Bacterial Soft Rot Research Articles

First Report of Bacterial Soft Rot of Potato Caused by Pectobacterium aroidearum strains in China.

Potato (Solanum tuberosum L.) is a globally important staple crop, and China has contributed more than 20% of the world's production. In Guangdong Province, potato has become one of the most important winter crops, increased the farmer income. However, the potato were seriously affected by soft rot disease caused by Pectobacterium spp. in the past decade. In February 2024, typical symptoms of potato soft rot were observed on the plants of cultivar "Daxiyang" in Enping City, Guangdong Province, where the disease incidences ranged 3%-5% in the investigated fields. The stems adjacent to the tubes showed typical inky black symptoms, and the plants appeared yellow. The interior of the tubes appeared water-soaked and had developed to overall decay, with an evident smell. Eleven symptomatic plants were collected and used to isolate the causal agents. Pieces of tube tissue (about 5×5 mm) were removed between the rot-symptomatic and non-symptomatic margins, and surface sterilized using 75% ethanol for 30 s and 2% NaClO for 1 min. The pieces were rinsed in sterile water for three times, and then plated on Luria-Bertani (LB) medium agar for 36 h at 30°C. The produced colonies were selected for hypersensitive test on tobacco, and the colonies with a positive reaction were subcultured three times. Two representative strains, EP51-2 and EP51-3, were processed for gene sequencing, including house-keeping genes, danX, leuS, and recA (Portier et al. 2019). The gene sequences (GenBank accessions no. PP870934 to PP870939) of these two representative strains were matched with Pectobacterium aroidearum strain NCPPB 929 (CP166097.1) with identities ranging from 97.15 to 100% and the coverage of 100%. The sequences of three genes were concatenated and used for phylogenetic analyses, along with representative strains from 19 other Pectobacterium species. Phylogenetic analyses supported that EP51-2 and EP51-3 were grouped into P. aroidearum with the representative strains. Koch's postulates were applied to determine the pathogenicity of P. aroidearum strain EP51-2. According to the external inoculation method on potato stems (Czajkowski et al. 2010), each of six pots of healthy potato plants was inoculated at the basal stem with 100 µl of bacterial suspension (108 CFU/ml), while six pots inoculated with LB served as controls. The plants were then kept at 30°C and maintained at 95% humidity. After 2 days, all six plants inoculated with the bacteria exhibited typical symptoms on the stems, which progressed to the collapse of the whole plant; the controls remained symptom-free during the observation. Single colonies reisolated from the symptomatic stem of the inoculated plants were confirmed using PCR and sequencing with recA primers as described above, and the causal agent was identified as P. aroidearum strains, fulfilling Koch's postulates. P. aroidearum has been reported on Chinese cabbage (Xie et al. 2018) and leaf mustard (Chu et al. 2024) in China, as well as on crops in the family Araceae, such as konjac (Wei et al. 2021), taro (Zhou et al. 2022), and Pinellia ternata (Du et al. 2024). To our knowledge, this is the first report of P. aroidearum causing potato soft rot in China. This pathogen has been prevalent in the taro fields in Guangdong Province and has caused severe losses. This report highlights an expansion of the host range for this pathogen. Attention should be focused on this newly emerging pathogen affecting potatoes, and immediate measures should be implemented to control its spread.

A Mitogen-Activated Protein Kinase Pathway Is Required for Bacillus amyloliquefaciens PMB05 to Enhance Disease Resistance to Bacterial Soft Rot in Arabidopsis thaliana.

