EVALUATION OF SUSCEPTIBILITY OF 'PERA' SWEET ORANGE GENOTYPES TO CITRUS CANKER UNDER FIELD AND GREENHOUSE CONDITIONS

During the development of a citrus fruit, many cycles of infection by Xanthomonas citri subsp. citri may occur leading to the development of a range of characteristics of citrus canker lesions scattered across the fruit surface. This study aimed to determine whether the size of the lesions, their distance from the peduncle, and the number and time of appearance of the lesions on fruit of sweet orange were associated with premature fruit drop. A multiple linear regression analysis revealed a negative relationship between the fruit detachment force and the lesion diameter, the proximity of the nearest lesion to the peduncle and the number of lesions. A survival analysis demonstrated that these characteristics significantly influenced the probability and the time that a cankered fruit remained attached to the tree. More than 90% of dropped fruit had large lesions (>5 mm) but not all fruit with large lesions dropped before harvest. Approximately 50% of the harvested fruit had lesions >5 mm. On the harvested fruit remaining on the tree, although large, the lesions had a smaller diameter, were located farther from the peduncle, and were less numerous than those observed on dropped fruit. Small canker lesions neither reduced the detachment force nor the survival of fruit in the tree. The earlier a fruit expressed canker symptoms, the higher the probability the fruit developed large lesions near the peduncle and/or developed lesions in greater numbers. This study provides a better understanding on the relationship between the time of appearance of lesions of citrus canker on fruit and premature fruit drop. This information defines the critical period for fruit protection and may be used to improve disease management.

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Relative contribution of windbreak, copper sprays and leafminer control for citrus canker management and prevention of crop loss in sweet orange trees

Plant Disease ◽

10.1094/pdis-10-20-2153-re ◽

2020 ◽

Author(s):

Franklin Behlau ◽

José Belasque Junior ◽

RUI LEITE ◽

Armando Bergamin-Filho ◽

Tim Gottwald ◽

...

Keyword(s):

Sweet Orange ◽

Disease Incidence ◽

Citrus Canker ◽

Control Measures ◽

Disease Suppression ◽

Additional Contribution ◽

Crop Losses ◽

Rangpur Lime ◽

Relative Contribution ◽

Orange Trees

The management of citrus canker, caused by Xanthomonas citri subsp. citri, has been widely studied in endemic areas due to the importance of the disease in several citrus producing countries. A set of control measures is well-established, but no study has investigated the efficiency of each measure individually and their combination for disease suppression. This study comprised a 3-year field study to assess the relative contribution of three measures for the control of citrus canker and reduction of crop losses. Windbreak (Wb), copper sprays (Cu), and leafminer control (Lc) were assessed in eight different combinations in a split-split plot design. The orchard was composed of ‘Valencia’ sweet orange trees grafted onto ‘Rangpur’ lime. Casuarina cunninghamiana trees were used as Wb. Cu and Lc sprays were performed every 21 days throughout the year. Individually, Cu showed the highest contribution for canker control, followed by Wb. Lc had no effect on reducing citrus canker. Wb+Cu showed the highest efficiency for control of the disease. This combination reduced the incidence of diseased trees by ~60%, and the incidence of diseased leaves and fruit by ≥ 90% and increased the yield in 2.0 to 2.6-fold in comparison with the unmanaged plots. Cu sprays were important for reducing disease incidence and crop losses, whereas Wb had an additional contribution in minimizing the incidence of cankered, non-marketable fruit. The results indicated that the adoption of these measures of control may depend on the characteristics of the orchard and destination of the production.

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Increased Resistance Against Citrus Canker Mediated by a Citrus Mitogen-Activated Protein Kinase

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-04-13-0122-r ◽

2013 ◽

Vol 26 (10) ◽

pp. 1190-1199 ◽

Cited By ~ 15

Author(s):

Maria Luiza Peixoto de Oliveira ◽

Caio Cesar de Lima Silva ◽

Valéria Yukari Abe ◽

Marcio Gilberto Cardoso Costa ◽

Raúl Andrés Cernadas ◽

...

