Articles
EFFECT OF THE CULTURE MEDIUM ON MERISTEM DIFFERENTIATION AND PLANT REGENERATION IN VITIS VINIFERA L.
Article number
652_56
Pages
425 – 432
Language
English
Abstract
Grapevine culture is of great importance in the world, particularly in Europe where it occupies more than 60% of the cultivated area.
In Portugal, the grapevine plays an important role in the agricultural economy.
However, grapevines are susceptible to several diseases due to different pathogens, namely Grapevine leafroll associated virus-3 (GLRaV-3). In vitro culture is a useful tool for rapid propagation of grapevines and the meristem-tip culture allows the obtaining of virus-free plants.
In this study we analysed the effect of the culture medium for meristem differentiation and plant regeneration, as well as in vitro meristem culture in the elimination of GLRaV-3 in three cultivars of Vitis vinifera (Alvarinho, Touriga Nacional and Moscatel Galego). Virus detection was done by DAS-ELISA. In a total of 27 plants studied, an infection of 29.6% was registered.
The meristems were cultured in sixteen culture media containing different concentrations of 6-Benzylaminopurine (BAP) (0.0; 1.0; 2.0 and 2.5mg.L-1) and of Naphtalene Acetic Acid (NAA) (0.0; 0.5; 1.0; 1.5 and 2.0mg.L-1). The meristem differentiation registered an average of 48.3%, with Touriga Nacional attaining the highest value of 65.9%. The medium supplemented with the highest concentration of BAP and without NAA gave the best meristem differentiation results.
For plant regeneration, the same media with BAP and without NAA were used.
Although the presence of auxin in the differentiation medium can induce callogenesis, the regeneration of plants was more successful when the meristems were differentiated in an auxin-supplemented medium (1.5mg.L-1 NAA). Before acclimatization, the plants were submitted to a further DAS-ELISA in order to evaluate virus eradication.
All the plants were virus-free, proving the efficiency of this method for GLRaV-3 eradication.
In Portugal, the grapevine plays an important role in the agricultural economy.
However, grapevines are susceptible to several diseases due to different pathogens, namely Grapevine leafroll associated virus-3 (GLRaV-3). In vitro culture is a useful tool for rapid propagation of grapevines and the meristem-tip culture allows the obtaining of virus-free plants.
In this study we analysed the effect of the culture medium for meristem differentiation and plant regeneration, as well as in vitro meristem culture in the elimination of GLRaV-3 in three cultivars of Vitis vinifera (Alvarinho, Touriga Nacional and Moscatel Galego). Virus detection was done by DAS-ELISA. In a total of 27 plants studied, an infection of 29.6% was registered.
The meristems were cultured in sixteen culture media containing different concentrations of 6-Benzylaminopurine (BAP) (0.0; 1.0; 2.0 and 2.5mg.L-1) and of Naphtalene Acetic Acid (NAA) (0.0; 0.5; 1.0; 1.5 and 2.0mg.L-1). The meristem differentiation registered an average of 48.3%, with Touriga Nacional attaining the highest value of 65.9%. The medium supplemented with the highest concentration of BAP and without NAA gave the best meristem differentiation results.
For plant regeneration, the same media with BAP and without NAA were used.
Although the presence of auxin in the differentiation medium can induce callogenesis, the regeneration of plants was more successful when the meristems were differentiated in an auxin-supplemented medium (1.5mg.L-1 NAA). Before acclimatization, the plants were submitted to a further DAS-ELISA in order to evaluate virus eradication.
All the plants were virus-free, proving the efficiency of this method for GLRaV-3 eradication.
Authors
S. Gomes, A.M. Pereira, O. Pinto-Carnide
Keywords
Micropropagation, in vitro culture, grapevine, virus eradication, growth regulators
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