The Development of Plant Regeneration System from Callus of Pineapple (Ananas comosus L.)
Malaysia’s production of canned pineapples has been decreasing since 1992. Two important factors that have been a hindrance to the progress of this industry are competition from other producers and the increasing demand for fresh pineapples. Current varieties need to undergo qualitative improveme...
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| Format: | Thesis |
| Language: | English English |
| Published: |
2005
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| Online Access: | http://psasir.upm.edu.my/id/eprint/38/1/1000548950_TFP2005_27_PT.1.pdf http://psasir.upm.edu.my/id/eprint/38/ |
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| Summary: | Malaysia’s production of canned pineapples has been decreasing since 1992. Two important
factors that have been a hindrance to the progress of this industry are competition from other
producers and the increasing demand for fresh pineapples. Current varieties need to undergo
qualitative improvements. Genetic modification, breeding and selection are some crop
improvement techniques that are not successful at the moment in developing varieties that
can replace current world varieties. Somaclonal variation is another technique for obtaining
desirable variants, which have been achieved in crops such as sugarcane, wheat and sorghum.
Highly stable variants that can be transmitted to progenies, and a more controlled change of
their characteristics than those of induced mutations were achieved. However, this technique
requires a plant regeneration system from callus cells. These cells have a tendency to mutate,
and more cells are mutated under prolonged culture and rapid proliferation, and so generate
more variants for selection. Therefore, the objectives of this project are to induce calli,
proliferate old calli and regenerate shoot from calli.
For calli induction, meristemic globular bodies (MGB) of Moris and Josapine, were cultured
in various levels of auxin naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic (2,4-
D). The highest percentage of MGB forming calli was observed in treatment NAA 14 and 10
mg/L for Moris and Josapine respectively, at the end of 16 weeks.
For calli proliferation, 18 month-old calli were cultured in various levels of NAA, 2,4-D, bnaphtoxyacetic
acid (BNOA) and p-chlorophenoxyacetic acid (4-CPA) auxins for 12 weeks,
and then in various levels of casein hydrolysate (CH) and coconut water (CW) in the
presence of NAA 6 mg/L for 12 weeks. Among the various levels of the four auxins, NAA 6
mg/L proliferated healthy and high mean calli fresh weight. However, NAA 6mg/l
supplemented with CW and CH also gave healthy and generally higher mean calli fresh
weight than NAA 6mg/L alone. NAA 6 mg/L alone was considered the most economical
treatment for calli proliferation, while NAA 6mg/L supplemented with 15%v/v CW and
300mg/L CH gave significantly highest mean calli fresh weight and was considered the best
treatment for rapid calli proliferation.
For shoot regeneration, 18 month-old calli were cultured in various levels of NAA, 2,4-D,
BNOA and 4-CPA auxins for 12 weeks, and then in combinations of various levels of auxins
(BNOA and 2,4-D) and cytokinins (Benzylaminopurine [BAP], Kinetin and Adenine) for 12
weeks. Among the various levels of the four auxins, 2,4-D at 1mg/L regenerated high number
of shoots, and was considered the best treatment for high shoot regeneration from calli that
were considered as high competency calli. However, regeneration response from these calli
were gradually decreasing, such that treatment BNOA 6mg/L combined with BAP 1mg/L
(with subculture) (that statistically gave highest number of regenerated shoots) and an
extended culture period of 12 weeks (without subculture), generated mean number of shoots
was considered as not satisfactory. Subsequently, the 18 month old-calli (now 27 months)
were cultured in various combination levels of CH and CW for 12 weeks, but these failed to
show any regenerated shoots from calli that were now considered low competency calli.
However, calli obtained from treatment NAA 6 mg/L + 300mg/l CH + 15%v/v CW (rapid
calli proliferation treatment, and now 27 months old), preserved calli competency and
regenerated highest mean number of shoots in treatment 10%v/v CW and 200mg/l CH, and
was considered the best treatment for regeneration of shoots from low competency calli. |
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