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Abstract Background Alternaria alternata is the primary pathogen of potato leaf spot disease, resulting in significant potato yield losses globally. Endophytic microorganism-based biological control, especially using microorganisms from host plants, has emerged as a promising and eco-friendly approach for managing plant diseases. Therefore, this study aimed to isolate, identify and characterize the endophytic fungi from healthy potato leaves which had great antifungal activity to the potato leaf spot pathogen of A. alternata in vitro and in vivo. Results An endophytic fungal strain SD1-4 was isolated from healthy potato leaves and was identified as Talaromyces muroii through morphological and sequencing analysis. The strain SD1-4 exhibited potent antifungal activity against the potato leaf spot pathogen A. alternata Lill, with a hyphal inhibition rate of 69.19%. Microscopic and scanning electron microscope observations revealed that the strain SD1-4 grew parallel to, coiled around, shrunk and deformed the mycelia of A. alternata Lill. Additionally, the enzyme activities of chitinase and β-1, 3-glucanase significantly increased in the hyphae of A. alternata Lill when co-cultured with the strain SD1-4, indicating severe impairment of the cell wall function of A. alternata Lill. Furthermore, the mycelial growth and conidial germination of A. alternata Lill were significantly suppressed by the aseptic filtrate of the strain SD1-4, with inhibition rates of 79.00% and 80.67%, respectively. Decrease of leaf spot disease index from 78.36 to 37.03 was also observed in potato plants treated with the strain SD1-4, along with the significantly increased plant growth characters including plant height, root length, fresh weight, dry weight, chlorophyll content and photosynthetic rate of potato seedlings. Conclusion The endophyte fungus of T. muroii SD1-4 isolated from healthy potato leaves in the present study showed high biocontrol potential against potato leaf spot disease caused by A. alternata via direct parasitism or antifungal metabolites, and had positive roles in promoting potato plant growth.

Abstract Background Kiwifruit bacterial canker, caused by Pseudomonas syringae pv. actinidiae (Psa), is a destructive disease worldwide. Resistance genes that respond to Psa infection urgently need to be identified for controlling this disease. Laccase is mainly involved in the synthesis of lignin in the plant cell wall and plays a prominent role in plant growth and resistance to pathogen infection. However, the role of laccase in kiwifruit has not been reported, and whether laccase is pivotal in the response to Psa infection remains unclear. Results We conducted a bioinformatics analysis to identify 55 laccase genes (AcLAC1–AcLAC55) in the kiwifruit genome. These genes were classified into five cluster groups (I–V) based on phylogenetic analysis, with cluster groups I and II having the highest number of members. Analysis of the exon–intron structure revealed that the number of exons varied from 1 to 8, with an average of 5 introns. Our evolutionary analysis indicated that fragment duplication played a key role in the expansion of kiwifruit laccase genes. Furthermore, evolutionary pressure analysis suggested that AcLAC genes were under purifying selection. We also performed a cis-acting element analysis and found that AcLAC genes contained multiple hormone (337) and stress signal (36) elements in their promoter regions. Additionally, we investigated the expression pattern of laccase genes in kiwifruit stems and leaves infected with Psa. Our findings revealed that laccase gene expression levels in the stems were higher than those in the leaves 5 days after inoculation with Psa. Notably, AcLAC2, AcLAC4, AcLAC17, AcLAC18, AcLAC26, and AcLAC42 showed significantly higher expression levels (p

