Soft Rot Research Articles

Comparative genomic and transcriptome analyses of two Pectobacterium brasiliense strains revealed distinct virulence determinants and phenotypic features.

Potato soft rot caused by Pectobacterium spp. are devastating diseases of potato which cause severe economic losses worldwide. Pectobacterium brasiliense is considered as one of the most virulent species. However, the virulence mechanisms and pathogenicity factors of this strain have not been fully elucidated. Here, through pathogenicity screening, we identified two Pectobacterium brasiliense isolates, SM and DQ, with distinct pathogenicity levels. SM exhibits higher virulence compared to DQ in inducing aerial stem rot, blackleg and tuber soft rot. Our genomic and transcriptomic analyses revealed that SM encodes strain specific genes with regard to plant cell wall degradation and express higher level of genes associated with bacterial motility and secretion systems. Our plate assays verified higher pectinase, cellulase, and protease activities, as well as fast swimming and swarming motility in SM. Importantly, a unique endoglucanase S specific to SM was identified. Expression of this cellulase in DQ greatly enhances its virulence compared to wild type strain. Our study sheds light on possible determinants causing different pathogenicity of Pectobacterium brasiliense species with close evolutionary distance and provides new insight into the direction of genome evolution in response to host variation and environmental stimuli.

A dual-branch selective attention capsule network for classifying kiwifruit soft rot with hyperspectral images

Kiwifruit soft rot is highly contagious and causes serious economic loss. Therefore, early detection and elimination of soft rot are important for postharvest treatment and storage of kiwifruit. This study aims to accurately detect kiwifruit soft rot based on hyperspectral images by using a deep learning approach for image classification. A dual-branch selective attention capsule network (DBSACaps) was proposed to improve the classification accuracy. The network uses two branches to separately extract the spectral and spatial features so as to reduce their mutual interference, followed by fusion of the two features through the attention mechanism. Capsule network was used instead of convolutional neural networks to extract the features and complete the classification. Compared with existing methods, the proposed method exhibited the best classification performance on the kiwifruit soft rot dataset, with an overall accuracy of 97.08% and a 97.83% accuracy for soft rot. Our results confirm that potential soft rot of kiwifruit can be detected using hyperspectral images, which may contribute to the construction of smart agriculture.

Response of Chinese cabbage (Brassica rapa subsp. pekinensis) to bacterial soft rot infection by change of soil microbial community in root zone.

Chinese cabbage, scientifically known as Brassica rapa subsp. pekinensis, is a highly popular vegetable in China for its delectable taste. However, the occurrence of bacterial soft rot disease poses a significant threat to its growth and overall development. Consequently, this study aimed to explore the defense mechanisms employed by Chinese cabbage against bacterial soft rot disease. Specifically, the investigation focused on understanding the relationship between the disease and the microbial communities present in the soil surrounding the roots of Chinese cabbage. Significant disparities were observed in the composition of microbial communities present in the root-zone soil of healthy Chinese cabbage plants compared to those affected by Pectobacterium brasiliense-caused soft rot disease. The analysis of 16S rRNA gene high-throughput sequencing results revealed a lower abundance of Proteobacteria (8.39%), Acidobacteriot (0.85), Sphingomonas (3.51%), and Vicinamibacteraceae (1.48%), whereas Firmicutes (113.76%), Bacteroidota (8.71%), Chloroflexi (4.89%), Actinobacteriota (1.71%), A4b (15.52%), Vicinamibacterales (1.62%), and Gemmatimonadaceae (1.35%) were more prevalent in healthy plant soils. Similarly, the analysis of ITS gene high-throughput sequencing results indicated a reduced occurrence of Chytridiomycota (23.58%), Basidiomycota (21.80%), Plectosphaerella (86.22%), and Agaricomycetes (22.57%) in healthy soils. In comparison, Mortierellomycota (50.72%), Ascomycota (31.22%), Podospora (485.08%), and Mortierella (51.59%) were more abundant in healthy plant soils. In addition, a total of 15 bacterial strains were isolated from the root-zone soil of diseased Chinese cabbage plants. These isolated strains demonstrated the ability to fix nitrogen (with the exception of ZT20, ZT26, ZT41, ZT45, and ZT61), produce siderophores and indole acetic acid (IAA), and solubilize phosphate. Notably, ZT14 (Citrobacter freundii), ZT33 (Enterobacter cloacae), ZT41 (Myroides odoratimimus), ZT52 (Bacillus paramycoides), ZT58 (Klebsiella pasteurii), ZT45 (Klebsiella aerogenes), and ZT32 (Pseudomonas putida) exhibited significant growth-promoting effects as determined by the plant growth promotion (PGP) tests. Consequently, this investigation not only confirmed the presence of the soft rot pathogen in Chinese cabbage plants in Hangzhou, China, but also advanced our understanding of the defense mechanisms employed by Chinese cabbage to combat soft rot-induced stress. Additionally, it identified promising plant-growth-promoting microbes (PGPMs) that could be utilized in the future to enhance the Chinese cabbage industry.