When a plant is infected by a pathogen, endogenous immune responses are initiated. When the initiation of these defense responses is induced by a pathogen-associated molecular pattern (PAMP) of a pathogen, it is called PAMP-triggered immunity (PTI). Previous studies have shown that Bacillus amyloliquefaciens PMB05 can enhance PTI signals and improve disease control of bacterial soft rot and wilt in Arabidopsis thaliana. In the context of controlling bacterial wilt disease, the involvement of a mitogen-activated protein kinase (MAPK) signaling pathway has been established. Nevertheless, it remains unclear whether this pathway is also required for B. amyloliquefaciens PMB05 in controlling bacterial soft rot. In this study, A. thaliana ecotype Columbia (Col-0) and its mutants on a MAPK pathway-related pathway were used as a model and established that the ability of B. amyloliquefaciens PMB05 to control soft rot requires the participation of the MAPK pathway. Moreover, the enhancement of disease resistance by PMB05 is highly correlated with the activation of reactive oxygen species generation and stomata closure, rather than callose deposition. The spray inoculation method was used to illustrate that PMB05 can enhance stomatal closure, thereby restricting invasion by the soft rot bacterium. This control mechanism has also been demonstrated to require the activation of the MAPK pathway. This study demonstrates that B. amyloliquefaciens PMB05 can accelerate stomata closure via the activation of the MAPK pathway during PTI, thereby reducing pathogen invasion and achieving disease resistance against bacterial soft rot.

A novel genus of Pectobacterium bacteriophages display broad host range by targeting several species of Danish soft rot isolates

The bacterial diseases black leg and soft rot in potatoes cause heavy losses of potatoes worldwide. Bacteria within the genus Pectobacteriaceae are the causative agents of black leg and soft rot. The use of antibiotics in agriculture is heavily regulated and no other effective treatment currently exists, but bacteriophages (phages) have shown promise as potential biocontrol agents. In this study we isolated soft rot bacteria from potato tubers and plant tissue displaying soft rot or black leg symptoms collected in Danish fields. We then used the isolated bacterial strains as hosts for phage isolation. Using organic waste, we isolated phages targeting different species within Pectobacterium. Here we focus on seven of these phages representing a new genus primarily targeting P. brasiliense; phage Ymer, Amona, Sabo, Abuela, Koroua, Taid and Pappous. TEM image of phage Ymer showed siphovirus morphotype, and the proposed Ymer genus belongs to the class Caudoviricetes, with double-stranded DNA genomes varying from 39 kb to 43 kb. In silico host range prediction using a CRISPR-Cas spacer database suggested both P. brasiliense, P. polaris and P. versatile as natural hosts for phages within the proposed Ymer genus. A following host range experiment, using 47 bacterial isolates from Danish tubers and plants symptomatic with soft rot or black leg disease verified the in silico host range prediction, as the genus as a group were able to infect all three Pectobacterium species. Phages did, however, primarily target P. brasiliense isolates and displayed differences in host range even within the species level. Two of the phages were able to infect two or more Pectobacterium species. Despite no nucleotide similarity with any phages in the NCBI database, the proposed Ymer genus did share some similarity at the protein level, as well as gene synteny, with currently known phages. None of the phages encoded integrases or other genes typically associated with lysogeny. Similarly, no virulence factors nor antimicrobial resistance genes were found, and combined with their ability to infect several soft rot-causing Pectobacterium species from Danish fields, demonstrates their potential as biocontrol agents against soft rot and black leg diseases in potatoes.

Underwood cropping method in preventing soft rot disease of Amorphophallus konjac—from soil properties to rhizosphere microbial communities

Underwood cropping methods have been considered an effective way to prevent bacterial soft rot in Amorphophallus konjac，however, the relationship between rhizosphere soil properties, microbial communities and konjac soft rot under different planting types is not clear. In this study, we assessed the differences in soil nutrients and rhizosphere microorganisms of single-cropping, intercropping, and underwood-cropping konjac plants in an experimental field. The results show that the soil nutrient abundance in the rhizosphere soil and leaves of konjac underwood cropping were the highest. Moreover, the abundance and alpha diversity of root-associated bacteria such as Actinobacteria, Acidobacteria, and Nitrospirae were the most abundant in underwood cropping sites. The presence of these bacteria was significantly positively correlated with soil phosphatase, catalase, organic matter, available nitrogen, and available potassium. All these characteristics greatly improved the konjac resistance to soft rot pathogenic bacteria. This research may provide support for konjac soft rot prevention.

Dickeya fangzhongdai was prevalent and caused taro soft rot when coexisting with the Pectobacterium complex, with a preference for Araceae plants.