Keyword(s):

Hydrogen Peroxide ◽

Sweet Orange ◽

Plant Immunity ◽

Gene Promoter ◽

Citrus Canker ◽

Hypersensitive Reaction ◽

Mitogen Activated Protein Kinase ◽

Defense Gene Expression ◽

Defense Gene ◽

Mitogen Activated Protein

Mitogen-activated protein kinases (MAPK) play crucial roles in plant immunity. We previously identified a citrus MAPK (CsMAPK1) as a differentially expressed protein in response to infection by Xanthomonas aurantifolii, a bacterium that causes citrus canker in Mexican lime but a hypersensitive reaction in sweet oranges. Here, we confirm that, in sweet orange, CsMAPK1 is rapidly and preferentially induced by X. aurantifolii relative to Xanthomonas citri. To investigate the role of CsMAPK1 in citrus canker resistance, we expressed CsMAPK1 in citrus plants under the control of the PR5 gene promoter, which is induced by Xanthomonas infection and wounding. Increased expression of CsMAPK1 correlated with a reduction in canker symptoms and a decrease in bacterial growth. Canker lesions in plants with higher CsMAPK1 levels were smaller and showed fewer signs of epidermal rupture. Transgenic plants also revealed higher transcript levels of defense-related genes and a significant accumulation of hydrogen peroxide in response to wounding or X. citri infection. Accordingly, nontransgenic sweet orange leaves accumulate both CsMAPK1 and hydrogen peroxide in response to X. aurantifolii but not X. citri infection. These data, thus, indicate that CsMAPK1 functions in the citrus canker defense response by inducing defense gene expression and reactive oxygen species accumulation during infection.

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First Report of Xanthomonas citri pv. citri-A Causing Asiatic Citrus Canker in Mayotte

Plant Disease ◽

10.1094/pdis-01-13-0128-pdn ◽

2013 ◽

Vol 97 (7) ◽

pp. 989-989

Author(s):

J. Hoarau ◽

C. Boyer ◽

K. Vital ◽

T. Chesneau ◽

C. Vernière ◽

...

Keyword(s):

Indian Ocean ◽

Sweet Orange ◽

Citrus Canker ◽

Negative Control ◽

Sterile Water ◽

Citrus Species ◽

Indian Ocean Region ◽

Southwest Indian Ocean ◽

Mexican Lime ◽

Ocean Region

Asiatic citrus canker, caused by Xanthomonas citri pv. citri, is a bacterial disease of major economic importance in tropical and subtropical citrus-producing areas. X. citri pv. citri pathotype A can cause severe infection in a wide range of citrus species and induces erumpent, callus-like lesions with water-soaked margins evolving to corky cankers and leading to premature fruit and leaf drop and twig dieback on susceptible/very susceptible cultivars. A chlorotic halo is typically visible around canker lesions on leaves and young fruit, but not on mature fruit and twigs. This quarantine organism can strongly impact both national and international citrus markets. Long distance dispersal is mainly through infected propagative material. Asiatic citrus canker occurs on most islands in the Southwest Indian Ocean region (Comoros, Mauritius, Reunion, Rodrigues, and Seychelles islands), but was not yet reported in Mayotte (EPPO-PQR available at http://www.eppo.int ). In May 2012, typical canker-like symptoms were observed on sweet orange (Citrus sinensis) groves on Mtsamboro islet and soon after on the main island of Mayotte, mostly on sweet oranges, but also on Tahiti limes (C. latifolia) and mandarins (C. reticulata). Eighty-one Xanthomonas-like strains were isolated using KC semi-selective medium (4) from disease samples collected from both commercial groves and nurseries on different Citrus species located all over the island. Sixteen Xanthomonas-like isolates were tentatively identified as X. citri pv. citri based on a specific PCR assay with 4/7 primers (3). All strains but the negative control, sterile water, produced an amplicon of the expected size similar to X. citri pv. citri strain IAPAR 306 used as positive control. Multilocus sequence analysis targeting six housekeeping genes (atpD, dnaK, efp, gltA, gyrB, and lepA) (1,2) fully identified three strains from Mayotte (LJ225-3, LJ228-1, and LJ229-11) as X. citri pv. citri (and not other xanthomonad pathovars pathogenic to citrus or host range-restricted pathotypes of pathovar citri), and more specifically as sequence type ST2 composed of pathotype A strains of X. citri pv. citri (2) (including all strains from the Southwest Indian Ocean region). Eight strains were inoculated by a detached leaf assay (2) to Mexican lime SRA 140 (C. aurantifolia), Tahiti lime SRA 58, sweet orange cv. Washington Navel, alemow SRA 779 (C. macrophylla), and tangor cv. Ortanique (C. reticulata × C. sinensis) and developed typical erumpent, callus-like tissue at wound sites for all Citrus species, fulfilling Koch's postulates. Xanthomonas-like yellow colonies were reisolated from symptoms produced by the eight strains inoculated on Mexican lime. Boiled bacterial suspensions were assayed by PCR with 4/7 primers (3) and produced the expected 468-bp amplicon in contrast with the negative control (sterile water). No lesions developed on the negative control consisting of inoculations by 10 mM tris buffer (pH 7.2). Citrus canker-free nurseries and grove sanitation should be implemented for decreasing the prevalence of Asiatic canker in this island territory. References: (1) N. F. Almeida et al. Phytopathology 100:208, 2010. (2) L. Bui Thi Ngoc et al. Int. J. Syst. Evol. Microbiol. 60:515, 2010. (3) J. S. Hartung et al. Phytopathology 86:95, 1996. (4) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005.

