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Unravelling the regulation of somatic embryogenesis by extracellular calcium and auxin efflux blockers in hypocotyl explants of albizzia lebbeck l.: similarity in action

Tanushri Kaul*, Babu Ram, Saurabh Pandey, Chinreddy Subramanyam Reddy, Malireddy K. Reddy

Abstract


Somatic embryogenesis involves developmental reprogramming and de-differentiation of somatic cells towards embryo development that is crucial for cellular totipotency in higher plants. It is a prototype for comprehension of the physiological, biochemical and molecular biological events transpiring during plant embryo development. In this study, we analyzed the role of exogenous calcium and auxin during somatic embryogenesis in hypocotyl explants of Albizzia lebbeck L. Light microscopic studies revealed that anterior and posterior ends of hypocotyl explants had entirely different patterns of differentiation. Scanning electron microscopic analysis of cultured hypocotyl explants exhibited globular-, heart- and torpedo-shaped somatic embryos developing from the anterior cut surface and gradually proceeding towards the other end. Surface texture of these somatic embryos showed uni-or bi-celled trichomes. Presence of exogenous calcium (1-4mM) in hormone-free B5 medium is prerequisite for induction of somatic embryos in hypocotyl explants derived from 7-d-old, light grown seedlings of Albizzia lebbeck L. A two-fold increase in calcium concentration from 2mM (B5 medium) to 4mM led to doubling of somatic embryogenic response whereas, embryogenic callus was induced in explants raised on medium containing 10 or 20mM calcium. Depletion or high dosage in the medium (20 mM) induce non-embryogenic callus in explants. A similar response induced on hypocotyl explants upon treatments with NPA and PCIB, (auxin transport blockers). Results indicate that polar auxin transport or basipetal movement of IAA triggers acropetal transport of calcium. This combination of inverse fluxes acts as the primary signal for embryogenic response at the cut ends proximal to shoot apex and callusing at the cut ends distal to shoot apex of explants. Moreover, a correlation exists between NPA and PCIB-induced disruption of basipetal IAA transport and the inhibitory effect of NPA and PCIB on calcium translocation in the induction of similar morphogenetic responses.

Keywords


calcium chloride; indole-3-acetic acid; morphogenesis; naphthylphthalamic acid; parachlorophenoxyisobutyric acid; somatic embryos; tree legume; trichomes.

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DOI: http://dx.doi.org/10.21746/ijbio.2014.11.0018

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