Barley stripe mosaic virus-induced gene silencing (BSMV-IGS) as a tool for functional analysis of barley genes potentially involved in nonhost resistance

Delventhal, Rhoda; Zellerhoff, Nina; Schaffrath, Ulrich

doi:10.4161/psb.6.6.15240

Cited by 8 publications

(7 citation statements)

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“…Once the correct target(s) have been recognized, different experimental approaches can be deployed to generate mlo loss‐of‐function genotypes. Barley stripe mosaic virus ‐induced gene silencing (BSMV‐VIGS) and antisense approaches have been used to downregulate Mlo expression in barley (Schweizer et al ., ; Delventhal et al ., ), wheat (Várallyay et al ., ) and peach (Jiwan et al ., ), demonstrating that targeting MLO genes via RNA interference can result in enhanced powdery mildew resistance. However, this approach relies on sustained transgene expression, which is often undesirable (e.g.…”

Section: Let It Be: Breeding For the Futurementioning

confidence: 99%

Magical mystery tour: MLO proteins in plant immunity and beyond

2014

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Summary Stable heritable restriction of the ubiquitous powdery mildew disease is a desirable trait for agri‐ and horticulture. In barley (Hordeum vulgare), loss‐of‐function mutant alleles of the Mildew resistance locus o (Mlo) gene confer broad‐spectrum resistance to almost all known isolates of the fungal barley powdery mildew pathogen, Blumeria graminis f.sp. hordei. Despite extensive cultivation of barley mlo genotypes, mlo resistance has been durable in the field. Mlo genes are present as small families in the genomes of all higher plant species. The presumed negative regulatory role of particular members in plant immunity is evolutionarily conserved, as powdery mildew resistant mlo mutants have also been described in Arabidopsis thaliana, tomato (Solanum lycopersicum) and pea (Pisum sativum). Barley Mlo encodes a plasma membrane‐localized seven‐transmembrane domain protein of unknown biochemical activity. Here, we review the known requirements for mlo‐mediated disease resistance in barley and Arabidopsis and reflect current views regarding Mlo function. We discuss additional mlo mutant phenotypes recently discovered in Arabidopsis and present a meta‐analysis of the phylogenetic relationships within the Mlo family. Finally, we consider the novel versatile tools for functional analysis and targeted genome modification that can be used to induce mlo‐based powdery mildew resistance in virtually any plant species.

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Section: Let It Be: Breeding For the Futurementioning

confidence: 99%

Magical mystery tour: MLO proteins in plant immunity and beyond

2014

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“…Our next aim was to functionally evaluate whether CYP96B22 has a crucial role in the defense of barley against different Magnaporthe isolates. For this purpose, we used a transient Barley stripe mosaic virus (BSMV)-induced gene silencing assay which was first described by Holzberg et al [ 24 ] and which we had already adapted for Magnaporthe infection assays [ 25 ]. We inserted a 234 bp fragment of the CYP96B22 coding sequence into the γ-subunit of BSMV.…”

Section: Resultsmentioning

confidence: 99%

“…We addressed the biological function of CYP96B22 in a transient assay using BSMV as a tool to knock-down gene expression (Figure 4 ). The usefulness of BSMV-mediated VIGS for functional genomics in wheat and barley has been demonstrated in several recent publications [ 37 , 38 ] and we already adopted the assay for the study of barley- Magnaporthe interactions [ 25 ]. Since it was shown for wheat that BSMV-infection prior to inoculation with M. oryzae host isolates may lead to decreased penetration [ 39 ], we used in all of our VIGS-experiments plants inoculated with BSMV without silencing construct as an internal control (Figure 4 ).…”

