Home⇛Annals of Tropical Research⇛vol. 30 no. 2 (2008)

Development of bacterial blight resistant Mestizohybrid maintainer and restorer lines throughmarker-aided backcrossing

Lucia M. Borines | Emilie O. Espejo | Robelyn T. Piamonte | Casiana N. Vera Cruz | Edilberto Redoña

 

Abstract:

A study was conducted to establish a reliable procedure for somatic embryogenesis and plant regeneration from callus cultures of purple food yam (Dioscorea alata L.). The procedure involved three steps: (1) culture of nodal stem segments from greenhouse-grown plants to generate in vitro plantlets; (2) induction of callus from the leaf, petiole, and nodal stem tissues; and (3) initiation of somatic embryos from callus. Results showed that agar-solidified Murashige and Skoog (MS) medium containing 30 g/L sugar, 0.1 g/L alpha-cysteine, 10 mg/L calcium pantothenic acid, 2.0 mg/L asparagine, 2.0 mg/L arginine, 80.0 mg/L adenine sulfate (AdSO4), and 0.1 mg/L naphthalene acetic acid (NAA) effectively broke dormancy of lateral buds of nodal stem cultures from both 'VU-2' and 'Kinampay' varieties. Multiple adventitious shoots were produced after transferring in vitro nodal pieces to the same medium supplemented with 1.0 mg/L benzylaminopurine (BAP) or to MSA medium. Callus was effectively induced from vegetative tissues on MS medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) or with picloram. Among the three types of explants, the nodal stem was the most suitable, producing purplish, nodular embryogenic callus. A higher percentage of nodal stem-derived calli produced globular embryos on MS medium containing 1.0 mg/L 2,4-D and 0.5 mg/L BAP, or 1.0 mg/L picloram and 0.5 mg/L BAP, compared to the plant growth regulator-free control medium. Embryo maturation was facilitated by a one-month culture in MS medium containing 0.1 mg/L abscisic acid (ABA) and 100 mg/L glutamine. This step improved somatic embryo germination in half-strength, PGR-free MS medium containing 100 mg/L glutamine (regeneration medium). All somatic embryo-derived plantlets were morphologically normal and established well in soil.



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