MzmL, a novel marine derived N-acyl homoserine lactonase from Mesoflavibacter zeaxanthinifaciens that attenuates Pectobacterium carotovorum subsp. carotovorum virulence.

Quorum sensing (QS) is a conserved cell-cell communication mechanism widely distributed in bacteria, and is oftentimes tightly correlated with pathogen virulence. Quorum quenching enzymes, which interfere with QS through degrading the QS signaling molecules, could attenuate virulence instead of killing the pathogens, and thus are less likely to induce drug resistance. Many Gram-negative bacteria produce N-acyl homoserine lactones (AHLs) for interspecies communication. In this study, we isolated and identified a bacterial strain, Mesoflavibacter zeaxanthinifaciens XY-85, from an Onchidium sp. collected from the intertidal zone of Dapeng Reserve in Shenzhen, China, and found it had strong AHL degradative activity. Whole genome sequencing and blast analysis revealed that XY-85 harbors an AHL lactonase (designated MzmL), which is predicted to have an N-terminal signal peptide and share the "HXHXDH" motif with known AHL lactonases belonging to the Metallo-β-lactamase superfamily. Phylogenetic studies showed MzmL was closest to marine lactonase cluster members, MomL and Aii20J, instead of the AiiA type lactonases. Ultra performance liquid chromatography-mass spectrometry analysis confirmed that MzmL functions as an AHL lactonase catalyzing AHL degradation through lactone hydrolysis. MzmL could degrade both short- and long-chain AHLs with or without a substitution of oxo-group at the C-3 position, and retained full bioactivity under a wide range of temperatures (28-100°C) and pHs (4-11). Furthermore, MzmL significantly reduced Pectobacterium carotovorum subsp. carotovorum virulence factor production in vitro, such as biofilm formation and plant cell wall degrading enzyme production, and inhibited soft rot development on potato slices. These results demonstrated that MzmL may be a novel type of AHL lactonase with good environmental stability, and has great potential to be developed into a novel biological control agent for bacterial disease management.

The biocontrol potentiality of Bacillus amyloliquefaciens against postharvest soft rot of tomatoes and insights into the underlying mechanisms

Tomatoes can easily be affected by soft rot, one of the main postharvest infections caused by a bacterium called Pectobacterium carotovorum. This study examined the soft rot control effectiveness by Bacillus amyloliquefaciens in tomatoes, and identify the underlying action mechanisms from the perspective of the inhibition effects of antibacterial metabolites derived from B. amyloliquefaciens on pathogen and the influences of this antagonist on reactive oxygen species (ROS) scavenging capability of the fruit. The results demonstrated that B. amyloliquefaciens effectively lessened postharvest soft rot occurrence in tomatoes. The non-volatile metabolites synthesized by B. amyloliquefaciens suppressed P. carotovorum subsp. brasiliense (Pcb) growth in vitro and in vivo. The antibacterial ingredients presented in methanol-soluble sediments obtained from non-volatile metabolites. UHPLC-ESI-MS spectra revealed that the primary antibacterial ingredients were C12-C15 surfactin analogues, along with C14-C16 iturins and C16-C17 fengycins. Moreover, B. amyloliquefaciens could raise the activity of enzymatic ROS scavenging system in tomatoes, including several antioxidases and the enzymes related to glutathione peroxidase (GPX) and ascorbate-glutathione (AsA-GSH) cycles. In addition, B. amyloliquefaciens increased the levels of non-enzymatic system for ROS scavenging, including AsA and GSH. Together, O2-·production rate, H2O2 and malondialdehyde (MDA) levels were reduced. It suggested that B. amyloliquefaciens enhanced the ROS scavenging ability of tomatoes, thereby improving their resistance to the pathogen.

