX infection and the accumulation of PVX RNAs, further supporting that MP could act as an RNA silencing suppressor (RSS). From the analysis of the RSS-defective turnip crinkle virus (TCV), we do not find local RSS activity for MP, suggesting a link between MP suppressor activity and the prevention of systemic silencing. In the analysis of local suppressive activity using the GFP-based agroinfiltration assay in Nicotiana benthamiana (16 c line), we do not identify local RSS activity for the five OFV RNA1-encoded proteins. However, when evaluating the small interfering RNA (siRNA) accumulation, we find that the expression of MP significantly reduces the accumulation of GFP-derived siRNA. Finally, we examine whether the MP can prevent systemic silencing in 16c plants. Our findings show that MP inhibits the long-distance spread of RNA silencing, but does not affect the short-distance spread. Together, our findings indicate that MP is part of OFV's counter-defence mechanism, acting mainly in the prevention of systemic long-distance silencing. This work presents the first report of a VSR for a member of the genus Dichorhavirus.

Molecular epidemiology of the citrus bacterial pathogen Xanthomonas citri pv. citri from the Arabian Peninsula reveals a complex structure of specialist and generalist strains.

Molecular epidemiology studies are essential to refine our understanding of migrations of phytopathogenic bacteria, the major determining factor in their emergence, and to understand the factors that shape their population structure. Microsatellite and minisatellite typing are useful techniques for deciphering the population structure of Xanthomonas citri pv. citri, the causal agent of Asiatic citrus canker. This paper presents a molecular epidemiology study, which has improved our understanding of the history of the pathogen's introductions into the Arabian Peninsula, since it was first reported in the 1980s. An unexpectedly high genetic diversity of the pathogen was revealed. The four distinct genetic lineages within X. citri pv. citri, which have been reported throughout the world, were identified in the Arabian Peninsula, most likely as the result of multiple introductions. No copper‐resistant X. citri pv. citri strains were identified. The pathogen's population structure on Mexican lime (their shared host species) was closely examined in two countries, Saudi Arabia and Yemen. We highlighted the marked prevalence of specialist pathotype A* strains in both countries, which suggests that specialist strains of X. citri pv. citri may perform better than generalist strains when they occur concomitantly in this environment. Subclade 4.2 was the prevailing lineage identified. Several analyses (genetic structure deciphered by discriminant analysis of principal components, RST‐based genetic differentiation, geographic structure) congruently suggested the role of human activities in the pathogen's spread. We discuss the implications of these results on the management of Asiatic citrus canker in the region.

Differential Chemical Profile of Metabolite Extracts Produced by the Diaporthe citri (G-01) Endophyte Mediated by Varying the Fermented Broth pH.

Endophytic microorganisms show great potential for biotechnological exploitation because they are able to produce a wide range of secondary compounds involved in endophyte–plant adaptation, and their interactions with other living organisms that share the same microhabitat. Techniques used to chemically extract these compounds often neglect the intrinsic chemical characteristics of the molecules involved, such as the ability to form conjugate acids or bases and how they influence the solubilities of these molecules in organic solvents. Therefore, in this study, we aimed to evaluate how the pH of the fermented broth affects the process used to extract the secondary metabolites of the Diaporthe citri strain G-01 endophyte with ethyl acetate as the organic solvent. The analyzed samples, conducted by direct-infusion electrospray-ionization mass spectrometry, were grouped according to the pH of the fermented broth (i.e., <7 and ≥7). A more extreme pH (i.e., 2 or 12) was found to affect the chemical profile of the sample. Moreover, statistical analysis enabled us to determine the presence or absence of ions of high importance; for example, ions at 390.7 and 456.5 m/z were observed mainly at acidic pH, while 226.5, 298.3, and 430.1 m/z ions were observed at pH ≥ 7. Extraction at a pH between 4 and 9 may be of interest for exploring the differential secondary metabolites produced by endophytes. Furthermore, pH influences the chemical phenotype of the fungal metabolic extract.

Genome Sequence Resource of Xanthomonas citri pv. citri from Formalin-Fixed Citrus Leaves Specimen Showing Canker Lesions Collected in 1982.

