Outbreaks of potato blackleg and soft rot caused by Pectobacterium species and more recently Dickeya species across the U.S. mid-Atlantic region have caused yield loss due to poor emergence as well as losses from stem and tuber rot. To develop management strategies for soft rot diseases, we must first identify which members of the soft rot Pectobacteriaceae are present in regional potato plantings. However, the rapidly expanding number of soft rot Pectobacteriaceae species and the lack of readily available comparative data for type strains of Pectobacterium and Dickeya hinder quick identification. This manuscript provides a comparative analysis of soft rot Pectobacteriaceae and a comprehensive comparison of type strains from this group using rep-PCR, MLSA and 16S sequence analysis, as well as phenotypic and physiological analyses using Biolog GEN III plates. These data were used to identify isolates cultured from symptomatic potato stems collected between 2016 and 2018. The isolates were characterized for phenotypic traits and by sequence analysis to identify the bacteria from potatoes with blackleg and soft rot symptoms in Pennsylvania potato fields. In this survey, P. actinidiae, P. brasiliense, P. polonicum, P. polaris, P. punjabense, P. parmentieri, and P. versatile were identified from Pennsylvania for the first time. Importantly, the presence of P. actinidiae in Pennsylvania represents the first report of this organism in the U.S. As expected, P. carotorvorum and D. dianthicola were also isolated. In addition to a resource for future work studying the Dickeya and Pectobacterium associated with potato blackleg and soft rot, we provide recommendations for future surveys to monitor for quarantine or emerging soft rot Pectobacteriace regionally. 

Multilocus sequence and phenotypic analysis of Pectobacterium and Dickeya type strains for identification of soft rot Pectobacteriaceae from symptomatic potato stems and tubers in Pennsylvania.

In summer of 2016, cankers were observed on 8-year-old pear trees (Pyrus communis L.) cv. Yali grafted on the Caleriana rootstock, in a commercial orchard in Monte Castelo, SC. Eight hundred out of 1200 trees died and the rest of the orchard was eradicated. Affected plants showed cankers on trunks and twigs with exudation; the branches turned brown and slowly died. Bacterial colonies were isolated on King’s B medium from infected tissues. Pathogenicity, biochemical tests were conducted and some isolates selected for whole genome sequencing using Ion Personal Genome Machine™ (PGM™) System (ThermoFisher Scientific). PCR with Y1/Y2 and PcaF/PcaR primers, specific for Pectobacterium carotovorum and Pectobacterium carotovorum subsp. actinidiae, respectively, were also carried out. The section of inoculated apple and pear branches showed longitudinal darkening, with wilt and leaf blight. Shoots of kiwifruit Actinidia deliciosa cv. Hayward inoculated with the bacterium showed no symptoms. The bacterium was re-isolated from all symptomatic tissues, satisfying Koch’s postulates. Selected isolates were gram-negative, facultative anaerobic, rod-shaped and exhibited a strong pectolytic activity in potato and carrot disks, and showed similar biochemical profile using a BIOLOG GEN III MicroPlate. The pathogen of the symptoms observed on pear trees in Brazil show highest genome (87.1%; DDH analysis) and proteome identity (99.8%; Patric analysis) with P. carotovorum subsp. actinidiae. A product of the expected size was obtained in both specific PCR (Y1/Y2 (434 bp) and PcaF/PcaR (343 bp) primer pairs). Results from the present study clearly show that P. carotovorum subsp. actinidiae was associated with canker on pear trees in Santa Catarina, Brazil.

Pectobacterium carotovorum subsp. actinidiae associated with canker on pear trees in Brazil

Pectobacterium spp. are a major cause of loss in vegetable and ornamental plant production. One of these species, Pectobacterium carotovorum, can cause soft rot disease on many plants, particularly potato. These diseases lead to significant economic loss and pose food security threats by reducing crop yields in the field, in transit, and during storage. The Gram-negative enterobacterium P. carotovorum WPP14 is a particularly virulent strain for which there is no available closed genome, limiting the molecular research for this important pathogen. Here, we report a high-quality complete and annotated genome sequence of P. carotovorum WPP14. The 4,892,225-bp genome was assembled with Nanopore reads and polished with Illumina reads, yielding 394× and 164× coverage, respectively. This closed genome provides a resource for research on improved detection and biology of P. carotovorum, which could translate into improved disease management.

/pdf/complete-genome-sequence-resource-for-the-necrotrophic-plant-2r1cr4u2x3.pdf

Complete Genome Sequence Resource for the Necrotrophic Plant-Pathogenic Bacterium Pectobacterium carotovorum WPP14

A bacterial pathogen strain was isolated from susceptible tissue of Hongyang variety kiwifruit in Zhongfeng Town, Ziyuan County, Guilin City, Guangxi, China. Due to the relatively single variety of kiwifruit in Guangxi, the control technology of fruit farmers is backward, and the climate is humid, which is suitable for the growth of pathogenic bacteria, resulting in frequent occurrence of diseases. In this study, the pathogen strain was identified based on morphological, physiological, and biochemical tests; 16S rRNA gene; PCR detection with specific primers; and Biolog analysis. The results showed that a tobacco allergic reaction could be induced by inoculation with the pathogenic bacteria. Additionally, brown necrotic plaques appeared on kiwifruit leaves, necrotic phloem lesions appeared, and wounds on kiwifruit branches turned brown. The characteristics identified by morphological, physiological, biochemical, and Biolog identification were similar to those caused by Pectobacterium sp. Through 16S rRNA gene sequence analysis and PCR identification with specific primers, bands with a size corresponding to target bands indicated that the pathogen was Pectobacterium carotovorum subsp. actinidiae. This is the first report of kiwifruit canker disease caused by P. carotovorum subsp. actinidiae in Guangxi, China. In addition, through this study, a preliminary understanding of the pathogen has been obtained, which will lay the foundation for the prevention and control of the disease in the future. 

A Report of Pectobacterium carotovorum subsp. actinidiae Causing Kiwifruit Bacterial Canker in Guangxi, China

Abstract During the sugar beet growing seasons of 2019 and 2020, symptomatic leaf samples displaying necrotic spots were collected for the isolation of pectinolytic bacteria. Seven bacterial isolates were obtained from these infected leaves on crystal violet pectate (CVP) medium. Two of these isolates exhibited the ability to macerate potato tuber slices and induced disease symptoms on petioles of sugar beet plants. Initially, their identity was determined to be Pectobacterium spp. through a polymerase chain reaction (PCR) that produced a 434 bp amplicon using Y1/Y2 primers. Subsequently, they generated a 550 bp PCR product with the EXPCCF/EXPCCR primers, classifying them as P. carotovorum . Further analysis of both strains involved the examination of the dna X and mdh genes to confirm their taxonomic status. BLASTn analysis revealed the highest sequence similarity with P. actinidiae strains in GenBank for dna X (99.39%–100%) and mdh (100%) genes. Phylogenetic analysis, based on sequences from dna X, mdh , and a concatenation of both genes, conducted alongside 21 species of Pectobacterium spp., unequivocally confirmed that strains Pact1 and Pact2 belong to P. actinidiae . This study marks the first‐ever report of the presence of P. actinidiae in sugar beet worldwide. 

Isolation, identification, and characterization of Pectobacterium actinidiae isolated from infected sugar beet leaves

First Report of Pectobacterium carotovorum subsp. actinidiae Causing Summer Canker on Kiwifruit in Zhejiang Province of China

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First Report of Pectobacterium carotovorum subsp. actinidiae Causing Summer Canker on Kiwifruit in Zhejiang Province of China