Abstract
Shoots of Cavalcade (Centrosema pascuorum cv. Cavalcade) and Stylo 184 (Stylosanthes guianensis CIAT 184) from in vitro germinated seeds were cultured on Murashige and Skoog (MS) medium supplemented with 0 to 7 mg L-1 N 6-benzyladenine (BA) in combination with 0 to 0.5 mg L-1 napthalene acetic acid (NAA) for shoot induction and MS supplemented with 0 to 0.5 mg L-1 indolebutyric acid (IBA) for root induction. For Cavalcade, the medium containing 1 mg L-1 BA produced the best shoot multiplication with an excess of six shoots produced from a single shoot (over four weeks) with a mean height 2.0 ± 0.01 cm. Adventitious shoot regeneration was obtained directly from stem axes. For Stylo 184, the maximum shoot regeneration (29.5 ± 1.0 cm shoots/explant) and height (1.5 ± 0.1 cm) was achieved using 7 mg L-1 BA and 0.01 mg L-1 NAA. Direct and indirect shoot regeneration was obtained on the medium containing 1 mg L-1 BA and 0.01 mg L-1 NAA. The regeneration of shoots from callus of Stylo 184 varied between different genotypes and was high (2.6 to 5.8 shoots/explant) even after maintenance in culture of over three years. Both Cavalcade and Stylo 184 shoots were rooted on media supplemented with IBA (0 to 0.5 mg L-1) and readily transferred to soil (Stylo 184). Key words: Callus, forage legume, micropropagation, organogenesis, root induction.
Highlights
The use of legumes in areas of livestock production provides considerable benefits as these plants contain high protein and increase the nitrogen content of soil (Batterham and Egan, 1986; Graham and Vance, 2003; Kabirizia et al, 2007; Mapiye et al, 2007; Rao and Northup, 2009; Lupwayi et al, 2011)
Cavalcade cultures were maintained on these media for a further four weeks, and three types of explants were transferred to Murashige and Skoog (MS) medium with 1, 3, 5 or 7 mg L-1 BA; the experiment was conducted with 30 replicates and regeneration percentage was recorded after five weeks
There was no evidence of adventitious shoot production from callus; multiple shoots grew from both axillary buds and stem axes (Figure 2d)
Summary
The use of legumes in areas of livestock production provides considerable benefits as these plants contain high protein and increase the nitrogen content of soil (Batterham and Egan, 1986; Graham and Vance, 2003; Kabirizia et al, 2007; Mapiye et al, 2007; Rao and Northup, 2009; Lupwayi et al, 2011). Shoot tips of Cavalcade and Stylo 184 were cultured on MS medium supplemented with combinations of NAA (0, 0.01, 0.1, and 0.5 mg L-1) and BA (0, 1, 3, 5 and 7 mg L-1). Cavalcade cultures were maintained on these media for a further four weeks, and three types of explants (yellow friable callus, green compact callus and green compact callus adjacent to stem axis) were transferred to MS medium with 1, 3, 5 or 7 mg L-1 BA; the experiment was conducted with 30 replicates and regeneration percentage was recorded after five weeks.
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