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Journal of Integrative Agriculture  2021, Vol. 20 Issue (9): 2463-2470    DOI: 10.1016/S2095-3119(20)63336-6
Special Issue: 植物病理合辑Plant Protection—Plant Pathology 植物病毒合辑Plant Virus
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Elimination of grapevine fleck virus and grapevine rupestris stem pitting-associated virus from Vitis vinifera 87-1 by ribavirin combined with thermotherapy
HU Guo-jun, DONG Ya-feng, ZHANG Zun-ping, FAN Xu-dong, REN Fan
Research Institute of Pomology, Chinese Academy of Agricultural Sciences, Xingcheng 125100, P.R.China
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摘要  

本研究以感染葡萄斑点病毒(grapevine fleck virus,GFkV)和沙地葡萄茎痘伴随病毒(grapevine rupestris stem pitting-associated virusGRSPaV)的87-1葡萄试管苗作为试验材料,研究了不同浓度的病毒醚处理(15和25 μg·mL-1; R15和R25)、热处理(37°C; T)以及病毒醚结合热处理(R15+T和R25+T)对葡萄试管苗中两种病毒的脱除效果。处理过程中发现,在R15R25病毒醚处理的过程中,葡萄试管苗均产生了药害,主要表现为植株长势弱。热处理对葡萄植株的生长具有促进作用。TR15+TR25+T三个高温处理中植株的株高显著高于CKR15R25三个常温处理的植株株高。此外,两种方法的结合可以降低高温对植株的热损伤,处理结束时TR15+TR25+T三个处理的植株死亡率分别为51.4%11.4%8.6%。处理结束后切取一定大小的茎尖进行再生,各处理再生植株的存活率均高于68.0%。同时发现,病毒醚的处理时间与再生茎尖的成活率和两种病毒的脱除效率有关。R15+T处理304050 d,植株存活率分别为97.3%90.7%74.4%3个时间段中GRSPaV的脱除率分别为55.6%84.6%93.8%。此外,病毒醚的浓度也与两种病毒的脱除效率有关。R25(35/44)处理对GFkV的脱除率比R15(25/45)处理高23.9%,对GRSPaV的脱除率比R15处理高7.0%。对各处理的脱除效率进行分析发现,热处理结合化学处理可以明显促进GFkVGRSPa的脱除,R25+T处理50天可完全脱除葡萄试管苗中的这两种病毒。



Abstract  
Vitis vinifera 87-1 plants infected by grapevine fleck virus (GFkV) and grapevine rupestris stem pitting-associated virus (GRSPaV) were used as the plant materials for virus elimination treatment.  This study evaluated the effects of ribavirin at different concentrations (15 and 25 μg mL–1; R15 and R25, respectively), thermotherapy (37°C; T), and the combination of ribavirin and thermotherapy (R15+T and R25+T) on eliminating viruses from grapevine plants in vitro.  Both R15 and R25 had phytotoxic effects and weakened plant growth.  Thermotherapy positively affected the growth of grapevine plants.  Plant height was significantly greater in T, R15+T, and R25+T than in CK, R15 and R25.  The proportion of dead plants after T, R15+T, and R25+T was 51.4, 11.4, and 8.6%, respectively.  The survival rates of regenerated plants after all treatments were >68.0%.  Ribavirin concentration and treatment time were related to the regeneration of shoot tips and elimination efficiencies of the two viruses.  The survival rates of plants after R15+T for 30, 40, and 50 days were 97.3, 90.7, and 74.4%, respectively.  The elimination rates of GRSPaV from plants in the three time quantum were 55.6, 84.6, and 93.8%, respectively.  The elimination rate of GFkV was 23.9% higher in R25 (35/44) than in R15 (25/45), and that of GRSPaV was 7.0% higher in R25 than in R15.  The combination of thermotherapy and chemotherapy was found to have a positive effect on the eradication of GFkV and GRSPaV, and R25+T for 50 days was able to completely eliminate the two viruses from in vitro grapevines.  