First Report of Postharvest Fruit Rot Caused by Neofusicoccum parvum on Prunus salicina (Nai Plum) in China.

Nai plum (Prunus salicina var. cordata cv. Younai) is one of the most popular fruit crop in South China. In July 2023, a fruit rot of nai plum with about 5 % disease incidence was observed in a fruit market of Changsha city, Hunan Province, China. Initially, small, brown lesions appeared randomly on the fruit surface, with disease progression, the lesions gradually expanded and developed into soft rot. To isolate possible fungi from rotten fruits, small pieces (2 × 2 mm) from the periphery of 10 infected fruits were surface-sterilized using 70% ethanol for 10 s, rinsed three times in sterile distilled water, air dried, and then placed onto potato dextrose agar (PDA) plates and incubated at 28℃ for three days. Emerging colonies were subcultured by hyphal tiptransfer on fresh PDA. A total of ten isolates with similar morphology were obtained. Fungal colonies were initially white, gradually turning gray and eventually becoming black, and aerial hyphae were dense and fluffy. Conidia were hyaline, single celled, ellipsoidal to fusiform, and range from 12.7 to 20.0 μm long (avg. 16.9 ± 2.39 μm) × 5.3 to 7.3 μm wide (avg. 6.3 ± 0.82 μm). These morphological characteristics of these isolates matched those of Neofusicoccum parvum (Phillips et al. 2013). To future confirmation of the identify, the internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (TEF1-a), and beta-tubulin TUB2) genes of two representative isolates (JXNP1 and JXNP2) were amplified and sequenced using primer sets ITS5/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999; Phillips et al. 2013), and BT2A/BT2B (Glass and Donaldson 1995), respectively. The sequences of both isolates were deposited in GenBank for the ITS (accession nos. OR899331 and OR899332), TEF1-a gene (accession nos. OR909890 and OR909891) and TUB2 gene (accession nos. OR909892 and OR909893). BLAST analysis showed 99-100% identity with the ex-type strain of N. parvum (CMW9081) for ITS, TEF1-a and TUB2. A maximum likelihood phylogenetic tree was constructed using IQtree web server based on combined ITS, TEF1-a and TUB2 data set. The phylogenetic tree revealed that two isolates clustered with N. parvum in a clade with 90% bootstrap support. Based on morphological and molecular data analysis, the isolates were identified as N. parvum. To confirm the pathogenicity, five healthy nai plum fruits were wounded by using a sterile needle after surface sterilization with 75% ethanol, then a 5-mm-diameter mycelial disc of isolate JXNP1 was taped to the wound, the control fruits were taped with sterile agar plugs. All fruits were incubated at 25 ℃ with 80% humidity. After five days, typical naturally occurring fruit rot symptoms appeared on the fruits which inoculated with N. parvum, whereas control fruits remained asymptomatic. To fulfill Koch's postulates, the pathogen was re-isolated from the inoculated fruits and comfirmed as N. parvum by morphological and molecular analysis. Previous studies reported that N. parvum caused fruit rot on various common fruits in China, including loquat, kiwifruit and citrus (Lei et al. 2013; Zhai et al. 2019; Zhou et al. 2013). To our knowledge, this is the first report of N. parvum causing postharvest fruit rot on nai plum in China. This finding provides critical insights for the management of the high-risk disease on plum in China.

Role of Volatile Organic Compounds Produced by Kosakonia cowanii Cp1 during Competitive Colonization Interaction against Pectobacterium aroidearum SM2.

