ДИАГНОСТИКА ВИРОИДА ЛАТЕНТНОЙ МОЗАИКИ ПЕРСИКА (PLMVD) МЕТОДОМ ОТ-ПЦР

Лейла Надирова; Гульшан Станбекова; Булат Искаков; Андрей Жигайлов

doi:10.37884/3-2024/13

Abstract

Viroids, the smallest plant pathogens, cause significant economic damage to agriculture worldwide. Currently, much attention is paid to their timely diagnostics in order to prevent their further spread. For the first time in the Republic of Kazakhstan, molecular diagnostics of the PLMVd viroid, the causative agent of latent mosaic of peach, included in the list of quarantine pathogens in the territory of the EAEU countries, was carried out. Samples of pear, cherry, peach and apricot leaves collected in the Almaty region were analyzed for the presence of the viroid. Reverse transcription followed by polymerase chain reaction (RT-PCR) in the presence of specific primers developed by us, revealed a DNA fragment of 338 nucleotide pairs in two peach samples growing in the city of Almaty and the Karasai district of the Almaty region. PCR products were cloned into a bacterial vector with subsequent determination of their nucleotide sequence. Sanger sequencing and BLAST analysis confirmed the presence of PLMVd nucleic acid in the samples. Comparative analysis of the nucleotide sequences of local PLMVd isolates showed 95.5% similarity between them and 81-96% similarity with published variants from other countries.

viroid, PLMVd, RT-PCR, cloning, sequencing, nucleotide analysis.

01 Introduction

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02 References

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7. Desvignes J.C. Different symptoms of the peach latent mosaic // Acta Phytopathologica Academia Sciencae Hungariae. – 1980. – Vol. 15. - P. 183-190. doi:17660/ActaHortic.1981.94.24.
8. Ambrós S., Llácer G., Desvignes J.C., Flores R. Peach latent mosaic and pear blister canker vi-roids: detection by molecular hybridization and relationships with specific maladies affecting peach and pear trees // Acta Horticulturae. – 1995. – Vol. 386. - P. 515–521. doi: 10.17660/actahortic.1995.386.74.
9. Ambrós S., Hernández C., Desvignes J.C., Flores R. Genomic structure of three phenotypically different isolates of peach latent mosaic viroid: implications of the existence of constraints limiting the heterogeneity of viroid quasispecies // Journal of Virology. – 1998. –Vol.7. – P. 397-406. doi: 10.1128/JVI.72.9.7397-7406.1998.
10. Ragozzino E., Faggioli F., Barba M. Development of a one tube-one step RT-PCR protocol for the detection of seven viroids in four genera: Apscaviroid, Hostuviroid, Pelamoviroid and Pospiviroid // J. Virol. Methods. – 2004. - Vol.121. – P. 25–29. doi:10.1016/j.jviromet.2004.05.012.
11. Parisi O., Lepoivre P., Jijakli M.H. Development of a quick quantitative real-time PCR for the in vivo detection and quantification of peach latent mosaic viroid // Plant Disease. – Vol. 95. – P. 137-142. doi: 10.1094/PDIS-07-10-0512.
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13. Luigi M., Faggioli F. Development of quantitative real-time RT-PCR for the detection and quantification of peach latent mosaic viroid // Eur. J. Plant Pathol. – 2011. Vol. 95. – P. doi: 137–142. 10.1007/s10658-010-9738-2.
14. Boubourakas I.N., Fukuta S., Kyriakopoulou P.E. Sensitive and rapid detection of peach latent mosaic viroid by the reverse transcription loop-mediated isothermal amplification // J. Virol. Methods. - 2009. Vol. 160. – P. 63–68. doi:10.1016/j.jviromet.2009.04.021.
15. Lee H.J., Kim H.J., Lee K., Jeong R.D. Rapid detection of peach latent mosaic viroid by reverse transcription recombinase polymerase amplification // Mol Cell Probes. – 2020. Vol. 53. –P. 101627. doi: 10.1016/j.mcp.2020.101627.
16. Zhang Y., Yin J., Jiang D., Xin Y., Ding F., Deng Z., Wang G., Ma X., Li F., Li G., Li M., Li S., Zhu S. A universal oligonucleotide microarray with a minimal number of probes for the detection and identification of viroids at the genus level // PLoS One. – 2013. – Vol. 8, 5 e64474. doi: 10.1371/journal.pone.0064474.
17. Долгих С., Кабылбекова Б. Перспективы производства сертифицированного посадочного материала плодовых культур в казахстане // Izdenister Natigeler. -2023.- №2 (98). - С. 133–143. doi: 10.37884/2-2023/1.
18. Li R., Mock R., Huang Q., Abad J., Hartung J., Kinard G. A reliable and inexpensive method of nucleic acid extraction for the PCR-based detection of diverse plant pathogens // J Virol Methods. – 2008. – Vol. 154. – P. 48-55. doi: 10.1016/j.jviromet.2008.09.008.
19. Zhou M-Y., Gomez-Sanchez C.E. Universal TA Cloning // Current Issues in Molecular Biology. – 2000. – Vol. 2. – P.1-7. doi.org/10.21775/cimb.002.001.