Bacterial soft rot caused by coinfection with Dickeya spp. and Pectobacterium spp. in hosts can cause successive changes in fields, and it is difficult to prevent the spread of and control the infection. Pectobacterium spp. are prevalent in the growing areas of tuberous crops, including taro and potato. Recently, Dickeya fangzhongdai has emerged as a virulent pathogen in taro. To determine the prevalence status of the causal agents and evaluate the potential spreading risks of D. fangzhongdai, screening and taxonomic classification were performed on phytopathogenic bacteria collected from different taro-growing areas in Guangdong Province, China, and biological and genomic characteristics were further compared among typical strains from all defined species. The causative agents were verified to be phytobacterial strains of D. fangzhongdai, Pectobacterium aroidearum and Pectobacterium colocasium. P. aroidearum and P. colocasium were found to form a complex preferring Araceae plants and show intensive genomic differentiation, indicating their ancestor had adapted to taro a long time prior. Compared with Pectobacterium spp., D. fangzhongdai was more virulent to taro corms under conditions of exogenous infection and more adaptable at elevated temperatures. D. fangzhongdai strains isolated from taro possessed genomic components of additional T4SSs, which were accompanied by additional copies of the hcp-vgrG genes of the T6SS, and these contributed to the expansion of their genomes. More gene clusters encoding secondary metabolites were found within the D. fangzhongdai strains than within the Pectobacterium complex; interestingly, distinct gene clusters encoding zeamine and arylpolyene were both most similar to those in D. solani that caused potato soft rot. These comparisons provided genomic evidences for that the newly emerging pathogen was potentially equipped to compete with other pathogens. Diagnostic qPCR verified that D. fangzhongdai was prevalent in most of the taro-growing areas and coexisted with the Pectobacterium complex, while the plants enriching D. fangzhongdai were frequently symptomatic at developing corms and adjacent pseudostems and caused severe symptoms. Thus, the emerging need for intensive monitoring on D. fangzhongdai to prevent it from spreading to other taro-growing areas and to other tuberous crops like potato; the adjustment of control strategies based on different pathopoiesis characteristics is recommended.

Native bacterial bioagents for management of potato soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum

BackgroundThe reduced rate of potato production may occur under different conditions such as field, transit, storage, and marketing. Potato cultivation is frequently affected by various pathogens, among which Pectobacterium carotovorum subsp. carotovorum is a notorious bacterial pathogen responsible for causing bacterial soft rot disease. This pathogen poses a significant threat to potato production worldwide, resulting in substantial economic losses and food security concerns. This study aimed to investigate the effectiveness of three bacterial bioagents, namely Brevundimonas bullata, Bacillus siamensis, and Bacillus velezensis, against P. carotovorum subsp. carotovorum, a notorious bacterium responsible for causing potato tuber soft rot disease.ResultsFifteen isolates were isolated from rhizosphere of potato plants. Out of 15 isolates, 3 isolates, No. 1, 11, and 12, showed highly antagonistic property to control the growth of P. carotovorum subsp. carotovorum in vitro. They were identified as B. bullata, B. velezensis, and B. siamensis, by using 16S rRNA nucleotide sequence analysis. Results from these experiments revealed that three bioagents exhibited notable inhibitory effects on the growth of the pathogenic bacterium. However, B. velezensis stood out as the most effective inhibitor among the tested bioagents, showcasing the highest level of bacteriostatic activity (2.6 cm). The findings from the in vitro experiments provided promising insights into the potential of these bacterial bioagents as natural antagonists against potato tuber soft rot disease. Building upon these promising results, the study recommended the utilization of all tested bioagents for controlling soft rot disease in potato tubers, especially during storage conditions where the risk of bacterial proliferation is heightened. Treatment with B. bullata, B. siamensis, and B. velezensis demonstrates varying degrees of efficacy in controlling the progression of soft rot disease. Notably, B. velezensis shows promising results with relatively low disease severity scores across all time points compared to the other treatments. B. velezensis demonstrates the lowest disease severity percentage at 7.00% compared to the other treatments.ConclusionsThe study underscores the significant potential of B. bullata, B. siamensis, and B. velezensis as effective biological control agents against potato tuber soft rot disease. By targeting the causative agent directly, these bioagents offer a promising avenue for mitigating the detrimental impact of soft rot disease on potato production, thereby contributing to improved crop yield and quality.

