Development of one-tube real-time qRT-PCR and evaluation of RNA extraction methods for the detection of Eggplant mottled dwarf virus in different species

Pappi, Polyxeni; Chaintoutis, Serafeim C.; Dovas, Chrysostomos Ι.; Efthimiou, K.; Katis, N. I.

doi:10.1016/j.jviromet.2014.11.001

Cited by 11 publications

(3 citation statements)

References 40 publications

(57 reference statements)

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“…Ten percent suspensions of each fecal sample were prepared in phosphate-buffered saline (PBS), and RNA was extracted according to RNA extraction method C described by Pappi et al (2015). RNA extracts were stored at −80°C until testing.…”

Section: Virological Analysismentioning

confidence: 99%

Evaluation of the in-field efficacy of oregano essential oil administration on the control of neonatal diarrhea syndrome in calves

Katsoulos

Karatzia

Dovas

et al. 2017

Research in Veterinary Science

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The objective of this study was to evaluate under field conditions, whether daily administration of oregano essential oil is effective in preventing and/or diminishing the severity of neonatal diarrhea syndrome in calves aged less than 15days. Ninety-one newborn calves from three dairy farms were assigned into two groups; "Eco" group (n=46) calves were drenched with Greek oregano (Origanum vulgare ssp. Hirtum) essential oil (ECODIAR® liquid 5%) at the dose of 12.5mg/kg body weight once per day until the age of 10days. "Conts" group (n=45) calves were left untreated and served as controls. All animals were monitored daily for the incidence of diarrhea until the age of 15days and their fecal score was recorded. Fecal samples were collected on days 3, 6 and 10 for microbiological and parasitological evaluation. Average fecal score throughout the experiment, incidence of diarrhea, duration and severity of diarrhea episodes were significantly lower in Eco group compared to the controls. Daily administration of oregano essential oil in calves for the first 10days of their life effectively diminishes the severity of naturally acquired diarrhea under field conditions and, under certain hygiene practices, possess a preventive effect against neonatal diarrhea syndrome.

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Section: Virological Analysismentioning

confidence: 99%

Evaluation of the in-field efficacy of oregano essential oil administration on the control of neonatal diarrhea syndrome in calves

Katsoulos

Karatzia

Dovas

et al. 2017

Research in Veterinary Science

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“…This virus is more polyphagous than PVY, since it has been reported in a range of solanaceous species (eggplant, tomato, potato, pepper) and other crops (e.g., cucumber and tobacco), ornamental and wild plants. EMDV is reportedly widespread in the Mediterranean basin [48,49]. To the best of our knowledge, this is the first report of EMDV in eggplant in Lebanon, a novelty that will require future monitoring for assessing the spread and the consequent damage that might occur due to this virus.…”

Section: Discussionmentioning

confidence: 82%

Occurrence and Distribution of Major Viruses Infecting Eggplant in Lebanon and Molecular Characterization of a Local Potato Virus X Isolate

et al. 2021

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This research was carried out in order to evaluate the presence and distribution of viral infections causing severe disease in eggplant plants collected from different districts in Bekaa valley, Lebanon. Most infected plants showed virus-like symptoms consisting predominantly of leaf blotch, mottling chlorotic and ring spots; leaf twisting and plant dwarf were also observed in the visited fields. Symptomatic and asymptomatic plants were collected and screened by ELISA test for the presence of several different pathogenic viruses potentially present in the area. Results showed that potato virus Y (PVY) was the most prevalent virus found by ELISA (detected in the 15.3% of the tested plants), followed by eggplant mottled dwarf virus (EMDV, 2.9%) and cucumber mosaic virus (CMV, 1.2%), while tomato spotted wilt virus (TSWV), alfalfa mosaic virus (AMV) and pepper mottle virus (PepMoV) were not detected. Biological indexing of symptomatic ELISA-negative plants, followed by electron microscopy, indicated the presence of virus-like particles of the genus Potexvirus, which was subsequently confirmed as potato virus X (PVX) by RT-PCR and Sanger sequencing. PVX was found in 35.3% of the tested plants, all sampled in the northern Bekaa area. In a phylogenetic analysis, the partial coat protein gene sequence of a selected Lebanese isolate, PVX-AK1, clustered together with other PVX isolates from Asia. Furthermore, the 124-aa sequence of PVX-AK1 shared 100% identity with PVX-UK3, an isolate which is known as avirulent in potato genotypes carrying either Nx or Rx resistance genes. This work revealed a picture of the previously uninvestigated phytosanitary status of eggplant crops in an important horticultural area of Lebanon.

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“…The qRT-PCR amplification conditions were as those described by Pappi et al 69 . The 25 μl qRT-PCR reaction consisted of buffer 5x, 1.5 μl of EVA Green, 0.2 mM of each dNTP, 4 mM DTT, 1 U of Superscript III RNaseH− Reverse Transcriptase, 1.5 mM additional MgSO 4 (Invitrogen-Life Technologies, Groningen, The Netherlands), 3 U of HotStartTaq DNA polymerase (Qiagen, Hilden, Germany) and 1 M of each primer.…”

Section: Discussionmentioning

confidence: 99%

Proteomic analysis upon peach fruit infection with Monilinia fructicola and M. laxa identify responses contributing to brown rot resistance

Papavasileiou

Tanou²,

Samaras

et al. 2020

Sci Rep

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Brown rot, caused by Monilinia spp., is a major peach disease worldwide. in this study, the response of peach cultivars Royal Glory (RG) and Rich Lady (RL) to infection by Monilinia fructicola or Monilinia laxa, was characterized. Phenotypic data, after artificial inoculations, revealed that 'RL' was relatively susceptible whereas 'RG' was moderately resistant to Monilinia spp. comparative proteomic analysis identified mesocarp proteins of the 2 cultivars whose accumulation were altered by the 2 Monilinia species. Functional analysis indicated that pathogen-affected proteins in 'RG' were mainly involved in energy and metabolism, while, differentially accumulated proteins by the pathogen presence in 'RL' were involved in disease/defense and metabolism. A higher number of proteins was differentiated in 'RG' fruit compared to 'RL'. Upon Monilinia spp. infection, various proteins were-down accumulated in 'RL' fruit. Protein identification by mass spectrometric analysis revealed that several defense-related proteins including thaumatin, formate dehydrogenase, S-formylglutathione hydrolase, cBS domaincontaining protein, HSP70, and glutathione S-transferase were up-accumulated in 'RG' fruit following inoculation. The expression profile of selected defense-related genes, such as major latex allergen, 1-aminocyclopropane-1-carboxylate deaminase and UDP-glycoltransferase was assessed by RT-PCR. This is the first study deciphering differential regulations of peach fruit proteome upon Monilinia infection elucidating resistance responses.

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Development of one-tube real-time qRT-PCR and evaluation of RNA extraction methods for the detection of Eggplant mottled dwarf virus in different species

Cited by 11 publications

References 40 publications

Evaluation of the in-field efficacy of oregano essential oil administration on the control of neonatal diarrhea syndrome in calves

Evaluation of the in-field efficacy of oregano essential oil administration on the control of neonatal diarrhea syndrome in calves

Occurrence and Distribution of Major Viruses Infecting Eggplant in Lebanon and Molecular Characterization of a Local Potato Virus X Isolate

Proteomic analysis upon peach fruit infection with Monilinia fructicola and M. laxa identify responses contributing to brown rot resistance

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