Section: Discussionmentioning

confidence: 99%

Ectoparasitic growth of Magnaporthe on barley triggers expression of the putative barley wax biosynthesis gene CYP96B22 which is involved in penetration resistance

et al. 2014

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BackgroundHead blast caused by the fungal plant pathogen Magnaporthe oryzae is an upcoming threat for wheat and barley cultivation. We investigated the nonhost response of barley to an isolate of the Magnaporthe species complex which is pathogenic on Pennisetum spp. as a potential source for novel resistance traits.ResultsArray experiments identified a barley gene encoding a putative cytochrome P450 monooxygenase whose transcripts accumulate to a higher concentration in the nonhost as compared to the host interaction. The gene clusters within the CYP96 clade of the P450 plant gene family and is designated as CYP96B22. Expression of CYP96B22 was triggered during the ectoparasitic growth of the pathogen on the outside of the leaf. Usage of a fungicidal treatment and a Magnaporthe mutant confirmed that penetration was not necessary for this early activation of CYP96B22. Transcriptional silencing of CYP96B22 using Barley stripe mosaic virus led to a decrease in penetration resistance of barley plants to Magnaporthe host and nonhost isolates. This phenotype seems to be specific for the barley-Magnaporthe interaction, since penetration of the adapted barley powdery mildew fungus was not altered in similarly treated plants.ConclusionTaken together our results suggest a cross-talk between barley and Magnaporthe isolates across the plant surface. Since members of the plant CYP96 family are known to be involved in synthesis of epicuticular waxes, these substances or their derivatives might act as signal components. We propose a functional overlap of CYP96B22 in the execution of penetration resistance during basal and nonhost resistance of barley against different Magnaporthe species.

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“…Numerous studies were aimed at maximizing the level of transient expression, with a view to establishing the most suitable conditions required to achieve stable transformation and the generation of transgenic plants. Examples in barley have included the analysis of the plant-pathogen interaction in the leaf epidermis (Zimmermann et al 2006;Delventhal et al 2011) and the assessment of gene function in the response to dehydration (Marzin et al 2008). Later, the biolistic method was perfected, in which metal particles coated with a plasmid harbouring the transgene were fired into the host explant.…”

Section: Transient Expression Systemsmentioning

confidence: 99%

Biotechnological Approaches to Barley Improvement

Kumlehn

Stein

2014

Biotechnology in Agriculture and Forestry

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This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. Exempted from this legal reservation are brief excerpts in connection with reviews or scholarly analysis or material supplied specifically for the purpose of being entered and executed on a computer system, for exclusive use by the purchaser of the work. Duplication of this publication or parts thereof is permitted only under the provisions of the Copyright Law of the Publisher's location, in its current version, and permission for use must always be obtained from Springer. Permissions for use may be obtained through RightsLink at the Copyright Clearance Center. Violations are liable to prosecution under the respective Copyright Law. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. While the advice and information in this book are believed to be true and accurate at the date of publication, neither the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may be made. The publisher makes no warranty, express or implied, with respect to the material contained herein.Printed on acid-free paper Springer is part of Springer Science+Business Media (www.springer.com) PrefaceBarley is deemed one of the first plant species to be domesticated. Its cultivation over a very broad range of environments and its versatile utilization for the production of raw material for feed and food render it a top-four crop plant worldwide. Representing the temperate cereals of the Triticeae tribe (wheat, rye, triticale), barley has been used as experimental model since the era of classical genetics and cytogenetics-a role that has been reinforced along the development of contemporary biology and crop plant research.The generation of comprehensive genome sequence data is regarded as a cornerstone with major impact into all areas of modern barley research. Thus, we are convinced of the timeliness and importance to survey and reflect the current state of biotechnologically oriented barley research in the present volume of the Springer series Biotechnology in Agriculture and Forestry. Besides genome sequencing, major progress was achieved by deep sequencing-based transcriptomics, the establishment of comprehensive mutant panels and TILLING platforms, the generation of high-resolution genetic maps, efficient site-directed mutagenesis using customizable endonucleases, as well as the establishment of automa...

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Barley stripe mosaic virus-induced gene silencing (BSMV-IGS) as a tool for functional analysis of barley genes potentially involved in nonhost resistance

Cited by 8 publications

References 20 publications

Magical mystery tour: MLO proteins in plant immunity and beyond

Magical mystery tour: MLO proteins in plant immunity and beyond

Ectoparasitic growth of Magnaporthe on barley triggers expression of the putative barley wax biosynthesis gene CYP96B22 which is involved in penetration resistance

Biotechnological Approaches to Barley Improvement

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