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Altered trichothecene biosynthesis in TRI6-silenced transformants of Fusarium culmorum influences the severity of crown and foot rot on durum wheat seedlings

Scherm, Barbara and Orrù, Marcella and Balmas, Virgilio and Spanu, Francesca and Azara, Emanuela and Delogu, Giovanna and Hammond, Thomas M. and Keller, Nancy P. and Migheli, Quirico (2011) Altered trichothecene biosynthesis in TRI6-silenced transformants of Fusarium culmorum influences the severity of crown and foot rot on durum wheat seedlings. Molecular Plant Pathology, Vol. 12 (8), p. 759-771. eISSN 1364-3703. Article.

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DOI: 10.1111/J.1364-3703.2011.00709.X

Abstract

An RNA silencing construct was used to alter mycotoxin production in the plant pathogenic fungus Fusarium culmorum, the incitant of crown and foot rot on wheat. The transformation of a wild-type strain and its nitrate reductase-deficient mutant with inverted repeat transgenes (IRTs) containing sequences corresponding to the trichothecene regulatory gene TRI6 was achieved using hygromycin B resistance as a selectable marker. Southern analysis revealed a variety of integration patterns of the TRI6 IRT. One transformant underwent homologous recombination with deletion of the endogenous TRI6 gene, whereas, in another transformant, the TRI6 IRT was not integrated into the genome. The TRI6 IRT did not alter the physiological characteristics, such as spore production, pigmentation or growth rate, on solid media. In most transformants, a high TRI6 amplification signal was detected by quantitative reverse transcriptionpolymerase chain reaction, corresponding to a TRI6-hybridizing smear of degraded fragments by Northern analysis, whereas TRI5 expression decreased compared with the respective nontransformed strain. Four transformants showed increased TRI5 expression, which was correlated with a dramatic (up to 28-fold) augmentation of deoxynivalenol production. Pathogenicity assays on durum wheat seedlings confirmed that impairment of deoxynivalenol production in the TRI6 IRT transformants correlated with a loss of virulence, with decreased disease indices ranging from 40% to 80% in nine silenced strains, whereas the overproducing transformants displayed higher virulence compared with the wild-type.

Item Type:Article
ID Code:6184
Status:Published
Refereed:Yes
Uncontrolled Keywords:Fusarium culmorum, TRI6 IRT virulence, TRI5, trichothecene
Subjects:Area 07 - Scienze agrarie e veterinarie > AGR/12 Patologia vegetale
Divisions:001 Università di Sassari > 01 Dipartimenti > Protezione delle piante
002 Altri enti e centri di ricerca del Nord Sardegna > CNR-Consiglio Nazionale delle Ricerche > Istituto di chimica biomolecolare, Sassari
002 Altri enti e centri di ricerca del Nord Sardegna > Istituto nazionale biostrutture e biosistemi, Unità di ricerca, Sassari
Publisher:Wiley / Blackwell
eISSN:1364-3703
Copyright Holders:© 2011 The Authors. Molecular Plant Pathology © 2011 BSPP and Blackwell Publishing LTD
Deposited On:23 Jun 2011 17:36

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