In vitro propagation of Vitis: The effects of organic substances on shoot multiplication

The effects of organic substances on shoot multiplication from subcultured shoots of the Vitis hybrid Remaily Seedless were investigated. Culture media contained the inorganic nutrients of MURASHIGE and SKOOG (1962).Benzylaminopurine (BAP) was most effective in promoting shoot multiplication at 5 μM. 60 % of the shoots had at least 3 nodes (1.5 cm total length), a size considered adequate for micropropagation.The addition of adenine sulfate from 2.5 to 20 x 10 - 4 M depressed shoot multiplication when BAP was optimal.Combinations of thiamine (2 and 4 μM) and inositol (50; 100; 500 μM) were tested. Shoot multiplication decreased with increasing inositol concentration and was higher with 4 μM of thiamine. Best multiplication was with 4 μM thiamine and 50 μM inositol and gave similar number of shoots per explant as obtained on our standard medium which contains in addition nicotinic acid and pyridoxine. However, twice _as many 3-node shoots were produced on the standard medium.The amino acids arginine, aspartate, asparagine, glutamate, glutamine and tyrosine at 2.5-40 x 10 - 4 M had no effect on shoot multiplication.With the culture conditions used in this research, an adequate set of organic constituents for shoot multiplication of grapevines is defined as 3 μM thiamine · HCl, 55.5 μM myo-inositol, 8 μM nicotinic acid, 5 μM pyridoxine · HCl and 5 μM benzylaminopurine. The addition of 4 μM of aspartate may be beneficial. It was found that optimization of vitamin concentration is important in shoot multiplication. The restricted Optimum cytokinin concentration for best shoot production suggests that its concentration may have to be adjusted for each cultivar

In vitro vegetative propagation of Vitis: Application of previously defined culture conditions to a selection of genotypes

A range of grapevine species, cultivars and hybrids was surveyed for their responses to in vitro micropropagation.21 genotypes were established in culture from shoot apices (apical dome plus 2- to 4-leaf primordia) with a method previously developed for the Vitis hybrid Rougeon. However, this method proved to be inadequate for the establishment of the hybrid Seyval.Shoots grew on for 21 genotypes using a medium previously devised for the Vitis hybrid Remaily Seedless. Cultural conditions were inadequate for 4 of these genotypes. 10-h days resulted in best shoot production for 3 genotypes and 16-h days for 1. In general, shoot production was better or equal with short days than it was with long days. Shoots were multiplied from 3- to 4-node shoots (1.5 cm long) obtained in vitro. For the first 5 subcultures, 17 genotypes were multiplied on the medium used for first shoot production. Best results were obtained with Remaily Seedless which produced 13 shoots of at least 3 nodes, per subculture shoot after 2 months in culture. The 6th subculture employed the C2D salts which had previously been devised to improve shoot multiplication of Remaily Seedless. Yields were increased from 31 % to 350 % for all genotypes except V. labruscana Catawba which died in culture. For the rooting of subcultured shoots, a medium which had previously been developed for that purpose on Remaily Seedless was used successfully on 15 genotypes.Multiplication végétative de la vigne in vitro:Application des conditions de culture définies auparavant pour une sélection de génotypesLa multiplication végétative in vitro a été réalisée pour une collection d'espèces, de cultivars et d'hybrides du genre Vitis. 21 génotypes furent mis en culture à partir d'apex de 2 à 4 ébauches foliaires, par une méthode que nous avions développé pour l'hybride Rougeon. Ces conditions de culture n'ont pas été favorables â la croissance de l'hybride Seyval.La production de pousses herbacées fut obtenue pour ces 21 génotypes sur un milieu nutritif que nous avions défini pour l'hybride Remaily Seedless. Pour 4 génotypes les conditions de cultures n'ont pas été favorables. L'influence de la photopériode sur la production de pousses fut étudiée également. Les journées de 10 h bénéficièrent 3 génotypes et les journées de 16 h un seul. En général, la production en jours courts s'est avérée meilleur qu'en jours longs. La multiplication des pousses fut éffectuée par repiquage de boutures terminales obtenues in vitro et comprenant 3 à 4 noeuds (1,5 cm). Pour les 5 premiers passages la multiplication de 17 génotypes fut continuée sur le milieu de culture utilisé pour la production des premières ·pousses. Les meilleurs résultats ont été obtenus pour Remaily Seedless. En 2 mois chaque bouture apicale de ce cultivar a produit en moyenne 13 pousses ayant au moins 3 noeuds. Le 6e repiquage a été fait sur un milieu, C2D, développé auparavant pour améliorer la multiplication de Remaily Seedless. La récolte fut augmentée ainsi de 31 % à 350 % selon le génotype sauf pour V. labruscana Catawba qui s'est nécrosé.Pour l'enracinement de boutures apicales obtenues en culture, un milieu défini à cet effet pour Remaily Seedless fut utilisé avec succès pour 15 génotypes

