Agrotransformation of Phytophthora nicotianae: a simplified and optimized method

• Main Authors: Ronaldo José Durigan Dalio, Paulo José Camargo dos Santos, Heros José Máximo, Pamela Ayumi Kawakami, Eduardo Goulin, Marcos Antônio Machado
• Format: Article
• Language: English
• Published: Grupo Paulista de Fitopatologia
• Series: Summa Phytopathologica
• Subjects: genetic transformation; DsRed; Agrobacterium tumefaciens
• Online Access: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-54052016000300254&lng=en&tlng=en

ABSTRACT Phytophthora nicotianae is a plant pathogen responsible for damaging crops and natural ecosystems worldwide. P. nicotianae is correlated with the diseases: citrus gummosis and citrus root rot, and the management of these diseases relies mainly on the certification of seedlings and eradication of infected trees. However, little is known about the infection strategies of P. nicotianae interacting with citrus plants, which rises up the need for examining its virulence at molecular levels. Here we show an optimized method to genetically manipulate P. nicotianae mycelium. We have transformed P. nicotianae with the expression cassette of fluorescence protein DsRed. The optimized AMT method generated relatively high transformation efficiency. It also shows advantages over the other methods since it is the simplest one, it does not require protoplasts or spores as targets, it is less expensive and it does not require specific equipment. Transformation with DsRed did not impair the physiology, reproduction or virulence of the pathogen. The optimized AMT method presented here is useful for rapid, cost-effective and reliable transformation of P. nicotianae with any gene of interest.