Potential Embryogenic Callus Induction Protocol Through Cell Suspension Culture For High Frequency Plant Regeneration Of Maspine Pineapple (Ananas comosus L.)
Abstract
To explore the potential for embryogenic callus induction protocol through cell suspension culture forhigh frequency plant regeneration of Maspine pineapple (Ananas comosus L.), eight different culture
media formulation were evaluated for their effects on the induction of somatic embryos from sucker
explants. Explants were cultured on MS medium supplemented with various media concentration
(NAA, Dicamba and BAP, Picloram, Kinetin and NAA, 2,4-D, TDZ, and TDZ and BAP).
Embryogenic callus induction percentage, color and texture of the callus were assessed after five
months of culture. The optimum medium for the proliferation of in vitro shoots from sucker explants
was MS medium supplemented with 3 mg/L BAP. Meanwhile, the optimum medium for the induction
of fastest and high percentage of embryogenic callus growth from in vitro leaf-based was MS medium
supplemented with Picloram. Results of mean comparison showed that 3 mg/L Picloram were more
effective on explants than 10 mg/L. Results of the double staining method proved that somatic
embryogenesis occurred in MS supplemented with 3 mg/L Picloram. Under microscopic observations,
the globular-stage of the embryos were revealed in callus cells which is relatively suitable for
suspension cells inoculums, indicating that the tested PGR were significantly effective for somatic
embryogenesis formation in this species. Most embryogenic callus from sucker explants was
yellowish-mucilaginous-wet-friable. The developed protocol potentially leads to the production of
embryogenic callus from sucker explants and plant regeneration through somatic embryogenesis.
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