【Objective】The research was carried out to clarify the field control effects of six agents on blossom blight of kiwifruit, and to screen efficient and safe agents for guiding the field production of kiwifruit.【Method】The field control experiment was carried out by using random block design, and the incidence rate and disease index of blossom blight of kiwifruit after the application of six agents were determined, and the control effects were evaluated by calculating the change values of incidence rate and disease index before and after treatment. The safety of each agent was evaluated by its effects on the growth situation of kiwifruit plants and the development characteristics of flowers and fruits.【Result】Six agents showed different control effects on blossom blight of kiwifruit, and all of them were safe to kiwifruit plants. The results of field trials showed that there was no significant difference in incidence rate and disease index between the treatment groups and the CK group before control. After two consecutive treatments, the incidence rates of treatment groups and CK group showed an increasing trend compared with those before control, and the change values of the disease situation were significantly different, among which the change value of the CK group was the largest, and the incidence rate and disease index increased by 13.29% and 6.84, respectively. The change values of kasumin • brothermothingonil and tetramycin • propyrithiazole were the smallest, with the increase of incidence rate less than 1.50% and the increase of disease index less than 1.00. The field control effect corresponded to the disease situation. After two consecutive treatments, the treatment of kasumin • brothermothingonil showed the highest control effect, followed by tetramycin • propyrithiazole, both of which were above75%. All agents had no phytotoxicity symptoms and other adverse effects on kiwifruit plants after the application of agents, and showed different degrees of flower protection, fruit protection and yield increase compared with the CK group. The results showed that among the six agents, kasumin • brothermothingonil and tetramycin • propyrithiazole were the most effective in delaying the transmission and spread of the pathogen of blossom blight of kiwifruit, reducing the disease severity, and controlling blossom blight of kiwifruit efficiently and safely.【Conclusion】Two effective and safe agents - kasumin • brothermothingonil and tetramycin • propyrithiazole are screened for blossom blight of kiwifruit, which can be promoted in field production of kiwifruit.

To find a precise, efficient, and safe method to control rice sheath blight disease (caused by Rhizoctonia solani), this study used the natural product magnolol as the fungicide. Based on the characteristics of cell wall-degrading enzymes (CWDEs) released by R. solani during the early stages of infection, a Mag@CMCS@CPSG (magnolol@carboxymethyl chitosan@carboxymethyl cellulose, pectin, sodium alginate, and glycerol) film was constructed to control the release of magnolol in response to CWDEs. The Mag@CMCS@CPSG film increased magnolol toxicity against R. solani, decreased EC50 from 7.33 μg/mL to 4.37 μg/mL, and effectively blocked the invasion of mycelia. Under the stimulation of CWDEs released during R. solani infection, the cumulative release of magnolol by the Mag@CMCS@CPSG film reached 75.62% after 96 h, and the disease index of rice treated with Mag@CMCS@CPSG film significantly decreased. The maximum field control effect of Mag@CMCS@CPSG film on rice sheath blight disease was 72.47%, which was not significantly different from that of the commercial preparation jinggangmycin; however, the holding period was better. The Mag@CMCS@CPSG film reduced the loss of rice yield and quality caused by infection with R. solani and did not cause phytotoxicity or residue. In conclusion, the Mag@CMCS@CPSG film can achieve accurate and green control of rice sheath blight.

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Soft rot causes significant economic losses in the kiwifruit industry. This study isolated strain CTXW 7-6-2 from healthy kiwifruit tissue; this was a gram-positive bacterium that produced the red pigment pulcherrimin. The phylogenetic tree based on 16S ribosomal RNA, gyrA, rpoB, and purH gene sequences identified CTXW 7-6-2 as a strain of Bacillus subtilis. CTXW 7-6-2 inhibited hyphal growth of pathogenic fungi that cause kiwifruit soft rot, namely, Botryosphaeria dothidea, Phomopsis sp., and Alternaria alternata, by 81.76, 69.80, and 32.03%, respectively. CTXW 7-6-2 caused the hyphal surface to become swollen and deformed. Volatile compounds (VOC) produced by the strain inhibited the growth of A. alternata and Phomopsis sp. by 65.74 and 54.78%, respectively. Whole-genome sequencing revealed that CTXW 7-6-2 possessed a single circular chromosome of 4,221,676 bp that contained 4,428 protein-coding genes, with a guanine and cytosine (GC) content of 43.41%. Gene functions were annotated using the National Center for Biotechnology Information (NCBI) non-redundant protein, Swiss-Prot, Kyoto Encyclopedia of Genes and Genomes, Clusters of Orthologous Groups of proteins, Gene Ontology, Pathogen–Host Interactions, Carbohydrate-Active enZYmes, and Rapid Annotations using Subsystem Technology databases, revealing non-ribosomal pathways associated with antifungal mechanisms, biofilm formation, chemotactic motility, VOC 3-hydroxy-2-butanone, cell wall-associated enzymes, and synthesis of various secondary metabolites. antiSMASH analysis predicted that CTXW 7-6-2 can produce the active substances bacillaene, bacillibactin, subtilosin A, bacilysin, and luminmide and has four gene clusters of unknown function. Quantitative real-time PCR (qRT-PCR) analysis verified that yvmC and cypX, key genes involved in the production of pulcherrimin, were highly expressed in CTXW 7-6-2. This study elucidates the mechanism by which B. subtilis strain CTXW 7-6-2 inhibits pathogenic fungi that cause kiwifruit soft rot, suggesting the benefit of further studying its antifungal active substances.