The competitive colonization of bacteria on similar ecological niches has a significant impact during their establishment. The synthesis speeds of different chemical classes of molecules during early competitive colonization can reduce the number of competitors through metabolic effects. In this work, we demonstrate for the first time that Kosakonia cowanii Cp1 previously isolated from the seeds of Capsicum pubescens R. P. produced volatile organic compounds (VOCs) during competitive colonization against Pectobacterium aroidearum SM2, affecting soft rot symptoms in serrano chili (Capsicum annuum L.). The pathogen P. aroidearum SM2 was isolated from the fruits of C. annuum var. Serrano with soft rot symptoms. The genome of the SM2 strain carries a 5,037,920 bp chromosome with 51.46% G + C content and 4925 predicted protein-coding genes. It presents 12 genes encoding plant-cell-wall-degrading enzymes (PCDEWs), 139 genes involved in five types of secretion systems, and 16 genes related to invasion motility. Pathogenic essays showed soft rot symptoms in the fruits of C. annuum L., Solanum lycopersicum, and Physalis philadelphica and the tubers of Solanum tuberosum. During the growth phases of K. cowanii Cp1, a mix of VOCs was identified by means of HS-SPME-GC-MS. Of these compounds, 2,5-dimethyl-pyrazine showed bactericidal effects and synergy with acetoin during the competitive colonization of K. cowanii Cp1 to completely reduce soft rot symptoms. This work provides novel evidence grounding a better understanding of bacterial interactions during competitive colonization on plant tissue, where VOC synthesis is essential and has a high potential capacity to control pathogenic microorganisms in agricultural systems.

Elicitation of salicylic acid and methyl jasmonate provides molecular and physiological evidence for potato susceptibility to infection by Erwinia carotovora subsp. carotovora

Among the range of severe plant diseases, bacterial soft rot caused by Erwinia carotovora is a significant threat to crops. This study aimed to examine the varying response patterns of distinct potato cultivars to the influence of E. carotovora. Furthermore, it seeks to highlight the potential role of salicylic acid (SA) and methyl jasmonate (MeJA) in stimulating the antioxidant defence system. We collected eight bacterial isolates from diseased and rotted tubers which were morphologically and physiologically identified as E. carotovora subsp. carotovora. We conducted a greenhouse experiment to analyse the antioxidant responses of three different potato cultivars (Diamont, Kara, and Karros) at various time intervals (2, 4, 6, 8, 12, and 24 h) after bacterial infection (hpi). We assessed the extent of disease damage by applying a foliar spray of 0.9 mM salicylic acid (SA) and 70 μM methyl jasmonate (MeJA). Inoculating with Ecc led to an increase in total phenolic levels, as well as the activities and gene expression of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO) and peroxidase (POX) as time progressed. Additionally, the application of SA and MeJA resulted in a further increase relative to the diseased treatments. The Karros cultivar, unlike the Diamont and Kara cultivars, demonstrated the highest expression levels of PAL, PPO and POX through inoculation, reflecting its higher levels of activity and resistance. Furthermore, the genetic response of potato cultivars to infection at 0 hpi varied depending on their susceptibility. The examination of the rate of PAL activity upregulation following SA or MeJA stimulation clarifies the cultivars' susceptibility over time. In conclusion, the study identified E. carotovora subsp. carotovora as the most virulent isolate causing soft rot disease in potato tubers. It further revealed that the Karros cultivar displayed superior resistance with high activities and gene expression of PAL, PPO and POX, while the cv. Diamont exhibited sensitivity. Additionally, foliar exposure to SA and MeJA induced antioxidant responses, enhancing the potato plants' resistance against Ecc pathogenesis and overall plant defence.

Characterization and identification of Burkholderia gladioli causing stalk rot of sweet corn in Thailand

AbstractStalk rot disease is becoming an important disease in sweet corn crops because it causes significant yield losses of the crop. In July 2022, typical symptoms of stem rot were observed in fields of Mae Hia Agricultural Research, Demonstration and Training Center in Chiang Mai province of Thailand. The symptoms exhibited by affected crop included circular water‐soaked lesions, rot and necrotic tissue at the base of the stalk. Koch's postulates were fulfilled with reisolation of the inoculated strain from the inoculated tissues, and confirmation was obtained by phenotypic characters and 16S rRNA sequences. The isolated colony on nutrient agar was cream‐coloured, circular, with a convex elevation, entire margin and smooth surface; the optical character was translucent and exuded a yellowish pigment that diffused in the nutrient agar, gram‐negative, lacked arginine dihydrolase, occurrence of the oxidase and catalase reaction. The strains were able to utilize d‐galactose and glucose but were unable to utilize d‐maltose, d‐cellobiose, d‐fructose, d‐mannose, mannitol, lactose and sucrose. The strains showed positive results for gelatine liquefaction test, citrate utilization and casein hydrolysis, but starch hydrolysis and indole production were negative result. These characteristics are the same as those described previously for Burkholderia gladioli. Disease severity in sweet corn was 43.8%, with infections showing spots with dark margins and soft rot on mature leaves and flag leaves. Disease severity in waxy corn and maize were 21.9% and 15.6%, respectively, showing a few light or dark brown spots at the infection sites. Therefore, this represents the first report of leaf stripe and stalk rot disease in sweet corn in Thailand.

