Efficient regeneration from hypocotyl explants in three cotton cultivars

Abstract

A high frequency in vitro shoot bud differentiation and multiple shoot production protocol from hypocotyl segments of 8 to 10-d-old seedlings of cotton has been developed. Murashige and Skoog (MS) basal medium with Nitsch and Nitsch vitamins was found to be optimal in shoot regeneration. A combination of 2 mg dm -3 thidiazuron and 0.05 mg dm -3 naphthaleneacetic acid was the most effective for shoot regeneration (76 %) and an average of 10.6 shoots per responding explant. Combination of the cytokinins benzylaminopurine and kinetin induced better regeneration response than their individual treatments. Supplementation of the culture medium with ethylene inhibitor silver nitrate and activated charcoal showed beneficial effects. Optimal rooting was obtained on half-strength MS medium supplemented with 1 mg dm -3 indolebutyric acid and activated charcoal. Scanning electron micrographs of in vitro cultured explants revealed that shoot primordia were formed de novo.

In vitro organogenesis of rangpur lime

“…Hypocotyl segments have been described as a very responsive explants for species other than citrus such as passion flower (FERNANDO et al, 2007;PINTO et al, 2010), cotton (DIVYA et al, 2008), and Eucommia ulmoides (CHEN et al, 2008). Hypocotyl segment attached to a cotyledon fragment was described by Ananthakrishnan et al (2003) as an efficient explant for improving squash in vitro organogenesis.…”

In vitro organogenesis of Citrus volkameriana and Citrus aurantium

“…Ananthakrishnan et al (2003) described an efficient explant for squash in vitro organogenesis consisting of a proximal segment of cotyledon attached to hypocotyl fragment. Hypocotyl segments have been described as very responsive explants in other species such as passion fruit (Fernando et al 2007), cotton (Divya et al 2008), and Eucommia ulmoides (Chen et al 2008). Also, Moreira-Dias et al (2001) described that the capacity of epicotyl segments regeneration decreases markedly by the cotyledonary node distancing.…”

Effect of EMS and SA on seed germination percentage in M1 generation of Psophocarpus tetragonolobus (L.) Dc.

“…and Divya et al, (2008) with shoot tip and hypocotyl explants respectively in the Indian elite cultivars with higher concentrations of TDZ. Similarly, Chakravarthy (2013) observed decrease in mean shoot number and percent response due to callus formation at the base of the cotyledonary node explants at higher concentration of TDZ.…”

“…Silver nitrate (5 g -l ) an anti ethylene factor has been used in the shoot induction medium to reduce hyper hydrosity nature of explant which has been well reported for the same in various species viz. Brassica sp (Palmer, 1992), Peanut (Pestana et al, 1995;Ozudogru et al, 2005) and Pomegranate (Naik and Chand, 2003) including Cotton (Tripathy et al, 2002;Abdellatef and Khalafalla, 2007;Divya et al, 2008). Further, woody plants especially cotton produces enormous amount of phenolics from the cut ends of the tissues, which causes deleterious effects on regenerating tissues, to minimize this effect plant tissue culture grade activated charcoal (1.0 g -l ) was used in this study in all the media excepting in (Fig.…”

“…To circumvent these problems, organogenesis was considered as one of the alternative potential methods in several species including cotton. Several reports in cotton through organogenesis have been published especially with cotyledonary nodal explants (Rauf et (Divya et al, 2008). However, the lower regeneration frequency of these reports ultimately affects the transformation frequency.…”