Micropropagation is a unique method for the production of uniform and disease free plants. This technique would facilitate to obtain large number of uniform plants irrespective of season and will serve as an alternative source of seed materials. In vitro preservation of germplasm is also a safe method to protect the species by reducing the risk of natural vagaries1.
The process of micropropagation using various phytohormones in different concentration will result in the production of large number of plantlets in short period of time
Generic nature of plant plays a major role in defining the quantity of active principles. Cultivation and production of secondary metabolites is constant under standard conditions. Hence production of plant by tissue culture method will be helpful in producing the plants of specific type of significant importance.
Estimation of active constituent content in invitro regenerated plant by tissue culture and comparing it with the content of natural propogated plant has not been yet reported. Thus the present work is an attempt to know whether the secondary cell constituent has increased in In vitro grown plant or not.
Considering the importance of above parameters it is concluded that research work on tissue culture studies of Alpinia galanga plant is of significance.
6.2: Review of literature.
Emerging buds of rhizome of Alpinia galanga produced shoots and roots simultaneously when cultured in MS medium supplemented with Kn 3.0mgl-1. Each explanted shoot bud produced 8 shoots in an average and roots simultaneously with in 8 weeks. Shoot proliferation was continued even after a year by transferring each divided shoot explants to the same media. Regenerated plants were successfully transferred to soil where they grew well within 10-12 weeks with 80% survivality1.
Alpinia galanga rhizome oils were screened for their anti-fungal activity against five dermatophytes (Trichophyton mentagrophytes, T. rubrum, Microsporum canis, M. nanum and Epidermophyton floccosum), three filamentous fungi (Aspergillus niger., A. fumigatus and Mucor sp.) and five strains of yeasts (Saccharomyces cerevisiae, Cryptococcus neoformans, Candida albicans, Candida tropicalis and Torulopsis glabrata). The anti-fungal testing was carried out by using broth microdilutions and disc gel diffusion methods. The rhizome oil of Alpinia galanga was reported to have anti-fungal activity against selected species of A.fumigates with a MIC value of 0.63µgμl-1. The chemical composition of the rhizome oil was investigated by GC and GCMS methods2.
Hypoglycemic activity was investigated in rhizome extracts of Alpinia galanga. In normal rabbits, powdered rhizomes and its methanol, aqueous extracts significantly lowered the blood glucose levels. In alloxan-diabetic rabbits, Alpinia galanga and its methanol, aqueous extracts did not produced significant reduction in blood glucose levels. The hypoglycemic effect of Alpinia galanga in normal rabbits was comparable with gliclazide. It was concluded that Alpinia galanga produced fall in blood glucose level in normal rabbits3.
Methanolic extract, aqueous extract and volatile oils of the fresh rhizomes of Alpinia galanga have been assessed for the free radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and cytotoxic activity against MCF7 (breast adenocarcinoma) and LS174T (adenocarcinoma) cell lines. The oils of Alpinia galanga (AGV) was analysed by GC/MS. The novel compound p-coumaryl-9-methyl ether, was isolated from methanolic extract of Alpinia galanga4.
Chloroform, methanol and aqueous extracts of Alpinia galanga was investigated for anti-bacterial activity against important pathogenic bacteria commonly associated with AIDS infection. Inhibition growth was tested using paper disc agar diffusion method. Minimum inhibitory concentration and Minimum bacterial concentration were determined by agar microdilution method and agar dilution method in petridishs with millipore filters. The gram +ve bacteria were proved to be susceptible to the chloroform extract of Alpinia galanga and also it showed greatest inhibition zones of 29.1 and 23.7mm against S.aureus and MRSA(Methicillin-resistant-Staphylococcus aureus). An active compound 1’-acetoxy-chavicol acetate, was identified with MIC values against MRSA and Staphylococcus aureus of 64 and 128μg/ml5.