Green synthesis of silver nanoparticles by sweet cherry and its application against cherry spot disease

Asia has a rich history of cultivating sweet cherries, a practice that has been carried out since ancient times. However, the effective management of Alternaria disease in sweet cherry crops has presented a formidable challenge, resulting in notable decreases in yield. Various attempts have been made to employ both chemical and biological treatments; however, their effectiveness has been restricted. In order to tackle this problem, an investigation was carried out, with the primary objective of isolating and identifying Alternaria isolates that are accountable for the occurrence of sweet cherry soft spot rot. Out of the twelve isolates examined, the CHM-4 isolate was found to be the most pathogenic. Its identification was achieved through the use of the ITS genomic region (ITS1 and ITS4), and the BLAST results revealed a 95 % similarity with Alternaria alternata (MG744381.1). The objective of the research was to explore the potential of silver nanoparticles (SNPs) synthesized by phytosynthesis as a novel antifungal agent to combat sweet cherry soft spot pathogenicity. The biosynthesis of SNPs was carried out using sweet cherry fruits kernel exudate, which served as an environmentally friendly source. The exudates exhibited the ability to produce nanoparticles with an average size of 24.97 nm. Analysis conducted using a transmission electron microscope (TEM) revealed the multifaceted structure of these nanoparticles. Furthermore, when tested at concentrations of 5, 10, 20, and 40 μg/ml, these biosynthetic nanoparticles demonstrated the capability to inhibit the growth of Alternaria fungi and effectively destroy fungal hyphae. It is advisable to utilize diverse components of sweet cherry for the synthesis of various nanoparticles owing to their compatibility with the surrounding environment.

First Report of Dickeya dadantii Causing Potato Blackleg in Fujian Province China.

Blackleg and soft rot are harmful diseases in potato (Solanum tuberosum) caused by Pectobacterium spp. and Dickeya spp. (Czajkowski et al. 2015). The occurrence of potato blackleg was serious in potato-producing areas around Xiapu County in Fujian Province, China, in 2021 (6 ha) and 2022 (7 ha), with an incidence of approximately 5%, which reached nearly 23%. Three diseased plants were collected to isolate the pathogen. Single colonies from each sampled plant were isolated and streaked onto fresh plates. DNA from three colonies from different plants was PCR amplified with primer pair 27F/1492R (Lane 1991) for the 16S rRNA gene. Since the sequences were identical, we selected strain M2-3 for further analysis. The strain M2-3 was gram-negative, pectolytic on CVP, grew at 37°C and 5% NaCl. The bacterium was positive for phosphatase activity, erythromycin sensitivity, indole production, gelatin liquefaction, malonic utilization, and acid production from, melibiose, raffinose, and arabinose. The bacterium was negative for sucrose, α-methyl glucoside, sorbitol, trehalose, lactose, and sodium citrate (Fujimoto et al. 2018;),although sucrose and lactose did not provide the expected results, there are exception in all species. The genome of strain M2-3 was sequenced and deposited in the NCBI database under accession numbers: CP077422. An Average Nucleotide Identity (ANI) analysis showed that M2-3 clustered with other D. dadantii strains and has a 98.39% identity with D. dadantii strain DSM 18020 (CP023467). The housekeeping genes (recA, dnaX, acnA, gapA, icd, mdh, mtlD and pgi) were amplified with primer pairs designed previously(Fujimoto et al. 2018; Ma et al. 2007) and sequenced. A multilocus sequence analysis (MLSA) was performed by concatenating the 8 gene sequences and constructing a maximum likelihood phylogenetic tree using PhyloSuite version 1.2.1 (Zhang et al. 2020) and IQ-tree version 1.6.8 (Nguyen et al. 2015) software. Strain M2-3 was clustered together with Dickeya dadantii. For the pathogenicity test, three plants per treatment, totaling nine plants, were used. Bacterial suspensions (1×10^8 CFU/mL) were made in a 10mM PBS buffer. 10 μL of M2-3, D. dadantii type strain 18020 (positive control), and buffer (negative control) were injected into the plant stems near the base. Water stains appeared at the site of inoculation after 2 days and they gradually became black and rotten. The leaves became yellow and wilted, and the petiole base rotted within 5 days of inoculation completing the Koch postulate. According to average nucleotide identity and housekeeping gene sequence analysis, strain M2-3 was identified as Dickeya dadantii. Previous studies have reported several pathogens that cause potato blackleg in China, including P. atrosepticum, P. carotovorum, P. brasiliense, P. parmentieri, P. polaris, and P. punjabense (Li-ping et al. 2020; Wang et al. 2021). To the best of our knowledge, this study is the first to report potato blackleg disease caused by Dickeya dadantii in Fujian Province, China. This finding suggests that this pathogen may cause a threat to potato production in Fujian Province.

