ريزازديادي و بررسي آلكالوئيدهاي دارويي گياه پراونش ﴿Catharanthus roseus﴾ در شرايط آزمايشگاهي

چكيده انگليسي :

Micropropagation and Detection of Medicinal Alkaloids of Periwinkle Catharanthus roseus in In Vitro Condition Forough Eatesam eatesam f@gmail com Date of submission April 6 2011 Department of Agricultural Biotechnology Isfahan University of Technology Isfahan 84156 83111 Iran Degree M Sc Language Farsi 1 B E Seyed Tabatabaei Prof Supervisor Sayedt@cc iut ac ir 2 M Talebi Assist Prof Supervisor mtalebi@cc iut ac irAbstract The herbaceous species Catharanthus roseus L G Don commonly known as MadagascarPeriwinkle belongs to the family Apocynaceae that native to tropical and subtropical region such assouthern india Indonesia and Madagascar but it is distributed worldwide now This plant accumulatesmore than 130 terpenoid indole alkaloids TIAs of which two dimeric alkaloids Vinblastine andVincristine have antineoplastic activity and useful for the treatment of various cancers Although thesedrugs are demanded highly the amount of these alkaloids are very low in the intact plant which is still theonly source for them Complexity and multiple step of biosynthetic pathway of these two alkaloids andexistence of MULTIPLE CHIRAL CENTERS spesific production of some biosynthetic pathwayprocedures of them in specialized tissue less effectiveness of semi synthetic medicine in comparsion withnatural ones and high demand for these two medicines are the most important reasons which drawresearcher s attention to use biotechnological methods and plant tissue culture in order to increase theproduction of these important alkaloids in plant In this research to indicate the best media formicropropagation and increase in amount of anti cancer alkaloids Vinblastine and Vincristine inPeriwinkle plant 25 plant hormones combination to produce callus from leaf explant 7 treatments forregeneration of calluses and 30 media supplemented with various concentration of plant hormones andactivated charcoal for direct regeneration of node explant are considered Then the content of Vinblastineand Vincristine alkaloids in regenerated plants are extracted by Hollow fiber based liquid phasemicroextraction HF LPME method and analyzed by ion pair liquid chromatography Results showedthat indirectly regeneration of leaf explant is not a suitable method to micropropagation of Periwinkleplant due to hard regeneration of callus probability of mutation and lower number of produced branchescompared with direct regeneration But using BAP in culture media containing 1 g l activated charcoal showed the best direct regeneration of node explants and shoot proliferation Also increasing of BAPconcentration and decreasing of activated charcoal the amont of Vinblastine and Vincristine increased Therefore although due to exist of many phenolic compounds in Periwinkle plant using of activatedcharcoal in culture media is necessary for plant growth its use in media is important because it hasnegative effect on adsorption of hormones and resulted in reducing the shoot proliferation and alkaloidsproduction so seems that concentration of 1 g l activated charcoal is a appropriate amount to make shootproliferation media Also transporting regenerated shoots to culture media containing NAA resulted inincreasing the length of shoots However it reduced the amount of dimeric alkaloids This reduction ofconcentration in Vincristine alkaloids are more than Vinblastine In HF LPME best extraction ofalkaloids was obtained in acetic acid 0 1 mol l as acceptor phase pH 11 5 for donor phase andphosphoric acid 0 1 mol l to preparation the plant extract Because of simultaneous elution of dimericalkalois and their monomers it seems that using negative ion pair Sodium 1 butanesulfonate C4H9NaO3S in HPLC buffer is a more suitable method to separation of Vinblastine and Vincristinealkaloids Keywords Periwinkle Micropropagation Vinblastine Vincristine HF LPME HPLC

اعتصام، فروغ

ريزازديادي و بررسي آلكالوئيدهاي دارويي گياه پراونش ﴿Catharanthus roseus﴾ در شرايط آزمايشگاهي

وين بلاستين , وين كريستين , ريز استخراج فاز مايع , كروماتوگرافي جفت يوني