Four isomers of acetoxycineoles,(trans and cis)-2-3-acetoxy-1,8-cineoles, were identified as the odorous components of the rhizomes of Alpinia galanga. Their structures were confirmed by comparing the retention indices by GC and Mass spectra with those of synthesized compound. The isomers presented individual odor features: the (trans and cis)-2-isomers exhibited woody and sweet aromas. While the (trans and cis)-3-isomers showed sweet floral and camphoraceous aromas6.
Chemical investigation of chloroform extract of rhizomes of Alpinia galanga yielded p-hydroxy cinnamaldehyde and [di-(p-hydroxy-cis-styryl)]methane. The former was isolated for the first time in nature and the latter is a new chemical component. These compounds were characterized from spectral studies and chemical reactions7.
The effect of 1’S-1’-acetoxy-chavicol acetate and related phenylpropanoids isolated from rhizomes of Alpinia galanga on ethanol induced gastric lesions in rat were examined. Among them 1’S-1’-acetoxy-chavicol acetate and 1’S-1’-acetoxyeugenol acetate markedly inhibited the ethanol induced gastric mucosal lesions (ED50=0.61 and ca. 0.90mg/kg)8.
6.3: Main objectives of the study:
Plant regeneration of Alpinia galanga by Tissue culture method.
To study the effect of hormones on micropropagated plantlets.
Study and quantify the secondary active constituent content from the in- vitro regenerated plants and comparing it with the constituents of naturally propogated plant.
To study the anti-microbial and anti-fungal activity using various extracts of Alpinia galanga rhizome.
Materials and methods:
7.1: Source of the data:
The required data will be obtained from:
Electronic data (Internet).
Published research papers.
Review articles from journals.
Library of IISC, Govt. College of Pharmacy and IIHR.
7.2 Methods of collection of the data (including sampling procedure if any)
Collection of the plant material and authentication.
Studies regarding the quantity of active constituent in plant regenerated through micropropagation method and comparing it with naturally propogated plant.
The extracts prepared from in vitro regenerated plants are screened for its anti-microbial and anti-fungal activities.
7.3: Does the study require any investigations or intervention to be conducted on patients other human or animals? If so, please describe briefly:
- No –
7.4: Has ethical clearance been obtained from your institute in case of 7.3
-- Not applicable—
List of references:
Borthakur M, Hazarika J, Singh RS. A protocol for micropropagation of Alpinia galanga. Plant cell, Tissue and Org cult. 1999;55:231-233.
Ibrahim bin Jantan, Mohd Salleh Mohd Yassin, Chen Bee Chin, Lau Lee Chen, Ng Lee Sim. Anti-fungal activity of the Essential Oils of Nine Zingiberaceae Species. Pharmaceutical Biology. 2003;41(5):392-397.
Akhtar MS, Khan MA, Malik MT. Hypoglycaemic activity of Alpinia galanga rhizome and its extracts in rabbits. Fitoterapia. 2002;73:623-628.
Sariga Zaeoung, Anuchit Plubrukarn, Niwat Keawpradub. Cytotoxic and free radical scavenging activities of Zingiberaceous rhizomes. J Sci. Technol. 2005;27(4):799-812.
Supayang piyawan Voravuthikunchai, Souwalak Phongpaichit, Sanan Subhadhirasakul. Evaluation of Anti-bacterial Activities of Medicinal plants widely used among AIDS Patients in Thailand. Pharmaceutical Biology. 2005;43(8):701-706.
Kikue Kubota, Kae Nakamura (Murayama), Akio Kobayashi. Acetoxy-1,8-cineoles as Aroma Constituents of Alpinia galangaWilld. J Agric. Food Chem 1998;46(12):5244-5247.
Barik BR, Kundu AB, Dey AK. Two phenolic constituents from Alpinia galanga rhizomes. Phytochemistry. 1987;26(7):2126-2127.
Hisashi Matsuda, Yutana Pongpiriyadacha, Toshio Morikawa, MomotaroOchi ,Masayuki Yoshikawa. Gastroprotective effects of Phenylpropanoids from the rhizomes of Alpinia galanga in rats: structural requirements and mode of action. European Journal of Pharmacology. 2003;471(1):59-67.