Kiwifruit bacterial canker caused by Pseudomonas syringae pv. actinidiae (Psa) is a major threat to kiwifruit worldwide, and effective control measures are still lacking. Sulfur, as a mineral, has been proved to improve plants' resistance to pathogens. It is of great significance to study the effect of sulfur on rhizosphere microorganisms in kiwifruit planting areas infected by Psa for controlling kiwifruit canker. In this study, the sulfur powder and organic fertilizer were mixed as base fertilizer to treat the soil in the area where kiwifruit bacterial canker occurs. We investigated the incidence of kiwifruit bacterial canker in 2018 and 2019 after sulfur application and the changes in microbial characteristics and community composition structure in the kiwifruit rhizosphere by using the plate-counting method and high-throughput sequencing technology. Fertilization treatments of kiwifruit roots with sulfur and organic fertilizer reduced kiwifruit bacterial canker severity. The diversity of soil microbial communities increased significantly after sulfur application in the range of 1.0~2.0 kg/m3. In particular, the bacterial genera level showed a high diversity after 2 years of sulfur application, reaching more than 516 genera. Furthermore, sulfur treatment resulted in a significant increase in specific microbial taxa, including members of the Acidothermus, norank_f__HSB_OF53-F07, and norank_f __Acidobacteriaceae__Subgroup_1. Moreover, the proportion of the dominant bacteria Acidothermus in the population showed an increasing trend. Altogether, the sulfur application is the key factor leading to microbial differences in kiwifruit rhizosphere soil. Appropriate sulfur can improve microbial structure characteristics of kiwifruit rhizosphere soil, increase bacterial diversity index, and change bacterial community composition structure.

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.

Botryosphaeria dothidea is the source of the deadly kiwifruit disease known as soft rot. In order to explore the role of melatonin in regulating the postharvest quality and disease resistance of kiwifruit at different growth and development stages, in this study, we applied melatonin at different concentrations to kiwifruit at the young fruit, expansion, and late expansion stages to assess its effect on fruit resistance to B. dothidea, minimize soft rot, and maintain postharvest fruit quality. The results showed that melatonin significantly suppressed the mycelial growth of B. dothidea, with 1.0 mmol/L melatonin inhibiting it by up to 50%. However, 0.1–0.3 mmol/L melatonin had the best control over soft rot. Furthermore, spraying MT during kiwifruit growth can successfully increase fruit weight; preserve postharvest fruit firmness; reduce respiration intensity in the early stages of storage; delay the rise in soluble solids, while maintaining a high titratable acid content to ensure suitable solid acid ratio; increase total phenol, flavonoid, chlorophyll, carotenoid, and ascorbic acid contents; and delay the rise in soluble sugar contents in the late stages of storage. These results have a positive effect on maintaining the nutritional composition of kiwifruit. However, the effects on weight loss, dry matter content, and soluble protein content were not significant. In addition, the results of the principal component analysis demonstrated that 0.3 mmol/L MT increased kiwifruit’s resistance to soft rot while preserving postharvest fruit quality.

Magnolol is a natural compound extracted from the traditional Chinese medicine Magnolia officinalis, which exhibits antimicrobial properties. However, magnolol is insoluble in water and consists of a phenolic hydroxyl group, which is volatile; these factors hinder its application. In this study, a safe and environmentally friendly method to improve the microbial resistance and storability of harvested fruits is developed using the water-soluble carrier carboxymethyl chitosan (CMCS) and magnolol. Magnolol was loaded on CMCS particles to form Magnolol@CMCS antimicrobial particles, a preservation coating agent. Magnolol@CMCS particles effectively solved the problems of water insolubility and agglomeration of magnolol and reduced the size distribution D50 value of magnolol from 0.749 to 0.213 μm. Magnolol@CMCS particles showed greater toxicity against Staphylococcus aureus, Escherichia coli, and Botryosphaeria dothidea than that of magnolol alone, with effective medium concentration (EC50) values of 0.9408, 142.4144, and 8.8028 μg/mL, respectively. Kiwifruit treated with the Magnolol@CMCS solution showed delayed changes in fruit hardness and soluble solid and dry matter contents and significantly higher ascorbic acid (vitamin C) and soluble total sugar contents and sugar:acid ratios compared with that of the control fruit. In addition, no disease spots were observed on fruit treated with the Magnolol@CMCS solution within 7 days after inoculation with B. dothidea. In conclusion, Magnolol@CMCS particles showed antimicrobial activity on harvested fruits, effectively delayed the hardness and nutritional changes of fruits during storage, and improved the storability of 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.

