Investigation of Chemical Properties of Green Tea Ethanolic Extract and Its Inhibitory and Lethal Effects on Aspergillus niger, Botrytis cinerea and Rhizopus stolonifer (Causing Rot in Strawberry and Grapes)

Strawberry and grapes are generally infected with pathogenic fungi (e.g., Aspergillus niger, Botrytis cinerea, Rhizopus stolonifera, etc.). Synthetic fungicides are commonly used as the first line of defense against post-harvest pathogens on packaging lines. However, disposal of toxic waste is a costly process and the hazardous waste causes serious environmental problems. In addition, fungal pathogens have shown a worrying trend of resistance to these fungicides, thus shortening the shelf life of products. Compounds that can be equally effective in controlling pathogens, but preventing or minimizing the waste problems will be inevitable. The large volume of internationally processed agricultural products, as well as the increasing demand for organically produced fruits, emphasizes the need to replace synthetic fungicides with safer and biodegradable alternatives. Natural plant-derived products effectively meet this criterion and have great potential to influence modern agricultural research. Catechins and other polyphenols in green tea show strong antioxidant activity. Also, the antimicrobial activity of green tea extract against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans has been reported. Therefore, the present study was performed to prepare the ethanolic extract of green tea and to determine the content of total phenol, total flavonoids, antioxidant activity, and its antifungal effect against Aspergillus niger, Botrytis cinerea, and Rhizopus stolonifer (causing rot in strawberry and grapes).

Fresh green tea leaves were dried at room temperature and then powdered. Then, ethanol (70%) was added to the powdered leaves (solvent to powder ratio of 10:1 v/w) and the mixture was refluxed for 120 min. The resulting mixture was filtered through a filter paper and then concentrated under vacuum and finally dried in an oven.Total phenol content (by Folin-Ciocalteu reagent at 756 nm), total flavonoid content (spectrophotometrically at 510 nm), antioxidant activity (by DPPH and ABTS radical scavenging methods), and antifungal effect (by disk diffusion agar, well diffusion agar, minimum inhibitory concentration, and minimum fungicidal concentration) of the extract were evaluated.

The extract contained 175.60 mg GAE /g total phenol and 47.53 mg QE/g total flavonoids and its antioxidant activity using DPPH and ABTS free radical assays was 78.89% and 86.57%, respectively. The results of antifungal activity showed that the diameter of the growth inhibition zone increased significantly with increasing the concentration of the extract, and Botrytis cinerea and Rhizopus stolonifer were the most sensitive and resistant fungal strains to the extract, respectively. The minimum fungicidal concentrations for the strains of Botrytis cinerea and Rhizopus stolonifer were 64 and 512 mg/ml, respectively.

The results of the present study showed that the ethanolic extract of green tea could be considered as potential source of natural antioxidant and antifungal agents. The presence of phenolic and flavonoid compounds may be responsible for the antifungal and antioxidant effects of the extract. However, due to the fact that this study was performed with the crude extract of green tea, it is difficult to identify compounds responsible for antifungal and antioxidant activity. On this point, only the separation of the components of the extract allows the detection of antifungal and antioxidant compounds. This study provides a basis for further researches, in particular the use of these antioxidants and antifungal compounds. Green tea extract is especially suitable for products with high sensitivity to lipid oxidation and infection with molds.