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.

Plum is commercially cultivated worldwide for the rich nutrient in its fruit. In May 2019, plum with symptoms of fruit rot were collected from fields located in Liuma town, Guizhou Province, China. The incidence of the disease varied from 10 to 20%, which was observed in 15 plum orchards (18 hectares) surveyed. Estimated yield loss was~5 to 10% for each field. Diseased fruits showed deformity, wilting and sunken lesions, and subsequenly became melanized and rotted. Diseased tissues were surface disinfected with 70% ethanol for 45 s and rinsed with sterile distilled water three times. Four morphologically similar colonies with white fluffy aerial mycelium and a reddish pigment were obtained after 3 days incubation on potato dextrose agar (PDA) at 25°C. Four single-spore isolates produced conidia with 1 to 2 septa that were sickle-shaped, thin-walled with a tapering and curved apical cell, measuring 15.6 to 29.6 × 4.8 to 8.7 μm (average 19.5×5.9 μm, n=50). Based on the cultural and conidial morphology, the isolates were identified as Fusarium (Mun et al. 2012; Leslie and Summerell 2006). DNA of two isolates was extracted using the Ezup Column Fungal Genomic DNA Extraction Kit (Sangon Bioengineering Shanghai, LTD.). To confirm the morphological diagnosis, DNA sequence data from three loci were obtained. PCR amplification was carried out with universal primers ITS1/ITS4 (White et al. 1990), translation elongation factor (EF-1α), EF1-H (5'-ATGGGTAAGGAAGACAAGAC-3') and EF2-T (5'-GGAAGTACCAGTGATCATGTT-3') (O'Donnell et al. 1998) and the second largest subunit of RNA polymerase II (RPB2), 5F2(5'-GGGGWGAYCAGAAGAAGGC-3') and 7cR (5'-CCCATRGCTTGYTTRCCCAT-3') (O'Donnell et al. 2007). Primers ITS1 and ITS4 produced a 559-bp amplicon (GenBank accession. MW085028). BLAST analysis showed 100% sequence identity to sequences of several species, deposited in GenBank, including F. fujikuroi. The EF-1α sequence (MW086868) was 100% identical to that of Fusarium fujikuroi (MN193860.1). The RPB2 primers amplified a fragment (MW086869) that was 99.9% identical to that of F. fujikuroi (MN193888.1). The BLASTn results based on the partial EF-1α and RPB2 sequences suggest isolate HJGF1 is F. fujikuroi. A pathogenicity assay was conducted using an agar disk inoculation method on plum. Fruits were stab inoculated with HJGF1 by piercing 1-mm at 3 points using a sterile needle, and fruits were mock inoculated with sterile PDA, each fruit was inoculated with three disks. (Fig. 1). The treated fruit were maintained in a growth chamber with 90% relative humidity at 25°C, and a daily 12-h photoperiod. After 5 days, the artificially inoculated fruit showed blotches with sunken lesions similar to those observed in the orchards, whereas no symptoms were observed on the control fruit. The experiment was repeated twice with similar results. F. fujikuroi was reisolated from infected tissues and confirmed by sequence analysis. To our knowledge, this is the first report of F. fujikuroi causing fruit blotch of plum in China. Considering the economic importance of plum in China and throughout the world, F. fujikuroi may be an emerging problem for plum cultivation. Thus, further study of fruit blotch of plum is warranted.

