Somatic embryogenesis from cell suspension cultures of banana (Musa sp. 'Lakatan') and somatic embryo germination and regeneration in banana and abaca (Musa textilis Nee 'Inosa') Phase V and Phase V-extension

Date

2015

Abstract

Meristematic buds (scalps) of banana 'Lakatan' were inoculated initially to full strength liquid MS + 1.11 mg/L 2,4-D + 0.22 mg/L zeatin. However, due to the delayed globule formation in scalps of 'Lakatan', another set of scalp cultures were transferred to full strength liquid MS + 1 mg/L 2,4-D + 1 mg/L biotin + 100 mg/L glutamine + 100 mg/L malt extract + 4.5% sucrose (M2) (Coteet al., 1996) to induce globule formation until sufficient number of cell suspension cultures were obtained. Cell suspension cultures were transferred to liquid embryogenesis media (EM) (1/2 MS + 2.20 mg/L zeatin and 1/2 MS + 2.20 mg/L zeatin + 2 mg/L BA). Cell clusters observed under the microscope have dense cytoplasm, indicative of embryogenic nature of the somatic embryo-like structures. Another set of globule cultures of 'Lakatan' were individually picked and plated on germination media composed of MS basal salts + N and N vitamins + 0.18 mg/L IAA + 100 mg/L glutamine + 100 mg/L malt extract and with one level of BA (5 mg/L). Globules formed root-like structure in germination medium without BA while white protrusions and h embryogenic complex were observed in germination media with 5 mg/L BA. For the induction of somatic embryogenesis in abaca 'Inosa', two media formulations were adopted for the embryogenic cell suspension cultures (i.e. GFMM-B and M2). GFMM-B derived cell cultures were observed under microscope to be elongated in shape with dense cytoplasm and were induced to undergo somatic embryogenesis in ½ MS + 2.20 mg/L zeatin + 2 mg/L BA. M2-derived cell cultures produced spherical shaped cells with dense cytoplasm and were continuously induced in the same medium formulation devoid of 2,4-D but with addition of 1000 mg/L myo-inositol. The putative somatic embryos were plated onto two different germination media: (A) MS basal salts N & N vitamins + 0.18 mg/L IAA + 100 mg/L glutamine + 100 mg/L malt extract + 2.5 mg/L BA produced moderately profuse greenish round proembryo masses. After 3 months in culture these proembryos were transferred to MS + 3 mg/L BA and turned black after 4 weeks of continuous incubation and (B) MS basal salts + N & N vitamins + 0.18 mg/L IAA + 100 mg/L glutamine + 100 mg/L malt extract + 5 mg/L BA, where faster proliferation and more profuse proembryos were observed than in (A). Development of shoot-like and root like structures started to regenerate 4 weeks after it was transferred in MS + 3 mg/ L BA. On the other hand, embryogenic calli were induced from shoot tips of banana 'Lakatan' in two semi-solid media formulations (Ganapathi et al., 2001) and were found most favorable in MS + 2 mg/L picloran than in SH + 2 mg/L 2, 4-D + 0.2 mg/L zeatin. These embryogenic calli were induced to undergo somatic embryogenes in two (2) types of media formulations consisting (A) semi-solid MS + 1 mg/l 2 ,4-D + 1 mg/L biotin + 100 mg/L glutamine + 100 mg/L malt extract which produced profuse pro-embryo masses and (B) MS + 1 mg/L 2,4-D + 0.2 mg/L zeatin + 1 mg/L biotin + 100 mg/L l-glutamine + 100 mg/L malt extract which produced less profuse proembryo masses showing less synergistic relationship between 2,4-D and zeatin in inducing somatic embryos in 'Lakatan'.

Language

English

Document Type

Article

Pages /Collation

90 leaves

En – AGROVOC descriptors

MUSA (BANANAS); VARIETIES; MUSA TEXTILIS; ABACA; SOMATIC EMBRYOGENESIS; GERMINATION; CELL CULTURE

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