Keywords:  in vitro grapevine        grapevine fleck virus (GFkV)        grapevine rupestris stem pitting-associated virus (GRSPaV)        chemotherapy        thermotherapy        virus elimination  
Received: 31 March 2020   Accepted:
Fund: This work was supported by the National Key R&D Program of China (2019YFD1001800) and the China Agricultural Research System of MOF and MARA (CARS-29).
Corresponding Authors:  Correspondence DONG Ya-feng, Tel: +86-429-3598278, E-mail: yfdong@163.com   
About author:  HU Guo-jun, E-mail: hugj3114@163.com;

Cite this article: 

HU Guo-jun, DONG Ya-feng, ZHANG Zun-ping, FAN Xu-dong, REN Fan. 2021. Elimination of grapevine fleck virus and grapevine rupestris stem pitting-associated virus from Vitis vinifera 87-1 by ribavirin combined with thermotherapy. Journal of Integrative Agriculture, 20(9): 2463-2470.

Al Rwahnih M, Daubert S, Golino D, Rowhani A. 2009. Deep sequencing analysis of RNAs from a grapevine showing Syrah decline symptoms reveals a multiple virus infection that includes a novel virus. Virology, 387, 395–401.
Alabi O J, Martin R R, Naidu R A. 2010. Sequence diversity, population genetics and potential recombination events in grapevine rupestris stem pitting-associated virus in Pacific North-West vineyards. Journal of General Virology, 91, 265–276.
Ashnayi M, Kharrazi M, Sharifi A, Mehrvar M. 2012. Carnation etched ring virus elimination through shoot tip culture. Journal of Biological and Environmental Sciences, 6,175–180.
Bota J, Cretazzo E, Montero R, Rossello J, Cifre J. 2014. Grapevine fleck virus (GFKV) elimination in a selected clone of “Vitis vinifera” L. cv. Manto Negro and its effects on photosynthesis. International Journal of Vine and Wine Sciences, 48, 11–19.
Buciumeanu E, Vi?oiu E. 2000. Elimination of grapevine viruses in Vitis vinifera L. cultivars. In: Proceedings of the 13th Meeting of the International Council for the Study of Viruses and Virus-Like Diseases of the Grapevine. Adelaide, Australia. pp. 165–166.
Chahardehi A M, Rakhshandehroo F, Mozafari J, Mousavi L. 2016. Efficiency of a chemo-thermotherapy technique for eliminating Arabis mosaic virus (ArMV) and Prunus necrotic ringspot virus (PNRSV) from in vitro rose plantlets. Journal of Crop Protection, 5, 497–506.
Chalak L, Elbeaino T, Elbitar A, Fattal T, Choueiri E. 2015. Removal of viruses from Lebanese fig varieties using tissue culture and thermotherapy. Phytopathologia Mediterranea, 54, 531–535.
Chien K W, Agrawal D C, Tsay H S, Chang C A. 2015. Elimination of mixed ‘Odontoglossum ringspot’ and ‘Cymbidium mosaic’ viruses from Phalaenopsis hybrid ‘V3’ through shoot-tip culture and protocorm-like body selection. Crop Protection, 67, 1–6.
Cretazzo E, Tomás M, Padilla C, Rosselló J, Medrano H, Padilla V, Cifre J. 2010. Incidence of virus infection in old vineyards of local grapevine varieties from Majorca: Implications for clonal selection strategies. Spanish Journal of Agricultural Research, 8, 409–418.
Crotty S, Cameron C, Andino R. 2002. Ribavirin’s antiviral mechanism of action: Lethal mutagenesis? Journal of Molecular Medicine, 80, 86–95.
Dziedzic E. 2008. Elimination of Prunus necrotic ring spot virus (PNRSV) from plum ‘Earliblue’ shoots through thermotherapy in vitro. Journal of Fruit and Ornamental Plant Research, 16, 101–109.
Eichmeier A, Kominkova M, Pecenka J, Kominek P. 2019. High-throughput small RNA sequencing for evaluation of grapevine sanitation efficacy. Journal of Virological Methods, 267, 66–70.
Gribaudo I, Gambino G, Cuozzo D, Mannini F. 2006. Attempts to eliminate Grapevine rupestris stem pitting-associated virus from grapevine clones. Journal of Plant Pathology, 88, 293–298.
