Transient transformation of Podosphaera xanthii by electroporation of conidia
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- 作者：David Vela-Corcía ; Diego Romero ; Juan Antonio Torés ; Antonio De Vicente…
- 关键词：Electrotransformation ; Hygromycin B resistance ; MBC resistance ; Powdery mildew fungi ; β ; tubulin
- 刊名：BMC Microbiology
- 出版年：2015
- 出版时间：December 2015
- 年：2015
- 卷：15
- 期：1
- 全文大小：1,800 KB
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Diego Romero (1)
Juan Antonio Torés (2)
Antonio De Vicente (1)
Alejandro Pérez-García (1)
1. Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora- Universidad de Málaga - Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Departamento de Microbiología, Universidad de Málaga, Bulevar Louis Pasteur 31 (Campus Universitario de Teatinos), 29071, Málaga, Spain
2. Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora- Universidad de Málaga - Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Estación Experimental “La Mayora- 29750 Algarrobo-Costa, Málaga, Spain- 刊物主题：Microbiology; Biological Microscopy; Fungus Genetics; Parasitology; Virology; Life Sciences, general;
- 出版者：BioMed Central
- ISSN：1471-2180
- 作者单位：David Vela-Corcía (1)
文摘
Background Powdery mildew diseases are a major phytosanitary issue causing important yield and economic losses in agronomic, horticultural and ornamental crops. Powdery mildew fungi are obligate biotrophic parasites unable to grow on culture media, a fact that has significantly limited their genetic manipulation. In this work, we report a protocol based on the electroporation of fungal conidia, for the transient transformation of Podosphaera fusca (synonym Podosphaera xanthii), the main causal agent of cucurbit powdery mildew. Results To introduce DNA into P. xanthii conidia, we applied two square-wave pulses of 1.7?kV for 1?ms with an interval of 5?s. We tested these conditions with several plasmids bearing as selective markers hygromycin B resistance (hph), carbendazim resistance (TUB2) or GFP (gfp) under control of endogenous regulatory elements from Aspergillus nidulans, Neurospora crassa or P. xanthii to drive their expression. An in planta selection procedure using the MBC fungicide carbendazim permitted the propagation of transformants onto zucchini cotyledons and avoided the phytotoxicity associated with hygromycin B. Conclusion This is the first report on the transformation of P. xanthii and the transformation of powdery mildew fungi using electroporation. Although the transformants are transient, this represents a feasible method for the genetic manipulation of this important group of plant pathogens.