2021
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; den Akker, Sebastian Eves-van
In: G3, vol. 11, no. 2, 2021, ISSN: 2160-1836.
Abstract | Links | BibTeX | Tags: genetic modification, germline, lipofection, plant-parasitic nematodes, transformation, transient expression
@article{kranse_toward_2021,
title = {Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles},
author = {Olaf Kranse and Helen Beasley and Sally Adams and Andre Pires-daSilva and Christopher Bell and Catherine J Lilley and Peter E Urwin and David Bird and Eric Miska and Geert Smant and Godelieve Gheysen and John Jones and Mark Viney and Pierre Abad and Thomas R Maier and Thomas J Baum and Shahid Siddique and Valerie Williamson and Alper Akay and Sebastian Eves-van den Akker},
url = {http://dx.doi.org/10.1093/g3journal/jkaa058},
doi = {10.1093/g3journal/jkaa058},
issn = {2160-1836},
year = {2021},
date = {2021-02-01},
journal = {G3},
volume = {11},
number = {2},
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.},
keywords = {genetic modification, germline, lipofection, plant-parasitic nematodes, transformation, transient expression},
pubstate = {published},
tppubtype = {article}
}
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.
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.