DETEKSI PENYAKIT BACTERIAL FRUIT BLOTCH PADA MELON MENGGUNAKAN ELISA (DETECTION OF BACTERIAL FRUIT BLOTCH OF MELON USING ELISA)

Bacterial fruit blotch (BFB) caused by Acidovorax citrulli is a serious seedborne disease in Cucurbitaceae causing 90-100% yield losses. The aim of this study was to explore BFB symptom on melon and also to detect A. citrulli infection in commercial seed and symptomatic fruits from the field in Yogyakarta Special Region province and its surrounding using DAS-ELISA method. Samples include melon from Sleman, Bantul, Kulon Progo, Gunung Kidul, Magelang, Purworejo regencies while commercial seeds i.e. Action 434, Glamour and Mai 116 were collected. DASELISA detection method used reagent set from Agdia. Based on the field observation, this study found melon commercial fruit shares similar symptom with BFB, which showed discrete oily dark green spots, while the netting failed to develop over necrotic areas, resulting in smooth sunken spots. DAS-ELISA detection revealed that samples collected from Jetak village, district of Mungkid, Magelang and from Bligo village, district of Ngluwar, Magelang and in commercial seed Mai 116 were positively infected byA. citrulli.Keywords: Acidovorax citrulli, bacterial fruit blotch, ELISA, melonBacterial fruit blotch (BFB) merupakan penyakit penting pada famili Cucurbitaceae yang disebabkan oleh Acidovorax citrulli. Penyakit ini dilaporkan dapat menurunkan hasil mencapai 90-100%. Tujuan dari penelitian ini adalah untuk mengetahui gejala penyakit BFB pada melon dan deteksiA. citrulli pada benih komersial dan sampel buah bergejala dengan metode DAS-ELISA di DIY dan sekitarnya. Pengambilan sampel dilakukan di kabupaten Sleman, Bantul, Kulon Progo, Gunung Kidul, Purworejo dan Magelang. Selain dari lapangan, diuji pula benih melon komersial yaitu Action-434, Glamour dan Mai 116. Metode deteksi dengan ELISA menggunakanreagent set dari Agdia. Dari hasil pengamatan di lapangan ditemukan buah melon dengan gejala yang mirip dengan gejala BFB yaitu adanya becak berwarna hijau tua kebasahan pada permukaan buah, jaring tidak terbentuk sempurna dan pada bagian daging buah di bawah becak tadi membusuk. Hasil deteksi dengan DAS-ELISAmengindikasikan bahwa A. citrulli terdeteksi pada sampel yang berasal dari desa Bligo, kecamatan Ngluwar dan desa Jetak, kecamatan Mungkid, kabupaten Magelang, serta pada benih komersial MAI 116.Kata kunci: Acidovorax citrulli, bacterial fruit blotch, ELISA, melon

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Development of Molecular Markers for Detection of Acidovorax citrulli Strains Causing Bacterial Fruit Blotch Disease in Melon

International Journal of Molecular Sciences ◽

10.3390/ijms20112715 ◽

2019 ◽

Vol 20 (11) ◽

pp. 2715 ◽

Cited By ~ 1

Author(s):

Md. Rafiqul Islam ◽

Mohammad Rashed Hossain ◽

Hoy-Taek Kim ◽

Denison Michael Immanuel Jesse ◽

Md. Abuyusuf ◽

...

Keyword(s):

Infected Leaf ◽

Bacterial Fruit Blotch ◽

Early Disease Detection ◽

Acidovorax Citrulli ◽

Seed Health ◽

Field Samples ◽

Whole Genomes ◽

Pathogenesis Related ◽

Primer Sets ◽

Inoculation Assay

Acidovorax citrulli (A. citrulli) strains cause bacterial fruit blotch (BFB) in cucurbit crops and affect melon significantly. Numerous strains of the bacterium have been isolated from melon hosts globally. Strains that are aggressively virulent towards melon and diagnostic markers for detecting such strains are yet to be identified. Using a cross-inoculation assay, we demonstrated that two Korean strains of A. citrulli, NIHHS15-280 and KACC18782, are highly virulent towards melon but avirulent/mildly virulent to the other cucurbit crops. The whole genomes of three A. citrulli strains isolated from melon and three from watermelon were aligned, allowing the design of three primer sets (AcM13, AcM380, and AcM797) that are specific to melon host strains, from three pathogenesis-related genes. These primers successfully detected the target strain NIHHS15-280 in polymerase chain reaction (PCR) assays from a very low concentration of bacterial gDNA. They were also effective in detecting the target strains from artificially infected leaf, fruit, and seed washing suspensions, without requiring the extraction of bacterial DNA. This is the first report of PCR-based markers that offer reliable, sensitive, and rapid detection of strains of A. citrulli causing BFB in melon. These markers may also be useful in early disease detection in the field samples, in seed health tests, and for international quarantine purposes.

