Effect of Benzyladenine Concentration and Double-Phase Culture System on In Vitro Multiplication of Adult Albariño Plants

Flow cytometry and microsatellite analyses were used to evaluate the trueness-to-type of somatic embryogenesis-regenerated plants from six important Spanish grapevine (Vitis vinifera L.) cultivars. Tetraploid plants were regenerated through somatic embryogenesis from all of the cultivars tested with the exception of 'Merenzao'. In addition, an octoploid plant was obtained in the cv. 'Albariño', and two mixoploids in 'Torrontés'. The most probable origin of these ploidy variations is somaclonal variation. The cv. 'Brancellao' presented significantly more polyploids (28.57%) than any other cultivar, but it must be noted that 50% of the adult field-grown 'Brancellao' mother plants analysed were mixoploid. Hence, it is probable that these polyploids originated either from somaclonal variation or by separation of genotypically different cell layers through somatic embryogenesis. Microsatellite analysis of somatic embryogenesis-regenerated plants showed true-totype varietal genotypes for all plants except six 'Torrontés' plants, which showed a mutant allele (231) instead of the normal one (237) at the locus VVMD5. There was not a clear relationship between the occurrence of the observed mutant regenerated plants and the callus induction media composition, the developmental stage of the inflorescences, the type of explant used for starting the cultures or the type of germination (precocious in differentiation medium or normal in germination medium) in any of the cultivars tested, except 'Torrontés'. The mutant plants described herein have been transplanted to soil for future evaluation of putative phenotypic traits of interest. These mutants can be useful both for breeding programs and for functional genomic approaches aimed at increasing knowledge of the biology of grapevine.

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Highly efficient in vitro tetraploid plant production via colchicine treatment using embryogenic suspension cultures in grapevine (Vitis vinifera cv. Mencía)

Acanda

Martı́nez

González

et al. 2015

Plant Cell Tiss Organ Cult

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In vitro chromosome doubling techniques have been used to produce polyploids in several species, including table grapes, but limited research in this field has been performed using winegrapes. An efficient procedure was established to induce tetraploid grapevine plants by colchicine treatment of embryogenic cell aggregates (ECAs) grown in winegrape cultivar (cv. Mencía) suspension cultures. Colchicine treatment caused a significant decrease in the survival and embryogenic potential of the ECAs compared with the control. However, plantlets were regenerated and assessed for ploidy level by flow cytometry. Almost all plants not treated with colchicine maintained their ploidy level, whereas colchicine treatment resulted in a high variation in ploidy level. Colchicine at 0.2 % was the most effective concentration for obtaining tetraploid plantlets (25 % tetraploids). No chimeric or mixoploid plantlets were detected in this study. Based on the unique cell origin of grapevine somatic embryos, our system prevented obtaining chimeric plants. This protocol allows an increase in grapevine genetic diversity and could be a valuable tool for improving this crop. To our knowledge, this is the first report on in vitro chromosome doubling by colchicine treatment of embryogenic cell aggregates growing in grapevine suspension cultures.

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Effect of Benzyladenine Concentration and Double-Phase Culture System on In Vitro Multiplication of Adult Albariño Plants

Cited by 6 publications

References 6 publications

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Hyperhydricity-Related Morphologic and Biochemical Changes in Vanilla (Vanilla planifolia)

Detection of somaclonal variants in somatic embryogenesis-regenerated plants of Vitis vinifera by flow cytometry and microsatellite markers

Highly efficient in vitro tetraploid plant production via colchicine treatment using embryogenic suspension cultures in grapevine (Vitis vinifera cv. Mencía)

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