Xanthomonas citri pv. citri is the causative agent of citrus canker, one of the most devastating diseases threatening global citrus production. Here, we present the genome sequence of X. citri pv. citri strain GD82 from a formalin-fixed citrus leaf specimen showing canker lesions collected in 1982 in Guangdong Province, China. The GD82 genome consisted of 5,197,217 bp with G+C content of 64.8%, along with four circular plasmids: pXAC33 (32,377 bp), pXAC64 (63,972 bp), pXAC47 (47,810 bp), and pGD82.1 (219,560 bp). This is the oldest X. citri pv. citri genome from historical citrus canker specimens in China, which will enrich the current X. citri pv. citri genome database and facilitate genomic evolution research of X. citri pv. citri.

Antioxidant and Anti-Cancer Effect of Ethanolic Extract of Citrus Fruits on Hep G2 and MCF-7 Cell Lines

Background/objective: Cancer is a complex genetic disease that occurs due to mutation in genes that control apoptosis and cell growth. Uncontrolled cell growth leads to the formation of tumors. Free radical causes mutation in genes and DNA sequence, however antioxidants can stabilize these harmful effects. Citrus fruits are the rich source of antioxidants. owing to this property this study was planned to evaluate the potential of the citrus fruits for the treatment of the cancer.&#x0D; Aim: This study aimed to evaluate antioxidant as well anticancer potential of five different citrus strains (Citrus deliciosa, Citrus maxima, Citrus limetta, Citrus sinensis and Citrus reticulata).&#x0D; Methods: Peel of all the citrus fruits were collected, grinded and ethanolic extracts were prepared separately, to evaluate radical scavenging ability by employing DPPH method followed by MTT assay of the cancer cell lines to explore the anticancer potential of the extracts.&#x0D; Results: It was observed that citrus peels exhibited good radical scavenging activity and inhibited tumor growth. Maximum effect was produced by Citrus reticulata, and least results were obtained with Citrus sinensis.&#x0D; Conclusion: It was concluded that antioxidant and anti-cancer effects of citrus peels may be due to be owing to the presence of antioxidants (ascorbic acid, flavonoids, phenols, limonene). This research might open new horizon in the treatment of cancer chemotherapy.

Comparative Genomic Analysis Uncovered Evolution of Pathogenicity Factors, Horizontal Gene Transfer Events, and Heavy Metal Resistance Traits in Citrus Canker Bacterium Xanthomonas citri subsp. citri.

Background: Worldwide citrus production is severely threatened by Asiatic citrus canker which is caused by the proteobacterium Xanthomonas citri subsp. citri. Foliar sprays of copper-based bactericides are frequently used to control plant bacterial diseases. Despite the sequencing of many X. citri strains, the genome diversity and distribution of genes responsible for metal resistance in X. citri subsp. citri strains from orchards with different management practices in Taiwan are not well understood.Results: The genomes of three X. citri subsp. citri strains including one copper-resistant strain collected from farms with different management regimes in Taiwan were sequenced by Illumina and Nanopore sequencing and assembled into complete circular chromosomes and plasmids. CRISPR spoligotyping and phylogenomic analysis indicated that the three strains were located in the same phylogenetic lineages and shared ∼3,000 core-genes with published X. citri subsp. citri strains. These strains differed mainly in the CRISPR repeats and pathogenicity-related plasmid-borne transcription activator-like effector (TALE)-encoding pthA genes. The copper-resistant strain has a unique, large copper resistance plasmid due to an unusual ∼40 kbp inverted repeat. Each repeat contains a complete set of the gene cluster responsible for copper and heavy metal resistance. Conversely, the copper sensitive strains carry no metal resistance genes in the plasmid. Through comparative analysis, the origin and evolution of the metal resistance clusters was resolved.Conclusion: Chromosomes remained constant among three strains collected in Taiwan, but plasmids likely played an important role in maintaining pathogenicity and developing bacterial fitness in the field. The evolution of pathogenicity factors and horizontal gene transfer events were observed in the three strains. These data suggest that agricultural management practices could be a potential trigger for the evolution of citrus canker pathogens. The decrease in the number of CRISPR repeats and pthA genes might be the result of adaptation to a less stressful environment. The metal resistance genes in the copper resistant X. citri strain likely originated from the Mauritian strain not the local copper-resistant X. euvesicatoria strain. This study highlights the importance of plasmids as ‘vehicles’ for exchanging genetic elements between plant pathogenic bacteria and contributing to bacterial adaptation to the environment.