POSTHARVEST EVOLUTION OF PECTINOLYTIC ERWINIA AND ERWINIA SOFT ROT DURING COLD STORAGE OF CONSUMPTION POTATOES PRODUCED IN ALGERIA

In Algeria, potatoes ranks second among the twenty main food crops. It’s characterized by periods of overproduction corresponding to seasonal crops. Except these periods, supply tends to fall. In order for the availability of potatoes to be constant throughout the non-harvest periods, it is imperative to store the surplus production to ensure a constant supply during periods of unavailability of a new harvest. The storage of the potato must maintain a satisfactory quality and minimize losses and the postharvest development of bacteria and fungi. This work aims to identify and follow the evolution of pectinolytic Erwinia and Erwinia soft rot during storage at 4 and 10° C of two varieties of potato grown in Algeria, i.e. Spunta and Désirée. The results indicated the presence of three species of Erwinia: E. carotovora subsp. carotovora, E. carotovora subsp. atroseptica and E. chrysanthemi. Initially, for both varieties, E. carotovora subsp. carotovora predominates followed by E. carotovora subsp. atroseptica and finally E. chrysanthemi. During storage, the three species evolve almost in the same way and their multiplication is more important at 10° C than at 4° C, particularly on the Spunta. About the total bacteria, the highest levels are recorded on Spunta, especially at 10° C. In addition, the Erwinia spp. / total bacteria ratio showed that Erwinia spp. are very most abundant compared to the total bacteria. The assessment of soft rot losses has also shown that they are greater in the Spunta variety whether at 4 or 10° C.

Surviving the Potato Stems: Differences in Genes Required for Fitness by Dickeya dadantii and Dickeya dianthicola.

Bacteria belonging to the genus Dickeya cause blackleg and soft rot symptoms on many plant hosts, including potato. Although there is considerable knowledge about the genetic determinants that allow Dickeya to colonize host plants, as well as the genes that contribute to virulence, much is still unknown. To identify the genes important for fitness in potato stems, we constructed and evaluated randomly barcoded transposon mutant (RB-TnSeq) libraries of Dickeya dadantii and Dickeya dianthicola. We identified 169 and 157 genes important for growth in D. dadantii and D. dianthicola in stems, respectively. This included genes related to metabolic pathways, chemotaxis and motility, transcriptional regulation, transport across membranes, membrane biogenesis, detoxification mechanisms, and virulence-related genes, including a potential virulence cluster srfABC, c-di-GMP modulating genes, and pectin degradation genes. When we compared the results of the stem assay with other datasets, we identified genes important for growth in stems versus tubers and in vitro conditions. Additionally, our data showed differences in fitness determinants for D. dadantii and D. dianthicola. These data provide important insights into the mechanisms used by Dickeya when interacting with and colonizing plants and thus might provide targets for management.

Pectobacterium araliae sp. nov., a pathogen causing bacterial soft rot of Japanese angelica tree in Japan.

