The Optimisation Of Growth Regulators, Precursors And Elicitors Supplementation For Maximum Limonene And Linalool Accumulation In Cell Cultures Of Citrus Grandis (L.) Osbeck

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