The International Conference on Agriculture, Forest, Food Sciences and Technologies(ICAFOF), Nevşehir, Turkey, 15 - 17 May 2017, pp.202
Bouganin antiviral protein (BAP) gene isolated from Bougainvillea spectabilis Willd. 8 was cloned, expressed and the antiviral and antimicrobial activities were investigated. 9 The fulllength bouganin antiviral protein gene was amplified by reverse transcription- 10 PCR using mRNA as template extracted from mature leaves. The coding region of 11 bouganin gene was cloned into prokaryotic expression vector pETDuet-1 after 12 amplification with end to end gene specific primers. The recombinant plasmid was 13 transformed into Escherichia coli cells BL21(DE3)pLysS and the expression of BAP 14 gene was induced by isopropyl β-D thiogalactopyranoside (IPTG). Bouganin antiviral 15 protein having a molecular mass of 28 kDa has been isolated from transformed bacterial 16 colonies. Antiviral activity of bouganin was assayed against Zucchini yellow mosaic 17 virus (ZYMV) by a mechanical inoculation test. The antifungal activity of purified 18 recombinant protein was tested against three types of pathogenic and non-pathogenic 19 fungi using disc diffusion method. The increased amount of antiviral protein reduced 20 the disease severity caused by ZYMV. The bouganin antiviral protein was inhibited the 21 growth of R. solani by 30.7% and of T. harzianum by 20% after 72 h compared to 22 control. No growth inhibition was observed for F. oxyporum. In vitro expressed BAP 23 protein showed that the all plants including controls treated with antiviral proteins 24 exhibited severe growth reduction compared with negative control (not treated) plants.