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

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...

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Autores principales: 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, Eves-van den Akker, Sebastian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8022973/
https://www.ncbi.nlm.nih.gov/pubmed/33585878
http://dx.doi.org/10.1093/g3journal/jkaa058
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author 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
Eves-van den Akker, Sebastian
author_facet 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
Eves-van den Akker, Sebastian
author_sort Kranse, Olaf
collection PubMed
description 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.
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spelling pubmed-80229732021-04-09 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 Eves-van den Akker, Sebastian G3 (Bethesda) Investigation 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. Oxford University Press 2021-02-15 /pmc/articles/PMC8022973/ /pubmed/33585878 http://dx.doi.org/10.1093/g3journal/jkaa058 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Genetics Society of America. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigation
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
Eves-van den Akker, Sebastian
Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles
title Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles
title_full Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles
title_fullStr Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles
title_full_unstemmed Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles
title_short Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles
title_sort toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mrna in second stage juveniles
topic Investigation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8022973/
https://www.ncbi.nlm.nih.gov/pubmed/33585878
http://dx.doi.org/10.1093/g3journal/jkaa058
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