Publications

4366

AbstractCandidatusLiberibacter solanacearum (CLso) is an economically important plant-pathogen of tomato (Solanum lycopersicum) crops in the United States, Mexico, Central America, and New Zealand. Currently, there are no reports of resistance toCLsoin tomato cultivars. Identification and development ofCLso-resistant cultivars may offer the most efficient way to manage this tomato disease. Resistance of 46 tomato landraces collected in different regions of Mexico, representing a wide range of genetic variability from this country was evaluated. Two assays were done in consecutively years to assess the resistance toCLsounder greenhouse conditions. Plants from both tests were inoculated withCLsothrough 20Bactericera cockerelliinsects per plant. In the first trial, landraces FC22 and FC44 showed a significantly higher proportion of resistant plants, less symptoms severity, and longer incubation time, followed by landraces FC40 and FC33 compared with the rest of the 42 landraces and 2 susceptible cultivars 60 days post inoculation (dpi). In the second assay, only landraces FC22 and FC44 had again significantly higher proportion of resistant plants, less symptoms severity, relative lowerCLsotiters, and longer incubation time in comparison with landraces FC40 and FC33 and the two susceptible cultivars 60 dpi, corroborating their resistance toCLso. Presence ofCLsoDNA in all resistant plants from both assays discards scape plants and indicates that the methodology used was adequate to discriminate between resistant and susceptible plants. These results confirm that landraces FC22 and FC44 are promising resistant sources for the development ofCLso-resistant cultivars of tomato.Author summaryThe bacterium “CandidatusLiberibacter solanacearum” (CLso) is an important plant-pathogen of tomato crops in the United States, Mexico, Central America, and New Zealand. Tomato growers are lacking of cultivars with resistance to this pathogen and the development of resistant cultivars of this crop would make a sustainable business for these growers and healthy tomato consumption for humans. Tomato landraces from countries that are center of domestication of cultivated crops like Mexico, are potentially sources of resistance to plant-pathogens. Therefore, two tests were done looking for resistance sources to this pathogen and we found two tomato landraces (FC22 and FC44) showing high level of resistance toCLsobecause they had significantly higher resistant plants, less symptoms severity, lowerCLsoDNA concentration, and delay of the first symptoms in the inoculated plants in comparison with the two commercial cultivars and 44 tomato landraces collected from Mexico 60 days post infection. These landraces are promising resistant sources for the development ofCLso-resistant cultivars of tomato.

AbstractCandidatusAccumulibacter phosphatis (CAP) and its sub-clades-level diversity has been associated and implicated in successful phosphorus removal performance in enhanced biological phosphorus removal (EBPR). Development of high-throughput untargeted methods to characterize clades of CAP in EBPR communities can enable a better understanding of Accumulibacter ecology at a higher-resolution beyond OTU-level in wastewater resource recovery facilities (WRRFs). In this study, for the first time, using integrated 16S rRNA gene sequencing, oligotyping and genome-resolved metagenomics, we were able to reveal clade-level differences in Accumulibacter communities and associate the differences with two different full-scale EBPR configurations. The results led to the identification and characterization of a distinct and dominant Accumulibacter oligotype - Oligotype 2 (belonging to Clade IIC) and its matching MAG (RC14) associated with side-stream EBPR configuration. We are also able to extract MAGs belonging to CAP clades IIB (RCAB4-2) and II (RC18) which did not have representative genomes before. This study demonstrates and validates the use of a high-throughput approach of oligotyping analysis of 16S rRNA gene sequences to elucidate CAP clade-level diversity. We also show the existence of a previously uncharacterized diversity of CAP clades in full-scale EBPR communities through extraction of MAGs, for the first time from full-scale facilities.

Huanglongbing (HLB) is a highly destructive citrus disease and is associated with a nonculturable bacterium, ‘Candidatus Liberibacter asiaticus’. ‘Ca. L. asiaticus’ in the United States was first found in Florida in 2005 and is now endemic there. In California, ‘Ca. L. asiaticus’ was first detected in Hacienda Heights in Los Angeles County in 2012 and has now been detected in multiple urban locations in southern California. Knowledge of ‘Ca. L. asiaticus’ strain diversity in California is important for HLB management. In this study, genomic diversity among 10 ‘Ca. L. asiaticus’ strains from six California locations were analyzed using a next-generation sequencing (NGS) (Illumina MiSeq and HiSeq) approach. Draft genome sequences of ‘Ca. L. asiaticus’ strains were assembled. Sequences of the 16S ribosomal RNA gene and nrdB confirmed ‘Ca. L. asiaticus’ identity. Prophages were detected in all ‘Ca. L. asiaticus’ strains. The California ‘Ca. L. asiaticus’ strains formed four prophage typing groups (PTGs): PTG1, with type 1 prophage only (strains from Anaheim, San Gabriel, and Riverside); PTG2, with type 2 prophage only (strains from Hacienda Heights); PTG1-3, with both type 1 and 3 prophages (a strain from Cerritos); and PTG1-2, with both type 1 and type 2 prophages (a strain from La Habra). Analyses of the terL sequence showed that all California ‘Ca. L. asiaticus’ strains were not introduced from Florida but likely from locations in Asia. Miniature inverted-repeat transposable elements were found in all ‘Ca. L. asiaticus’ strains, yet, a jumping-out event was detected in the ‘Ca. L. asiaticus’ strain from Cerritos. Altogether, this study demonstrated that the NGS approach focusing on prophage variation was sensitive and effective in revealing diversity of ‘Ca. L. asiaticus’ strains in California.