Gu?? I C, Buciumeanu E C, Vi?oiu E. 2014. Elimination of Grapevine fleck virus by in vitro chemotherapy. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 42, 115–118.
Han Y T, Shi X H, Yang G S, Wang X R, Liu K Y, Xu F, Ni J J. 2011. Shoot tip tissue virus-free culture and RT-PCR detection of GLRaV-3 in Red globe grape. Chinese Agricultural Science Bulletin, 27, 198–202. (in Chinese)
Hansen A J, Hildebrandt A C. 1966. The distribution of tobacco mosaic virus in plant callus cultures. Virology, 28, 15–21.
Hu G J, Dong Y F, Zhang Z P, Fan X D, Ren F. 2019. Elimination of apple necrosis mosaic virus from potted apple plants by thermotherapy combined with shoot-tip grafting. Scientia Horticulturae, 252, 310–315.
Hu G J, Dong Y F, Zhang Z P, Fan X D, Ren F, Li Z N. 2018a. Effect of pre-culture on virus elimination from in vitro apple by thermotherapy coupled with shoot tip culture. Journal of Integrative Agriculture, 17, 2015–2023.
Hu G J, Dong Y F, Zhang Z P, Fan X D, Ren F, Li Z N, Zhang S N. 2018b. Elimination of Grapevine rupestris stem pitting-associated virus from Vitis vinifera ‘Kyoho’ by an antiviral agent combined with shoot tip culture. Scientia Horticulturae, 229, 99–106.
Hu G J, Dong Y F, Zhang Z P, Fan X D, Ren F, Zhou J. 2015. Virus elimination from in vitro apple by thermotherapy combined with chemotherapy. Plant Cell, Tissue and Organ Culture, 121, 435–443.
Hu G J, Hong N, Wang L P, Hu H J, Wang G P. 2012. Efficacy of virus elimination from in vitro-cultured sand pear (Pyrus pyrifolia) by chemotherapy combined with thermotherapy. Crop Protetion, 37, 20–25.
Kidulile C E, Miinda Ateka E, Alakonya A E, Ndunguru J C. 2018. Efficacy of chemotherapy and thermotherapy in elimination of East African cassava mosaic virus from Tanzanian cassava landrace. Journal of Phytopathology, 166, 739–745.
Komínek P, Jandurová O M. 2011. Thermotherapy sanitation of two grapevine cultivars. Acta Virologica, 55, 89–90.
Komínek P, KomÍnková M, Jandová B. 2016. Effect of repeated Ribavirin treatment on grapevine viruses. Acta Virologica, 60, 400–403.
Kostadinovska E, Mitrev S, Bianco P A, Casati P, Bulgari D. 2014. First report of Grapevine virus A and Grapevine fleck virus in the former Yugoslav republic of macedonia. Plant Diseases, 98, 1747–1747.
K?i?an B, Ondrušiková E, Holleinova V, Moravcova K, Blahova L. 2009. Elimination of Grapevine fanleaf virus in grapevine by in vivo and in vitro thermotherapy. Horticultural Science, 36, 105–108.
Li B Q, Feng C H, Hu L Y, Wang M R, Wang Q C. 2016. Shoot tip culture and cryopreservation for eradication of Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) from apple rootstocks ‘M9’ and ‘M26’. Annals of Applied Biology, 168, 142–150.
Lunden S, Meng B, Avery J, Qiu W. 2010. Association of Grapevine fanleaf virus, Tomato ringspot virus and Grapevine rupestris stem pitting-associated virus with a grapevine vein-clearing complex on var. Chardonnay. European Journal of Plant Pathology, 126, 135–144.
Maliogka V I, Skiada F G, Eleftheriou E P, Katis N I. 2009. Elimination of a new ampelovirus (GLRaV-Pr) and Grapevine rupestris stem pitting associated virus (GRSPaV) from two Vitis vinifera cultivars combining in vitro thermotherapy with shoot tip culture. Scientia Horticulturae, 123, 280–282.
Martelli G P. 2012. Grapevine virology highlights: 2010–2012. In: Proceedings of the 17th Congress of International Council for the Study of Virus and Virus-like Diseases of the Grapevine (ICVG). University of California-Davis, California, USA. pp. 13–31.