Genome-wide association study identifies candidate genes for bacterial soft rot resistance in cucumber seedlings

Bacterial soft rot (BSR) caused by Pectobacterium carotovorum subsp. brasiliense (Pcb) is a serious bacterial disease which negatively impact yield and quality in cucumber. However, the genetic mechanism of BSR resistance in cucumber has not been reported. Here, we investigated the BSR resistance of 119 cucumber core germplasm worldwide at the seedling stage and identified 26 accessions highly resistant to BSR. A total of 1 642 740 single-nucleotide polymorphisms (SNPs) were used to conduct GWAS, and five loci associated with BSR resistance were detected on four chromosomes: gBSR2.1, gBSR2.2, gBSR3.1, gBSR4.1 and gBSR5.1. Based on haplotype analysis, sequence polymorphisms, functional annotation and qRT-PCR analysis, six candidate genes were identified within the five loci. CsaV3_2G014450, CsaV3_2G014490, CsaV3_2G016000, CsaV3_3G000850, CsaV3_4G033150, and CsaV3_5G000390 each had nonsynonymous SNPs, and were significantly up-regulated in the resistant genotypes after inoculation. And CsaV3_5G000390 in the susceptible genotype was significantly up-regulated after inoculation. The identification of these candidate genes lays a foundation for understanding the genetic mechanism of BSR resistance in cucumber. Generally, our study mined genes associated with BSR resistance in cucumber seedlings and will assist the breeding of BSR-resistant cucumber cultivars.

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.

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.

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 .

Differential Resistance of Radish Cultivars against Bacterial Soft Rot Caused by Pectobacterium carotovorum subsp. carotovorum.

Bacterial soft rot caused by Pectobacterium carotovorum subsp. carotovorum (Pcc) is one of the most severe diseases in radish cultivation. To control this plant disease, the most effective method has been known to cultivate resistant cultivars. Previously, we developed an efficient bioassay method for investigating resistance levels with 21 resistant and moderately resistant cultivars of radish against a strain Pcc KACC 10421. In this study, our research expanded to investigate the resistance of radish cultivars against six Pcc strains, KACC 10225, KACC 10421, ATCC 12312, ATCC 15713, LY34, and ECC 301365. To this end, the virulence of the six Pcc strains was determined based on the development of bacterial soft rot in seedlings of four susceptible radish cultivars. The results showed that the Pcc strains exhibited different virulence in the susceptible cultivars. To explore the race differentiation of Pcc strains corresponding to the resistance in radish cultivars, we investigated the occurrence of bacterial soft rot caused by the six Pcc strains on the 21 resistant and moderate resistant cultivars. Our results showed that the average values of the area under the disease progress curve were positively correlated with the virulence of the strains and the number of resistant cultivars decreased as the virulence of Pcc strains increased. Taken together, our results suggest that the resistance to Pcc of the radish cultivars commercialized in Korea is more likely affected by the virulence of Pcc strains rather than by race differentiation of Pcc.

Control of bacterial soft rot disease of Chinese cabbage by geraniol, thymol, and o-vanillin and the residue characteristics in cabbage

Control of bacterial soft rot disease of Chinese cabbage by geraniol, thymol, and <i>o</i>-vanillin and the residue characteristics in cabbage

First Report of Pectobacterium aroidearum Causing Soft Rot of Pinellia ternata in China.

