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

Frontiers in Plant Science
Athanasios DalakourasMichael Wassenegger

Abstract

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...Continue Reading

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Citations

May 11, 2019·International Journal of Molecular Sciences·Alexandra S Dubrovina, Konstantin V Kiselev
Nov 20, 2019·Pest Management Science·Clauvis Nji Tizi TaningGuy Smagghe
Mar 24, 2020·BioTechniques·Shuo YanJie Shen
Jul 17, 2020·Frontiers in Plant Science·Protiva Rani Das, Sherif M Sherif
Sep 6, 2019·International Journal of Molecular Sciences·Chiara Pagliarani, Giorgio Gambino
May 30, 2020·Plants·Athanasios Dalakouras, Kalliope K Papadopoulou
Nov 12, 2019·Frontiers in Plant Science·Deise CagliariMoisés João Zotti
Jul 1, 2020·Frontiers in Plant Science·Michael BennettAlberto Iandolino
Jan 5, 2021·Frontiers in Plant Science·Veli Vural UsluMichael Wassenegger
Mar 29, 2021·The New Phytologist·Aline Koch, Michael Wassenegger
May 5, 2021·Plant Biotechnology Journal·Gaetano GiudiceWalter Chitarra
May 22, 2021·Journal of Experimental Botany·Athanasios Dalakouras, Dimitrios Vlachostergios
Jul 3, 2021·International Journal of Molecular Sciences·Konstantin V KiselevAlexandra S Dubrovina
Jul 4, 2021·Current Opinion in Biotechnology·Dongdong NiuHailing Jin
Oct 16, 2021·Nature Plants·Federico BettiPierdomenico Perata

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Methods Mentioned

BETA
transgenic
Fluorescence microscopy
Confocal microscopy

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