Botryosphaeria dothidea is one of the most frequent pathogens of soft rot disease in kiwifruit. The aim of this study was to investigate the role of oligosaccharins used together with tebuconazole to control soft rot and their influences on kiwifruit’s disease resistance, growth and quality. The results show that tebuconazole displayed a toxicity against B. dothidea RF-1 with 0.87 mg kg−1 of EC50 value. Oligosaccharins used together with tebuconazole effectively managed soft rot with 84.83% of the field management effect by spraying tebuconazole + oligosaccharins (0.5:0.5, m/v) as a 5000-fold dilution liquid, which significantly (p < 0.01) exceeded the 72.05%, 52.59%, 62.17% and 33.52% effect of tebuconazole 2500-, oligosaccharins 2500-, tebuconazole 5000- and oligosaccharins 5000-fold liquids, respectively. Simultaneously, co-application of tebuconazole and oligosaccharins was more effective for enhancing the resistance, growth and quality of kiwifruit compared with tebuconazole or oligosaccharins alone. This work highlights that oligosaccharins used together with tebuconazole can be proposed as a practicable measure for managing kiwifruit soft rot and reducing the application of chemical synthetic fungicides.

Bacterial canker disease has become the largest threat to kiwifruit cultivation and production. A monomorphic subpopulation of Pseudomonas syringae pv. actinidiae biovar 3 (Psa3) is responsible for the pandemic worldwide. Diversity in pathogenicity has been found in the pandemic subpopulation and in other Psa3 subpopulations causing epidemics in China. However, the genetic bases have not yet been elucidated. In this study, 117 Psa3 isolates were identified by Psa- and Psa3-specific primers, and evaluated for pathogenicity. Three isolates G4, G40, and S2 are not pathogenic to kiwifruit and do not elicit hypersensitivity responses (HRs) in non-host Nicotiana benthamiana leaves. Two isolates, G25 and G35, exhibited attenuated HR-eliciting activity in non-host N. benthamiana, but they exhibited greatly and slightly reduced pathogenicity in host plants, respectively. The genomes of the five isolates were sequenced and compared with closely related isolates revealed by MLVA and whole-genome typing methods. The candidate genetic loci responsible for the changes in pathogenicity and HR elicitation, were further evaluated by allele replacement experiments. We found that the three non-pathogenic isolates were formed due to the independent, identical insertion events of ISPsy36 transposon in the hrpR gene, encoding a key regulator of type III secretion system (T3SS) and type III effectors (T3Es). In the symptomatic sample from which G4 was isolated, 27% HR negative isolates were detected. In isolate G25, transposon insertion of ISPsy32 at the non-coding sequence upstream of the hrpR gene was detected, similar to a previously reported low-virulent Psa3 strain M227. In isolate G35, we detected disruptions of T3Es hopBB1-1 and hopBB1-2, which induce HR in N. benthamiana leaves revealed by Agrobacterium tumefaciens infiltration. These phenotype-changed isolates were formed at low frequencies during the course of pathogen infection in host plants, supported by the binding assay of ISPsy32 and the non-coding DNA sequences upstream of the hrpR gene, the co-isolation of the virulent isolates belonging to the same MLVA clade, and the low levels of transcription of the transposon genes. Taken together, in terms of short-term field evolution, transposon insertions in the T3SS-related genes resulted in the formation of non-pathogenic and low-virulent Psa3 isolates.

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.