Micropropagação do porta-enxerto de videira '420-A' Micropropagation of '420-A' grapevine rootstock

O objetivo deste trabalho foi estabelecer um protocolo para a micropropagação do porta-enxerto de videira 420-A. O estabelecimento das culturas foi realizado com segmento nodal, cuja fonte dos explantes foram brotações de estacas lenhosas armazenadas sob refrigeração. No cultivo inicial, foram testados: o efeito de 6-benzilaminopurina e cinetina nas concentrações de 0; 1; 5 e 10 &micro;M, diferentes meios de cultura (MS, NN e WPM) e diluições do meio básico (MS, MS/2, MS/4 e MS/8). Na fase de alongamento e multiplicação, os meios de cultura testados foram MS, MS/2, NN e WPM. No enraizamento, foram testados: o meio de cultura MS/2 sem e com carvão ativado (1gL-1). Na aclimatização, foram testados vermiculita, Plantmax® e casca de arroz carbonizada como substrato. A cinetina não apresentou efeito sobre a brotação e o crescimento dos segmentos nodais. Já o BAP promoveu um aumento no número de brotos por explante. O aumento na concentração de BAP reduziu o número de folhas emitidas por explante e aumentou os sintomas de vitrificação, sendo os melhores resultados obtidos com 1 &micro;M de BAP. No cultivo inicial, o meio de cultura MS, com a concentração normal de sais, permitiu o maior crescimento das brotações. As diluições do meio MS em 1/4 e 1/8 mostraram-se prejudiciais ao desenvolvimento do porta-enxerto '420-A', afetando o crescimento das brotações após o primeiro subcultivo. Durante a multiplicação o meio MS/2 foi o que proporcionou melhores resultados. O enraizamento ocorreu naturalmente durante a multiplicação, sendo desnecessário o uso de carvão ativado no meio de cultura. A aclimatização foi realizada com sucesso em câmara de nebulização, com substrato vermiculita (95,8%) e Plantmax® (87%). Conclui-se que o porta-enxerto '420-A' pode ser micropropagado pelo cultivo inicial de segmentos nodais em meio de cultura MS + 1 &micro;M de BAP, alongamento das brotações e multiplicação pelo seccionamento das mesmas em meio MS/2 e aclimatização em substrato vermiculita ou Plantmax®.<br>The objective of this work was to establish a protocol for the rootstock of 420-A micropropagation. The establishment of the cultures was accomplished with nodal segments, whose source of explants was the budding of woody stakes stored under refrigeration. In the initial cultivation were tested: the effect of 6-benzilaminopurine and kinetin with 0, 1, 5 and 10 &micro;M concentrations, different culture medium (MS, NN and WPM) and dilutions of the basic medium (MS, MS/2, MS/4 and MS/8). In the alongation and multiplication steps were tested the following culture medium: MS, MS/2, NN and WPM. In the rooting were tested: the MS/2 culture medium with or without activated coal (1gL-1). In the acclimatization were tested the substrate vermiculite, Plantmax and carbonized rice hulls. The kinetin didn't present effect on the budding and the growth of the nodal segments. BAP already promoted an increase in the number of sprouts per explant. The increase in the concentration of BAP reduced the number of leaves emitted by explant and increased the vitrification symptoms, being the best results obtained with 1 &micro;M of BAP. In the initial cultivation the MS medium culture with the normal concentration of salts allowed the largest growth of the buddings. The dilutions of the MS medium in &frac14; and 1/8 showed very harmful to the development of the rootstock '420-A', being quite harmed the growth of the shoots after the first subcultivation. During the multiplication the medium MS/2 showed more appropriate. The roots happened naturally during the multiplication, being unnecessary the use of activated coal in the culture medium. The acclimatization was accomplished with success in a misty camera, being obtained high survival rates in vermiculite (95,8%) and Plantmax® (87%). It is concluded that the rootstock '420-A' can be micropropagated by initial cultivation of nodal segments in a culture medium of MS+1&micro;M of BAP, multiplication by sectionalizing the shoots in MS/2 medium and acclimatization in vermiculite substratum or Plantmax®