Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole Absorption.
Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole Absorption.
复制标题
DOI：
10.3389/fpls.2018.01253
复制
发表时间：
2018
影响因子：
5.6
通讯作者：
Wassenegger M
中科院分区：
文献类型：
--
作者：
Dalakouras A;Jarausch W;Buchholz G;Bassler A;Braun M;Manthey T;Krczal G;Wassenegger M
自发现以来，RNA干扰已被广泛应用于作物保护。最近，已经报道了基于具有在植物中触发RNAi的能力的RNA分子的外源应用的无转基因程序。然而，将此类RNA分子有效递送到植物，特别是树木，提出了重大的技术挑战。在这里，我们描述了基于树干注射和/或叶柄吸收的将发夹RNA（hpRNA）和小干扰RNA（siRNA）有效递送到苹果、葡萄属和本氏烟草的简单方法。所施加的RNA分子被有效地吸收并全身转运。在顶端叶中，RNA在施用后1天（dpa）就已经可检测到，并且根据施用方法，至少可检测到10 dpa。共聚焦显微镜显示，摄取和系统转运的RNA分子严格限制在木质部和质外体，这可以说明为什么所施加的hpRNA不被植物DICER-LIKE（DCL）内切核酸酶加工成siRNA。这些创新方法可能会对害虫管理产生巨大影响，以对抗咀嚼和/或木质部汁液为食的载体以及驻留在木质部中的真核病原体。
Since its discovery, RNA interference has been widely used in crop protection. Recently, transgene-free procedures that were based on exogenous application of RNA molecules having the capacity to trigger RNAi in planta have been reported. Yet, efficient delivery of such RNA molecules to plants and particularly to trees poses major technical challenges. Here, we describe simple methods for efficient delivery of hairpin RNAs (hpRNAs) and small interfering RNAs (siRNAs) to Malus domestica, Vitis vinifera, and Nicotiana benthamiana that are based on trunk injection and/or petiole absorption. The applied RNA molecules were efficiently taken up and systemically transported. In apical leaves, the RNA was already detectable 1 day post-application (dpa) and could be detected at least up to 10 dpa, depending on the method of application. Confocal microscopy revealed that the uptaken and systemically transported RNA molecules were strictly restricted to the xylem and apoplast which may illustrate why the applied hpRNAs were not processed into siRNAs by plant DICER-LIKE (DCL) endonucleases. These innovative methods may have great impact in pest management against chewing and/or xylem sap-feeding vectors and eukaryotic pathogens that reside in the xylem.
登录
查看更多内容
DOI：
10.1261/rna.048116.114
发表时间：
2015-05
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Ivashuta S;Zhang Y;Wiggins BE;Ramaseshadri P;Segers GC;Johnson S;Meyer SE;Kerstetter RA;McNulty BC;Bolognesi R;Heck GR
通讯作者：
Heck GR
影响因子：
3.7
作者：
Ghosh SK;Hunter WB;Park AL;Gundersen-Rindal DE
通讯作者：
Gundersen-Rindal DE
对植物防御刺吸式和螟虫害虫的 RNAi 方法的新见解
影响因子：
7.3
作者：
Li, Haichao;Guan, Ruobing;Miao, Xuexia
通讯作者：
Miao, Xuexia
影响因子：
9.2
作者：
Melnyk, Charles W.;Molnar, Attila;Baulcombe, David C.
通讯作者：
Baulcombe, David C.
影响因子：
6.2
作者：
Gan, Defang;Zhang, Jiao;Cheng, Beijiu
通讯作者：
Cheng, Beijiu