6954

6496

كارشناسي ارشد

بيوتكنولوژي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1390

چهارده، 73ص.: مصور، جدول، نمودار

ص.ع. به فارسي و انگليسي

مجيد طالبي، احمد يامچي

محمد سراجي، بدرالدين ابراهيم سيد طباطبائي

24/5/91

بهرام شريف نبي، مهدي رحيم ملك

1396/09/18

كتابنامه

كشاورزي

مهندسي كشاورزي

ID6496

به فارسي وانگليسي: قابل رويت در نسخه ديجيتالي

Measuring Gene Expression Change for Producing of Medicinal Alkaloids in Periwinkle Catharanthus roseus in Response to Phyto hormones by Real Time PCR Abolfazl Pakniyat abps 2500@yahoo com Date of submission February 5 2012 Department of Plant biotechnology Isfahan University of Technology Isfahan 84156 83111 Iran Degree M Sc Language Farsi 1 M talebi Assist Professor Supervisor mtalebi@cc iut ac ir 2 A yamchi Assist Professor Supervisor yam12001@yahoo comAbstract The Catharanthus roseus L G Don commonly known as madagascar periwinkle is a medicinally important plantwhich is of interest because it contains more than 130 terpenoid indole alkaloids TIAs some of which exhibitstrong pharmacological activities Two of these bioactive compounds vinblastine and vincristine from C roseushave been used extensively as anticancer drugs in the treatment of lymphoma and leukemia like These compoundsare produced in small quantities in the shoot tissues of periwinkle and expensive to harvest and purify Severalattempts has been performed to increase TIAs production by media optimization precursor feeding and geneticmanipulations of pathway genes Despite significant efforts heretofore limited success has been brought both in cellsuspension and hairy root cultures of C roseus Increase in global demand for these two medicines have drawnattention to use biotechnological methods to increase the production of these important alkaloids in intact plant TIApathway is highly regulated so that changes in precursor concentration or protein concentration are probablycontrolled by the plant phyto hormones such as jasmonic acid JA and salicylic acid SA Therefore the aim ofpresent study was to investigate the inducible effect of JA and SA hormones on expression of dat d4h str and g10hgenes in TIAs pathway in shoot and root tissues and changes in accumulation of vinblastine and vincristine alkaloidswhich are products of this pathway For do this the total RNA was extracted from roots and leaves of plants at 0 6 24 and 72 hours after treatment with 50mM JA and 100mM SA hormones cDNA was synthesized using specificprimers and gene expression pattern was monitored by real time PCR technic The result of this study showed noproduction of vinblastine and vincristine in the roots which can be due to non expression of dat and d4h genes in thistissue These genes seem to have an important role in production of these alkaloids str and g10h genes wereexpressed in the root and shoot but the expression in the root is more than shoot Analytical measurements indicatedthat both JA and SA hormones positively affect the expression of these four genes though the effect of SA wassignificantly more than JA Vinblastine and Vincristine alkaloids of treated plants were extracted by hollow fiber based liquid phase microextraction HF LPME method and analyzed by high performance liquid chromatography HPLC Data analysis revealed an increase in vinblastine and vincristine alkaloids content after the treatment ofhormones which SA showed more pronounced effect The most increase in both gene expression and alkaloidsproduction was observed at 72 h after treatment of SA and JA Based on the results of real time PCR and HPLCexperiments it can be concluded that phyto hormone treatment can increase expression of genes involved in TIAspathway and consequently affect the production of vinblastine and vincristine anticancer alkaloids Key words Periwinkle Vinblastin Vincristin jasmonic acid salicylic acid Real Time PCR HPLC

مجيد طالبي، احمد يامچي

محمد سراجي، بدرالدين ابراهيم سيد طباطبائي

بهرام شريف نبي، مهدي رحيم ملك