Identification of Embryogenic Callus and in vitro Somatic Embryo Formation in Gerbera jamesonii Bolus ex. Hook f.

Hasbullah, Nor Azlina; Taha, Rosna; Awal, Asmah

Identification of Embryogenic Callus and in vitro Somatic Embryo Formation in Gerbera jamesonii Bolus ex. Hook f.

Document Type : Original Article

Authors

¹ Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia

² Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.

³ MARA University of Technology, Negeri Sembilan Branch, 72000 Kuala Pilah, Negeri Sembilan, Malaysia

Abstract

In vitro cultures of Gerbera jamesonii Bolus ex. Hook f. were initiated from leaf explants. White
friable callus was formed after 4 weeks at an average of 70% onMurashige and Skoog (MS) medium
supplemented with 2, 4 dichlorophenoxyacetic acid (2,4-D). The identification of somatic embryos at
early stages was done by treatment with 2% Acetocarmine and 0.5% Evan’s Blue (double staining
method). The embryogenic head cells stained bright red (acetocarmine) and suspensor cells stained
blue in an embryogenic mass. In the mass of non-embryogenic callus, cells did not show any
organization of heads and suspensors and will stain blue with Evan’s Blue. White-cream friable
embryogenic callus of Gerbera jamesonii was formed after 4 weeks when leaf explants were cultured
on MS medium supplemented with 0.1 mg/l-2.0 mg/l 2, 4-D, 3.0% sucrose and solidified with 0.8%
agar. The embryogenic callus was transferred into MS suspension culture medium supplemented with
1.0 mg/l 2, 4-D and 0.1 mg/l Naphthalene acetic acid (NAA) and was subcultured at 10 days interval
for 1 month. Subsequent withdrawal of 2, 4-D from induction medium resulted in the induction and
growth of somatic cells. Somatic embryos formed at globular phase were then sieved and transferred
into maturation medium. Heart, torpedo and cotyledon phases of somatic embryo were identified.

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