Abstract
In vitro culture of apricot cultivars ‘Qaysi’, ‘Shams’, and ‘Ordubad’, infected with Apple chlorotic leaf spot virus (ACLSV), Apple mosaic virus (ApMV), and Tobacco ring spot virus (TRSV) was used in this study. We investigated whether electrotherapy (0, 30, 40, 50, and 100 mA for 15 min), combining thermotherapy (38 °C for 7 days) with shoot tip culture, chemotherapy (ribavirin at 0, 25, 50, 75, and 100 mg L−1 for 30 days) with shoot-tip culture, or cryotherapy (incubating shoot-tips in plant vitrification solution-2 (PVS2) for 40 min) could effectively eradicate ACLSV, ApMV, and TRSV from virus-infected cultures. After treatments, shoot tips were cultured in vitro for 4 months before testing for the presence of viruses, and virus-free plants were grown in greenhouse for 18 months prior to retesting for the presence of the viruses. The highest virus elimination rates of 90–100% were obtained in combining thermotherapy with shoot tip culture (1.1–2.0 mm). Chemo (25 mg L−1)—and electro (40 mA)-treatments were also effective for producing virus-free planting material with frequencies of 75–100% and 40–60%, respectively. Cryotherapy treatment resulted in rates of 60–80%.The highest efficiency of virus eradication was detected in chemotherapy (58.6–93.3%), cryotherapy (60–80%), and thermotherapy (54–80%), respectively. According to our results, production of virus-free regenerants would be feasible in Prunus armeniaca L. providing proper therapeutic methods are adopted as regards the type of infecting virus and the host variety.
Key message
Apricot cultivars have been successfully treated with electro-, thermo-, chemo-, and cryotherapy for the first time with the eradication rates of 40–100% to produce virus-free regenerants.
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Data availability
The data that support the findings of this study are openly available under reasonable request. All data generated or analyzed during this study are included in this published article.
Abbreviations
- ACLSV:
-
Apple chlorotic leafspot virus
- ApMV:
-
Apple mosaic virus
- ArMV:
-
Arabis mosaic virus
- PDV:
-
Prune dwarf virus
- PNRSV:
-
Prunus necrotic ringspot virus
- PPV:
-
Plum pox virus
- ToRSV:
-
Tomato ringspot virus
- TRSV:
-
Tobacco ringspot virus
- RT-PCR:
-
Reverse transcription- polymerase chain reaction
- BAP:
-
6-Benzylaminopurine
- Fe-EDDHA:
-
Ethylenediamine di-2-hydroxyphenyl acetate ferric
- GA3 :
-
Gibberellic acid
- IBA:
-
Indole-3-butyric acid
- PVS2:
-
Plant vitrification solution-2
- DMSO:
-
Dimethyl sulfoxide
- NaCl:
-
Sodium chloride
References
Adil S, Singh V, Anjum A, Quraishi A (2022) A mini-review on electrotherapeutic strategy for the plant viral elimination. Plant Cell Tiss Org Cult 150:41–55. https://doi.org/10.1007/s11240-022-02265-w
Bădărău CL, Chiru N, Guţă IC (2014) The effect of some therapies on potato virus Y and potato virus X infected Solanum tuberosum L plantlets (cv ‘Roclas’). Analele Univ Din Oradea Fasc 21(1):24–32
Barba M, Ilardi V, Pasquini G (2015) Control of pome and stone fruit virus diseases. Adv Vir Res 91:47–83. https://doi.org/10.1016/bs.aivir.2014.11.001
Bayati Sh, Shams-Bakhsh M, Moieni A (2011) Elimination of grapevine virus A (GVA) by cryotherapy and electrotherapy. J Agr Sci Tech 13:443–450
Ben Mahmoud K, Najar A, Jemai N, Jemmali A (2017) Advances in sanitation methods for fruit tree species through in vitro technologies: possibilities and limits. J New Sci 45(4):2483–2495
Bettoni JC, Costa MD, Souza JA, Volk GM, Nickel O, da Silva FN, Kretzschmar AA (2018) Cryotherapy by encapsulation-dehydration is effective for in vitro eradication of latent viruses from ‘Marubakaido’ apple rootstock. J Biotecnol 269:1–7. https://doi.org/10.1016/j.jbiotec.2018.01.014
Bettoni JC, Souza JA, Volk GM, Dalla Costa M, da Silva FN, Kretzschmar AA (2019) Eradication of latent viruses from apple cultivar ‘Monalisa’ shoot tips using droplet-vitrification cryotherapy. Sci Hortic 250:12–18. https://doi.org/10.1016/j.scienta.2019.02.033
Bettoni JC, Mathew L, Pathirana R, Wiedow C, Hunter DA, McLachlan A, Khan S, Tang J et al (2022a) Eradication of Potato Virus S, Potato Virus A, and Potato Virus M from infected in vitro-grown potato shoots using in vitro therapies. Front Plant Sci 13:878733. https://doi.org/10.3389/fpls.2022.878733
Bettoni JC, Fazio G, Carvalho Costa L, Hurtado-Gonzales OP, Rwahnih MA, Nedrow A, Volk GM (2022b) Thermotherapy followed by shoot tip cryotherapy eradicates latent viruses and Apple Hammerhead Viroid from in vitro apple rootstocks. Plants 11:582. https://doi.org/10.3390/plants11050582
Brison M, Boucaud MT, Pierronnet A, Dosba F (1997) Effect of cryopreservation on the sanitary state of a cv Prunus rootstock experimentally contaminated with Plum Pox Potyvirus. Plant Sci 123:189–196
Faccioli G (2001) Control of potato viruses using meristem and stem-cuttings cultures, thermotherapy and chemotherapy. In: Loebenstein G, Berger PH, Brunt AA, Lawson RH (eds) Virus and virus-like diseases of potato and production of seed-potatoes. Kluwer, Dordrecht, pp 365–390
FAO (2020) Food and agriculture organization of the United Nations. www.fao.org.
