Publications

4046

Lethal wilt (LW), also known as “Marchitez Letal (ML)” in Colombia, is an endemic disease affecting oil palms (Elaeis guineensis Jacq.) and is a leading cause of crop loss. The disease is characterized by the drying of leaflets from the tip to the base, primarily impacting the lower third of the plant and progressively moving upward. This progression leads to physiological disturbances, including necrosis at the tips of immature inflorescence bracts (spines) and the detachment of bunch fruits, ultimately causing wilting. As a phytosanitary measure, infected palms are eradicated to prevent further spread of the disease. The primary goal of this research was to identify the bacteria associated with LW and to validate a molecular detection method. A 16S amplicon-based analysis was employed to identify and compare the microbial diversity in LW-affected palm tissues with those of healthy plants. Among the 16 OTUs corresponding to different bacterial genera found in all LW samples, taxonomic classification and symptomatology suggested that the bacteria closely associated with LW belong to the genus Candidatus Liberibacter. Further phylogenetic analysis indicated that these bacteria are part of the Rhizobiaceae family, grouping closely with other species of the genus Candidatus. Liberibacter. The concentration of the pathogen in different oil palm tissues was determined using droplet digital PCR (ddPCR) and quantitative PCR (qPCR), expressed in copies/µL in the LW samples. This study represents the first report of ‘Candidatus Liberibacter sp’. being associated with lethal wilt in oil palms of the Arecaceae family in Colombia. The findings from this research have the potential to contribute significantly to the development of effective management strategies to prevent crop losses.

The Asian citrus psyllid (ACP) is the vector of ‘Candidatus Liberibacter asiaticus’ (CLas), the causal agent of citrus greening or Huanglongbing (HLB), one of the most devastating citrus diseases worldwide. The citrus industry in Georgia (U.S.A.) is in the process of a rapid expansion, and based on experiences with HLB in Florida, there is great concern about the potential impacts of HLB on this emerging industry. Prior to 2023, ACP had been identified in residential citrus trees in isolated Georgia counties, but little to no testing of psyllids for CLas had occurred. However, in 2023, one individual psyllid collected from Chatham County was confirmed positive for CLas by PCR and sequencing. Furthermore, during 2023, ACP adults and nymphs were identified for the first time in a Georgia commercial citrus grove. The finding of ACP in a commercial planting represents a significant risk for CLas dissemination and thereby has the potential to stall the rapid expansion of Georgia’s citrus industry. In the coming years, surveillance and testing of ACP from commercial groves will be essential for the early detection and management of HLB and its vector to reduce HLB spread within Georgia’s commercial groves.

Phytoplasmas cause infections in numerous plants in agricultural ecosystems, causing significant yield and quality losses in products. In recent years, it has been known that diseases caused by phytoplasmas cause economic losses in eggplant (Solanum melongena L.) cultivation. In Turkey, research on infections caused by phytoplasmas in eggplant growing areas is quite limited. This study was carried out to detect phytoplasma infections symptomatologically and molecularly in eggplant production areas in Şanlıurfa province. Fourteen samples were collected from eggplants exhibiting symptoms such as witches’ broom, flower abnormalities (virescence, phyllody), elongation of the pedicle, arising of new shoots from flower parts, yellowing and proliferation. Phytoplasma infection was detected in 8 symptomatic samples using 16S rRNA-specific primers, P1/P7 and R16F2n/R16R2, by direct and nested PCR. Sequence information of fragments obtained as a result of molecular studies was extracted and BLAST analyses were performed. According to nucleotide sequence similarity in the 16S rRNA gene region, it was determined that the genetic group of phytoplasma causing infection in eggplant was related to ‘Candidatus Phytoplasma solani’ (CaPsol) belonging to 16SrXII-A subgroup with 98% sequence identity. To our best knowledge, this study suggests comprehensive symptomatic diagnosis of CaPsol infecting eggplants in Türkiye.

Marine heterotrophic bacteria in coastal waters respond to the influx of carbon from natural and anthropogenic sources. We identified two nearly identical, (99.9% average nucleotide identity; 100% amino acid identity; same DNA G + C content of 52.3 mol%) high-quality (≥99% CheckM completeness and ≤ 1.3% contamination) draft metagenome-assembled genomes (MAGs; SJ0813 and SJ0972) from seawater microbiomes of a southern island of Singapore that is in a protected marine park. The MAGs were only assigned to the Cellvibrionaceae family according to Genome Taxonomy Database. Overall genome related indices to Pseudomaricurvus alkylphenolicus KU41GT as the closest phylogenetic relative revealed no more than 70.45% average nucleotide identity (ANIcutoff < 95%), below the 50% percentage of conserved proteins (POCPcutoff = 43.54%) for genera cutoff and low digital DNA–DNA hybridization values (DDH = 20.6 and 20.8%). The major respiratory quinone is predicted to be ubiquinone-9 from the annotation of 3-demethylubiquinone-9 3-methyltransferase (ubiG, K00568) involved in the last step of the ubiquinone biosynthesis pathway (M00117), which differed from the ubiquinone-8 utilized by known members of Cellvibrionaceae. Both MAGs contained a complete pathway for dissimilatory nitrate reduction to ammonia, which increases bioavailability of nitrogen in seawater. An identical choline dehydrogenase found in both MAGs have a low amino-acid identity (≤64.47%) compared to existing GMC family oxidoreductases, expanding on the diversity of this family of enzymes. The MAGs meet nearly all the minimum requirements but lack a 16S rRNA gene of sufficient length required for the proposed novel genus and species under SeqCode. Nevertheless, phylogenetic trees based on core-genome and RpoB as an alternative phylogenetic marker are congruent with the taxon standing as a monophyletic clade to other taxa of the order Cellvibrionales. Taken together, the MAGs (SJ0813 and SJ0972) represent an uncultured, undescribed genus and species in which we tentatively propose the name Candidatus Pelagadaptatus aseana gen. nov., sp. nov. and strain SJ0813TS (=BAABNI000000000.1TS) as type sequence. Phylogenetic inference from core-genome and RpoB phylogenetic trees placed Umboniibacter marinipuniceus KMM 3891T outside Cellvibrionaceae. We, therefore, propose the transfer of the genus Umboniibacter from the family Cellvibrionaceae to a new family Umboniibacteraceae according to the International Code of Nomenclature of Prokaryotes.