Citation Links

[1]2024. DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR. Izdenister natigeler. 3(103) (Sep. 2024), 114–123. DOI:https://doi.org/10.37884/3-2024/13.

(1)DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR. Izdenister natigeler 2024, No. 3(103), 114-123. https://doi.org/10.37884/3-2024/13.

DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR. (2024). Izdenister Natigeler, 3(103), 114-123. https://doi.org/10.37884/3-2024/13

DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR. Izdenister natigeler, [S. l.], n. 3(103), p. 114–123, 2024. DOI: 10.37884/3-2024/13. Disponível em: https://natsus.kaznaru.edu.kz/index.php/research/article/view/687. Acesso em: 15 sep. 2026.

“DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR”. 2024. Izdenister Natigeler, no. 3(103) (September): 114-23. https://doi.org/10.37884/3-2024/13.

“DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR” (2024) Izdenister natigeler, (3(103), pp. 114–123. doi:10.37884/3-2024/13.

[1]“DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR”, Izdenister natigeler, no. 3(103), pp. 114–123, Sep. 2024, doi: 10.37884/3-2024/13.

“DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR”. Izdenister Natigeler, no. 3(103), Sept. 2024, pp. 114-23, https://doi.org/10.37884/3-2024/13.

“DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR”. Izdenister natigeler, no. 3(103) (September 30, 2024): 114–123. Accessed September 15, 2026. https://natsus.kaznaru.edu.kz/index.php/research/article/view/687.

1.DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR. Izdenister natigeler [Internet]. 2024 Sep. 30 [cited 2026 Sep. 15];(3(103):114-23. Available from: https://natsus.kaznaru.edu.kz/index.php/research/article/view/687

1.DIAGNOSIS OF PEACH LATENT MOSAIC VIROID (PLMVD) BY RT-PCR. Izdenister natigeler. 2024;(3(103):114-123. doi:10.37884/3-2024/13

[1] 1. Tsagris E.M., Martínez de Alba A.E., Gozmanova M., Kalantidis K. Viroids. Cell Microbiology. – 2008. - Vol. 10. – P. 2168-79. doi: 10.1111/j.1462-5822.2008.01231.x.

[2] 2. Ortolá B., Daròs J.-A. Viroids: Non-Coding Circular RNAs Able to Autonomously Replicate and Infect Higher Plants // Plants. Biology (Basel). – 2023. – Vol. 12. – P. 172. doi: 10.3390/biology12020172.

[3] 3. Надирова Л.Т., Станбекова Г.Э., Бейсенов Д.К., Искаков Б.К. Молекулярная диагностика вироида веретеновидности клубней картофеля в Республике Казахстан // Eurasian Journal of Applied Biotechnology. – 2018. – №3. doi: 10.11134/btp.3.2016.5.

[4] 4. Надирова Л.Т., Станбекова Г.Э., Искаков Б.К. Молекулярная диагностика вироидов плодовых растений // Материалы междунар. конференции «Современные проблемы молекулярной и клеточной биологии, биохимии и биотехнологии». г. Алматы, 2009 г. – С. 46.

[5] 5. Bussiere F., Quellet J., Cote F., Levesque D. and Perreault J.P. Mapping in solution shows the peach latent mosaic viroid to possess a new pseudoknot in a complex, branched secondary structure // J. Virology. – 2000. – Vol. 74. – P. 2647–2654. doi: 10.1128/jvi.74.6.2647-2654.2000.

