Diagnosis and Control of Brown Leaf Spot of Kiwi (Actinidia deliciosa) Using Biochar-Zinc Oxide Nanocomposite (MB-ZnO) as a Non-Toxic Bio-Fungicides

Abstract

Kiwi is one of the best natural sources of vitamin C and has wide applications. During October–November 2021, small brown spots were examined on the Kiwi leaves. The diseased leaf samples were collected and placed on potato dextrose agar nutrient media for diagnosis. Morphological, anatomical, and molecular studies revealed this disease-causing agent to be Rhizopus oryzae. Molecular characterizations of the isolated pathogen were performed by using actin translation elongation factor (EF-1α) and ribosomal deoxyribose nucleotide inter transcribed sequence (rDNA ITS ITS1/ITS4) and elongation factors (EFl-F/EFl-R) primers. A BLAST study of the resultant ITS1/ITS4 sequence showed > 99% resemblance with R. oryzae (MT603964.1), while the EF-1α sequence revealed 100% similarity with translation elongation factor-1α gene of R. oryzae (MK510718.1). The obtained ITS1/ITS4 sequence was submitted to NCBI (MW603842.1). Koch’s postulates established the pathogenicity of isolated R. oryzae and proved it to be the brown spot pathogen of Kiwi. For the environmentally-friendly management of Kiwi leaf spot, maize biochar-Zinc Oxide (MB-ZnO) nanocomposite was used. The prepared nanocomposite was characterized by Fourier transform infrared (FTIR) spectroscopy, thermo gravitational analysis (TGA), X-ray diffraction (XRD) analysis, scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) analysis. After successful preparation, MB-ZnO was assessed for its possible antifungal potential against R. oryzae. MB-ZnO displayed substantial growth inhibition, and the highest growth inhibition (79%) was observed at a 19 mg/mL dose rate of nanoparticles. These excellent findings propose that Ball-milled synthesis is a fast, economical, and environmentally friendly method for nanocomposite in the near future. The nanocomposite is used as a nominal substitute for chemical fungicides.