Soft rot disease caused by Botryosphaeria dothidea and Phomopsis sp. is the most serious fungal disease of the kiwifruit production area in southwest China. In this work, the role of the co-application of tetramycin and matrine in the resistance of kiwifruit fruits against soft rot disease and its effects on development, quality and amino acids of kiwifruit fruits were investigated. The results indicate that matrine exhibited an outstanding toxicity against B. dothidea RF-1 and Phomopsis sp. RF-2 with EC50 values of 0.442 and 0.332 mg kg−1. The foliar co-application of 0.3% tetramycin aqueous solutions (AS) 5000-fold liquid + 0.5% matrine AS 1000-fold liquid could effectively control soft rot disease with a control efficacy of 82.68%, which was significantly (p < 0.05) higher than 75.19% of 0.3% tetramycin AS 5000-fold liquid and significantly (p < 0.01) higher than 68.50% of 0.5% matrine AS 500-fold liquid. Moreover, the co-application of tetramycin and matrine was more effective than tetramycin or matrine alone in improving disease resistance, quality and amino acids of kiwifruit fruits. This study highlights that the co-application of tetramycin and matrine can be used as a practicable, cost-effective and environmentally friendly candidate or alternative approach for controlling soft rot disease of kiwifruit.

Leaf spot disease caused by Lasiodiplodia theobromae and Alternaria tenuissima is a seriously fungal disease in kiwifruit production. In this study, the co-application of tetramycin and chitosan against leaf spot disease in kiwifruit and its effects on the disease resistance, photosynthesis, quality and amino acids of kiwifruit were investigated. The results show that tetramycin exhibited an excellent antifungal activity against L. theobromae and A. tenuissima with EC50 values of 2.37 and 0.16 mg kg−1. In the field, the foliar co-application of tetramycin and chitosan could effectively control leaf spot disease with control efficacy of 89.44% by spraying 0.3% tetramycin aqueous solutions (AS) 5000 time liquid + chitosan 100 time liquid, which was significantly (ANOVA, p < 0.01) higher than 79.80% of 0.3% tetramycin AS 5000 time liquid and 56.61% of chitosan 100 time liquid. Simultaneously, the co-application of tetramycin and chitosan was more effective than tetramycin or chitosan alone in enhancing the disease resistance and photosynthesis of kiwifruit leaves, as well as improving the quality and amino acids of kiwifruit fruits. This work highlights that chitosan is a practicable, cost-effective and eco-friendly adjuvant of tetramycin for controlling leaf spot disease of kiwifruit, enhancing resistance and photosynthesis of leaves and improving quality and amino acids of fruits.

Leaf spot disease caused by Lasiodiplodia theobromae is one of the most serious fungal diseases of kiwifruit production. In this work, the co-application of isopyrazam·azoxystrobin and chitosan against leaf spot disease in kiwifruit and its effects on disease resistance, photosynthesis, yield, quality, and amino acids of kiwifruit were investigated. The results show that isopyrazam·azoxystrobin exhibited a superior bioactivity against L. theobromae with an EC50 value of 0.1826 mg kg−1. The foliar application of chitosan could effectively enhance isopyrazam·azoxystrobin against leaf spot disease with a field control efficacy of 86.83% by spraying 29% isopyrazam·azoxystrobin suspension concentrate (SC) 1500 time + chitosan 100-time liquid, which was significantly (p < 0.05) higher than 78.70% of 29% isopyrazam·azoxystrobin SC 1000-time liquid. The co-application of isopyrazam·azoxystrobin and chitosan effectively enhanced soluble protein, resistance enzymes’ activity in kiwifruit leaves, and reduced their malonaldehyde (MDA), as well as reliably improved their photosynthetic characteristics. Simultaneously, their co-application was more effective in promoting growth, quality, and amino acids of kiwifruit fruits compared to isopyrazam·azoxystrobin or chitosan alone. This study highlights that the co-application of isopyrazam·azoxystrobin and chitosan can be used as a green, safe, and efficient approach for controlling leaf spot disease of kiwifruit and reducing the application of chemical fungicides.

Kiwifruit is a nutritious and economically important fruit that is widely cultivated in China. In 2021, leaf spot disease of kiwifruit was discovered in the main kiwifruit-producing area of Xifeng County, Guizhou Province, China. Leaf spot disease weakens plant photosynthesis and reduces nutrient synthesis, thereby affecting plant growth. We studied the morphological characteristics and performed a combined analysis of EF-1α, RPB2, and TUB2 genes of Fusarium fujikuroi, a fungus associated with leaf spot disease. The pathogenicity of F. fujikuroi followed Koch’s hypothesis, confirming that this fungus is the cause of kiwifruit leaf spot disease. The sensitivity of seven natural antifungal agents against F. fujikuroi was measured using the mycelial growth rate method. Honokiol, cinnamaldehyde, and osthol showed good antifungal effects against F. fujikuroi, with EC50 values of 18.50, 64.60, and 64.86 μg/mL, respectively. The regression coefficient of cinnamaldehyde was the largest at 2.23, while that of honokiol was the smallest at 0.408. Fusarium fujikuroi was the most sensitive to cinnamaldehyde.