Phylogenetic analysis of the genus Aureimonas, utilizing both 16S rRNA gene sequences and comprehensive whole-genome data, revealed its polyphyletic nature, necessitating a revision to accommodate phylogenetically distinct species. Based on established threshold values for genus demarcation – specifically, 16S rRNA gene similarity, Average Amino Acid Identity and Percentage of Conserved Proteins – a notably substantial divergence was observed within the genus Aureimonas, and the division of Aureimonas into four distinct genera is strongly supported. To address this, we propose the establishment of four new genera: Dennerimonas gen. nov., Mesocryomonas gen. nov., Rathsackimonas gen. nov. and Plantimonas gen. nov. These classifications accommodate species that are significantly divergent from the type species of Aureimonas, thereby more accurately reflecting their distinct evolutionary lineages. Additionally, Aureimonas glaciistagni is proposed to be reclassified within the genus Jiella as Jiella glaciistagni comb. nov. based on phylogenetic evidence indicating a closer evolutionary relationship to Jiella species than to other members of Aureimonas. Our analysis, which included assessments of 16S rRNA gene similarity, Average Nucleotide Identity, and digital DNA–DNA hybridization values exceeding species delineation thresholds, further supports the reclassification of Jeongeupella Jiang et al. 2024 as a later heterotypic synonym of Antarcticirhabdus Du et al. 2023.

Surveys for exotic plant pests conducted during July and August of 2023 and 2024 across 20 vineyards in 12 counties throughout Minnesota, USA, revealed that less than 2% of the approximatively 3000 vines inspected (Vitis spp., hybrid grape varieties) exhibited symptoms suggestive of phytoplasma yellows disease. Observed symptoms included yellowing of leaf lamina, downward rolling of leaf margins, and necrosis of leaf margins. To investigate a potential association between these symptoms and phytoplasmas, two symptomatic plants were selected in August of 2024 from two distinct vineyards for further analysis. Petiole tissue DNA extracts were initially tested using a phytoplasma-specific real-time PCR assay (Hodgetts et al. 2009), which confirmed the presence of phytoplasma DNA in both symptomatic plants. Subsequently, the samples underwent semi-nested PCR amplification targeting the phytoplasma 16S rRNA gene, using primers P1/16S-SR followed by P1A/16S-SR (Deng and Hiruki 1991; Lee et al. 2004). The resulting amplicons were cloned and sequenced. No amplicon was produced from an asymptomatic grapevine DNA sample included as a test control. Analysis of the 16S rRNA gene sequences revealed that each plant was infected by one of two distinct phytoplasma strains, designated MN450 and MN466. Representative sequences obtained from the two samples exhibiting interoperon heterogeneity were deposited in the GenBank database under accession numbers: PQ889195 (rrnA) and PQ889196 (rrnB) for strain MN450, and PQ889197 (rrnA) and PQ889198 (rrnB), for strain MN466. Sequence analysis using the online phytoplasma classification tool iPhyClassifier (Zhao et al. 2009) showed that the 16S rRNA gene sequences from MN450 shared 1491/1495 bp (99.73%) and 1492/1495 bp (99.80%) identity (rrnA and rrnB, respectively) with that of the ‘'Ca. Phytoplasma pruni' rrnA’ reference strain (GenBank JQ044393), identifying MN450 as a ‘'Ca. Phytoplasma pruni' rrnA’-related strain and possibly representing a new subgroup within the 16Sr group III. For MN466, the 16S rRNA gene sequences shared 1471/1482 bp (99.26%) and 1469/1482 bp (99.12%) identity (rrnA and rrnB, respectively) with that of the ‘Ca. Phytoplasma asteris’ reference strain (GenBank M30790), and identifying the phytoplasma as a ‘Ca. P. asteris’-related strain belonging to subgroup 16SrI-A. To further characterize the two phytoplasma strains, the secY gene was PCR amplified from the DNA extracts as described by Lee et al. (2010). The obtained amplicons were cloned and sequenced, and representative secY gene sequences were deposited in GenBank under accession numbers PQ899621 (MN450) and PQ899622 (MN466). BLASTn searches against the NCBI core nucleotide database revealed that the secY gene sequence for MN450 exhibited 100% identity (1614/1614 bp) with ‘Ca. P. pruni’ (GenBank KM268860). Similarly, the sequence for MN466 showed 100% identity (1287/1287 bp) with ‘Ca. P. asteris’ (GenBank CP000061). In North America, grapevine yellows disease associated with strains related to 'Ca. P. asteris' and 'Ca. P. pruni' has previously been reported in various states and referred to as North American grapevine yellows (NAGY) by Davis et al. (2015). To our knowledge, this is the first report of NAGY in Minnesota, confirming that the two phytoplasma species detected are consistently associated with the disease in U.S. vineyards. While further research is needed to evaluate the potential impacts of NAGY in Minnesota, this study highlights the broader distribution of the disease across the United States.