Micropropagation technique of Kacip Fatimah (Labisia Pumila (BL.) F. Vill.)

Hartinie Marbawi, (2007) Micropropagation technique of Kacip Fatimah (Labisia Pumila (BL.) F. Vill.). PhD thesis, Universiti Malaysia Sabah.

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Abstract

Micropropagation technique was established for Labisia pumila (BL.) F. Vill.(Myrsinaceae), an important and potential medicinal herb species in Malaysia. Development of techniques for in vitro seed germination, callus induction and plant regeneration were successfully achieved by manipulation of plant growth factors such as basal medium, carbon source and plant growth regulator. Studies were conducted to investigate the effect of these factors on in vitro growth and development of this species. The results of this study showed that culture medium, carbon sources and plant growth regulators markedly influenced in vitro seeds germination, callus induction and plantlet regeneration of L pumila. Seeds obtained from mature fruits were surface sterilized and germinated on Murashige & Skoog (MS) basal media with or without 6-Benzylaminopurine (BAP) (1 - 3 µM) and supplied with 3% (w/V) of sucrose. High percentage of germination (up to 90%) was achieved on all treatments after 16 weeks of culture. Addition of BAP into germination medium promoted direct shoot regeneration (8.1 ± 1.8 shoots per explant) from newly developed epicotyls. On various cytokinins containing-media, shoot elongation and leaf formation of in vitro seedlings were enhanced with addition of 1 µM Zeatin (ZEA) into the medium. MS basal medium and 3% (w/V) of sucrose was the best treatment among basal media and carbon sources tested for development of shoot from shoot tips culture of L pumila. The maximum percentage (up to 100%) of callus formation was obtained on in vitro leaf segments cultured on MS medium supplemented with 20 1 µM 2,4-Dichlorophenoxyacetic acid (2,4-D) after 4 weeks of culture. The combination of 2,4-D with BAP had a reduced callus induction percentage (Cip) whereas combination of 2,4-D with ZEA did not show any significant effect. In basal medium selection, callus induction was affected by the types of media used but not the nutrient medium concentration. Full strength MS medium was the most suitable for callus induction while 3% (w/v) of sucrose was the best source of carbon compared to other carbon sources tested. Shoot regeneration was achieved either through indirect organogenesis from leaf explants (83.3%) in the presence of 2.5:2.5 1 µM 2,4-D and BAP combination in MS medium or via direct organogenesis from stem node explants (88.9%) cultured on MS medium supplemented with 18 µM ZEA. The regeneration ability and determination of the types of organ regenerated from explants were highly dependent on auxin: cytokinin ratiO, concentrations and combinations of plant growth regulators that were supplied into the medium. Regenerated shoots rooted well on MS basal medium with or without NAA. However, the presence of 2.5 µM NAA in the medium enhanced the number of root (6.7 roots per shoot) on shoots. Micropropagated plantlets grew healthy when acclimatized on peat moss or soil and successfully survived after directly transferred to soil condition. The protocol described for micropropagation of L pumila using various culture techniques offer a reliable method for propagation of this species and solution for over collection of the plant materials for herbal industry in this country.

Item Type:Thesis (PhD)
Uncontrolled Keywords:Micropropagation technique, Medicinal herb in Malaysia, Labisia pumila (BL) F. Vill. (Myrsinaceae), Kacip Fatimah
Subjects:?? QK495 ??
Divisions:SCHOOL > School of Science and Technology
ID Code:3239
Deposited By:IR Admin
Deposited On:08 Dec 2011 11:07
Last Modified:08 Dec 2011 11:07

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