Pinelliae rhizoma is the dried tuber of Pinellia ternata (Thunb.) Breit., and has been used for thousand of years in traditional Chinese medicine as an antivomit, anticough, and analgesic (Ying et al. 2007). In September 2022, P. ternata planted in Bijie, Guizhou Province, showed severe soft rot symptoms with incidence of about 50%. The diseased plants showed water-soaked symptoms and produced a foul soft rot smell, and finally the whole plant collapsed. Lesions were first observed at the tip of a leaf or wound, and symptoms of the disease spread rapidly, with the entire plant collapsing and dying within a week. The tissue sections of six plants with typical symptoms from the diseased field were disinfected with 75% ethanol for 30 seconds and 0.3% NaClO for 3 minutes. The tissue sections were then washed with sterile water for three times. A small piece of tissue (5x5mm) was removed from the edge of the lesion and mashed in a 1.5 ml centrifuge tube containing 20 μl of sterile water. The tissue liquid was then diluted 100 times with prepared sterile water. The bacteria were streaked on LB (tryptone/yeast extract/NaCl) AGAR medium and cultured at 37°C for 48 h (Kravitz, 1962). Isolated colonies were streaked on Luria-Bertani (LB) AGAR medium to obtain single colonies for further identification. A total of 13 representative isolates were selected for PCR amplification using primers targeting the conserved region of the 16S rDNA gene, which were in turn analyzed via the BLASTn search engine on the NCBI website. The results of the analysis revealed that seven of the isolates were similar to P. aroidearum strain SCRI 109 (GenBank accession no. NR_159926), with strain BX13 exhibiting the highest similarity to P. aroidearum (99.93% similarity), and therefore, this strain was selected for further investigation. The strain BX13 was incubated on LB solid medium for 24 h at 37 °C, and the single colonies were creamy white, translucent and round, slightly elevated in the center, with smooth surfaces and neat edges (Figure S1 B1). Then,the Scanning Electron Microscope revealed that the thalli of strain BX13 were short rod-shaped and somewhat blunt round at both ends (Figure S1 B2). The steward genes (icdA, gapA, proA) of BX13 were amplified and sequenced for further identification. The sequences of the amplified fragments were all deposited in GenBank 16S rDNA (OQ874505,) icdA (OQ954122)，gapA (OQ954123), proA (OQ954124). Sequence analysis using the BLASTn program at the NCBI revealed gene icdA, gapA, and proA had 100% identity to P. aroidearum strain QJ002 (GenBank accession no. CP090597).. Meanwhile, a maximum likelihood phylogenetic tree was constructed based on multigene sequence analysis of BX13 16S rDNA and steward genes (gapA, icdA, proA) by MEGA X (Liang et al. 2022). Phylogenetic results also showed that BX13 and P. aroidearum strain QJ002 gathered in the same clade(Figure S2). Accordingly, the morphological and molecular characteristics of strain BX13 indicate that it is P. aroidearum. (Nabhan S., et al.2013,Xu et al. 2020). In order to confirm the pathogenicity of strain BX13, a bacterial suspension containing 107 CFU/ml (10 ml/ inoculation point) was injected into the base of a one-week-old P. ternata stems, control seedlings were inoculated with sterile water, inoculated and control seedlings (each of six plants) were kept in a growth chamber maintained at 26°C with a relative humidity range of 70% to 80%. Plants were watered as needed. After 3 days, the stem base of the plants inoculated with bacteria solution showed water-soaked necrosis and stems began to rot, while the plants inoculated with water did not show this symptom. The strains were then successfully re-isolated from the symptomatic P. ternata. Then the strain re-isolated was identified using the BLASTn program at the NCBI and found that it has the same 16S rDNA, icdA, gapA, and proA sequences as strain BX13, thus completing the Koch's postulates. To our knowledge, this is the first report of P. aroidearum causing P. ternata soft rot in China, which expands its known host range. Accordingly, this study provides essential information for the breeding of P. ternata resistant to bacterial soft rot and the development of control measures in China.

Targeted metabolite profiling of Salvia rosmarinus Italian local ecotypes and cultivars and inhibitory activity against Pectobacterium carotovorum subsp. carotovorum.