Phytopathogenic bacteria (MAFF 302110T and MAFF 302107) were isolated from lesions on Japanese angelica trees affected by bacterial soft rot in Yamanashi Prefecture, Japan. The strains were Gram-reaction-negative, facultatively anaerobic, motile with peritrichous flagella, rod-shaped, and non-spore-forming. The genomic DNA G+C content was 51.1 mol % and the predominant cellular fatty acids included summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C16 : 0, summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), summed feature 2 (comprising any combination of C12 : 0 aldehyde, an unknown fatty acid with an equivalent chain length of 10.928, C16 : 1 iso I, and C14 : 0 3OH), and C12 : 0. Phylogenetic analyses based on 16S rRNA and gyrB gene sequences, along with phylogenomic analysis utilizing whole-genome sequences, consistently placed these strains within the genus Pectobacterium. However, their phylogenetic positions did not align with any known species within the genus. Comparative studies involving average nucleotide identity and digital DNA-DNA hybridization with the closely related species indicated values below the thresholds employed for the prokaryotic species delineation (95-96 % and 70 %, respectively), with the highest values observed for Pectobacterium polonicum DPMP315T (92.10 and 47.1 %, respectively). Phenotypic characteristics, cellular fatty acid composition, and a repertoire of secretion systems could differentiate the strains from their closest relatives. The phenotypic, chemotaxonomic, and genotypic data obtained in this study show that MAFF 302110T/MAFF 302107 represent a novel species of the genus Pectobacterium, for which we propose the name Pectobacterium araliae sp. nov., designating MAFF 302110T (=ICMP 25161T) as the type strain.

Genome Sequence Resource of a Taro Bacterial Soft Rot Pathogen, Dickeya fangzhongdai ZXC1

Dickeya fangzhongdai ZXC1 is a newly identified highly virulent causal agent of taro bacterial soft rot disease isolated from a taro sample showing typical soft rot symptoms in Shaoguan, Guangdong Province, China. The pathogen produces plant cell wall–degrading enzymes that macerate the taro tube tissues. In this study, we report the whole-genome sequencing analysis of strain ZXC1. The results showed that strain ZXC1 has one circular DNA chromosome of 5,129,951 bp with a 56.59% G+C content. In addition to sharing a conserved zms gene cluster that encodes the genes for biosynthesis of phytotoxin zeamines and 11 copies of predicted pectate lyase genes, the strain ZXC1 genome contains more prophage loci and a higher number of type IV secretion system and type IV secretion system gene clusters than the D. fangzhongdai strains isolated from pears or phalaenopsis, which may account for its strong virulence phenotype. The data from this study present a valuable resource for elucidation of the pathogenic mechanisms of D. fangzhongdai and may aid in developing new disease control approaches to safeguard taro production. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Unraveling disease ecology: insights from soft rot *Pectobacteriaceae* co-infections

Unraveling disease ecology: insights from soft rot *Pectobacteriaceae* co-infections

First report of Pseudomonas marginalis causing tuber soft rot of potato in Iran

In 2016, potato tuber soft rot symptoms were observed in major potato production fields in Jiroft county (southern Kerman province, Iran). Bacteria, which appeared as white to creamy colonies, were isolated from diseased tissues and shown to be pathogenic on potato tubers after inoculation and re-isolation of the isolates. Based on the assessment of colony morphology, biochemical characteristics, and analysis of 16S rRNA and gyrB gene sequences, the pathogen was identified as Pseudomonas marginalis. To our knowledge, this is the first report of P. marginalis causing potato tuber soft rot disease in Iran.

Plant growth-promoting and biocontrol traits of endophytic Bacillus licheniformis against soft rot causing Pythium myriotylum in ginger plant.

