Plants were regenerated from hypocotyl and anther explains of berseem clover (Trifolium alexandrinum L.) on Murashige and Skoog (MS) medium containing various combinations of plant growth regulators. The most efficient production of plants from hypocotyl explants involved:
callus induction on MS medium with 1.0 mg/liter of naphthaleneacetic acid (NAA) and 1.5 mg/liter 6‐furfurylaminopurine (KIN);
callus increase on MS medium with 2.0 mg/liter of NAA and 0.1 mg/liter of N6‐(Δ2‐isopentenyl) adenine (2iP);
induction of shoots on MS medium with 0.5 mg/liter each of NAA and KIN followed by induction of roots on MS medium with 1.0 mg/liter of indoleacetic acid (IAA) and 0.1 mg/liter of 6‐benzylaminopurine (BAP). Suspension cultures in liquid MS medium containing 2.0 mg/liter of NAA and 0.2 mg/liter of 2iP provided filterable cell preparations with 45% viable cells, 4% of which gave rise to colonies within 3 weeks after transfer to agar plates. Shoot development was observed when callus from the colonies was cultured on MS medium with 0.5 mg/liter of NAA and KIN. Twenty percent of uncontaminated anthers from a single plant cultured on MS medium containing 1.0, 0.1, and 0.01 mg/liter of NAA, 2,4‐D, and 2iP, respectively, produced callus which could be maintained and from which plants could be regenerated by culturing on MS medium with 0.5 mg/liter each of NAA and KIN. Preliminary results indicate that cells of root tips from hypocotyl‐ and antherderived callus have the expected diploid and haploid number of chromosomes (2n = 16 and n = 8, respectively).
Incorporating activated charcoal (AC) in culture media has been shown to affect growth and development of various organisms. Since AC stimulates the development of tobacco haploid plantlets from cultured anthers, research was conducted to determine the effect of activated charcoal on pith-derived callus growth and shoot development in Nicotiana tabacum cv. Wisconsin 38. Our results indicate that the hormones required for callues growth and shoot development in Wisconsin-38 tobacco are adsorbed by AC, thereby inhibiting callus growth and prohibiting shoot development. This effect was observed even when AC was removed from the medium by filtration prior to culturing the callus.
Callus was induced in different somatic organs of Oryza sativa L. Specific minimum 2,4‐dichlorophenoxyacetic acid (2,4‐D) concentrations in the medium were necessary for the induction of callus from different organs while high levels of 2,4‐D (6–10 mg/l) induced callus formation in each organ tested. The optimum 2,4‐D concentration for callus induction and growth for root‐derived calli was 2 mg/l and for leaf‐derived 6 mg/l. Root and shoot organogenesis were induced in both root‐ and leaf‐derived calli by sub‐culturing to a medium lacking 2,4‐D. Root organogenesis occurred at a higher frequency than shoot organogenesis. Shoot organogenesis rarely occurred in calli without differentiated roots. Increased age of callus cultures almost completely inhibited shoot development. The addition of the cytokinin 6‐γ,γ‐dimethylallyl‐amino purine partially restored the potential for shoot organogenesis. Whole plants were easily recovered from the calli and grown to maturity with some plants exhibiting phenotypic abnormalities.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.