Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole Absorption

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 p...

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Main Authors: Athanasios Dalakouras, Wolfgang Jarausch, Guenther Buchholz, Alexandra Bassler, Mario Braun, Thorsten Manthey, Gabi Krczal, Michael Wassenegger
Format: Article
Language:English
Published: Frontiers Media S.A. 2018-08-01
Series:Frontiers in Plant Science
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fpls.2018.01253/full
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author Athanasios Dalakouras
Athanasios Dalakouras
Wolfgang Jarausch
Guenther Buchholz
Alexandra Bassler
Mario Braun
Thorsten Manthey
Gabi Krczal
Michael Wassenegger
Michael Wassenegger
author_facet Athanasios Dalakouras
Athanasios Dalakouras
Wolfgang Jarausch
Guenther Buchholz
Alexandra Bassler
Mario Braun
Thorsten Manthey
Gabi Krczal
Michael Wassenegger
Michael Wassenegger
author_sort Athanasios Dalakouras
collection DOAJ
description 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.
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spelling doaj.art-b895cb77c421471f8033009ca682eb582022-12-22T00:12:22ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2018-08-01910.3389/fpls.2018.01253392749Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole AbsorptionAthanasios Dalakouras0Athanasios Dalakouras1Wolfgang Jarausch2Guenther Buchholz3Alexandra Bassler4Mario Braun5Thorsten Manthey6Gabi Krczal7Michael Wassenegger8Michael Wassenegger9RLP AgroScience GmbH, AlPlanta - Institute for Plant Research, Neustadt, GermanyInstitute of Plant Breeding and Genetic Resources ELGO-DEMETER, Thessaloniki, GreeceRLP AgroScience GmbH, AlPlanta - Institute for Plant Research, Neustadt, GermanyRLP AgroScience GmbH, AlPlanta - Institute for Plant Research, Neustadt, GermanyRLP AgroScience GmbH, AlPlanta - Institute for Plant Research, Neustadt, GermanyRLP AgroScience GmbH, AlPlanta - Institute for Plant Research, Neustadt, GermanyRLP AgroScience GmbH, AlPlanta - Institute for Plant Research, Neustadt, GermanyRLP AgroScience GmbH, AlPlanta - Institute for Plant Research, Neustadt, GermanyRLP AgroScience GmbH, AlPlanta - Institute for Plant Research, Neustadt, GermanyCentre for Organismal Studies Heidelberg, Heidelberg, GermanySince 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.https://www.frontiersin.org/article/10.3389/fpls.2018.01253/fullRNAidsRNAsiRNAstrunkpetioleMalus domestica
spellingShingle Athanasios Dalakouras
Athanasios Dalakouras
Wolfgang Jarausch
Guenther Buchholz
Alexandra Bassler
Mario Braun
Thorsten Manthey
Gabi Krczal
Michael Wassenegger
Michael Wassenegger
Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole Absorption
Frontiers in Plant Science
RNAi
dsRNA
siRNAs
trunk
petiole
Malus domestica
title Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole Absorption
title_full Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole Absorption
title_fullStr Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole Absorption
title_full_unstemmed Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole Absorption
title_short Delivery of Hairpin RNAs and Small RNAs Into Woody and Herbaceous Plants by Trunk Injection and Petiole Absorption
title_sort delivery of hairpin rnas and small rnas into woody and herbaceous plants by trunk injection and petiole absorption
topic RNAi
dsRNA
siRNAs
trunk
petiole
Malus domestica
url https://www.frontiersin.org/article/10.3389/fpls.2018.01253/full
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