Toward genetic modification of plant-parasitic nematodes: Delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles

Kranse, Olaf, Beasley, Helen, Adams, Sally, Pires-daSilva, Andre, Bell, Christopher, Lilley, Catherine J., Urwin, Peter E., Bird, David, Miska, Eric, Smant, Geert, Gheysen, Godelieve, Jones, John, Viney, Mark, Abad, Pierre, Maier, Thomas R., Baum, Thomas J., Siddique, Shahid, Williamson, Valerie, Akay, Alper and Eves-van den Akker, Sebastian (2021) Toward genetic modification of plant-parasitic nematodes: Delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles. G3: Genes, Genomes, Genetics, 11 (2). ISSN 2160-1836

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Abstract

Plant-parasitic nematodes are a continuing threat to food security, causing an estimated 100 billion USD in crop losses each year. The most problematic are the obligate sedentary endoparasites (primarily root knot nematodes and cyst nematodes). Progress in understanding their biology is held back by a lack of tools for functional genetics: forward genetics is largely restricted to studies of natural variation in populations and reverse genetics is entirely reliant on RNA interference. There is an expectation that the development of functional genetic tools would accelerate the progress of research on plant-parasitic nematodes, and hence the development of novel control solutions. Here, we develop some of the foundational biology required to deliver a functional genetic tool kit in plant-parasitic nematodes. We characterize the gonads of male Heterodera schachtii and Meloidogyne hapla in the context of spermatogenesis. We test and optimize various methods for the delivery, expression, and/or detection of exogenous nucleic acids in plant-parasitic nematodes. We demonstrate that delivery of macromolecules to cyst and root knot nematode male germlines is difficult, but possible. Similarly, we demonstrate the delivery of oligonucleotides to root knot nematode gametes. Finally, we develop a transient expression system in plant-parasitic nematodes by demonstrating the delivery and expression of exogenous mRNA encoding various reporter genes throughout the body of H. schachtii juveniles using lipofectamine-based transfection. We anticipate these developments to be independently useful, will expedite the development of genetic modification tools for plant-parasitic nematodes, and ultimately catalyze research on a group of nematodes that threaten global food security.

Item Type: Article
Additional Information: © The Author(s) 2021. Published by Oxford University Press on behalf of Genetics Society of America.
Uncontrolled Keywords: genetic modification,germline,lipofection,plant-parasitic nematodes,transformation,transient expression,molecular biology,genetics,genetics(clinical) ,/dk/atira/pure/subjectarea/asjc/1300/1312
Faculty \ School:
Faculty of Science > School of Biological Sciences
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 25 Feb 2021 00:58
Last Modified: 10 Sep 2021 00:19
URI: https://ueaeprints.uea.ac.uk/id/eprint/79326
DOI: 10.1093/g3journal/jkaa058

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