Upward movement of Verticillium dahliae from soil to olive plants detected by qPCR

Ceccherini, Maria Teresa; Luchi, Nicola; Pantani, Ottorino‐Luca; Ascher, Judith; Capretti, P.; Pietramellara, Giacomo

doi:10.1007/s11274-013-1342-0

Cited by 6 publications

(2 citation statements)

References 25 publications

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“…The use of real‐time PCR for detection and quantification of V. dahliae in samples from diseased olive trees has been reported by several authors (e.g. Mercado‐Blanco et al ., ; Lievens et al ., ; Markakis et al ., ; Ceccherini et al ., ; Gramaje et al ., ).…”

Section: Introductionmentioning

confidence: 99%

Reliable detection of unevenly distributed Verticillium dahliae in diseased olive trees

et al. 2016

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Verticillium wilt caused by Verticillium dahliae is one of the most threatening diseases of olive worldwide. For preplanting and post-planting control of verticillium wilt in olive trees, availability of a rapid, reliable and non-destructive method for detection of V. dahliae is essential. For such a method, suitable and easily performed sampling and efficient processing of samples for extraction of DNA are necessary. In this study, the suitability of young twig and leaf samples of olive trees, which are easy to collect and extract DNA from, were assessed for the detection of V. dahliae in routine procedures. The lower (about 50 cm from the tip) and top parts (about 5 cm from the tip) of twigs, as well as leaves from infected olive trees were screened for V. dahliae infection and distribution using real-time PCR. The biomass of V. dahliae detected in individual twigs was highly variable, but there was no significant difference between mean quantities of V. dahliae DNA detected in top and lower parts of twigs. Furthermore, it was demonstrated that analysis of combined samples containing DNA extracted from five twigs of an infected tree accurately detected the presence of the pathogen. Similarly, testing combined samples of 5-10 leaves enabled reliable detection of the pathogen in an infected tree. The development of this assay enables reliable detection of V. dahliae in infected olive trees that can aid in management decisions for the implementation of integrated disease management.

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Section: Introductionmentioning

confidence: 99%

Reliable detection of unevenly distributed Verticillium dahliae in diseased olive trees

et al. 2016

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“…Culture-independent methods have rapidly been recognized as a valuable alternative to culture-dependent methods (Agrimonti et al 2013;Ceccherini et al 2013;De Medici et al 2015;Ilabaca et al 2014;Ke et al 2014;Reitschuler et al 2014;Rodriguez-Lazaro et al 2015;Ruiz et al 2014). These methods can be based on the direct analysis of DNA extracted from the food matrix (Achilleos and Berthier 2013;Rodríguez et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Comparison of real-time PCR assay and plate count for Lactobacillus paracasei enumeration in yoghurt

et al. 2015

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Lactobacillus paracasei is a mesophilic lactic acid bacterium technologically active in food fermentation. Culture-independent methods have rapidly been recognized as a valuable alternative to culture-dependent methods for lactic acid bacteria enumeration. In the present work, the efficacy of different protocols to extract DNA from yoghurt were compared, real-time PCR (qPCR) targeting tuf gene for L. paracasei enumeration was evaluated, and qPCR and plate counts of L. paracasei in yoghurt samples were compared. Total DNA concentrations from commercial yoghurts were higher using DNAzol method 2 than using the other tested methods. Standard curves presented suitable mean efficiency values of 91 % (pure L. paracasei strain CTT 7501), 95 % (pure L. paracasei strain FNU), and 103 % (yoghurt with L. paracasei strain FNU). Limit of detection is 3 log DNA copy number, corresponding to 2.78 log CFU, a suitable range of CFU enumeration for probiotic bacteria in yoghurt samples, considering that they should be present in large amounts. The L. paracasei (CFU) enumerated by qPCR were compared to culturable L. paracasei enumerated by plate counts at 7, 14, 21, and 28 days of yoghurt manufacture. Differences between qPCR and plate counts were observed only 28 days after yoghurt preparation, counts were similar at 7, 14, and 21 days. In conclusion, this qPCR assay is a useful and rapid tool to enumerate L. paracasei in yoghurt, although it does not distinguish dead and viable cells.

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Powerful qPCR assays for the early detection of latent invaders: interdisciplinary approaches in clinical cancer research and plant pathology

Luchi

Capretti

Pazzagli

et al. 2016

Appl Microbiol Biotechnol

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Latent invaders represent the first step of disease before symptoms occur in the host. Based on recent findings, tumors are considered to be ecosystems in which cancer cells act as invasive species that interact with the native host cell species. Analogously, in plants latent fungal pathogens coevolve within symptomless host tissues. For these reasons, similar detection approaches can be used for an early diagnosis of the invasion process in both plants and humans to prevent or reduce the spread of the disease. Molecular tools based on the evaluation of nucleic acids have been developed for the specific, rapid, and early detection of human diseases. During the last decades, these techniques to assess and quantify the proliferation of latent invaders in host cells have been transferred from the medical field to different areas of scientific research, such as plant pathology. An improvement in molecular biology protocols (especially referring to qPCR assays) specifically designed and optimized for detection in host plants is therefore advisable. This work is a cross-disciplinary review discussing the use of a methodological approach that is employed within both medical and plant sciences. It provides an overview of the principal qPCR tools for the detection of latent invaders, focusing on comparisons between clinical cancer research and plant pathology, and recent advances in the early detection of latent invaders to improve prevention and control strategies.

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Upward movement of Verticillium dahliae from soil to olive plants detected by qPCR

Cited by 6 publications

References 25 publications

Reliable detection of unevenly distributed Verticillium dahliae in diseased olive trees

Reliable detection of unevenly distributed Verticillium dahliae in diseased olive trees

Comparison of real-time PCR assay and plate count for Lactobacillus paracasei enumeration in yoghurt

Powerful qPCR assays for the early detection of latent invaders: interdisciplinary approaches in clinical cancer research and plant pathology

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