Kulaveerasingam, Harikrishna

Mahmood, Marziah

Md. Sukari, Mohd Aspollah

Mohd Ghazali, Hasanah

Universiti Putra Malaysia Institutional Repository

UPM Research Report 1998: Volume 2OPTIMISATION OF THE PRODUCTIONCONDITIONS OF FLAVOUR COMPOUNDS FROM'KARl' MURRA·YAKOENIGII{L.) SPRENG.Hasanah Mohd. Ghazali, Marziah Mahmood,Aspollah bin Sukari and K. HarikrishnaFaculty of Food Science and BiotechnologyUniversiti Putra Malaysia, 43400 UPM, Serdang, Selangor,MalaysiaKeywords: Curry flavour, plant cell culture, Murrayakoenigii, callus.IntroductionCurry tree or Murraya koenigii (L.) Spreng. belongs to thefamily of Rutaceae and is found in South East Asia, TropicalAustralia and New Caledonia (Salikutty and Peter, 1985).Curry leaves are used extensively in Indian and also Malaycooking and form one of the major leafy spices. Researchshows that volatile component such as cyrophylene, pineneand pinene are found in the leaves. Other compounds thathave been identified include maharnnimbine from the stems,koenoline methoxy-3-hydroxy-methyl-carbazole from theroot and bark and mukonicine, antibacterial agents and anti-oxidants from the leaves. Plant metabolites such as the fla-vour compounds are generally produced in highly differenti-ated tissues under specific growth conditions. Since plantcalli are undifferentiated cells, the production of these me-tabolites is naturally more difficult. Several studies haveshown that synthesis of similar or greater quantities of vola-tiles in the calli as in the original plant materials is possible(Bhuyan et al. 1997; Wee et al. 1997).1t has also been shownthat some of the compounds synthesised were different orabsent from the original plants. The objective of this projectwas to optimise the production of curry flavour by callusculture of Murraya koenigii.Materials and MethodsVarious explants from the curry plant including seeds wereused to induce callus formation. The seeds were excised fromcurry fruits that have been sterilised. Also used as explantswere parts of plantlets generated from curry seeds germi-nated under sterile condition on solid Murashige and Skoog'smedium supplemented with 3% sucrose and 1 mg/L BAPunder a 16-hour photoperiod growth regime. Where neces-sary, the explants were first sterilised before their culture onsolid media to which phytohormones at known concentra-tions and combinations were added. Stem and petiole ex-plants were cultured in both dark and dark-light. The hor-mones used were Picloram, Dicamba, NAA,!AA, 2,4-D andBAP. Callus obtained from suitable 'kari' explants werepropagated, and was maintained by regular sub-culture onfresh medium. Various growth regimes including liquid sus-196pension .culture are being evaluated for the ability of thecurry callus to produce flavour compounds.Results and DiscussionResults show that stem explant was the best in terms of easeand frequency of callusing when cultured on MS mediumsupplemented with 3% sucrose, 2-4 mg/L Dicamba and 4mg/L NAA. Higher concentrations of hormones slowed cal-lus growth, and may cause the callus to become brown anddie. Cotyledon and embryo explants took similar time as thestem explant to produce callus. However, the frequency ofcallus formed was lower (50-80%) and especially whenlower (2 mg/L) of Dicamba was used compared to 4 mg/L,Leaf explant had a very low (1%) success rate of callus for-mation. It failed to respond to most phytohormones eitheradded singly or in combination to the growth media, andconditions of culture. The flowers failed to produce any cal-lus under the conditions of this study. Generally, callus thatwere formed were friable and white/green and whitelbrownin colour. Combinations of BAP and 2,4-D were less effec-tive, and using BAP alone was inadequate to induce callus.MS medium was better than LS medium. Dark incubationwas more effective in inducing callus from both stem andpetiole compared to dark-light incubation. Addition of char-coal to the growth media improved induction of callus. Cal-lus that developed was sub-cultured and maintained on freshgrowth medium. To date, callus grown on solid medium hasfailed to produce flavour compounds similar to those presentin Murraya koenigii leaves. Induction of hairy root or crowngall formation through infection with Agrobacterium tumefa-ciens was unsuccessful. Currently, studies are being carriedout to induce flavour compound synthesis using callus sus-pension culture.ConclusionWhile callu~ of Murraya koenigii could be formed by usingthe appropnate explants, induction of flavour compoundformation by the callus is difficult. More experiments willhave to be conducted to find the right combination and con-centrations of hormones, culture conditions to achieve suc-cess.ReferencesBhuyan, A.K., Pattnaik, S and Chand, P.K 1997. Micropropagationof c~ ~eaf tr~e [~urraya koenigii (L.) Spreng.] by axillary~~~.iferal1onusing mtact seedlings. Plant Cell Reports 16: 779-Salikutty, .J and ~~ter, K.V. 1985. Curry leaf (Murraya koenigii),~~enrual, nutnl:!ous, leafy vegetable. Economic Botany. 39: 68-Wee, C.Y., ~arziah, M., Ghazali, H.M and Aspollah, S. 1998. Cal-lus from different explants ofMurraya koenigii L. In Proceedings~~~e 10th National Biotechnology Seminar, Shah Alam. p. 236-Supported by IRPA Grant 09-02-04-0035

Optimisation of the production conditions of flavour compounds from 'kari' Murraya koenigii (L.) Spreng

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Optimisation of the production conditions of flavour compounds from 'kari' Murraya koenigii (L.) Spreng

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