Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles.
Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles.
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DOI：
10.1093/g3journal/jkaa058
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发表时间：
2021-02-09
期刊：
影响因子：
--
通讯作者：
Eves-van den Akker S
中科院分区：
文献类型：
--
作者：
Kranse O;Beasley H;Adams S;Pires-daSilva A;Bell C;Lilley CJ;Urwin PE;Bird D;Miska E;Smant G;Gheysen G;Jones J;Viney M;Abad P;Maier TR;Baum TJ;Siddique S;Williamson V;Akay A;Eves-van den Akker S
植物寄生线虫是对粮食安全的持续威胁，估计每年造成1000亿美元的作物损失。最有问题的是专性定居体内寄生虫（主要是根结线虫和囊线虫）。由于缺乏功能遗传学的工具，理解它们生物学的进展受到阻碍：正向遗传学在很大程度上限于研究种群的自然变异，而反向遗传学完全依赖于RNA干扰。人们期望功能性遗传工具的开发将加速植物寄生线虫研究的进展，从而促进新型控制解决方案的开发。在这里，我们开发了一些提供植物寄生线虫功能遗传工具包所需的基础生物学。我们的特点男性的生殖腺的线虫schachtii和北方根结线虫的精子发生的背景下。我们测试和优化各种方法的交付，表达，和/或检测外源核酸在植物寄生线虫。我们证明了将大分子传递到孢囊和根结线虫雄性生殖系是困难的，但是可能的。同样，我们证明了寡核苷酸的交付，根结线虫配子。最后，我们开发了一个植物寄生线虫的瞬时表达系统，通过展示在整个H.使用基于脂质体的转染的schachtii青少年。我们预计这些发展将是独立有用的，将加快植物寄生线虫的遗传修饰工具的开发，并最终催化对一组威胁全球粮食安全的线虫的研究。
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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影响因子：
6.7
作者：
Kikuchi T;Cotton JA;Dalzell JJ;Hasegawa K;Kanzaki N;McVeigh P;Takanashi T;Tsai IJ;Assefa SA;Cock PJ;Otto TD;Hunt M;Reid AJ;Sanchez-Flores A;Tsuchihara K;Yokoi T;Larsson MC;Miwa J;Maule AG;Sahashi N;Jones JT;Berriman M
通讯作者：
Berriman M
影响因子：
12.3
作者：
Eves-van den Akker S;Laetsch DR;Thorpe P;Lilley CJ;Danchin EG;Da Rocha M;Rancurel C;Holroyd NE;Cotton JA;Szitenberg A;Grenier E;Montarry J;Mimee B;Duceppe MO;Boyes I;Marvin JM;Jones LM;Yusup HB;Lafond-Lapalme J;Esquibet M;Sabeh M;Rott M;Overmars H;Finkers-Tomczak A;Smant G;Koutsovoulos G;Blok V;Mantelin S;Cock PJ;Phillips W;Henrissat B;Urwin PE;Blaxter M;Jones JT
通讯作者：
Jones JT
影响因子：
3.3
作者：
Guo Y;Fudali S;Gimeno J;DiGennaro P;Chang S;Williamson VM;Bird DM;Nielsen DM
通讯作者：
Nielsen DM
影响因子：
1.5
作者：
O'Halloran, Damien M.
通讯作者：
O'Halloran, Damien M.
影响因子：
4.6
作者：
Adams, Sally;Pathak, Prachi;Pires-daSilva, Andre
通讯作者：
Pires-daSilva, Andre