Bacterial endophytes from plants harbor diverse metabolites that play major roles in biocontrol and improve plant growth. In this study, a total of 12 endophytic bacteria were isolated from the ginger rhizome. The strain K3 was highly effective in preventing mycelia growth of Pythium myriotylum (78.5 ± 1.5% inhibition) in dual culture. The cell-free extract (2.5%) of endophyte K3 inhibited 76.3 ± 4.8% mycelia growth, and 92.4 ± 4.2% inhibition was observed at a 5% sample concentration. The secondary metabolites produced by Bacillus licheniformis K3 showed maximum activity against Pseudomonas syringae (24 ± 1 mm zone of inhibition) and Xanthomonas campestris (28 ± 3 mm zone of inhibition). The strain K3 produced 28.3 ± 1.7 IU mL-1 protease, 28.3 ± 1.7 IU mL-1 cellulase, and 2.04 ± 0.13 IU mL-1 chitinase, respectively. The ginger rhizome treated with K3 in the greenhouse registered 53.8 ± 1.4% soft rot incidence, and the streptomycin-treated pot registered 78.3 ± 1.7% disease incidence. The selected endophyte K3 improved ascorbate peroxidase (1.37 ± 0.009 µmole ASC min-1 mg-1 protein), catalase (8.7 ± 0.28 µmole min-1 mg-1 protein), and phenylalanine ammonia-lyase (26.2 ± 0.99 Umg-1) in the greenhouse. In addition, K3 treatment in the field trial improved rhizome yield (730 ± 18.4 g) after 180 days (p < 0.01). The shoot length was 46 ± 8.3 cm in K3-treated plants, and it was about 31% higher than the control treatment (p < 0.01). The lytic enzyme-producing and growth-promoting endophyte is useful in sustainable crop production through the management of biotic stress.

Impact of Parvodontia austrosinensis on Lansium domesticum in Northern Thailand

ABSTRACT Lansium domesticum (Osbeck) Sahni and Bennet (Longkong) is a highly important and valuable fruit crop in Thailand. The mycobiota of L. domesticum has been understudied. In our survey of fungal diseases affecting fruit crops in Thailand, we observed unusual mycelial development in organically cultivated L. domesticum in Uttaradit Province. The mycelia initially appear as cords on the upper half of the young trees and spread towards the crown. However, the adverse growth of mycelial cords shows invasive development over the tree and is associated with fruits showing soft rot. The trees covered with mycelia cords eventually decay and die, and the effect was observed to be irreversible. Taxonomic and phylogenetic studies identified the mycelial cord-forming fungal species as Parvodontia austrosinensis. Pathogenicity studies on detached leaves and fruit bunches showed field symptoms confirming P. austrosinensis as the mycelial cord-forming fungi on L. domesticum and pathogen associated with fruit soft rot. This study presents the first geographic and host record of P. austrosinensis in Thailand on L. domesticum. Moreover, it is the first report of P. austrosinensis associated with fruit soft rot and plantation decay. Furthermore, this study is the first report on the disease potential of Cystostereaceae, which was previously known for a saprobic lifestyle.

Control of postharvest soft rot of green peppers by Bacillus subtilis through regulating ROS metabolism

Soft rot caused by Pectobacterium carotovorum is one of the most serious postharvest bacterial diseases of green peppers. The present research aimed to determine the biocontrol effect of Bacillus subtilis on postharvest soft rot of green peppers and to further delve into biocontrol mechanisms by analyzing the metabolism of reactive oxygen species (ROS) in green peppers. As key results, soft rot occurrence of green peppers was alleviated with the increase of the concentration of B. subtilis, and the disease incidence and index of the fruits treated with 1 × 109 CFU/mL of B. subtilis for 4 d were 28.3% and 12.7%, respectively, which were significantly decreased compared with that of the control. B. subtilis could significantly increase NADPH oxidase (NOX) activity in green peppers to trigger ROS outbreaks, which induced defense response of the fruits. Furthermore, it promoted the enzymatic activities of the ROS scavenging system, including antioxidant enzymes (superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)), ascorbic acid-glutathione (AsA-GSH) cycle-related enzymes (ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR)), glutathione peroxidase (GPX) cycle-related enzymes (GPX, GR and glutathione-s-transferase (GST)). Besides, B. subtilis increased the non-enzymatic ROS scavenging system contents, including ascorbate (AsA) and glutathione (GSH) in green peppers. Importantly, the levels of O2−·, H2O2 and malondialdehyde (MDA) in green peppers were reduced, thereby enhancing green pepper disease resistance. This study offers novel insights to advanced strategies in preventing and controlling postharvest soft rot in green peppers.