Farhadi-Tooli S, Ghanbari A, Jafarkhani Kermani M, Zeinalabedini M, Bettoni JC, Naji AM, Kazemi N (2021) Droplet-vitrification cryotherapy and thermotherapy as efficient tools for the eradication of apple chlorotic leaf spot virus and apple stem grooving virus from virus infected quince in vitro cultures. Eur J Plant Pathol 162:31–43. https://doi.org/10.1007/s10658-021-02400-x
Fuchs M, Abavi GS, Marsella-Herrick P, Cox R, Cox KD, Carroll JE, Martin RR (2010) Occurrence of Tomato ringspot virus and Tobacco ringspot virus in highbush blueberry in New York State. J Plant Pathol 92:451–459
Grieco F, Alkowni R, Saponari M, Savino V, Martelli GP (2000) Molecular detection of olive viruses. Bull OEPP 30:469–473. https://doi.org/10.17660/ActaHortic.2002.586.158
Guţă IC, Buciumeanu EC, Tătaru LD, Topală CM (2017) Regeneration of grapevine virus-free plants by in vitro chemotherapy. Acta Hortic 1188:319–322. https://doi.org/10.17660/ActaHortic.2017.1188.42
Guţă IC, Buciumeanu EC, Tătaru LD, Oprescu B, Topală CM (2019) New approach of electrotherapy for grapevine virus elimination. Acta Hortic 1242:697–702. https://doi.org/10.17660/ActaHortic.2019.1242.103
Hassan M, Myrta A, Polak J (2006) Simultaneous detection and identification of four pome fruit viruses by one-tube pentaplex RT-PCR. J Virol Method 133:124–129. https://doi.org/10.1016/j.jviromet.2005.11.002
Hauptmanova A, Polak J (2011) The elimination of Plum pox virus in plum cv. Bluefree and apricot cv. Hanita by chemotherapy of in vitro cultures. Hort Sci 38:49–53. https://doi.org/10.17221/10/2010-HORTSCI
Helliot B, Madur D, Dirlewanger E, Boucaud DE (2001) Evaluation of genetic stability in cryopreserved prunus. In Vitro Cell Dev Biol-Plant 38:493–500. https://doi.org/10.1079/IVP2002325
Hilaire J, Tindale S, Jones G, Pingarron-Cardenas G, Bačnik K, Ojo M, Frewer LJ (2022) Risk perception associated with an emerging agri-food risk in Europe: plant viruses in agriculture. Agric Food Secur 11:21. https://doi.org/10.1186/s40066-022-00366-5
Howell WE, Eastwell KC, Li TSC (2001) Heat treatment, chemo-therapy and hydroponic culture for obtaining virus-free trees of sweet cherry. Acta Hortic 550:455–457. https://doi.org/10.17660/ActaHortic.2001.550.68
Hu G, Dong Y, Zhang Z, Fan X, Ren F, Zhou J (2015) Virus elimination from in vitro apple by thermotherapy combined with chemotherapy. Plant Cell Tiss Org Cult 121:435–443. https://doi.org/10.1007/s11240-015-0714-6
Hu G, Dong Y, Zhang Z, Fan X, Ren F (2022) Inefficiency of ribavirin to eliminate apple scar skin viroid from apple plants. Plant Cell Tiss Org Cult 151(1):189–197. https://doi.org/10.1007/s11240-022-02344-y
Jarošová J, Kundu JK (2010) Simultaneous detection of stone fruit tree viruses by one-step multiplex RT-PCR. Sci Hortic 125:68–72. https://doi.org/10.1016/j.scienta.2010.02.011
Jiang F, Zhang J, Wang S, Yang L, Luo Y, Gao S, Zhang M, Wu S, Hu S, Sun H, Wang Y (2019) The apricot (Prunus armeniaca L) genome elucidates Rosaceae evolution and beta-carotenoid synthesis. Hortic Res 6(128):1–12. https://doi.org/10.1038/s41438-019-0215-6
Kaur C, Raj R, Kumar S, Purshottam DK, Agarwal L, Chauhan PS, Raj SK (2019) Elimination of Bean yellow mosaic virus from infected cormels of three cultivars of gladiolus using thermo-, electro- and chemotherapy. 3-Biotech 9(154):1–10. https://doi.org/10.1007/s13205-019-1684-x