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Bacteriophage Biocontrol of Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch

Proceedings ◽

10.3390/proceedings2020050010 ◽

2020 ◽

Vol 50 (1) ◽

pp. 10

Author(s):

Aryan Rahimi-Midani ◽

Tae-Jin Choi

Keyword(s):

Crop Production ◽

Germination Rate ◽

Causal Agent ◽

Head Size ◽

Genotype I ◽

Bacterial Fruit Blotch ◽

Double Stranded Dna ◽

Acidovorax Citrulli ◽

Group I ◽

Next Generation Sequencing Ngs

Bacterial fruit blotch caused by Acidovorax citrulli is known to be the major threat to cucurbit crop production worldwide. The pathogen can penetrate into seed coat and cause disease symptoms at any stage of plant growth, which results in fruit loss. Two main genotypes (genotype I and II) are reported in A. citrulli, in which genotype II is the main cause of Bacterial Fruit Blotch (BFB) in watermelon and group I is known to be a causal agent of BFB in melon. To date, there are no commercially available cultivars resistant to BFB, and available strategies are not able to completely manage the disease. In this study, we aim to isolate bacteriophages to control BFB. Samples collected from watermelon, melon, and pumpkin were used to isolate bacteriophages. All isolated bacteriophages were tested against 42 strains of A. citrulli, among which two phages with the ability to lyse a greater number of hosts were selected and characterized. Bacteriophage ACP17 from the Myoviridae family, with a head size of 100 ± 5 nm and tail of 150 ± 5 nm, infected 29 strains of A. citrulli mostly belonging to genotype group I, whereas the second isolated bacteriophage, ACPWH from Siphoviridae, with a head size of 60 ± 5 nm and tail of 180 ± 5 nm, infected 39 A. citrulli strains. Genome analysis of both bacteriophages using Next generation Sequencing (NGS) showed that ACP17 and ACPWH have double-stranded DNA with sizes of 156,972 kb and 424,299 kb, respectively. Watermelon seeds coated with ACPWH showed a germination rate of up to 90% in the presence of A. citrulli in contrast to untreated seed, which showed no germination or germinated juveniles with BFB symptoms in the presence of A. citrulli. The results of this study show that the use of bacteriophages of A. citrulli represents a potential biocontrol method for controlling BFB.

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Short communication: Pepino mosaic virus, a new threat for Serbia’s tomatoes

Spanish Journal of Agricultural Research ◽

10.5424/sjar/2020184-16244 ◽

2020 ◽

Vol 18 (4) ◽

pp. e10SC05

Author(s):

Ivana Stankovic ◽

Ana Vucurovic ◽

Katarina Zecevic ◽

Branka Petrovic ◽

Danijela Ristic ◽

...

Keyword(s):

Mosaic Virus ◽

Rt Pcr ◽

Tomato Production ◽

Pepino Mosaic Virus ◽

Tomato Plants ◽

Rapid Spread ◽

Infected Tomato ◽

Dark Green ◽

Relationship Of ◽

Das Elisa

Aim of study: To report the occurrence of Pepino mosaic virus (PepMV) on tomato in Serbia and to genetically characterize Serbian PepMV isolates.Area of study: Tomato samples showing virus-like symptoms were collected in the Bogojevce locality (Jablanica District, Serbia).Material and methods: Collected tomato samples were assayed by DAS-ELISA using antisera against eight economically important or quarantine tomato viruses. Three selected isolates of naturally infected tomato plants were mechanically transmitted to tomato ‘Novosadski jabučar’ seedlings. For confirmation of PepMV infection, RT-PCR was performed using specific primers PepMV TGB F/PepMV UTR R. Maximum-likelihood phylogenetic tree was constructed with 47 complete CP gene sequences of PepMV to determine the genetic relationship of Serbian PepMV isolates with those from other parts of the world.Main results: The results of DAS-ELISA indicated the presence of PepMV in all tested samples. Mechanically inoculated ‘Novosadski jabučar’ seedlings expressed yellow spots and light and dark green patches, bubbling, and curled leaves. All tested tomato plants were RT-PCR positive for the presence of PepMV. The CP sequence analysis revealed that the Serbian PepMV isolates were completely identical among themselves and shared the highest nucleotide identity of 95.1% (99.2% aa identity) with isolate from Spain (FJ263341). Phylogenetic analysis showed clustering of the Serbian PepMV isolates into CH2 strain, but they formed separate subgroup within CH2 strain.Research highlights: This is the first data of the presence of PepMV in protected tomato production in Serbia. Considering increased incidence and rapid spread in Europe, the presence of PepMV on tomato could therefore represent serious threat to this valuable crop in Serbia.

