Zakaria, Zarina
(2005)
The Optimisation Of Growth Regulators,
Precursors And Elicitors Supplementation
For Maximum Limonene And Linalool Accumulation
In Cell Cultures Of Citrus Grandis (L.) Osbeck.
PhD thesis, Universiti Sains Malaysia.
Abstract
The study on Citrus grandis (L.) Osbeck encompasses tissue culture production
and the extraction of limonene and linalool. The tissue culture methodology comprises
of sterilisation, induction and maintenance of callus on media favouring cell growth, to
favour production of limonene. Explants sterilisation attained high yields of sterile
explants (~ 90%) in treatment with 20% Chlorox® (I% sodium hypochloride). Callus
gro\\rth is favourable from explants originated from 5-week-old fruits and cultured on
modified MS media with 510 mg/1 phosphate, 3 mg/1 each 2,4-D and kinetin and 0.2
mg/1 ABA. Limonene was first detected from 7- month-old callus up to 10-month-old
cultures which yields the highest limonene concentration before slightly down at the
11th month. The amount of linalool extracted was highest from 7-month-old callus and
decreased as the age of callus cultures increased. Media 6 with supplementation of 51 0
mg/1 phosphate, 0.2 mg/1 ABA and 3 mg/1 2,4-D and kinetin respectively, gave
maximum concentration of limonene, which is 0.0031 mg/g fresh weight of callus. The
effects of supplementation various concentrations of exogenous mevalonic acid (MY A)
and linalool to C. grandis callus cultured at varying periods were studied. Callus grov,rth
was found to be proportional to the concentrations of MY A and linalc~_ol especially
during the first 6 weeks.
Kajian ke atas Citrus grandis (L.) Osbeck melibatkan penghasilan kultur tisu
dan pengekstrakan limonena dan linalool. Kaedah kultur tisu terdiri daripada
pensterilan, induksi dan pengekalan kalus dalam media yang menyokong pertumbuhan
sel, yang juga menyokong penghasilan limonena. Pertumbuhan kalus adalah paling
sesuai menggunakan eksplan yang terdiri daripada buah yang berumur lima minggu
yang dikultur dalam media MS yang diubahsuai dengan 510 mg/1 fosfat, 2,4-D dan
kinetin yang masing-masing 3 mg/1 dan ABA sebanyak 0.2 mg/1. Limonena dikesan
bermula pada kalus berumur 7 bulan sehingga bulan ke-1 0 yang memberikan
penghasilan limonena yang tertinggi sebelum menurun sedikit pada bulan ke-11.
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