Production of watermelon (Citrullus lanatus) in Malaysia was 150,000 mt in 2020 (Malaysian Department of Agriculture, 2021). In November 2019, nine locally produced watermelon fruit (red flesh, seedless) from five local stores in the states of Kelantan, Terengganu, and Penang exhibited sunken, circular, brown lesions that enlarged to1.5 to 10 cm in diameter with scattered orange masses of conidia. Lesions coalesced to cover approximately 50% of the fruit surface. Lesions were surface sterilized by spraying 70% alcohol onto the fruit followed by drying with sterilized paper towels. A total of 153 tissue segments (1×1 cm) were excised from the rind, immersed in 1% sodium hypochlorite for 3 min, rinsed twice for 1 min in sterilized distilled water, air-dried, transferred to potato dextrose agar (PDA) plates, and incubated at 25±1°C for 7 days. Single-spore transfers produced pure cultures, resulting in 12 isolates. Colonies on PDA were initially white and turned pale gray with age. Conidia were hyaline, one end round and the other narrowly acute, aseptate, smooth-walled, straight, cylindrical to clavate, 10.5-16.5 µm × 3-4.5 μm (n = 30). Observed morphological characters matched published description of Colletotrichum spp. (Damm et al. 2012). Internal transcribed spacer (ITS) and glyceraldehyde-phosphate dehydrogenase (GAPDH) genes were amplified using primer sets ITS1/ITS4 and GDF1/GDF2, respectively. All sequences were deposited in GenBank (MW856808 for ITS; MZ219296 for GAPDH). A BLASTn search of both sequences on GenBank showed 99% identity with C. scovillei along with other closely related Colletotrichum species. Phylogenetic analysis of ITS and GAPDH alignments, using maximum likelihood along with reference strains of closely related species from Mycobank, confirmed species identity as C. scovillei. A pathogenicity test was conducted on two healthy watermelon fruit (red flesh, seedless). A 6-mm-diameter mycelial plug of a colony on PDA was positioned on a 0.5-cm-long wound on each fruit; a sterile PDA plug placed on a similar wound on the opposite side served as a control. Fruit were incubated at 25±1°C for 7 days in plastic-wrapped trays above distilled water to maintain high humidity. Small, sunken, circular brown lesions appeared and expanded at inoculation sites within 7 days. Symptoms were identical to those produced by natural infections, and the controls were asymptomatic. Isolates from the lesions at the inoculation sites were confirmed as C. scovillei based on morphological characteristics, fulfilling Koch's postulates. The pathogenicity test was conducted four times with a total of eight fruit. Many species in the C. orbiculare complex cause watermelon anthracnose (Keinath, 2018). To our knowledge, this is the first report of C. scovillei (C. acutatum species complex; Damm et al. 2012) causing anthracnose on watermelon in Malaysia. Anthracnose caused by C. scovillei has been confirmed on other crops such as pepper (Toporek and Keinath, 2021), banana (Zhou et al., 2017), and chili (Oo et al., 2017). This insight will inform efforts to improve management of watermelon anthracnose in Malaysia.
In this study, the fabrication of titanium dioxide/reduced graphene oxide (TiO2/rGO) utilising banana peel extracts (Musa paradisiaca L.) as a reducing agent for the photoinactivation of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) was explored. The GO synthesis was conducted using a modified Tour method, whereas the production of rGO involved banana peel extracts through a reflux method. The integration of TiO2 into rGO was achieved via a hydrothermal process. The successful synthesis of TiO2/rGO was verified through various analytical techniques, including X-ray diffraction (XRD), gas sorption analysis (GSA), Fourier-transform infrared (FT-IR) spectroscopy, ultraviolet-visible diffuse reflectance spectroscopy (UV-Vis DRS), scanning electron microscope-energy dispersive X-ray (SEM-EDX) and transmission electron microscopy (TEM) analyses. The results indicated that the hydrothermal-assisted green synthesis effectively produced TiO2/rGO with a particle size of 60.5 nm. Compared with pure TiO2, TiO2/rGO demonstrated a reduced crystallite size (88.505 nm) and an enhanced surface area (22.664 m2/g). Moreover, TiO2/rGO featured a low direct bandgap energy (3.052 eV), leading to elevated electrical conductivity and superior photoconductivity. To evaluate the biological efficacy of TiO2/rGO, photoinactivation experiments targeting E. coli and S. aureus were conducted using the disc method. Sunlight irradiation emerged as the most effective catalyst, achieving optimal inactivation results within 6 and 4 h.