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Comparison between Protoplast Transformation and Co-transformation in `Hamlin' Sweet Orange [Citrus sinensis (L.) Osbeck]

HortScience ◽

10.21273/hortsci.41.4.972d ◽

2006 ◽

Vol 41 (4) ◽

pp. 972D-973

Author(s):

Ahmad A. Omar ◽

Wen-Yuan Song ◽

James H. Graham ◽

Jude W. Grosser

Keyword(s):

Transgenic Plants ◽

Citrus Sinensis ◽

Fluorescent Protein ◽

Sweet Orange ◽

Citrus Canker ◽

Yield Potential ◽

High Yield ◽

Pcr Analysis ◽

Dna Encoding ◽

Early Maturity

Citrus canker disease caused by the bacterial pathogen Xanthomonas axonopodis pv. citri is becoming a worldwide problem. Xa21 gene is a member of the Xa21 gene family of rice, which provides broad spectrum Xanthomonas resistance in rice. `Hamlin' sweet orange [Citrus sinensis (L.) Osbeck) is one of the leading commercial cultivars in Florida because of its high yield potential and early maturity. `Hamlin' also has a high regeneration capacity from protoplasts and is often used in transformation experiments. Since the citrus canker pathogen is in the same genus, this gene may have potential to function against canker in citrus. The wild-type Xa21 gene contains an intron, and there are some questions whether dicot plants can process genes containing monocot introns (the cDNA is intron-free). Plasmids DNA, encoding the non-destructive selectable marker EGFP (Enhanced Green Fluorescent Protein) gene and the cDNA of the Xa21 gene were transformed or co-transformed into `Hamlin' orange protoplasts using polyethylene glycol. More than 200 transgenic embryoids were recovered. More than 400 transgenic plants were developed from 75 independent transgenic events. PCR analysis revealed the presence of the cDNA of the Xa21 and the GFP genes in the transgenic plants. Some of the plants have the GFP only. Southern analysis is showing integration of the cDNA into different sites ranges from one to five sites. Western analysis is showing the expression of the cDNA of the Xa21 gene in the transgenic citrus plants. This is the first time that a gene from rice has been stably integrated and expressed in citrus plants. Canker challenge assay is in progress.

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Reaction of detached leaves from different varieties of sweet orange to inoculation with Xanthomonas citri subsp. citri

Summa Phytopathologica ◽

10.1590/0100-5405/2143 ◽

2016 ◽

Vol 42 (2) ◽

pp. 125-133 ◽

Cited By ~ 2

Author(s):

Aline Maria Orbolato Gonçalves-Zuliani ◽

Kátia Aparecida Kern Cardoso ◽

José Belasque Junior ◽

Carlos Alexandre Zanutto ◽

Heraldo Takao Hashiguti ◽

...

Keyword(s):

Defense Mechanism ◽

Sweet Orange ◽

Citrus Canker ◽

Resistant Variety ◽

Xanthomonas Citri ◽

Lesion Development ◽

Sem Images ◽

Detached Leaf ◽

Detached Leaves ◽

Resistance To Infection

ABSTRACT Citrus genotypes show a broad range of resistance to infection with Xanthomonas citri subsp citri (Xcc), the causal agent of Asiatic citrus canker (ACC). The aim of this study was to evaluate a detached leaf inoculation method to compare nine different citrus genotypes that exhibit varied resistance levels to ACC, and to evaluate infection, colonization and symptom development of the varieties by using Scanning Electron Microscopy (SEM). Inoculation of Xcc in the leaves was carried out by producing injuries with needles. Lesion diameters were measured at 7, 10 and 13 days after inoculation (DAI). The variety Pera IAC was most resistant to the pathogen and had lesions with smaller diameters, while the variety Washington was most susceptible and had the largest lesions, indicating low resistance to Xcc. SEM images were taken from the lesion development, and sections were obtained from the disease lesions in the tested varieties. The most resistant variety, Pera IAC, had smaller stomatal openings and produced greater quantity of foliar exudates, compared with the variety Washington. We hypothesize that the exudates may contain Xcc toxic compounds and thus may be a defense mechanism against infection by Xcc, but further research is required to establish any antibiotic effects of the exudates against citrus canker.

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