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AGRICULTURE, AGROCHEMICAL, FEED PRODUCTION, AGROECOLOGY

AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES

Karlygash Aubakirova 1 , Zhibek Bakytzhanova 1 , Laura Yerbolova 1 , Akbota Rakhatkyzy 1 , Nurbol Galiakparov 1

1 M.A.Aitkhozhin Institute of Molecular Biology and Biochemistry

doi.org/10.37884/3-2024/12 pp. 108-114 Admitted 31.07.2024 Published 30.09.2023

Abstract

Efficient isolation of high-quality RNA is an important step in the detection of plant RNA viruses by molecular methods. RNA isolation from grape, apple and other fruit leaves is problematic due to the presence of polysaccharides, polyphenols and other compounds that bind to or co-precipitate with RNA. In this work, a cetyltrimethylammonium bromide (CTAB)-based protocol for RNA isolation from grape, apple, apricot, plum and walnut leaves was optimized. The concentrations of PVP, 2-mercaptоethanоl and CTAB were increased. Incubation times with CTAB extraction buffer at 65 °C were optimized for each culture. For grape and apple leaves, the optimal incubation time is 40 minutes, for apricot leaves – 20 minutes, for plum and walnut leaves – 30 minutes. A comparative study of three different RNA extraction methods showed that the optimized protocol provides the highest quality and quantity of isolated RNA, as confirmed by electrophoresis and spectrophotometric analysis. RNA isolated using our protocol is suitable for subsequent molecular analyses, such as cDNA synthesis and PCR, demonstrating its suitability for the detection of viruses in various fruit tree species, including grape, apple, apricot, plum and walnut. The modified method is recommended for use in further studies on the detection of plant RNA viruses by PCR.

grape, apple, apricot, plum, walnut, RNA extraction, viruses of fruit plants, CTAB.

01 Introduction

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Citation Links

[1]2023. AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES. Izdenister natigeler. 3(103) (Sep. 2023), 108–114. DOI:https://doi.org/10.37884/3-2024/12.
(1)AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES. Izdenister natigeler 2023, No. 3(103), 108-114. https://doi.org/10.37884/3-2024/12.
AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES. (2023). Izdenister Natigeler, 3(103), 108-114. https://doi.org/10.37884/3-2024/12
AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES. Izdenister natigeler, [S. l.], n. 3(103), p. 108–114, 2023. DOI: 10.37884/3-2024/12. Disponível em: https://agrosoil.kaznaru.edu.kz/index.php/research/article/view/691. Acesso em: 15 sep. 2026.
“AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES”. 2023. Izdenister Natigeler, no. 3(103) (September): 108-14. https://doi.org/10.37884/3-2024/12.
“AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES” (2023) Izdenister natigeler, (3(103), pp. 108–114. doi:10.37884/3-2024/12.
[1]“AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES”, Izdenister natigeler, no. 3(103), pp. 108–114, Sep. 2023, doi: 10.37884/3-2024/12.
“AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES”. Izdenister Natigeler, no. 3(103), Sept. 2023, pp. 108-14, https://doi.org/10.37884/3-2024/12.
“AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES”. Izdenister natigeler, no. 3(103) (September 30, 2023): 108–114. Accessed September 15, 2026. https://agrosoil.kaznaru.edu.kz/index.php/research/article/view/691.
1.AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES. Izdenister natigeler [Internet]. 2023 Sep. 30 [cited 2026 Sep. 15];(3(103):108-14. Available from: https://agrosoil.kaznaru.edu.kz/index.php/research/article/view/691
1.AN IMPROVED METHOD OF RNA ISOLATION FOR MOLECULAR DIAGNOSTICS OF FRUIT AND BERRY VIRUSES. Izdenister natigeler. 2023;(3(103):108-114. doi:10.37884/3-2024/12