Genome analysis of Spiroplasma citri strains from different host plants and its leafhopper vectors

BackgroundSpiroplasma citri comprises a bacterial complex that cause diseases in citrus, horseradish, carrot, sesame, and also infects a wide array of ornamental and weed species. S. citri is transmitted in a persistent propagative manner by the beet leafhopper, Neoaliturus tenellus in North America and Circulifer haematoceps in the Mediterranean region. Leafhopper transmission and the pathogen’s wide host range serve as drivers of genetic diversity. This diversity was examined in silico by comparing the genome sequences of seven S. citri strains from the United States (BR12, CC-2, C5, C189, LB 319, BLH-13, and BLH-MB) collected from different hosts and times with other publicly available spiroplasmas.ResultsPhylogenetic analysis using 16S rRNA sequences from 39 spiroplasmas obtained from NCBI database showed that S. citri strains, along with S. kunkelii and S. phoeniceum, two other plant pathogenic spiroplasmas, formed a monophyletic group. To refine genetic relationships among S. citri strains, phylogenetic analyses with 863 core orthologous sequences were performed. Strains that clustered together were: CC-2 and C5; C189 and R8-A2; BR12, BLH-MB, BLH-13 and LB 319. Strain GII3–3X remained in a separate branch. Sequence rearrangements were observed among S. citri strains, predominantly in the center of the chromosome. One to nine plasmids were identified in the seven S. citri strains analyzed in this study. Plasmids were most abundant in strains isolated from the beet leafhopper, followed by strains from carrot, Chinese cabbage, horseradish, and citrus, respectively. All these S. citri strains contained one plasmid with high similarity to plasmid pSci6 from S. citri strain GII3–3X which is known to confer insect transmissibility. Additionally, 17 to 25 prophage-like elements were identified in these genomes, which may promote rearrangements and contribute to repetitive regions.ConclusionsThe genome of seven S. citri strains were found to contain a single circularized chromosome, ranging from 1.58 Mbp to 1.74 Mbp and 1597–2232 protein-coding genes. These strains possessed a plasmid similar to pSci6 from the GII3–3X strain associated with leafhopper transmission. Prophage sequences found in the S. citri genomes may contribute to the extension of its host range. These findings increase our understanding of S. citri genetic diversity.

PthAW1, a Transcription Activator-Like Effector of Xanthomonas citri subsp. citri, Promotes Host-Specific Immune Responses.

Citrus canker disease caused by Xanthomonas citri subsp. citri is one of the most destructive diseases in citrus. X. citri subsp. citri pathotypes display different host ranges. X. citri subsp. citri strain A (XccA) causes canker disease in most commercial citrus varieties, whereas strain AW (XccAW), which is genetically similar to XccA, infects only lime and alemow. Understanding the mechanism that determines the host range of pathogens is critical to investigating and utilizing host resistance. We hypothesized that XccAW would undergo mutations in genes that restrict its host range when artificially inoculated into incompatible citrus varieties. To test this hypothesis, we used an experimental evolution approach to identify phenotypic traits and genetic loci associated with the adaptation of XccAW to incompatible sweet orange. Repeated inoculation and reisolation cycles improved the ability of three independent XccAW strains to colonize sweet orange. Adapted XccAW strains displayed increased expression of type III secretion system and effector genes. Genome sequencing analysis indicated that two of the adapted strains harbored mutations in pthAW1, a transcription activator-like effector (TALE) gene, that corresponded to the removal of one or two repeats from the central DNA-binding repeat region. Introduction of the original but not the adapted pthAW1 variants into XccA abolished its ability to cause canker symptoms in sweet orange, Meyer lemon, and clementine but not in other XccAW-resistant citrus varieties. The original pthAW1, when expressed in XccA, induced ion leakage and the expression of pathogenesis-related genes but had no effect on CsLOB1 expression in sweet orange. Our study has identified a novel host-specific avirulence TALE and demonstrated active adaptive rearrangements of the TALE repeat array during host adaptation.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Comparing bacterial properties in relation to the virulence factors of Xanthomonas citri subsp. citri strains and evaluating resistance of subtribe Citrinae cultivars to the most virulent strain