The development of agriculture in terms of sustainability and low environmental impact is, at present, a great challenge, mainly in underdeveloped and marginal geographical areas. The Salvia rosmarinus "Eretto Liguria" ecotype is widespread in Liguria (Northwest Italy), and farmers commonly use it by for cuttings and for marketing. In the present study, this ecotype was characterized in comparison with other cultivars from the same geographical region and Campania (Southern Italy), with a view to application and registration processes for the designation of protected geographical indications. Moreover, the possibility of using the resulting biomass after removing cuttings or fronds as a source of extracts and pure compounds to be used as phytosanitary products in organic farming was evaluated. Specifically, the potential of rosemary extracts and pure compounds to prevent soft rot damage was then tested. A targeted NMR metabolomic approach was employed, followed by multivariate analysis, to characterize the rosemary accessions. Bacterial soft rot assay and disk diffusion test were carried out to evaluate the activity of extracts and isolated compounds against Pectobacterium carotovorum subsp. carotovorum. Enzymatic assay was performed to measure the in vitro inhibition of the pectinase activity produced by the selected pathogen. Molecular docking simulations were used to explore the possible interaction of the selected compounds with the pectinase enzymes. The targeted metabolomic analysis highlighted those different geographical locations can influence the composition and abundance of bioactive metabolites in rosemary extracts. At the same time, genetic factors are important when a single geographical area is considered. Self-organizing maps (SOMs) showed that the accessions of "Eretto Liguria" appeared well characterized when compared to the others and had a good content in specialized metabolites, particularly carnosic acid. Soft rotting Enterobacteriaceae belonging to the Pectobacterium genus represent a serious problem in potato culture. Even though rosemary methanolic extracts showed a low antibacterial activity against a strain of Pectobacterium carotovorum subsp. carotovorum in the disk diffusion test, they showed ability in reducing the soft rot damage induced by the bacterium on potato tissue. 7-O-methylrosmanol, carnosol and isorosmanol appeared to be the most active components. In silico studies indicated that these abietane diterpenoids may interact with P. carotovorum subsp. carotovorum pectate lyase 1 and endo-polygalacturonase, thus highlighting these rosemary components as starting points for the development of agents able to prevent soft rot progression.

Lytic phages isolated from Egypt for biocontrol of potato soft rot caused by Pectobacterium carotovorum

Pectobacterium carotovorum is an economically important phytopathogen causing destructive bacterial soft rot disease in many ornamental plants and fruit and vegetable crops worldwide, including potato. Phage therapy is a promising and environmentally safe alternative to combat bacterial diseases either in the field or during storage. In this study, we characterized two novel P. carotovorum-specific phages, designated PcaP1EGY and PcaP2EGY, isolated from soil in Egypt. We also sequenced the genomes of the two phages, which are the first two complete genome sequences determined for P. carotovorum-infecting podoviruses from Egypt. Phages PcaP1EGY and PcaP2EGY have a high phage titer of 1012 PFU/mL and growth characteristics of an exclusively lytic life cycle. Both phages displayed specificity to and visible lysis activity against all four tested strains of P. carotovorum. They both had a latent period of 30 min and a high burst size of approximately 599 PFU/infected cell for PcaP1EGY and 570 for PcaP2EGY. Additionally, they exhibited remarkable survivability under a wide range of pH, temperature, salt concentration and UV exposure conditions. Both phages, when used alone or in combination, significantly reduced in vitro growth of P. carotovorum after 4 h at all tested multiplicity of infection. Phages PcaP1EGY and PcaP2EGY offered significant protection for both potato tubers and potato plants against soft rot disease caused by P. carotovorum in both our tuber maceration test and greenhouse pot experiment as compared to tubers/plants inoculated with P. carotovorum without receiving any phage treatment. Our results suggest that phages PcaP1EGY and PcaP2EGY have great biocontrol potential against potato soft rot disease caused by P. carotovorum either in the field or during storage after harvest.

