Micropropagation and pigment extraction of Echinocereus cinerascens / Hashimah Elias
Protocols were successfully established for in vitro regeneration and coloured callus production of Echinocereus cinerascens. Investigation mainly focused on the effects of plant growth regulators in rapid production of this endangered species and the optimum production of coloured callus. Additi...
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| Format: | Thesis |
| Published: |
2017
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| Online Access: | http://studentsrepo.um.edu.my/7785/1/All.pdf http://studentsrepo.um.edu.my/7785/9/hashimah.pdf http://studentsrepo.um.edu.my/7785/ |
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| Summary: | Protocols were successfully established for in vitro regeneration and coloured callus
production of Echinocereus cinerascens. Investigation mainly focused on the effects of
plant growth regulators in rapid production of this endangered species and the optimum
production of coloured callus. Additional assessments studied concerning the
production of synthetic seeds, extraction of natural pigments and detection of
somaclonal variation of the regenerants. Rapid production through direct in vitro
regeneration gave the highest mean number of shoots, 4.37 ± 0.27, observed in MS
medium supplemented with 2.0 mg/l Kinetin + 1.0 mg/l IBA which promoted the
highest production of shoots after 4 months, 131 shoots in total. Nevertheless, through
indirect in vitro regeneration, somatic embryos of Echinocereus cinerascens were
successfully developed in two treatments of liquid medium including MS medium
supplemented with 0.5 mg/l 2,4-D + 0.1 mg/l BAP + 0.5 mg/l thiamine HCl and MS
medium supplemented with 0.5 mg/l 2,4-D + 0.5 mg/l BAP + 0.5 mg/l thiamine HCl, as
both media promoted 100% total mean production of somatic embryos (globular, heart,
torpedo-shaped and cotyledonary stage) after 4 months. Practically, the production of
ideal synthetic seeds was successfully established whereby, micro shoots as the most
responsive propagule were encapsulated in 3% of sodium alginate hardened in 100 mM
of calcium chloride dehydrate solution for 30 minutes gave 100% of germination rate
after 4 months. Complete plantlets were successfully acclimatized with the highest
survival rate of 90% observed in sand, the most suitable planting substrate which
possessed 3 major elements such as SiO2, Al2O3 and CaO that play important roles to
support the growth of Echinocereus cinerascens. Basically, the present study indicated
that 100% production of green, yellow and pink callus was obtained after 2 months in
several treatments of MS medium supplemented with 2,4-D + BAP + thiamine HCl
applied in combination. Interestingly, the occurrence of dramatic changes in the production of coloured callus was clearly observed in a conversion of green to pink
callus within 2 months. Moreover, pigment extraction analysis through UV-VIS
spectroscopy discovered that both regenerants and callus possessed chlorophyll a and b
as the major pigment while carotenoids as the minor pigment. Meanwhile, HPLC
analysis revealed individual carotenoids present in in vitro plantlets namely, neoxanthin,
β-carotene, lutein and violaxanthin, whereas in callus were β-carotene and lutein only.
Analysis of cytological studies clarified that there were no significant differences in cell
organization and behaviour of in vitro plantlets and ex vitro plants. The evidence
verified that Echinocereus cinerascens regenerated normally in vitro and grown
vigorously after being transferred to the natural environment |
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