Passion fruit (Passiflora edulis) is a tropical and subtropical plant that is widely cultivated in China due to its high nutritional value, unique flavor and medicinal properties. In August 2020, typical anthracnose symptoms with light brown and water-soaked lesions on Passiflora edulis Sims were observed, which result in severe economic losses. The incidence of this disease was approximately 30%. The pathogens from the infected fruit were isolated and purified by the method of tissue isolation. Morphological observations showed that the colony of isolate BXG-2 was gray to celadon and grew in concentric circles. The orange conidia appeared in the center after 14 days of incubation. The pathogenicity was verified by Koch’s postulates. The internal transcribed spacer (ITS), chitin synthase (CHS-1), actin (ACT), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were amplified by relevant PCR programs. The multi-gene (ITS, GAPDH, ACT, CHS-1) phylogeny analysis confirmed that isolate BXG-2 belongs to Colletotrichum fructicola. The inhibitory effect of six synthetic fungicides on the mycelial growth of the pathogen was investigated, among which difenoconazole 10% WG showed the best inhibitory effect against C. fructicola with an EC50 value of 0.5579 mg·L−1. This is the first report of anthracnose on Passiflora edulis Sims caused by Colletotrichum fructicola in China.

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.

Drought, low nutrition, and weeds have become the major limiting factors of young kiwifruit orchards. In this study, the effects of intercropping Vicia sativa L. on the moisture, microbe community, enzyme activity, and nutrients in rhizosphere soils of young kiwifruit plants and their growth were investigated. The results show that intercropping V. sativa could effectively enhance soil moisture by 1.39–1.47 folds compared with clean tillage. Moreover, intercropping V. sativa could significantly (p < 0.01) increase the microbial community, enzyme activity and nutrient of kiwifruit rhizosphere soils, and improve plant height, stem girth, leaf number, maximum leaf length, maximum leaf width, and chlorophyll content of young kiwifruit plants by 43.60%, 18.68%, 43.75%, 18.09%, 21.15%, and 67.57% compared to clean tillage, respectively. The moisture, microbial quantity, enzyme activity, and nutrients in rhizosphere soils of young kiwifruit plants exhibited good correlations with their plant height, stem girth, leaf number, maximum leaf length, maximum leaf width, and chlorophyll content. This study highlights that intercropping V. sativa in young kiwifruit orchard can be used as an effective, labor-saving, economical and sustainable practice to improve the moisture, microbial community, enzyme activity, and nutrient of soils, and enhance kiwifruit plant growth and control weeds.

In this study, the co–application of chitosan and tetramycin against kiwifruit soft rot and its effects on the disease resistance, growth, quality and aroma of kiwifruit were investigated. The results show that chitosan could effectively enhance tetramycin against soft rot of kiwifruit with the field control efficacy of 85.33% for spraying chitosan 100 time + 0.3% tetramycin AS 5000–time dilution liquid, which was higher than 80.99% for 0.3% tetramycin AS 5000–time dilution liquid and significantly (p < 0.01) higher than 40.66% for chitosan 100–time dilution liquid. Chitosan could significantly (p < 0.05) improve the promoting effects of tetramycin on total phenolics, total flavonoids, SOD activity of kiwifruit compared to tetramycin during storage for 0–28 days and enhance the disease resistance of kiwifruit. Moreover, the co–application of chitosan and tetramycin was more effective than tetramycin or chitosan alone in enhancing fruit growth, improving fruit quality and increasing fruit aroma. This study highlights that chitosan can be used as an adjuvant to enhance tetramycin against soft rot of kiwifruit and promote tetramycin’s improvement for the single fruit volume and weight, vitamin C, soluble sugar, soluble solid, dry matter, soluble protein, titratable acidity and aroma of kiwifruit.

