Detection and quantification of Leveillula taurica growth in pepper leaves

Article

Zheng, Zheng, Nonomura, Teruo, Boka, Karoly, Matsuda, Yoshinori, Visser, Richard G. F., Toyoda, Hideyoshi, Kiss, Levente and Bai, Yuling. 2013. "Detection and quantification of Leveillula taurica growth in pepper leaves." Phytopathology: International Journal of the American Phytopathological Society. 103 (6), pp. 623-632. https://doi.org/10.1094/PHYTO-08-12-0198-R
Article Title

Detection and quantification of Leveillula taurica growth in pepper leaves

ERA Journal ID2639
Article CategoryArticle
AuthorsZheng, Zheng (Author), Nonomura, Teruo (Author), Boka, Karoly (Author), Matsuda, Yoshinori (Author), Visser, Richard G. F. (Author), Toyoda, Hideyoshi (Author), Kiss, Levente (Author) and Bai, Yuling (Author)
Journal TitlePhytopathology: International Journal of the American Phytopathological Society
Journal Citation103 (6), pp. 623-632
Number of Pages10
Year2013
PublisherAmerican Phytopathological Society
Place of PublicationUnited States
ISSN0031-949X
1943-7684
Digital Object Identifier (DOI)https://doi.org/10.1094/PHYTO-08-12-0198-R
Web Address (URL)http://apsjournals.apsnet.org/doi/pdf/10.1094/PHYTO-08-12-0198-R
Abstract

Leveillula taurica is an obligate fungal pathogen that causes powdery mildew disease on a broad range of plants, including important crops such as pepper, tomato, eggplant, onion, cotton, and so on. The early stage of this disease is difficult to diagnose and the disease can easily spread unobserved; for example, in pepper and tomato production fields and greenhouses. The objective of this study was to develop a detection and quantification method of L. taurica biomass in pepper leaves with special regard to the early stages of infection. We monitored the development of the disease to time the infection process on the leaf surface as well as inside the pepper leaves. The initial and final steps of the infection taking place on the leaf surface were consecutively observed using a dissecting microscope and a scanning electron microscope. The development of the intercellular mycelium in the mesophyll was followed by light and transmission electron microscopy. A pair of L. taurica-specific primers was designed based on the internal transcribed spacer sequence of L. taurica and used in real-time polymerase chain reaction (PCR) assay to quantify the fungal DNA during infection. The specificity of this assay was confirmed by testing the primer pair with DNA from host plants and also from another powdery mildew species, Oidium neolycopersici, infecting tomato. A standard curve was obtained for absolute quantification of L. taurica biomass. In addition, we tested a relative quantification method by using a plant gene as reference and the obtained results were compared with the visual disease index scoring. The realtime PCR assay for L. taurica provides a valuable tool for detection and quantification of this pathogen in breeding activities as well in plant-microbe interaction studies.

Keywordsfungal DNA; Ascomycetes; isolation and purification; microbiology; pepper; physiology; plant disease; plant leaf; real time polymerase chain reaction; transmission electron microscopy; ultrastructure; Ascomycota; capsicum; DNA; fungal; transmission electron microscopy; plant diseases; plant leaves; Real-Time Polymerase Chain Reaction
ANZSRC Field of Research 2020310599. Genetics not elsewhere classified
310705. Mycology
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Byline AffiliationsWageningen University, Netherlands
Kindai University, Japan
Eotvos Lorand University (ELTE), Hungary
Hungarian Academy of Sciences, Hungary
Institution of OriginUniversity of Southern Queensland
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Limits of nuclear ribosomal DNA internal transcribed spacer (ITS) sequences as species barcodes for Fungi
Kiss, Levente. 2012. "Limits of nuclear ribosomal DNA internal transcribed spacer (ITS) sequences as species barcodes for Fungi." University of Adelaide. 109 (27), p. 1811. https://doi.org/10.1073/pnas.1207143109
Host phenology and geography as drivers of differentiation in generalist fungal mycoparasites
Pintye, Alexandra, Ropars, Jeanne, Harvey, Nick, Shin, Hyeon Dong, Leyronas, Christel, Nicot, Philippe C., Giraud, Tatiana and Kiss, Levente. 2015. "Host phenology and geography as drivers of differentiation in generalist fungal mycoparasites." PLoS One. 10 (3). https://doi.org/10.1371/journal.pone.0120703
Does the parasite follow its host? Occurrence of morphologically barely distinguishable powdery mildew anamorphs on Oenothera spp. in different parts of the world
Bereczky, Zsolt, Pintye, Alexandra, Csontos, Peter, Braun, Uwe and Kiss, Levente. 2015. "Does the parasite follow its host? Occurrence of morphologically barely distinguishable powdery mildew anamorphs on Oenothera spp. in different parts of the world." Mycoscience. 56 (3), pp. 267-272. https://doi.org/10.1016/j.myc.2014.08.001
First detection of tomato powdery mildew caused by Oidium neolycopersici in South Africa
Lebeda, A., Mieslerova, B., Jankovics, T., Kiss, L. and van der Linde, E. J.. 2015. "First detection of tomato powdery mildew caused by Oidium neolycopersici in South Africa." South African Journal of Botany. 99, pp. 153-157. https://doi.org/10.1016/j.sajb.2015.03.196
Refining a method for ascospore viability testing in overwintering chasmothecia of Erysiphe necator
Vagi, Pal, Caffi, Tito, Vaczy, Kalman Zoltan, Nemeth, Mark Z. and Kiss, Levente. 2016. "Refining a method for ascospore viability testing in overwintering chasmothecia of Erysiphe necator." European Journal of Plant Pathology. 144 (4), pp. 799-802. https://doi.org/10.1007/s10658-015-0797-2
Sporulation rate in culture and mycoparasitic activity, but not mycohost specificity, are the key factors for selecting Ampelomyces strains for biocontrol of grapevine powdery mildew (Erysiphe necator)
Legler, Sara Elisabetta, Pintye, Alexandra, Caffi, Tito, Gulyas, Szilvia, Bohar, Gyula, Rossi, Vittorio and Kiss, Levente. 2016. "Sporulation rate in culture and mycoparasitic activity, but not mycohost specificity, are the key factors for selecting Ampelomyces strains for biocontrol of grapevine powdery mildew (Erysiphe necator)." European Journal of Plant Pathology. 144 (4), pp. 723-736. https://doi.org/10.1007/s10658-015-0834-1
Molecular identification and pathogenicity assessment of a rust fungus infecting common ragweed (Ambrosia artemisiifolia) in its native North American range
Kassai-Jager, Edit, Seier, Marion K., Evans, Harry C. and Kiss, Levente. 2016. "Molecular identification and pathogenicity assessment of a rust fungus infecting common ragweed (Ambrosia artemisiifolia) in its native North American range." European Journal of Plant Pathology. 145 (1), pp. 81-87. https://doi.org/10.1007/s10658-015-0815-4