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      The status of RNAi-based transgenic research in plant nematology

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          Abstract

          With the understanding of nematode-plant interactions at the molecular level, new avenues for engineering resistance have opened up, with RNA interference being one of them. Induction of RNAi by delivering double-stranded RNA (dsRNA) has been very successful in the model non-parasitic nematode, Caenorhabditis elegans, while in plant nematodes, dsRNA delivery has been accomplished by soaking nematodes with dsRNA solution mixed with synthetic neurostimulants. The success of in vitro RNAi of target genes has inspired the use of in planta delivery of dsRNA to feeding nematodes. The most convincing success of host-delivered RNAi has been achieved against root-knot nematodes. Plant-mediated RNAi has been shown to lead to the specific down-regulation of target genes in invading nematodes, which had a profound effect on nematode development. RNAi-based transgenics are advantageous as they do not produce any functional foreign proteins and target organisms in a sequence-specific manner. Although the development of RNAi-based transgenics against plant nematodes is still in the preliminary stage, they offer novel management strategy for the future.

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          Top 10 plant-parasitic nematodes in molecular plant pathology.

          John Jones, Annelies Haegeman, Etienne G. J. Danchin … (2013)
          The aim of this review was to undertake a survey of researchers working with plant-parasitic nematodes in order to determine a 'top 10' list of these pathogens based on scientific and economic importance. Any such list will not be definitive as economic importance will vary depending on the region of the world in which a researcher is based. However, care was taken to include researchers from as many parts of the world as possible when carrying out the survey. The top 10 list emerging from the survey is composed of: (1) root-knot nematodes (Meloidogyne spp.); (2) cyst nematodes (Heterodera and Globodera spp.); (3) root lesion nematodes (Pratylenchus spp.); (4) the burrowing nematode Radopholus similis; (5) Ditylenchus dipsaci; (6) the pine wilt nematode Bursaphelenchus xylophilus; (7) the reniform nematode Rotylenchulus reniformis; (8) Xiphinema index (the only virus vector nematode to make the list); (9) Nacobbus aberrans; and (10) Aphelenchoides besseyi. The biology of each nematode (or nematode group) is reviewed briefly. © 2013 BSPP AND JOHN WILEY & SONS LTD.
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            Engineering broad root-knot resistance in transgenic plants by RNAi silencing of a conserved and essential root-knot nematode parasitism gene.

            Guozhong Huang, Rex Allen, Eric L. Davis … (2006)
            Secreted parasitism proteins encoded by parasitism genes expressed in esophageal gland cells mediate infection and parasitism of plants by root-knot nematodes (RKN). Parasitism gene 16D10 encodes a conserved RKN secretory peptide that stimulates root growth and functions as a ligand for a putative plant transcription factor. We used in vitro and in vivo RNA interference approaches to silence this parasitism gene in RKN and validate that the parasitism gene has an essential function in RKN parasitism of plants. Ingestion of 16D10 dsRNA in vitro silenced the target parasitism gene in RKN and resulted in reduced nematode infectivity. In vivo expression of 16D10 dsRNA in Arabidopsis resulted in resistance effective against the four major RKN species. Because no known natural resistance gene has this wide effective range of RKN resistance, bioengineering crops expressing dsRNA that silence target RKN parasitism genes to disrupt the parasitic process represents a viable and flexible means of developing novel durable RKN-resistant crops and could provide crops with unprecedented broad resistance to RKN.
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              Ingestion of double-stranded RNA by preparasitic juvenile cyst nematodes leads to RNA interference.

              Catherine J Lilley, William H. Atkinson, Peter E. Urwin (2002)
              RNA interference is of value in determining gene function in many organisms. Plant parasitic nematodes are refractory to microinjection as a means of introducing RNA and do not show any oral uptake until they are within plants. We have used octopamine to stimulate uptake by preparasitic second stage juveniles of two cyst nematodes, Heterodera glycines and Globodera pallida. This new technique was used to facilitate uptake of double stranded RNA (dsRNA) together with fluoroscein isothiocyanate as a visual marker. Targeting cysteine proteinases did not reduce the number of parasites but caused a shift from the normal female/male ratio of 3:1 to 1:1 by 14 days postinfection (dpi). Exposure of H. glycines to dsRNA corresponding to a newly characterized protein with homology to C-type lectins did not affect sexual fate, but 41% fewer parasites were recovered from the plants. As expected, treatment with dsRNA corresponding to the major sperm protein (MSP) had no effect on either parasite development or sexual fate over 14 days. Northern analysis showed lower transcript abundance for the two targeted mRNAs that occur in J2, plus a later inhibition for MSP transcripts when males developed sperm at 15 dpi. These findings establish a procedure for RNAi of plant parasitic nematodes.
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                Author and article information

                Contributors
                Journal
                Journal ID (nlm-ta): Front Microbiol
                Journal ID (iso-abbrev): Front Microbiol
                Journal ID (publisher-id): Front. Microbiol.
                Title: Frontiers in Microbiology
                Publisher: Frontiers Media S.A.
                ISSN (Electronic): 1664-302X
                Publication date (Electronic): 12 January 2015
                Publication date Collection: 2014
                Volume: 5
                Electronic Location Identifier: 760
                Affiliations
                Division of Nematology, Indian Agricultural Research Institute New Delhi, India
                Author notes

                Edited by: Peter Moffett, Université de Sherbrooke, Canada

                Reviewed by: Brad Day, Michigan State University, USA; Peter Moffett, Université de Sherbrooke, Canada; Julio Vega-Arreguin, National Autonomous University of Mexico, Mexico

                *Correspondence: Tushar K. Dutta, Scientist, Division of Nematology, Indian Agricultural Research Institute, New Delhi 110012, India e-mail: nemaiari@ 123456gmail.com

                This article was submitted to Plant-Microbe Interaction, a section of the journal Frontiers in Microbiology.

                Article
                DOI: 10.3389/fmicb.2014.00760
                PMC ID: 4290618
                PubMed ID: 25628609
                SO-VID: c21a6747-68e5-4b61-8a7b-03b3f414505d
                Copyright © Copyright © 2015 Dutta, Banakar and Rao.
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                Date received : 08 September 2014
                Date accepted : 12 December 2014
                Page count
                Figures: 1, Tables: 1, Equations: 0, References: 43, Pages: 7, Words: 5221
                Categories
                Subject: Plant Science
                Subject: Mini Review Article

                ScienceOpen disciplines: Microbiology & Virology
                Keywords: gene silencing,dsrna,sirna,root-knot nematodes,cyst nematodes,host-delivered rnai
                Data availability:
                ScienceOpen disciplines: Microbiology & Virology
                Keywords: gene silencing, dsrna, sirna, root-knot nematodes, cyst nematodes, host-delivered rnai

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