In situ localization of Grapevine fanleaf virus and phloem-restricted viruses in embryogenic callus of Vitis vinifera

Gambino, Giorgio; Vallania, R.; Gribaudo, Ivana

doi:10.1007/s10658-010-9619-8

Cited by 9 publications

(10 citation statements)

References 27 publications

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“…Differential distribution within the calli has also been previously reported for GFLV and two phloemlimited viruses and was proposed to be related to mechanisms of viral movement in tissues, possibly due to the characteristics and evolution of the callus after several months of culture (Gambino et al 2010). However, factors leading to the differential distribution of HSVd and GYSVd-1 observed here remain unknown, and additional studies are needed to investigate the distribution and movement of these viroids within infected plants and to formulate specific hypotheses.…”

Section: Discussionsupporting

confidence: 65%

“…Detailed histological observations of embryogenic cultures have been reported elsewhere (Gambino et al 2010). However, it could be of interest here to remark that no vascular connection between embryoids or between grapevine somatic embryos and the parent tissues was observed.…”

Section: Resultsmentioning

confidence: 99%

“…Paraffin-embedding was performed as previously described (Gambino et al 2010). Probes for detecting the plus strands of GYSVd-1 and HSVd were generated by in vitro transcription from previously described plasmid templates (Navarro et al 2009) using the Digoxigenin RNA labelling kit (Roche Applied Science).…”

Section: Methodsmentioning

confidence: 99%

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Somatic embryogenesis efficiently eliminates viroid infections from grapevines

Gambino¹,

Navarro²,

Vallania³

et al. 2011

Eur J Plant Pathol

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Indirect somatic embryogenesis is effective at eliminating the most important viruses affecting grapevines. Accordingly, this technique was tested as a method for eradicating two widespread viroids, Grapevine yellow speckle viroid 1 (GYSVd-1) and Hop stunt viroid (HSVd), from four grapevine cultivars. Both viroids were detected by RT-PCR in grapevine floral explants used for initiating embryogenic cultures, as well as in undifferentiated cells of embryogenic and non-embryogenic calli from anthers and ovaries. In contrast, somatic embryos differentiated from these infected calli were viroid-free, and viroids were not detected in embryo-derived plantlets even 3 years after their transfer to greenhouse conditions. A wider spatial distribution of HSVd than GYSVd-1 within proliferating calli was revealed by in situ hybridization, whereas no hybridization signal was detected in the somatic embryos. In addition, GYSVd-1 and HSVd were localised in the nucleus of infected cells, conclusively showing the nuclear accumulation of representative members of Apscaviroid and Hostuviroid genera, which has been only an assumption so far. Somatic embryogenesis was compared to in vitro thermotherapy, a technique routinely used for virus eradication. After thermotherapy, HSVd and GYSVd-1 were detected in all in vitro plantlets of the cultivar Roussan, and in all lines analysed after 3 years of culture in greenhouse. The high efficiency with which somatic embryogenesis may eliminate viroids and viruses from several infected grapevine cultivars, should allow the availability of virus-and viroid-free material, which would be useful not only for sanitary selection but also for basic research on plant-virus and plant-viroid interactions in grapevine.

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Section: Discussionsupporting

confidence: 65%

Section: Resultsmentioning

confidence: 99%

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Somatic embryogenesis efficiently eliminates viroid infections from grapevines

Gambino¹,

Navarro²,

Vallania³

et al. 2011

Eur J Plant Pathol

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“…Thermotherapy, even in association with other techniques such as tissue culture, represents the most fre- Goussard & Wiid, 1992;Popescu et al, 2003Popescu et al, , 2010Gambino et al, 2006Gambino et al, , 2010Gribaudo et al, 2006;Borroto-Fernández et al, 2009;Youssef et al, 2009Ramos & Zamora, 1999D'Onghia et al, 2001;Golino et al, 2007;Quainoo et al, 2008Fitch et al, 2001Parmessur et al, 2002;Ramgareeb et al, 2010Ebi et al, 2000Ayabe & Sumi, 2001;Ramírez-Malagón et al, 2006Truskinov & Rogozina, 1997Mahmoud et al, 2009Wangai & Bock, 1996Morris et al, 1997;S edivá et al, 2006;Kumar et al, 2009;Kabir et al, 2010 quently used technique for sanitation over the last 20 years, while applications of methods such as chemotherapy and tissue culture represent the main topic in plant sanitation research.…”

Section: Comparison Of Techniquesmentioning

confidence: 99%

Review. Elimination of viruses in plants: twenty years of progress

Panattoni

Luvisi

Triolo

2013

Span. j. agric. res.

