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Quantitative Effect of Hygromycin on Regeneration of Banana Shoot Clumps

Study on regeneration potential of banana shoot clumps under varied hygromycin concentrations, showcasing slow shoot bud elongation at higher doses. Data confirms stable integration of uidA gene in transgenic bananas.

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Quantitative Effect of Hygromycin on Regeneration of Banana Shoot Clumps

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  1. B C A D E F G H I Supplementary Fig. 1.Quantitative effect of hygromycin concentration (0-40 mg/l; Athrough I, top panel, bar =1.7 cm) on regeneration potential of multiple shoot clump (MSC) explants of banana [Musaacuminata cv. Matti (AA)]. The MSC explants were cultured on shoot initiation medium (MS medium supplemented with 3% sucrose, 20 mg/l ascorbic acid, 22.5 mg/l BAP and 0.4 mg/l IAA and hygromycin, as indicated). Note: Although proliferation of MSCs initiated and the explants remained green on 10 mg/lhygromycin (C), the adventitious shoot buds elongated at a very slow pace in comparison with the control explants. The data were scored 1 month after culture, and represent average (± S.D.) of three independent experiments each with 25-30 explants. Regeneration frequency (%) 0 5 10 15 20 25 30 35 40 Hygromycin concentration (mg l-1)

  2. Supplementary Fig. 2 (A) GenomicPCR, and (B) Southern blot analyses confirming stable integration and expression of uidA in young fully expanded leaves of transgenic banana [Musaacuminata cv. Matti (AA)] plants transformed with Agrobacterium strain EHA105 harboring the binary plasmid pCAMBIA-1301. Ten µg genomic DNA was digested overnight with NcoI and Eco721, size-fractionated on a 0.8% agarose gel and hybridized with radiolabeled 947 bp PCR-amplified uidAprobe. M, 1 kb DNA ladder; P, pCAMBIA-1301 plasmid, see Materials and methods; WT, wild-type untransformed control; G1 through G10, independently transformed primary transgenic lines of banana.

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