Summary Enhanced biological phosphorus removal (EBPR) exploits the metabolism of polyphosphate‐accumulating organisms (PAOs) to remove excess phosphorus (P) from wastewater treatment. Candidatus Accumulibacter phosphatis (Accumulibacter) is the most abundant and well‐studied PAO in EBPR systems. In a previous study, we detected polyphosphates throughout peripheral bay sediments, and hypothesized that an estuary is an ideal setting to evaluate PAOs in a natural system, given that estuaries are characterized by dynamic dissolved oxygen fluctuations that potentially favour PAO metabolism. We detected nucleotide sequences attributable to Accumulibacter (16S rRNA, ppk1 ) in sediments within three peripheral bays of the Columbia River estuary at abundances rivalling those observed in conventional wastewater treatment plants (0.01%–2.6%). Most of the sequences attributable to Accumulibacter were Type I rather than Type II, despite the fact that the estuary does not have particularly high nutrient concentrations. The highest diversity of Accumulibacter was observed in oligohaline peripheral bays, while the greatest abundances were observed at the mouth of the estuary in mesohaline sediments in the spring and summer. In addition, an approximately 70% increase in polyphosphate concentrations observed at one of the sites between dawn and dusk suggests that PAOs may play an important role in P cycling in estuary sediments.

ABSTRACT We present the results of a study of mesophilic anoxygenic phototrophic Chloroflexota bacteria from Mechigmen hot spring (the Chukotka Peninsula) and Siberia. According to 16S rRNA phylogenetic analysis, these bacteria belong to Oscillochloris trichoides. However, sequencing the draft genome of the bacterium from the Chukotka and analysis of the average nucleotide identity, as well as in silico DNA-DNA hybridization, reveal that this bacterium belongs to a novel species within the Oscillochloris genus. We, therefore, propose ‘Candidatus Oscillochloris fontis’ as a novel taxon to represent this mesophilic alkaliphilic anaerobic anoxygenic phototrophic bacterium. Spectrophotometry and high-performance liquid chromatography analysis show that the bacterium possesses bacteriochlorophylls c and a, as well as lycopene, β-carotene and γ-carotene. In addition, transmission electron microscopy shows the presence of chlorosomes, polyhydroxyalkanoate- and polyphosphate-like granules. The genome of ‘Ca. Oscillochloris fontis’ and the Siberian strains of Oscillochloris sp. possess the key genes for nitrogenase complex (nifH) and ribulose-1,5-bisphosphate carboxylase/oxygenase (cbbL), as previously described for O. trichoides DG-6. The results presented here, and previously published data, show that Oscillochloris bacteria from different aquatic environments have the potential for CO2 and N2 fixation. Additionally, we describe a new primer system for the detection of RuBisCo form I.

Huanglongbing (HLB), a destructive citrus disease, is associated with ‘Candidatus Liberibacter asiaticus’, which is transmitted by the Asian citrus psyllid Diaphorina citri. Both ‘Ca. L. asiaticus’ and its vector manipulate the host metabolism for their benefit, to meet their nutritional needs and neutralize the host defense responses. We used a targeted gas chromatography-mass spectrometry–based method to explore the connection between the tricarboxylic acid (TCA) cycle, γ-aminobutyric acid (GABA) shunt, and polyamines (PAs) pathways in citrus. ‘Ca. L. asiaticus’ and D. citri accelerated the conversion of α-ketoglutarate to glutamate, then to GABA, causing an accumulation of GABA in the cytosol. In silico analysis showed that the citrus genome possesses a putative GABA permease that connects the GABA shunt with the TCA cycle and supports the accumulation of succinate, fumarate, and citrate. Additionally, the PAs biosynthetic pathway might be connected directly to the TCA cycle, through the production of fumarate, or indirectly, via enhancement of GABA shunt. Taken together, we suggest that GABA shunt and PAs pathways are alternative pathways that contribute to the flux toward succinate rather than an intact TCA cycle in citrus. Both ‘Ca. L. asiaticus’ and its vector enhance these pathways. This study provides more insights into citrus responses to the HLB pathosystem and could be a further step toward clues for understanding the nutritional needs of ‘Ca. L. asiaticus’, which could help in culturing ‘Ca. L. asiaticus’.