Alternaria alternata is the main pathogenic species of various crops, including kiwifruit (Actinidia cinensis). In this study, an antagonistic fungus, J-1, with high antifungal activity against A. alternata was isolated from A. cinensis “Hongyang.” The strain J-1 was identified as Fusicolla violacea via morphological identification and DNA sequencing. This study aimed to evaluate the antifungal activity and potential mechanism of the strain J-1 against A. alternata. The strain J-1 exhibited antifungal activity against A. alternata, with an inhibition rate of 66.1% in vitro. Aseptic filtrate (AF) produced by the strain J-1 could suppress the mycelial growth and conidia germination of A. alternata at the inhibition rates of 66.8% and 80%, respectively, as well as suppress the spread of Alternaria rot in fresh kiwifruit. We observed that many clusters of spherical protrusions appeared at the mycelial tips of A. alternata after treatment with 200 mL L−1 AF of J-1. Scanning electron microscopy analysis results showed that the mycelial structures were bent and/or malformed and the surfaces were rough and protuberant. Variations in temperature, pH, and storage time had little effect on the antifungal activity of the AF. Moreover, the AF could damage the integrity of cell membranes and cause intracellular content leakage. Meanwhile, the chitinase and β-1,3-glucanase enzyme activities increased significantly, indicating that the function of A. alternata cell wall was seriously injured. Eleven antimicrobial metabolites were identified by gas chromatography–mass spectrometry (GC–MS). The strain J-I and its AF exhibited well broad-spectrum antifungal activity against Diaporthe eres, Epicoccum sorghinum, Fusarium graminearum, Phomopsis sp., and Botryosphaeria dothidea, with inhibition rates ranging from 34.4% to 75.1% and 42.7% to 75.2%, respectively. Fusicolla violacea J-1 is a potential biocontrol agent against A. alternata and other fungal phytopathogens.
Kiwifruit (Actinidia chinensis) is an important commercial crop in China, and the occurrence of diseases may cause significant economic loss in its production. In the present study, a new pathogen that causes brown leaf spot disease on kiwifruit was reported. The fungus was isolated from an infected sample and identified as Fusarium graminearum based on morphological and molecular evaluation. Koch’s postulates were confirmed when the pathogen was re-isolated from plants with artificially induced symptoms and identified as F. graminearum. Based on the biological characteristics of the pathogen, it was determined that: its optimal growth temperature was 25 °C; optimal pH was 7; most suitable carbon source was soluble starch; most suitable nitrogen source was yeast powder; and best photoperiod was 12 h light/12 h dark. Further investigations were conducted by determining 50% effective concentrations (EC50) of several active ingredients of biological fungicides against F. graminearum. The results showed that among the studied fungicides, tetramycin and honokiol had the highest antifungal activity against this pathogen. Our findings provide a scientific basis for the prevention and treatment of brown leaf spot disease on kiwifruit.
‘Hongyang’ kiwifruit (Actinidia chinensis, cultivar ‘Hongyang’) black spot disease is caused by the fungal pathogen Didymella glomerata, and is a serious disease, causing considerable losses to the kiwifruit industry during growth of the fruit. Hence, we aimed to identify a potential biocontrol agent against D. glomerata. In this study, bacterial isolates from the rhizosphere soil of kiwifruit were tested for their potential antifungal activity against selected fungal pathogens. Based on a phylogenetic tree constructed using sequences of 16S rDNA and the gyrA gene, BQ-33 with the best antifungal activity was identified as Bacillus mojavensis. We evaluated the antagonistic activity and inhibitory mechanism of BQ-33 against D. glomerata. Confrontation experiments showed that both BQ-33 suspension and the sterile supernatant (SS) produced by BQ-33 possessed excellent broad-spectrum antifungal activity. Furthermore, the SS damaged the cell membrane and cell wall of the mycelia, resulting in the leakage of a large quantity of small ions (Na+, K+), soluble proteins and nucleic acids. Chitinase and β-1,3-glucanase activities in SS increased in correlation with incubation time and remained at a high level for several days. An in vivo control efficacy assay indicated that 400 mL L−1 of SS completely inhibited kiwifruit black spot disease caused by D. glomerata. Therefore, BQ-33 is a potential biocontrol agent against kiwifruit black spot and plant diseases caused by other fungal pathogens. To our knowledge, this is the first report of the use of a rhizosphere microorganism as a biocontrol agent against kiwifruit black spot disease caused by D. glomerata.
Kiwifruit is moderately sweet and sour and quite popular among consumers; it has been widely planted in some areas of the world. In 2019, the crown gall disease of kiwifruit was discovered in the main kiwifruit-producing area of Guizhou Province, China. This disease can weaken and eventually cause the death of the tree. The phylogeny, morphological and biological characteristics of the bacteria were described, and were related to diseases. The pathogenicity of this species follows the Koch hypothesis, confirming that A. fabacearum is the pathogen of crown gall disease of kiwifruit in China. In this study, Loop-mediated isothermal amplification (LAMP) analysis for genome-specific gene sequences was developed for the specific detection of A. fabacearum. The detection limit of the LAMP method is 5 × 10−7 ng/μL, which has high sensitivity. At the same time, the amplified product is stained with SYBR Green I after the reaction is completed, so that the amplification can be detected with the naked eye. LAMP analysis detected the presence of A. fabacearum in the roots and soil samples of the infected kiwifruit plant. The proposed LAMP detection technology in this study offers the advantages of ease of operation, visibility of results, rapidity, accuracy and high sensitivity, making it suitable for the early diagnosis of crown gall disease of kiwifruit.