Bacterial canker caused by Pseudomonas syringae pv. actinidiae (Psa) has led to considerable losses in all major kiwifruit-growing areas. There are no commercial products in the market to effectively control this disease. Therefore, the defense resistance of host plants is a prospective option. In our previous study, sulfur could improve the resistance of kiwifruit to Psa infection. However, the mechanisms of inducing resistance remain largely unclear. In this study, disease severity and protection efficiency were tested after applying sulfur, with different concentrations in the field. The results indicated that sulfur could reduce the disease index by 30.26 and 31.6 and recorded high protection efficiency of 76.67% and 77.00% after one and two years, respectively, when the concentration of induction treatments was 2.0 kg/m³. Ultrastructural changes in kiwifruit stems after induction were demonstrated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), and the activities of phenylalanine ammonia-lyase (PAL), peroxidase (POD) and polyphenol oxidase (PPO), and the accumulation of lignin were determined by biochemical analyses. Our results showed that the morphological characteristics of trichomes and lenticels of kiwifruit stem were in the best defensive state respectively when the sulfur concentration was 3.0 kg/m³ and 1.5 kg/m³. Meanwhile, in the range of 0.5 to 2.0 kg/m³, the sulfur could promote the chloroplast and mitochondria of kiwifruit stems infected with Psa to gradually return to health status, increasing the thickness of the cell wall. In addition, sulfur increased the activities of PAL, POD and PPO, and promoted the accumulation of lignin in kiwifruit stems. Moreover, the sulfur protection efficiency was positively correlated with PPO activity (p < 0.05) and lignin content (p < 0.01), which revealed that the synergistic effect of protective enzyme activity and the phenolic metabolism pathway was the physiological effect of sulfur-induced kiwifruit resistance to Psa. This evidence highlights the importance of lignin content in kiwifruit stems as a defense mechanism in sulfur-induced resistance. These results suggest that sulfur enhances kiwifruit canker resistance via an increase in phenolic components and morphology structure modification in the kiwifruit stems. Therefore, this study could provide insights into sulfur to control kiwifruit canker caused by Psa.

Sulfur has been previously reported to modulate plant growth and exhibit significant anti-microbial activities. However, the mechanism underlying its diverse effects on plant pathogens has not been elucidated completely. The present study conducted the two-year field experiment of sulfur application to control kiwifruit canker from 2017 to 2018. For the first time, our study uncovered activation of plant disease resistance by salicylic acid after sulfur application in kiwifruit. The results indicated that when the sulfur concentration was 1.5–2.0 kg m⁻³, the induced effect of kiwifruit canker reached more than 70%. Meanwhile, a salicylic acid high lever was accompanied by the decline of jasmonic acid. Further analysis revealed the high expression of the defense gene, especially AcPR-1, which is a marker of the salicylic acid signaling pathway. Additionally, AcICS1, another critical gene of salicylic acid synthesis, was also highly expressed. All contributed to the synthesis of increasing salicylic acid content in kiwifruit leaves. Moreover, the first key lignin biosynthetic AcPAL gene was marked up-regulated. Thereafter, accumulation of lignin content in the kiwifruit stem and the higher deposition of lignin were visible in histochemical analysis. Moreover, the activity of the endochitinase activity of kiwifruit leaves increased significantly. We suggest that the sulfur-induced resistance against Pseudomonas syringae pv. actinidiae via salicylic activates systemic acquired resistance to enhance plant immune response in kiwifruit.

Plum bacterial shot-hole caused by Pantoea agglomerans (P. agglomerans) is one of the primary bacterial diseases in plum tree planting areas, resulting in abnormal growth of plum trees and severe economic losses. Early diagnosis of P. agglomerans is crucial to effectively control plant diseases. In this study, loop-mediated isothermal amplification (LAMP) analysis for genome-specific gene sequences was developed for the specific detection of P. agglomerans. We designed the LAMP primers based on the gyrB gene of P. agglomerans. The best reaction system was 0.2 μmol·L⁻¹ for outer primer F3/B3 and 1.6 μmol·L⁻¹ for inner primer FIP/BIP. The LAMP reaction was optimal at 65°C for 60 min based on the color change and gel electrophoresis. This technology distinguished P. agglomerans from other control bacteria. The detection limit of the LAMP technology was 5 fg·μl⁻¹ genomic DNA of P. agglomerans, which is 1,000 times that of the traditional PCR detection method. The LAMP technology could effectively detect the DNA of P. agglomerans from the infected leaves without symptoms after indoor inoculation. Furthermore, the LAMP technology was applied successfully to detect field samples, and the field control effect of 0.3% tetramycin after LAMP detection reached 82.51%, which was 7.90% higher than that of conventional control. 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 plum bacteria shot-hole disease.