Advanced Search
MyIDEAS: Login

Silencing of Mg -pat-10 and Mg -unc-87 in the Plant Parasitic Nematode Meloidogyne graminicola Using siRNAs

Contents:

Author Info

  • Joseph Nsengimana

    ()
    (Department of Molecular Biotechnology, Ghent University, 653 Coupure links, 9000 Gent, Belgium
    Current Address: Department of Applied Biology, Kigali Institute of Science and Technology (KIST), 3900 Avenue de la Paix, 250 Kigali, Rwanda;)

  • Lander Bauters

    ()
    (Department of Molecular Biotechnology, Ghent University, 653 Coupure links, 9000 Gent, Belgium)

  • Annelies Haegeman

    ()
    (Department of Molecular Biotechnology, Ghent University, 653 Coupure links, 9000 Gent, Belgium)

  • Godelieve Gheysen

    ()
    (Department of Molecular Biotechnology, Ghent University, 653 Coupure links, 9000 Gent, Belgium)

Registered author(s):

    Abstract

    Until recently, the standard method for RNA interference (RNAi)-based reverse genetics in plant parasitic nematodes (PPNs) was based on the use of long double-stranded RNA (dsRNA). This increased the chance of off-target gene silencing through interactions between different short interfering RNAs (siRNAs) and non-cognate mRNA targets. In this work, we applied gene-specific knockdown of Mg-pat-10 and Mg-unc-87 of the root knot nematode Meloidogyne graminicola , using discrete 21 bp siRNAs. The homologue of Mg-pat-10 in C. elegans encodes body wall troponin C, which is essential for muscle contraction, whereas the homologue of Mg-unc-87 encodes two proteins involved in maintenance of the structure of myofilaments in the body wall muscle of C. elegans . The knockdown at the transcript level, as seen by semi-quantitative RT-PCR analysis, indicates that the Mg-pat-10 gene was silenced after soaking the nematodes in a specific siRNA for 48 h. At 72 h post-soaking, the Mg-pat-10 mRNA level was similar to the control, indicating the recovery of expression between 48 h and 72 h post-soaking. For Mg-unc-87 the nematodes started to recover from siRNA silencing 24 h after thorough washing. A migration assay showed that for the nematodes that were soaked in the control (siRNA of β-1,4-endoglucanase), 77% of the nematodes completed migration through the column in a 12 h period. By comparison with the control, nematodes incubated in the siRNA of pat-10 or unc-87 were significantly inhibited in their motility. After 12 h, only 6.3% of the juveniles incubated in the Mg-pat-10 siRNA and 9.3% of those incubated in Mg-unc-87 siRNA had migrated through the column, representing 91.8% and 87.9% inhibition respectively compared to the control. In the present work, we demonstrated that M. graminicola is readily susceptible to siRNAs of two genes involved in nematode motility. This is an important contribution to the progressive use of siRNA for functional analysis. Moreover, the application of RNAi in PPNs opens the way for environmentally friendly control of M. graminicola .

    Download Info

    If you experience problems downloading a file, check if you have the proper application to view it first. In case of further problems read the IDEAS help page. Note that these files are not on the IDEAS site. Please be patient as the files may be large.
    File URL: http://www.mdpi.com/2077-0472/3/3/567/pdf
    Download Restriction: no

    File URL: http://www.mdpi.com/2077-0472/3/3/567/
    Download Restriction: no

    Bibliographic Info

    Article provided by MDPI, Open Access Journal in its journal Agriculture.

    Volume (Year): 3 (2013)
    Issue (Month): 3 (September)
    Pages: 567-578

    as in new window
    Handle: RePEc:gam:jagris:v:3:y:2013:i:3:p:567-578:d:28897

    Contact details of provider:
    Web page: http://www.mdpi.com/

    Related research

    Keywords: short interfering RNA (siRNA); Meloidogyne graminicola ( Mg ); Mg-pat-10 ; Mg-unc-87 ; migration assay; expression analysis;

    Find related papers by JEL classification:

    References

    No references listed on IDEAS
    You can help add them by filling out this form.

    Citations

    Lists

    This item is not listed on Wikipedia, on a reading list or among the top items on IDEAS.

    Statistics

    Access and download statistics

    Corrections

    When requesting a correction, please mention this item's handle: RePEc:gam:jagris:v:3:y:2013:i:3:p:567-578:d:28897. See general information about how to correct material in RePEc.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: (XML Conversion Team).

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    If references are entirely missing, you can add them using this form.

    If the full references list an item that is present in RePEc, but the system did not link to it, you can help with this form.

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your profile, as there may be some citations waiting for confirmation.

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.