Martelli G P. 1993. Graft-Transmissible Diseases of Grapevines: Handbook for Detection and Diagnosis. FAO Publication Division, Rome, Italy.
Martelli G P, Jelkmann W. 1998. Foveavirus, a new plant virus genus. Archives of Virology, 143, 1245–1249.
Martelli G P, Sabanadzovic S, Ghanem-Sabanadzovic N A, Saldarelli P. 2002. Maculavirus, a new genus of plant viruses. Archives of Virology, 147, 1847–1853.
Meng B, Gonsalves D. 2007. Grapevine rupestris stem pitting-associated virus: A decade of research and future perspectives. Plant Viruses, 1, 52–62.
Meng B Z, Johnson R, Peressini S, Forsline P L, Gonsalves D. 1999. Rupestris stem pitting associated virus-1 is consistently detected in grapevines that are infected with rupestris stem pitting. European Journal of Plant Pathology, 105, 191–199.
Morelli M, Minafra A, Boscia D, Martelli G P. 2011. Complete nucleotide sequence of a new variant of grapevine rupestris stem pitting-associated virus from southern Italy. Archives of Virology, 156, 543–546.
Nakaune R, Inoue K, Nasu H, Kagokawa K, Nitta H, Imada J, Nakano M. 2008. Detection of viruses associated with rugose wood in Japanese grapevines and analysis of genomic variability of Rupestris stem pitting-associated virus. Journal of General Plant Pathology, 74, 156–163.
Panattoni A, Triolo E. 2010. Susceptibility of grapevine viruses to thermotherapy on in vitro collection of Kober 5BB. Scientia Horticulturae, 125, 63–67
Poojari S, Lowery T, Rott M, Schmidt A M, DeLury N, Boulé J, Úrbez-Torres J R. 2016. First report and prevalence of Grapevine fleck virus in grapevines (Vitis vinifera) in Canada. Plant Diseases, 100, 1028.
Simpkins I, Walkey D G A, Neely H A. 1981. Chemical suppression of virus in cultured plant tissues. Annals of Applied Biology, 99, 161–169.
Skiada F G, Grigoriadou K, Maliogka V I, Katis N I, Eleftheriou E P. 2009. Elimination of Grapevine Leafroll-associated virus 1 and Grapevine Rupestris Stem Pitting-associated virus from grapevine cv. Agiorgitiko, and a micropropagation protocol for mass production of virus-free plantlets. Journal of Plant Pathology, 91, 175–182.
Verma N, Ram R, Zaidi A A. 2005. In vitro production of Prunus necrotic ringspot virus-free begonias through chemo- and thermotherapy. Scientia Horticulturae, 103, 239–247.
Vivek M, Modgil M. 2018. Elimination of viruses through thermotherapy and meristem culture in apple cultivar ‘Oregon Spur-II’. Virus Disease, 29, 75–82.
Wang H Y, Liu F C, Xue R G, Yang Z Y, Zhang Z P, Hong N, Wang G P. 1991. Study on the virus elimination from mother trees of apple. China Fruits, 4, 15–17. (in Chinese)
Wang J, Xi D, Liu J, Chen K, Li H, Liu X, Yan S, Ercisli S, Lin H. 2012. Genetic variability in Grapevine virus A from Vitis vinifera L.×Vitis labrusca L. in Sichuan, China. Turkish Journal of Biology, 36, 542–551.
Wang M R, Li B Q, Feng C H, Wang Q C. 2016. Culture of shoot tips from adventitious shoots can eradicate Apple stem pitting virus but fails in Apple stem grooving virus. Plant Cell, Tissue and Organ Culture, 125, 283–291.
Youssef S A, Al-Dhaher M M A, Shalaby A A. 2009. Elimination of Grapevine fanleaf virus (GFLV) and Grapevine leaf roll-associated virus-1 (GLRaV-1) from infected grapevine plants using meristem tip culture. International Journal of Virology, 5, 89–99.
Zhang Y, Uyemoto J K, Golino D, Rowhani A. 1998. Nucleotide sequence and RT-PCR detection of a virus associated with grapevine rupestris stem-pitting disease. Phytopathology, 88, 1231–1237.
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