Detection and quantification of Leveillula taurica growth in pepper leaves

Zheng, Zheng and Nonomura, Teruo and Boka, Karoly and Matsuda, Yoshinori and Visser, Richard G. F. and Toyoda, Hideyoshi and Kiss, Levente and Bai, Yuling (2013) Detection and quantification of Leveillula taurica growth in pepper leaves. Phytopathology, 103 (6). pp. 623-632. ISSN 0031-949X

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.


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Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Additional Information: Permanent restricted access to Published version due to publisher copyright policy. However, the published version is freely accessible from the publisher's website at: http://apsjournals.apsnet.org/doi/abs/10.1094/PHYTO-08-12-0198-R
Faculty / Department / School: No Faculty
Date Deposited: 25 May 2017 06:15
Last Modified: 25 May 2017 06:26
Uncontrolled Keywords: fungal 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
Fields of Research : 06 Biological Sciences > 0604 Genetics > 060499 Genetics not elsewhere classified
06 Biological Sciences > 0605 Microbiology > 060505 Mycology
Socio-Economic Objective: D Environment > 96 Environment > 9604 Control of Pests, Diseases and Exotic Species > 960403 Control of Animal Pests, Diseases and Exotic Species in Farmland, Arable Cropland and Permanent Cropland Environments
Identification Number or DOI: 10.1094/PHYTO-08-12-0198-R
URI: http://eprints.usq.edu.au/id/eprint/31885

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