Maryam Varasteh-kojourian; Ali Ganjeali; Javad Asili; Saeid Malekzadeh-Shafaroudi; Akram Taleghani
Abstract
Endophytic fungi are often producing host plant metabolites. Tanshinones are secondary metabolites of the Salvia genus which are also produced by some endophytic fungi. Efficient secondary metabolite production in endophytic fungi drops significantly after sequential subcultures. 5-azacytidine (5-AC) ...
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Endophytic fungi are often producing host plant metabolites. Tanshinones are secondary metabolites of the Salvia genus which are also produced by some endophytic fungi. Efficient secondary metabolite production in endophytic fungi drops significantly after sequential subcultures. 5-azacytidine (5-AC) is an analog of the naturally occurring pyrimidine nucleoside cytidine and a DNA methyltransferase inhibitor. In this relation, 5-AC is an effective tool to induce the expression of silenced secondary metabolite genes in fungi. We isolated 4 endophytic fungi from the roots of Salvia abrotanoides which produced tanshinone. Cryptotanshinone and tanshinone IIA were produced by Penicillium canescens, Penicillium nodositatum, and Penicillium pinophilum, while Paraphoma radicina only produced tanshinone IIA. The maximum amount of tanshinones was extracted from P. pinophilum culture with 130.826 mg cryptotanshinone /g of dry weight and 50.155 mg Tanshinone IIA/g of dry weight. These amounts were significantly more than tanshinones produced in plant roots (0.55 mg cryptotanshinone/g of dry weight, 1.3 mg Tanshinone IIA/g of dry weight). In the third subculture, tanshinone production decreased significantly. 5-azacytidine as an epigenetic modifier retrieved tanshinone production in the third subculture of P. pinophilum. Also, 5- azacytidine treatment made a big jump in Tanshinone IIA production in P. radicina (63.176 mg TIIA/g of dry weight) besides increasing Tanshinone IIA production in P. nodositatum cultures. This is the first report using 5- azacytidine to improve tanshinone production in endophytic fungi. Our results confirm that 5- azacytidine is an efficient, easy, and quick chemical to felicitate secondary metabolite production in endophytic fungi.
Narges Fazili; Zahra-Soheila Soheili; Saeid Malekzadeh-Shafaroudi; Shahram Samiei; Shamila D.Alipoor; Nasrin Moshtaghi; Abouzar Bagheri
Abstract
Royal jelly (RJ) from queen honeybee larva as a traditional medicine agent has a variety of pharmacological benefits. In the present study, the effect of Royal jelly was investigated on the urinary bladder cancer cell line (HTB-9 5637). To determine the cell viability in different concentrations ...
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Royal jelly (RJ) from queen honeybee larva as a traditional medicine agent has a variety of pharmacological benefits. In the present study, the effect of Royal jelly was investigated on the urinary bladder cancer cell line (HTB-9 5637). To determine the cell viability in different concentrations of Royal jelly, MTT assay was performed. An in vitro wound healing assay was applied to investigate the effect of RJ on cell migration. The activity and gene expression level of matrix metalloproteinase 2 and 9 was assessed by zymography and Real time PCR respectively. R.J.S at the concentration of 0.7 mg/ml had a significant effect on reducing the proliferation rate of 5637 cells after 72h (p < 0.009). R.J.S significantly decreased cell migration and induced a significant decrease in the transcriptional level of MMP9 after 72h (0.5x; P < 0.049). However R.J.S did not impose any effect on the expression level and activity of matrix metalloproteinase 2. These results indicate the potential of RJ as a promised natural anti-proliferative and anti-metastatic drug in combination with advanced therapy methods for cancer treatment. Royal jelly has the potential to be more focused as an anti-metastatic drug to control tumor growth and can be considered as a more effective alternative to the current chemotherapy drugs.
Maziar Habibi-Pirkoohi; Saeid Malekzadeh-Shafaroudi; Hasan Marashi; Saeid Zibaee; Afsaneh Mohkami; Saba NejatizaXeh
Abstract
An Agrobacterium-mediated transient gene expression assay was carried out in alfalfa (Medicago sativa) leaves for expression of a chimeric gene encoding a part of capsid protein of Foot and Mouth Disease virus called VP1. The plant leaves were transformed via agroinfiltration procedure. The presence ...
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An Agrobacterium-mediated transient gene expression assay was carried out in alfalfa (Medicago sativa) leaves for expression of a chimeric gene encoding a part of capsid protein of Foot and Mouth Disease virus called VP1. The plant leaves were transformed via agroinfiltration procedure. The presence of the foreign gene and its expression in transformed plants were evaluated by polymerase chain reaction (PCR), real time PCR, protein Dot blot and ELISA. Moreover, gene expression in the transformed leaves was quantified by ELISA method. The results obtained in this investigation indicated high level of gene expression in alfalfa leaves, showing that transient gene expression can be applied as an effective and time-saving procedure for the production of recombinant proteins. The procedures for transformation, detection of recombinant protein and its application for molecular experiments are described in the study.