Citrus Bacterial Canker disease, caused by Xanthomonas citri subsp. citri (Xcc), is one of the most devastating diseases that attacks citrus, especially in the southern areas of Iran. The objective of this investigation was to analyze several characteristics involved in Xcc virulence in relation to strain aggressiveness. To achieve this, swarming motility, biofilm formation, resistance to H2O2, and production of xanthan were evaluated considering 44 local strains causing citrus bacterial canker disease in Iran. All strains showed differential swarming motilities, biofilm formation abilities, and xanthan production. The most virulent strain, Xcc-KVXCC1, exhibited the greatest capability to form biofilm on solid surfaces, xanthan production, and pathogenicity on detached leaves of Citrus aurantifolia as well as positive chemotaxis toward C. aurantifolia extract. In this study, a total of 14 common and commercial citrus genotypes were evaluated for resistance to Xcc-KVXCC1. Genotypes were categorized into susceptible and resistant groups based on lesion number per inoculation site. Based on our results, C. aurantifolia was the most susceptible citrus cultivar to Xcc-KVXCC1. In contrast, C. aurantium, C. jumbhori, and C. reticulata cv. ponkan demonstrated high and moderately high resistance against Xcc-KVXCC1.

Inhibition of the Citrus Canker Pathogen Using a Photosensitizer Assisted by Sunlight Irradiation.

Citrus canker, induced by bacterial infection, seriously affects the growth and productivity of citrus around the world and has attracted strong research interest. The current treatment for this disease uses copper salts to inactivate the pathogenic bacteria: Xanthomonas citri subsp. citri (Xcc) strain. However, copper salts may have a negative impact on the environment or plant. In this work, we identify a chemical compound, 2,6-diiodo-1,3,5,7-tetramethyl-8-(P-benzoic acid)-4,4′-difluoroboradiazaindacene (DIBDP), to inactivate the pathogenic Xcc strain (29-1). DIBDP is activated by sunlight and generates reactive oxygen species to kill the bacteria. In order to overcome the degradation of DIBDP under sunlight, an adjuvant agent was identified to limit the photodegradation of DIBDP by forming a photosensitizer complex (PSC). This complex demonstrated significant antimicrobial activity to Xcc 29-1, which was 64-fold more potent than the copper biocides. The antimicrobial efficacy of PSC on citrus leaves infected by Xcc 29-1 also was much stronger than copper agent and, at the same time, the PSC was safe to the host exposed to sunlight. Thus, this PSC is a promising antibacterial agent to control citrus canker disease.

Phylogenetic, genetic, and phenotypic diversity of Pseudomonas syringae pv. syringae strains isolated from citrus blast and black pit in Tunisia

AbstractCitrus blast and black pit caused by Pseudomonas syringae pv. syringae (Pss) is the only bacterial disease reported in Tunisian Citrus orchards. The phylogenetic relationship between Pss strains was studied based on multilocus sequence analysis (MLSA), using partial sequences of housekeeping genes rpoD, rpoB, gyrB, cts, and pfk for 14 representative Pss Citrus strains, including the reference strain LMG5496. The MLSA revealed that the studied Tunisian Citrus strains are closely related to LMG5496 and cluster in phylogroup 02. Based on the cts gene, the majority of Citrus strains clustered in clades “a” and “b”. However, five strains were placed in a newly defined clade “g”. We describe the presence of six different type III secreted effectors (T3SEs). These were found with frequencies of 100% for the effector hopAN1 and the helper hrpK1, 65% for hopT1‐2, and 14% for hopN1, hopR1, and hopQ1‐2. Investigation of copper resistance showed that 67% of our Pss Citrus strains from Tunisia are resistant to copper sulphate in vitro, and the copper resistance genes copABCDR were detected in 23% of the strains. Our results present new data concerning the genetic diversity and phylogeny, presence of T3SEs, and copper resistance within the Pss populations that affect Citrus in Tunisia.