[6] 6. Di Serio F., Flores R., Verhoeven J. T. J., Li S.-F., Pallás V., Randles J. W., Owens R. A. Current status of viroid taxonomy // Archives of Virology. - 2014. – Vol. 159. – P. 3467–3478. doi: 10.1007/s00705-014-2200-6.

[7] 7. Desvignes J.C. Different symptoms of the peach latent mosaic // Acta Phytopathologica Academia Sciencae Hungariae. – 1980. – Vol. 15. - P. 183-190. doi:17660/ActaHortic.1981.94.24.

[8] 8. Ambrós S., Llácer G., Desvignes J.C., Flores R. Peach latent mosaic and pear blister canker vi-roids: detection by molecular hybridization and relationships with specific maladies affecting peach and pear trees // Acta Horticulturae. – 1995. – Vol. 386. - P. 515–521. doi: 10.17660/actahortic.1995.386.74.

[9] 9. Ambrós S., Hernández C., Desvignes J.C., Flores R. Genomic structure of three phenotypically different isolates of peach latent mosaic viroid: implications of the existence of constraints limiting the heterogeneity of viroid quasispecies // Journal of Virology. – 1998. –Vol.7. – P. 397-406. doi: 10.1128/JVI.72.9.7397-7406.1998.

[10] 10. Ragozzino E., Faggioli F., Barba M. Development of a one tube-one step RT-PCR protocol for the detection of seven viroids in four genera: Apscaviroid, Hostuviroid, Pelamoviroid and Pospiviroid // J. Virol. Methods. – 2004. - Vol.121. – P. 25–29. doi:10.1016/j.jviromet.2004.05.012.

[11] 11. Parisi O., Lepoivre P., Jijakli M.H. Development of a quick quantitative real-time PCR for the in vivo detection and quantification of peach latent mosaic viroid // Plant Disease. – Vol. 95. – P. 137-142. doi: 10.1094/PDIS-07-10-0512.

[12] 12. Приходько Ю.Н., Живаева Т.С., Лозовая Е.Н., Шнейдер Ю.А., Каримова Е.В. Разработка методов диагностики вироида латентной мозаики персика (PLMVd) // Фитосанитария. Карантин растений – 2021. – №3. – С.47-57. doi:10.69536/FKR.2021.27.95.001.

[13] 13. Luigi M., Faggioli F. Development of quantitative real-time RT-PCR for the detection and quantification of peach latent mosaic viroid // Eur. J. Plant Pathol. – 2011. Vol. 95. – P. doi: 137–142. 10.1007/s10658-010-9738-2.

[14] 14. Boubourakas I.N., Fukuta S., Kyriakopoulou P.E. Sensitive and rapid detection of peach latent mosaic viroid by the reverse transcription loop-mediated isothermal amplification // J. Virol. Methods. - 2009. Vol. 160. – P. 63–68. doi:10.1016/j.jviromet.2009.04.021.

[15] 15. Lee H.J., Kim H.J., Lee K., Jeong R.D. Rapid detection of peach latent mosaic viroid by reverse transcription recombinase polymerase amplification // Mol Cell Probes. – 2020. Vol. 53. –P. 101627. doi: 10.1016/j.mcp.2020.101627.

[16] 16. Zhang Y., Yin J., Jiang D., Xin Y., Ding F., Deng Z., Wang G., Ma X., Li F., Li G., Li M., Li S., Zhu S. A universal oligonucleotide microarray with a minimal number of probes for the detection and identification of viroids at the genus level // PLoS One. – 2013. – Vol. 8, 5 e64474. doi: 10.1371/journal.pone.0064474.

[17] 17. Долгих С., Кабылбекова Б. Перспективы производства сертифицированного посадочного материала плодовых культур в казахстане // Izdenister Natigeler. -2023.- №2 (98). - С. 133–143. doi: 10.37884/2-2023/1.

[18] 18. Li R., Mock R., Huang Q., Abad J., Hartung J., Kinard G. A reliable and inexpensive method of nucleic acid extraction for the PCR-based detection of diverse plant pathogens // J Virol Methods. – 2008. – Vol. 154. – P. 48-55. doi: 10.1016/j.jviromet.2008.09.008.

[19] 19. Zhou M-Y., Gomez-Sanchez C.E. Universal TA Cloning // Current Issues in Molecular Biology. – 2000. – Vol. 2. – P.1-7. doi.org/10.21775/cimb.002.001.