Adventitious plant regeneration on leaf explants from adult male kiwifruit and AFLP analysis of genetic variation

Abstract

The present work reports on a study of plant regeneration carried out with callus from the leaf blades and petioles of field-grown male adult kiwifruit plants (Actinidia deliciosa (Chev.) Liang and Ferguson). The cultivars used were 'Tomuri' and clone A, a selected male plant grown in north western Spain. The best shoot induction conditions were obtained in 'Tomuri' leaf blades cultured in K(h) medium in the presence of 23 lM Zeatin and 0.1 lM NAA. Under these conditions, more than 80% of organogenic callus induction was observed, with an average of 14 new shoots in the second subculture. The initial length of the shoots affected shoot elongation, which was accomplished by culturing isolated shoots in K(h) medium with half-strength salts, supplemented with 0.4 lM Zeatin and 0.1 lM NAA. A possible detrimental long-term effect of cytokinins on shoot elongation can account for the results, since elongation was not observed until 1 month of culture in elongation medium.For rooting, shoots (1 cm in length) were basally immersed in a 5 mM IBA solution for 15 s, and transferred to half-strength K(h) basal medium. Regenerated plants were acclimated in a sterile peat:perlite substrate for 10 days, and then transferred to soil. AFLP analysis was accomplished with 15 primer combinations from which 13 showed reproducible and well-resolved bands, producing a total of 1321 fragments from which 1281 were polymorphic (97%). A dendrogram was constructed using both monomorphic and polymorphic bands, showing genetic variation among field-grown plants and tissue culture-derived regenerants.

An evaluation of a new approach to the regeneration of Helichrysum italicum (Roth) G. Don, and the molecular characterization of the variation among sets of differently derived regenerants

“…[8], Carya illinoiensis [9,48], Syngonium podophyllum [49], Agave fourcroydes Lem. [50], Elaeis guineensis [51] Actinidia deliciosa [52], and Phoenix dactylifera [53]. In Arabidopsis, 66.7% of the analysed regenerated plants showed at least one polymorphism [4], while for barley, the literature data is controversial, since Ruiz et al [6] found that somaclonal variation did not appear to be a very frequent event, while Bednarek et al [54] reported that all the regenerated plants showed at least one polymorphism with respect to the control plant.…”

Somaclonal Variation in Tissue Culture: A Case Study with Olive

“…A close relationship between the type of explants, regeneration system, and the presence of somaclonal variation was detected in shoots regenerated from leaf callus [99].…”

In vitro organogenesis and plant regeneration from leaves ofActinidia erianthaBenth. cv White (kiwifruit)

“…Most studies have used female kiwifruit, but there is one report for male kiwifruit 'Tomuri' (Prado et al 2007). That research showed an organogenic callus induction rate of !80%, with an average of 14 new shoots in the second subculture from leaf explants cultured in media supplement with zeatin and NAA.…”