Somatic embryogenesis from chickpea (Cicer arietinum L.) inmature cotyledons: The effect of zeatin, gibberellic acid and indole-3-butyric acid

Abstract

Studies were conduced to test the effects of various cytokinins on somatic embryogenesis from chickpea (Cicer arietinum L.) immature cotyledons. Zeatin (13.7 lamol) added, to B5 basal medium, supplemented with 1.5 % sucrose and 0.2 tJmol indole-3-acetic acid, was the most effective cytokinin. Lobular structures obtained from cotyledons cultures were transferred to B5 basal medium supplemented with gibberellic acid and indole-3-butyric acid at different concentrations. The most effective treatment was B5 medium containing 14.4 pmol gibberellic acid plus 1.0 pmol indole-3-butyric acid in which 42.8 % of lobular strnctures cultured formed normal somatic embryos. High conversion of embryos into plantlets (61.0-65.2 % embryos regenerated plants) was observed when germinated embryos were placed on plant development medium. List of abbreviations."BAP, N 6 -b e n z y l a m i n opurine; B5, Gamborg medium; CLS, cotyledon-like structures; 2,4-D, 2,4-dichlorophenoxyacetic acid; GA3, gibberellic acid; IAA, indole-3-acetic acid; IBA, indole-3-butyric acid; KIN, kinetin; MS, Murashige and Skoog medium; 2,4,5-T, 2,4,5trichlorophenoxyacetic acid; Z, zeatin.

Gibberellin Is Essentially Required for Carrot (Daucus carotaL.) Somatic Embryogenesis: Dynamic Regulation of Gibberellin 3-Oxidase Gene Expressions

“…Crude extract was incubated with cofactors, such as 2-oxoglutaric acid (5 mM for GA 20-oxidase and GA 3-oxidase, 4 mM for GA 2-oxidase), ascorbic acid (5 mM for GA 20-oxidase and GA 3-oxidase, 4 mM for GA 2-oxidase) and FeSO4 (0.5 mM), and substrate GAs (200 ng each of GA12W 53, GA9W 20 or GA1W 4 for GA 20-oxidase, GA 3-oxidase or GA 2-oxidase, respectively). The reaction mixture (100 ml) was incubated for 1 h at 309 C for GA 20-oxidase or GA 2-oxidase or at 379 C for GA 3-oxidase, respectively.…”

“…The abnormalities caused by uniconazole treatment were almost prevented by concomitant treatment with bioactive GA, GA1 (Fig. 2, A-Uni＋GA1, and B-Uni＋GA1) or GA 4 (Fig. 2, A-Uni＋GA 4 , and B-Uni＋GA 4 ), although the ratio of heart-shaped embryos was not completely normal with exogenous GA4 (Table 1, Uni ＋GA 4 ).…”

“…Especially, the involvement of gibberellin (GA) is not conclusive, because its eŠects vary in diŠerent tissues and species. For example, exogenously applied GA3 stimulated embryogenesis in immature cotyledon cultures from chickpea, 4) geranium hypocotyls cultures, 5) and Medicago sativa tissue cultures. 6) In the geranium and Medicago cultures, GA biosytnthesis inhibitors such as paclobutrazol inhibited embryo production.…”

Genes and Trans-Factors Underlying Embryogenic Transition in Plant Soma-Cells

“…For example, exogenous application of a subtype of gibberellin GA3 in culture medium inhibited SE in carrot (Fujimura and Komamine 1975 ;Tokuji and Kuriyama 2003 ), citrus (Kochba et al 1978 ) and Geranium (Hutchinson et al 1997 ). Whereas exogenous application of GA3 stimulated embryogenesis in immature cotyledon cultures of Cicer (Hita et al 1997 ) and petiole derived cultures of Medicago (Ruduś et al 2002 ).…”

Callus induction via different growth regulators from cotyledon explants of indigenous chick pea (Cicer arietinum L.) cultivars KK-1 and Hassan-2K

“…Organogenic regeneration via callus induction has also been attempted using cotyledons, mature and immature embryo axes, leaflets, distal and proximal cotyledons and hypocotyl explants (Anju and Chawla, 2005). The effect of zeatin, GA3 on regeneration from immature cotyledons of chickpea has been studied by Hita and Gaerra, (1997). This study was undertaken to test the effect of different concentrations and combinations of growth regulators on chick pea callus induction and standardize the protocol for callus induction for two indigenous cultivars of chick pea widely grown in Pakistan, which would be efficient and suitable for the investigation of induced somaclonal variation and successful application of gene transfer technique.…”