Influence of some chemicals and solvents on the lytic activity and the adsorption of bacteriophages on Pectobacterium carotovoroum Subsp. carotovorum

Recently, bacteriophages have been used to control hazardous bacterial soft rot disease on crops. However, agricultural plants are frequently treated with different chemicals (fertilizers, pesticides and solvents), so we assessed the effect of some commonly used chemicals and solvents on the lytic activity of tested bacteriophages and their adsorption potential. This study reports the isolation of three specific phages against the Pectobacterium carotovorum subsp. carotovorum DSM 30170 strain, designated as ?PC1, ?PC2 and ?PC3, then partially characterized using electron microscopy and genome size. The 3 isolated phages belong to the Myoviridae family. The results obtained were based on the plaque-forming unit observed after incubation. By increasing the chemical concentrations (from 0.1 to 0.5 mM), calcium chloride (CaCl2) and potassium chloride (KCl) showed a significant increase in the lytic activity of the phages. Copper sulphate (CuSO4) and copper chloride (CuCl2) showed a substantial decrease in the activity of ?PC3; however, such a decrease was insignificant for ?PC1 and ?PC2. By increasing the solvent concentrations (from 30 % v/v to 70 % v/v), propanol, ethanol and methanol showed a significant decrease in the count of the three isolated phages, ?PC1, ?PC2 and ?PC3, compared to the control. Chloroform was the only solvent that did not reduce the phage titer. Our findings offer significant information for developing a strategy to combat the P. carotovorum subsp. carotovorum caused bacterial soft rot disease. avoiding copper compounds and alcoholic solvents such as propanol, ethanol and methanol in plots where phages are applied seems advisable.

Cauli-Det: enhancing cauliflower disease detection with modified YOLOv8.

Cauliflower cultivation plays a pivotal role in the Indian Subcontinent's winter cropping landscape, contributing significantly to both agricultural output, economy and public health. However, the susceptibility of cauliflower crops to various diseases poses a threat to productivity and quality. This paper presents a novel machine vision approach employing a modified YOLOv8 model called Cauli-Det for automatic classification and localization of cauliflower diseases. The proposed system utilizes images captured through smartphones and hand-held devices, employing a finetuned pre-trained YOLOv8 architecture for disease-affected region detection and extracting spatial features for disease localization and classification. Three common cauliflower diseases, namely 'Bacterial Soft Rot', 'Downey Mildew' and 'Black Rot' are identified in a dataset of 656 images. Evaluation of different modification and training methods reveals the proposed custom YOLOv8 model achieves a precision, recall and mean average precision (mAP) of 93.2%, 82.6% and 91.1% on the test dataset respectively, showcasing the potential of this technology to empower cauliflower farmers with a timely and efficient tool for disease management, thereby enhancing overall agricultural productivity and sustainability.

Multifunctional halloysite nanotube–polydopamine agro-carriers for controlling bacterial soft rot disease

Halloysite nanotube-polydopamine core–shell nano-carriers loaded with thymol offer a sustainable alternative to existing pesticide formulations while providing enhanced efficacy, stability, and crop safety.

Isolation, identification, and pathogenicity of Pseudomonas glycinae causing ginseng bacterial soft rot in China

By tissue separation method, tie-back experiment, and hypersensitive response test in potato, strain XJFL-1 was isolated and identified as the pathogen of ginseng bacterial soft rot in Liaoning Provence, China. The morphological characteristics of XJFL-1 were conformed to the Pseudomonads genus. Microbial fatty acid identification showed the principal cellular fatty acid traits of XLFJ-1 corresponded with Pseudomonas spp. API 50CH test results allowed the differentiation of strain XJFL-1 and MS586T from other closely related Pseudomonas species. The molecular identification, including 16S rRNA analysis and multilocus sequence typing (MLST) analysis, showed that XJFL-1 was in the same branch as P. glycinae MS586T.The genome of XJFL-1 was 6,296,473 bp, with an average guanine/cytosine (G + C) content of 60.72 %. Comparative genomics analysis using ANIb and GGDC algorithms indicated that the maximum value was observed between XJFL-1 and P. glycinae MS586T.The above morphological, cell morphology, and molecular biological identification results supported to identification of XJFL-1 as P. glycinae.This is the first report of P. glycinae as the plant pathogen causing ginseng bacterial root rot in China, which complements the biological significance of the species to a certain extent, enriches the pathogens of ginseng bacterial soft rot, and provides a theoretical basis for further investigation.