Callus Induction and Somatic Embryogenesis
in Nypa fruticans Wurmb Zygotic Embryo

 Georgianna Kae R. Oguis1, Cyrose Suzie C. Silvosa1 and Gilda C. Rivero1,2*

1College of Science and Mathematics
University of the Philippines Mindanao, Davao City, Davao del Sur
1,2Institute of Biology, College of Science,
University of the Philippines Diliman, Quezon City

corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.



Nypa fruticans has been reported to be a potential source of ethanol. To date, there is no available protocol on tissue culture suitable for N. fruticans to produce superior and uniform planting materials for future wide-scale production. This paper investigated the influence on callus induction of various concentrations of 2,4-D and activated charcoal as well as the effects of three orientations of the explant on the media. Sixty-four percent of the explants cultured in full-strength MS media, supplemented with various combinations of different concentrations of 2,4-D and activated charcoal, produced protocorm-like structures in cultures with lower concentrations of 2,4-D combined with high concentrations of activated charcoal. Fifty-two percent of protocorm-like structures in T18 cultures significantly (p<0.05) developed white, compact, nodular calli and soft, white, friable calli eight weeks after inoculation. Twenty percent of the explants in the T18 cultures developed somatic embryos sixteen weeks after inoculation. Explants in cultures with higher 2, 4-D concentrations combined with activated charcoal significantly (p<0.05) formed brown, hard, compact calli. Treatments devoid of activated charcoal did not induce callus and did not produce somatic embryos. The absence of activated charcoal in the MS media also led to browning of explants within the eighth week of incubation. The orientation of explants on the culture media resulted in callus initiation only in the T18 cultures, although protocorm-like structures were observed in all cultures regardless of the explant orientation.



Nypa fruticans Wurmb, the nipa palm, remains uncultivated and grows mainly as natural stands near estuaries and rivers (Rasco 2010; Tsuji et al. 2011) throughout the Philippines. Its large pinnate leaves are traditionally harvested for use as roof thatches and stabilizers of coastal areas, while its sap is regularly harvested for small-scale vinegar and vodka production (Rasco 2010; Rasco et al. 2012). N. fruticans sap contains as much as 60% sugar and its annual yield was reported to be much more than those from sweet potatoes, coconut, tapioca, and sugarcane (Hamilton and Murphy 1988). The depleting petroleum resources and increasing demand for fuel brought on great interest . . . . . read more


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