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First Report of Bacterial Fruit Blotch of Watermelon Caused by Acidovorax avenae subsp. citrulli in Illinois

Plant Disease ◽

10.1094/pdis.2002.86.4.443a ◽

2002 ◽

Vol 86 (4) ◽

pp. 443-443 ◽

Cited By ~ 3

Author(s):

M. Babadoost ◽

N. Pataky

Keyword(s):

Citrullus Lanatus ◽

Test Tube ◽

Green Water ◽

Enzyme Linked Immunosorbent Assay ◽

Distilled Water ◽

Mature Fruit ◽

American Phytopathological Society ◽

Bacterial Fruit Blotch ◽

Dark Green ◽

Fruit Surface

In August 2001, symptomatic mature fruit of watermelon (Citrullus lanatus) cvs. Crimson Sweet and Royal Sweet were collected from six fields (48 ha) in Iroquois, Kankakee, and Mason counties in Illinois. The fruit surface had dark green, water-soaked lesions, and the rind's epidermis was cracked. Sections of infected rind were examined with a light microscope, and bacterial streaming was observed. Rind tissue was disinfested by rubbing the fruit surface with a paper towel moistened with 70% ethanol. Pieces of rind from the margins of lesions were placed in a test tube containing sterile-distilled water. The tube was shaken, and the suspension was streaked on King's medium B agar in petri plates. The plates were incubated at 26°C for 3 days. A nonfluorescent, gramnegative, anaerobic, oxidase-positive, rod-shaped bacterium was isolated that produced creamy white colonies on nutrient agar. The bacterium was identified as Acidovorax avenae subsp. citrulli based on an enzyme-linked immunosorbent assay test specific for A. avenae subsp. citrulli (kit PSA 14,800; Agdia Inc., Elkhart, IN). The disease was diagnosed as bacterial fruit blotch (1,2). The occurrence of bacterial fruit blotch of watermelon in Illinois is important because (i) watermelon is widely grown in Illinois, (ii) the pathogen also infects melons and pumpkins, and (iii) Illinois grows approximately 1,000 ha of melons and 8,000 ha of pumpkins. References: (1) R. X. Latin. Bacterial fruit blotch. Pages 34-35 in: Compendium of Cucurbit Diseases. The American Phytopathological Society, St. Paul, MN,1996. (2) G. S. Somodi et al. Plant Dis. 75:1053, 1991.

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First Report of Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch, on Melon (Cucumis melo) in Guadeloupe (France)

Plant Disease ◽

10.1094/pdis-10-18-1825-pdn ◽

2019 ◽

Vol 103 (5) ◽

pp. 1017-1017 ◽

Cited By ~ 1

Author(s):

A. Cunty ◽

C. Audusseau ◽

S. Paillard ◽

V. Olivier ◽

C. François ◽

...

Keyword(s):

Cucumis Melo ◽

Causal Agent ◽

First Report ◽

Bacterial Fruit Blotch ◽

Acidovorax Citrulli

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Acidovorax citrulli is sensitive to elevated temperatures during early stages of watermelon seed germination

Seed Science and Technology ◽

10.15258/sst.2020.48.1.02 ◽

2020 ◽

Vol 48 (1) ◽

pp. 11-20

Author(s):

Mei Zhao ◽

Ron Walcott

Keyword(s):

Elevated Temperature ◽

Seed Germination ◽

Elevated Temperatures ◽

Transmitted Disease ◽

Effect Of Temperature ◽

Crop Species ◽

Inoculum Concentration ◽

Bacterial Fruit Blotch ◽

Acidovorax Citrulli ◽

Germinating Seeds

Bacterial fruit blotch (BFB), caused by Acidovorax citrulli, is a seed-transmitted disease of cucurbit crop species. During seed-to-seedling transmission of BFB, A. citrulli initially grows as a saprophyte on germinating seeds and subsequently switches to a pathogenic mode. We investigated the effect of temperature on A. citrulli colonisation of germinating watermelon seeds. Seeds were vacuum-infiltrated with 106 CFU/ml A. citrulli, germinated at 28°C and 100% relative humidity, and transferred to 40°C at different times. Mean BFB incidence was significantly lower for seeds that were sown at 28°C and transferred to 40°C three days after sowing (DAS), compared with seeds incubated constantly at 28°C. Seeds showed reduced mean BFB transmission percentages when transferred from 28 to 40°C at 3 DAS, regardless of initial A. citrulli concentration. The effect of increased temperature on BFB seedling transmission was reversible regardless of the initial A. citrulli inoculum concentration. Furthermore, the A. citrulli population on germinating watermelon seedlings that were transferred from 28 to 40°C at 3 DAS was significantly lower than seedlings maintained at 28°C. We conclude that A. citrulli cells associated with germinating watermelon seeds are more sensitive to elevated temperature during the first 3 DAS relative to the later days.

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