Micropropagation in banana using high levels of cytokinins does not involve any genetic changes as revealed by RAPD and ISSR markers

Abstract

Use of high levels of growth regulators during micropropagation results in undesirable clonal variability in important commercial crops such as banana. The present study investigated the effects of high levels of cytokinins on micropropagation in banana (genotype AAB), and the genetic stability of plantlets was assessed using RAPD and ISSR markers. Cytokinins, such as BA and kinetin were added to the routine shoot multiplication medium at concentrations up to 10 mg l -1 . After 12 weeks of culture involving three subcultures, the maximum number of shoot buds were produced in cultures receiving either 5 mg l -1 BA (80 shoot buds) or 4 mg l -1 kinetin (62 shoot buds). Certain morphological abnormalities observed during proliferation of shoot buds in vitro were not observed during acclimatization ex vitro. To check the genetic stability, RAPD and ISSR profiles of micropropagated plantlets obtained from different cytokinin-treatments were compared with control microplants maintained on MS medium as well as the field-grown mother plant. A total of 50 RAPD and 12 ISSR primers resulted in 625 distinct and reproducible bands. Thus a total of 17,400 bands were generated showing homogeneous RAPD and ISSR patterns. Band intensity histogram of each gel confirmed their monomorphic nature with no genetic variation in all the plantlets analysed. Based on these results a protocol for high rate shoot multiplication was worked out leading to uniform shoot production.

The Effects of Auxins and Cytokinin on Growth and Development of (<i>Musa</i> sp.) Var. “Yangambi” Explants in Tissue Culture

“…Other studies by [12,20,21] observed that 5mg/l BAP was the most efficient concentration for in vitro bud proliferation of many banana cultivars. In a review on banana cell and tissue culture, Strosse et al [22] indicated that for multiplication of propagules, a medium containing a range of concentration 0.1 -20 mg/ of BA is added to the media.…”

Analysis of Genetic Stability through Intersimple Sequence Repeats Molecular Markers in Micropropagated Plantlets of Anoectochilus formosanus HAYATA, a Medicinal Plant

“…46,47) Among the various DNA markers often used for the detection of somaclonal variation, ISSR markers have been successfully utilized for the analysis of genetic fidelity within lines of some plants, such as cauliflower, almond, banana, Swertia chirayita and Platanus acerifolia. 19,20,34,48,49) So in this study, due to the somaclonal variation and on the basis of some virtue of ISSR analysis compared to other molecular techniques comprising RFLP, RAPD, AFLP, we utilized the ISSR primers to investigate the genetic stability of A. formosanus propagated in vitro for a period of more than 5 years.…”

Assessment of genetic fidelity of micropropagated date palm (Phoenix dactylifera L.) plants by RAPD and ISSR markers assay

“…ISSR technique is also a very simple, fast, cost-effective, highly discriminative and reliable (Reddy et al, 2002). RAPD and ISSR markers have been successfully applied to detect the genetic similarities or dissimilarities in micropropagated material in various plants (Venkatachalam et al, 2007;Joshi and Dhawan, 2007;Santos et al, 2008;Ahmed et al, 2009;Huang et al, 2009). The objective of the present study was to monitor the genetic stability of long term (168 weeks) micropropagated plants of date palm using the RAPD and ISSR markers.…”