Kaya E (2021) Comparison of three different techniques for eradication of Apple mosaic virus (ApMV) from hazelnut (Corylus avellana L.). J Plant Prot Res 61(1):11–19. https://doi.org/10.24425/jppr.2021.136275
Kazemi N, Nahandi FZ, Habashi AA, Masoomi-Aladizgeh F (2020) Comparing the efficiency of conventional and novel methods of virus elimination using molecular techniques. Eur J Plant Pathol 157(4):887–897. https://doi.org/10.1007/s10658-020-02048-z
Koubouris GC, Maliogka VI, Efthimiou K, Katis NI, Vasilakakis MD (2007) Elimination of Plum pox virus through in vitro thermotherapy and shoot tip culture compared to conventional heat treatment in apricot cultivar Bebecou. J Gen Plant Pathol 73:370–373. https://doi.org/10.1111/j.1365-2338.2000.tb00931.x
Laimer M, Barba M (2011) Elimination of systemic pathogens by thermotherapy, tissue culture, or in vitro micrografting. In: Hadidi A, Barba M, Candresse T, Jelkmann M (eds) Virus and virus-like diseases of pome and stone fruits. American Phytopathological Society, St. Paul, pp 389–393
Leyssen P, Balzarini J, De Clrecq E, Neyts J (2005) The predominant mechanism by which ribavirin exerts its antiviral activity in vitro against flavivirus and apramyxoviruses is mediated by inhibition of IMP dehydrogenase. J Virol 79:1943–1947. https://doi.org/10.1128/JVI.79.3.1943-1947.2005
Lozoya-Saldana H, Abelló F, García GDLR (1996) Electrotherapy and shoot tip culture eliminate potato virus X in potatoes. Amer Potato J 73:149–153. https://doi.org/10.1007/BF02853073
Mahfouze SA, El-Dougdoug KA, Allam EK (2010) Production of potato spindle tuber viroid-free potato plant materials in vitro. J Am Sci 6(12):1570–1577
Maliogka VI, Olmos A, Pappi PG, Lotos L, Efthimiou K, Grammatikaki G, Candresse T, Katis NI, Avgelis AD (2015) A novel grapevine badnavirus is associated with the Roditis leaf discoloration disease. Virus Res 91:175–227. https://doi.org/10.1016/j.virusres.2015.03.003
Mathew L, Tiffin H, Erridge Z, McLachlan A, Hunter D, Pathirana R (2021) Efficiency of eradication of Raspberry bushy dwarf virus from infected raspberry (Rubus idaeus) by in vitro chemotherapy, thermotherapy and cryotherapy and their combinations. Plant Cell Tiss Org Cult 144:133–141. https://doi.org/10.1007/s11240-020-01829-y
Meier V, Bürger E, Mihm S, Saile B, Ramadori G (2003) Ribavirin inhibits DNA, RNA, and protein synthesis in PHA—stimulated human peripheral blood mononuclear cells: possible explanation for therapeutic efficacy in patients with chronic HCV infection. J Med Vir 69(1):50–58. https://doi.org/10.1002/jmv.10264
Menzel W, Jelkmann W, Maiss E (2002) Detection of four apple viruses by multiplex RT-PCR assays with coamplification of planty mRNA as internal control. J Virol Method 99:81–92. https://doi.org/10.1016/S0166-0934(01)00381-0
Meybodi DA, Mozafari J, Babaeiyan N, Rahimian H (2011) Application of electrotherapy for the elimination of potato Potyviruses. J Agr Sci Tech 13:921–927
Mirzaei L, Yadollahi A, Jafarkhani Kermani M, Naderpour M. Zeinanloo AA (2021) Screening and the possibility of providing virus-free stock plants of some commercial olive cultivars. PhD dissertation, Tarbiat Modares University, Tehran, Iran.
