Shoot proliferation, embryogenic callus induction, and plant regeneration in Lepturus repens (G. Forst.) R. Br.

2021 
Lepturus repens (G. Forst.) R. Br. is a perennial grass that grows on coral reef outcrops and sandy beaches and has drought tolerance and salt resistance. In this study, the internodes of L. repens were used as explants and cultured on Murashige and Skoog (MS) medium supplemented with 1.13 mg L−1 6-benzyladenine (BA) to successfully induce axillary shoots. Additional experimental results showed that cytokinins were a key factor for the successful induction and proliferation of shoots. BA induced 7.2 axillary shoots per explant, significantly more than kinetin (KIN), which induced 3.5 axillary shoots per explant. Shoots were proliferated more successfully on medium supplemented with 1.13 mg L−1 BA and a low concentration of 0.09 mg L−1 1-naphthaleneacetic acid (NAA) than BA alone. Shoot bases were used as explants for embryogenic callus induction and subsequent plant regeneration, which was possible on MS medium supplemented with 0.11 mg L−1 2,4-D, 0.09 mg L−1 NAA, and 0.22 mg L−1 TDZ. Separately, callus differentiated into adventitious shoot buds on MS medium containing 1.13 mg L−1 BA or 1.10 mg L−1 KIN singly or combined with 0.09 mg L−1 NAA. On MS medium without plant growth regulators (PGR-free) or only containing 0.09 mg L−1 NAA, callus induced both adventitious shoots and somatic embryos that developed a radical on PGR-free medium. All shoots formed roots on ½MS medium containing a low concentration (0.19 to 0.37 mg L−1) of NAA or IBA within 20 d. The survival percentage of plantlets with 3 to 4 shoot clusters exceeded 99% on a substrate consisting of yellow mud, organic fertilizer, and perlite (10:3:1, v:v:v). After transplanting, plantlets with one or two shoot clusters showed 68 to 86% survival within 1 mo.
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