145

112

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To shed light on trends about elimination of viruses from plants, a bibliographic research was conducted to identify thermotherapy, chemotherapy and tissue culture trials published from 1991 through 2010. Among woody plants, grapevine, apple and peach are the most frequent targets of sanitation protocols because their health status is strictly regulated. Even if thermotherapy represents the preferred method for the host, grapevine viruses can also be eliminated with chemotherapy and tissue culture; apple viruses respond to chemotherapy as well. Although a similar trend was reported among herbaceous plants, chemotherapy was the most frequently used technique in potato. With regard to virus, thermotherapy was successfully applied against viruses belonging to 13 families and an unassigned genus. Instead, chemotherapy and tissue culture techniques eradicated viruses belonging to fewer families (nine). An interpretation of thermotherapy effects considers the new metabolic “pathways” triggered by the natural antiviral response emitted by the infected plant, with particular reference to virus-induced gene silencing. With regard to chemotherapy, several groups of antiviral drugs belong to inosine monophosphate dehydrogenase inhibitors, S-adenosylhomocysteine hydrolase inhibitors, neuraminidase inhibitors. Tissue culture, usually adopted to regenerate plantlets in biotechnological breeding programs, represents the less used tool for eliminate viruses from plants.

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“…The observed difference in the eradication efficiency of different viruses in SCs was most likely related to the ability of the viruses to invade plant tissues and calli; for instance, phloem‐limited viruses, such as GLRaV‐3 and GVA, had more difficulty in invading callus cells than GFLV (Gambino et al ., 2010 ). This could justify the persistence of GFLV in some SCs, as reported elsewhere (Gambino et al ., 2009 ; San Pedro et al ., 2017 ; Malenica et al ., 2020 ).…”

Section: Resultsmentioning

confidence: 99%

Grapevine virome and production of healthy plants by somatic embryogenesis

Nuzzo

Moine

Nerva

et al. 2022

Microbial Biotechnology

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Summary Grapevine ( Vitis spp.) is a widespread fruit tree hosting many viral entities that interact with the plant modifying its responses to the environment. The production of virus‐free plants is becoming increasingly crucial for the use of grapevine as a model species in different studies. Using high‐throughput RNA sequencing, the viromes of seven mother plants grown in a germplasm collection vineyard were sequenced. In addition to the viruses and viroids already detected in grapevine, we identified 13 putative new mycoviruses. The different spread among grapevine tissues collected in vineyard, greenhouse and in vitro conditions suggested a clear distinction between viruses/viroids and mycoviruses that can successfully be exploited for their identification. Mycoviruses were absent in in vitro cultures, while plant viruses and viroids were particularly accumulated in these plantlets. Somatic embryogenesis applied to the seven mother plants was effective in the elimination of the complete virome, including mycoviruses. However, different sanitization efficiencies for viroids and grapevine pinot gris virus were observed among genotypes. The absence of mycoviruses in in vitro plantlets, associated with the absence of all viral entities in somaclones, suggested that this regeneration technique is also effective to eradicate endophytic/epiphytic fungi, resulting in gnotobiotic or pseudo‐gnotobiotic plants.

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In situ localization of Grapevine fanleaf virus and phloem-restricted viruses in embryogenic callus of Vitis vinifera

Cited by 9 publications

References 27 publications

Somatic embryogenesis efficiently eliminates viroid infections from grapevines

Somatic embryogenesis efficiently eliminates viroid infections from grapevines

Review. Elimination of viruses in plants: twenty years of progress

Grapevine virome and production of healthy plants by somatic embryogenesis

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