Naderpour M, Shahbazi R (2014) Investigation on viral diseases of pome fruit orchards in Western Azarbaijan and Khorasan Razavi provinces to introduce healthier orchards for stone fruit propagation industry. Final Report, Center for Agriculture Information and Scientific Documentation, AREEO, Ministry of Agriculture, Iran, 47282:69
Nerway ZAA, Duhoky MMS, Kassim NA (2020) In vitro elimination of Dahlia mosaic virus by using meristem culture electrotherapy and chemotherapy. Iraq J Agric Sci 51(2):665–674. https://doi.org/10.36103/ijas.v51i2.994
Ohta S, Kuniga T, Nishikawa F, Yamasaki A, Endo T, Iwanami T, Yoshioka T (2011) Evaluation of novel antiviral agents in the elimination of Satsuma dwarf virus (SDV) by semi-micrografting in citrus. J Japan Soc Hort Sci 80(2):145–149. https://doi.org/10.2503/jjshs1.80.145
Parštein F, Sedlák J, Svobodová L, Polák J, Gadiou S (2013) Results of in vitro chemotherapy of apple cv. Fragrance. Hort Sci 40:186–190
Paunovic S, Ruzic D, Vujovic T, Milenkovic S, Jevremovic D (2007) In vitro production of Plum pox virus– free plums by chemotherapy with ribavirin. Biotechnol Biotech 4:417–421. https://doi.org/10.1080/13102818.2007.10817486
Raj R, Kaur C, Agrawal L, Kumar S, Chauhan PS, Raj SK (2022) Development of a protocol for the elimination of Cyrtanthus elatus virus-A from Narcissus tazetta by in vitro chemotherapy in combination with electrotherapy. J Vir Method. https://doi.org/10.1016/j.jviromet.2021.114368
Retheesh S, Bhat A (2010) Simultaneous elimination of Cucumber mosaic virus and Cymbidium mosaic virus infecting Vanilla planifolia through meristem culture. Crop Prot 29:1214–1217. https://doi.org/10.1016/j.cropro.2010.05.017
Sanchez-Navarro JA, Aparicio F, Herranz MC, Minafra A, Myrta A, Pallas V (2005) Simultaneous detection and identification of eight stone fruit viruses by one-step RT-PCR. Eur J Plant Pathol 111:77–84. https://doi.org/10.1007/s10658-004-1422-y
Sastry KS, Zitter TA (2014) Management of virus and viroid diseases of crops in the tropics. Plant virus and viroid diseases in the tropics. Springer, Dordrecht
Sedlák J, Paprštein F, Suchá J (2019) Influence of chemotherapy on development and production of virus free in vitro strawberry plants. Hort Sci 46:53–56. https://doi.org/10.17221/249/2017-HORTSCI
Şeker MG, Süzerer V, Elibuyuk IO, Çiftçi YÖ (2015) In vitro elimination of PPV from infected apricot shoot tips via chemotherapy and cryotherapy. Int J Agric Biol 17:1065–1070. https://doi.org/10.17957/IJAB/15.0024
Sharma S, Singh B, Rani G, Zaidi AA, Hallan V, Nagpal A, Virk GS (2007) Production od Indian citrus ringspot virus free plants of Kinnow employing chemptherapy coupled with shoot tip grafting. J Cent Eur Agric 8(1):1–8
Singh B (2016) Effect of antiviral chemicals on in vitro regeneration response and production of PLRV-free plants of potato. J Crop Sci Biotech 18:341–348. https://doi.org/10.1007/s12892-015-0069-x
Singh B, Kaur A (2016) In vitro production of PLRV and PSTVd-free plants of potato using electrotherapy. J Crop Sci Biotech 19(4):285–294. https://doi.org/10.1007/s12892-016-0028-1
Te HS, Randall G, Jensen DM (2007) Mechanism of action of ribavirin in the treatment of chronic hepatitis C. Gastroenterol Hepatol 3(3):218–225
Thomas E, Ghany MG, Liang TJ (2012) The application and mechanism of action of ribavirin in therapy of hepatitis C. Antivir Chem Chemother 23(1):1–12. https://doi.org/10.3851/IMP2125
Thresh J (2006) Control of tropical plant virus diseases. Adv Virus Res 67:245–295
Vivek M, Modgil M (2018) Elimination of viruses through thermotherapy and meristem culture in apple cultivar ‘Oregon Spur-II. Virus- Dis 29(1):75–82. https://doi.org/10.1007/s13337-018-0437-5
Wang Q, Valkonen JP (2009) Cryotherapy of shoot tips: novel pathogen eradication method. Trends Plant Sci 14(3):119–122. https://doi.org/10.1016/j.tplants.2008.11.0