Tetramycin, a novel polyene agriculture antibiotic, has excellent antimicrobial activity against many plant pathogens. In this study, the antimicrobial activities of tetramycin and conventional antibiotics on eight common pathogens and their field control efficacies against four serious diseases in kiwifruit were investigated. The results show that 0.3% tetramycin aqueous solutions (AS) exhibited the superior antibacterial and antifungal activity against Pseudomonas syringae pv. actinidiae, Pseudomonas fulva, Agrobacterium tumefaciens, Botryosphaeriadothidea, Phomopsis sp., Alternaria tenuissima, Armillariella mellea and Phytophthora cactorum of kiwifruit pathogens with EC50 values of 1.21, 1.24, 0.72, 0.14, 0.09, 0.16, 0.06 and 0.17 mg kg−1, respectively. These EC50 values of tetramycin were much higher than those of conventional kasugamycin, zhongshengmycin or polyoxin. Meanwhile, 0.3% tetramycin AS possessed the good field control efficacies for canker, soft rot, blossom blight and brown spot disease of kiwifruit with 74.45, 83.55, 84.74 and 89.62%. Moreover, 0.3% tetramycin AS application notably increased fruit resistance substances contents, activated fruit superoxide dismutase and polyphenoloxidase activities, as well as remarkably enhanced fruit growth, improved fruit quality and storability. This study highlights that tetramycin can be used as a preferred alternative to conventional antibiotics in kiwifruit production.

Soft rot caused by Botryosphaeria dothidea and Phomopsis sp. is a critical disease in kiwifruit. In order to efficiently control soft rot, a 28.6% chitosan composite film (CCF) containing chitosan, dextrin, ferulic acid, calcium, and auxiliaries was successfully developed. The results showed that CCF had a strong inhibitory effect on mycelia growth of B. dothidea and Phomopsis sp., with mycelial EC50 values of 68.11 and 50.34 mg L−1, respectively. The concentration of 0.71−1.42 g L−1 CCF had noticeably preventive and curative effects against soft rot. The spray of CCF before pathogen infection effectively reduced the incidence of soft rot, remarkably increased the content of resistance compounds, and activated the activity of defense enzymes. Moreover, it notably enhanced the yield and quality and prolonged the shelf life of kiwifruit. Therefore, the excellent control effects of CCF against soft rot might be associated with its film-forming property and antifungal activity, which prevent infection and induce plant defense mechanisms. The concentration of 0.71−1.42 g L−1 CCF was optimal for the field application before the onset of disease symptoms in plants with B. dothidea and Phomopsis sp.

[Objective] This study aimed to investigate the resistance to different fungicides in Phytophthora parasitica var. nicotianae. [Method] Under indoor incubation conditions, the resistance to dimethomorph, metalaxyl-mancozeb, propamocarb and ovraclostrobin-dimethomorph in P. parasitica strain isolated from Zhenyuan County in Qiandongnan State was analyzed with colony growth measurement method. [Result] P. parasitica exhibited different levels of sensitivity to four fungicides. To be specific, P. parasitica exhibited the highest resistance to dimethomorph, and EC50 reached 1.19 pg/ml. [Conclusion] In Zhenyuan tobacco-growing area, long-term single use of dimethomorph possesses certain resistance risk in prevention and control of black shank disease. [ABSTRACT FROM AUTHOR]

The type 111 secretion system (T3SS) is a key factor in the pathogenesis Moreof Pseudomonas syringae pv. actinidiae biovar 3( Ps£13 ). the causal agent of a global kiwifruit bacterial canker pandemic. To monitor the T3SS expression levels in Ps£,3, we constructed a luciferase reporter plasmid-expressing HrpApm, 3-NLuc fusion protein. The expression of HrPA-NLuc was induced in h, p-inducing conditions whereas the level of luciferase naactivity correlated with the expression of h,-p/h·c· genes in Ps£13 confirmed the reliability of the reporter construct. Based on the readout of the NLuc reporter construct, three small molecule compounds 4-methoxy-cinnamic acid, sulforaphane, and ferulic acid were determined as T3SS inhibitors in Psal whereas sodium acetate was determined to be a T3SS inducer. Moreof over, the aqueous extract of fruit inhibited the accumulation of HrpA - NLuc iii Psa3 in medium and in planta. Additionally, the T3SS inhibitors suppress Psa3 vii·ulence, whereas the T3SS inducer promotes Ps£13 virulence on kiwifruit. Thus, our findings may provide clues to why the fi-uit is not infected by P.,·a3. and the Psa3 TJSS inhibitors have potential as alter naactivity tives to current nonspecific antimicrobials for disease management. [ABSTRACT FROM AUTHOR]