Wang L, Wang G, Hong N, Tang R, Deng X, Zhang H (2006) Effect of thermotherapy on elimination of apple stem grooving virus and apple chlorotic leaf spot virus for in vitro-cultured pear shoot tips. Hort Sci 41(3):729–732. https://doi.org/10.21273/HORTSCI.41.3.729
Wang Q, Cuellar WJ, Rajamaki ML, Hirata Y, Valkonen JPT (2008) Combined thermotherapy and cryotherapy for efficient virus eradication: relation of virus distribution, subcellular changes, cell survival and viral RNA degradation in shoot tips. Mol Plant Pathol 9(2):237–250. https://doi.org/10.1111/j.1364-3703.2007.00456.x
Wang MR, Li BQ, Feng CH, Wang QC (2016) Culture of shoot tips from adventitious shoots can eradicate apple stem pitting virus but fails in apple stem grooving virus. Plant Cell Tiss Org Cult 125:283–291. https://doi.org/10.1007/s11240-016-0948-y
Wang LY, Li YD, Sun HY, Liu HG, Tang XD, Wang QC, Zhang ZD (2017) An efficient droplet-vitrification cryopreservation for valuable blueberry germplasm. Sci Hort 219:60–69. https://doi.org/10.1016/j.scienta.2017.03.007
Wang MR, Cui ZH, Li JW, Hao XY, Zhao L, Wang QC (2018) In vitro thermotherapy-based methods for plant virus eradication. Plant Method 14:1–18. https://doi.org/10.1186/s13007-018-0355-y
Wang MR, Bi W-L, Bettoni JC, Zhang D, Volk GM, Wang Q-C (2022) Shoot tip cryotherapy for plant pathogen eradication. Plant Pathol 71:1241–1254. https://doi.org/10.1111/ppa.13565
Wu H, Qu X, Dong Z, Luo L, Shao C, Forner J, Lohmann JU, Su M, Xu M, Liu X, Zhu L (2020) WUSCHEL triggers innate antiviral immunity in plant stem cells. Science 370(6513):227–231. https://doi.org/10.1111/ppa.13565
Yi J-Y, Seo H-W, Choi Y-M, Park Y-E (2003) Ribavirin, electric current, and shoot-tip culture to eliminate several potato viruses. J Plant Biotechnol 5(2):101–105
Zare Khafri A, Solouki M, Zarghami R, Fakheri B, Mahdinezhad N, Naderpour M (2020) In vitro propagation of three Iranian apricot cultivars. In Vitro Cell Dev Biol Plant 57:102–117. https://doi.org/10.1007/s11627-020-10112-w
Zarghami R, Ahmadi B (2022) Production of Plum Pox Virus-Free and Prunus Necrotic Ringspot Virus-free regenerants using thermotherapy and meristem-tip culture in Prunus persica L. Erwerbs-Obstbau. https://doi.org/10.1007/s10341-022-00731-5
Zhang L, Zhang Z, Lin S, Zheng T, Yang X (2013) Evaluation of six methods for extraction of total RNA from Loquat. Not Bot Hort Agrobot 41(1):313–316. https://doi.org/10.15835/nbha4118233
Zhao L, Wang MR, Cui ZH, Chen L, Volk GM, Wang QC (2018) Combining thermotherapy with cryotherapy for efficient eradication of apple stem grooving virus from infected in vitro-cultured apple shoots. Plant Dis 102:1574–1580. https://doi.org/10.1094/PDIS-11-17-1753-RE
Zhao L, Wang M, Li J, Cui Z, Volk GM, Wang Q (2019) Cryobiotechnology: a double-edged sword for obligate plant pathogens. Plant Dis 103:1058–1067. https://doi.org/10.1094/PDIS-11-18-1989-FE
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This study is supported by the Agricultural Biotechnology Research Institute of Iran to Reza Zarghami.
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RZ and MN conceived the idea and designed the experiments. AZK conducted the research; LM, MN, and BA assisted in data analysis and inscribing the manuscript. All authors discussed the results and contributed to the final manuscript.
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Zare Khafri, A., Zarghami, R., Naderpour, M. et al. Assessment of virus eradication methods from infected in vitro-grown apricot cultures. Plant Cell Tiss Organ Cult 156, 52 (2024). https://doi.org/10.1007/s11240-023-02621-4
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DOI: https://doi.org/10.1007/s11240-023-02621-4