Publications

4364

The genus ‘Candidatus Phytoplasma’ was proposed to accommodate cell wall-less bacteria that are molecularly and biochemically incompletely characterized, and colonize plant phloem and insect vector tissues. This provisional classification is highly relevant due to its application in epidemiological and ecological studies, mainly aimed at keeping the severe phytoplasma plant diseases under control worldwide. Given the increasing discovery of molecular diversity within the genus ‘Ca. Phytoplasma’, the proposed guidelines were revised and clarified to accommodate those ‘Ca. Phytoplasma’ species strains sharing >98.65 % sequence identity of their full or nearly full 16S rRNA gene sequences, obtained with at least twofold coverage of the sequence, compared with those of the reference strain of such species. Strains sharing <98.65 % sequence identity with the reference strain but >98.65 % with other strain(s) within the same ‘Ca. Phytoplasma’ species should be considered related strains to that ‘Ca. Phytoplasma’ species. The guidelines herein, keep the original published reference strains. However, to improve ‘Ca. Phytoplasma’ species assignment, complementary strains are suggested as an alternative to the reference strains. This will be implemented when only a partial 16S rRNA gene and/or a few other genes have been sequenced, or the strain is no longer available for further molecular characterization. Lists of ‘Ca. Phytoplasma’ species and alternative reference strains described are reported. For new ‘Ca. Phytoplasma’ species that will be assigned with identity ≥98.65 % of their 16S rRNA gene sequences, a threshold of 95 % genome-wide average nucleotide identity is suggested. When the whole genome sequences are unavailable, two among conserved housekeeping genes could be used. There are 49 officially published ‘Candidatus Phytoplasma’ species, including ‘Ca. P. cocostanzaniae’ and ‘Ca. P. palmae’ described in this manuscript.

El trabajo se llevó a cabo en tres localidades, Salache, Machachi y Saquisilí, donde se determinó el comportamiento y hábitos del psílido (Bactericera cockerelli.), responsable de la transmisión del fitoplasma conocido como punta morada en el cultivo de papa (Solanum tuberosum). Se observó el desarrollo del insecto, alimentándolo con follaje fresco de plantas de papa, mantenidos en cámaras artesanales de cría; para Salache la temperatura promedio fue de 21 ºC y una humedad relativa del 68% con un ciclo total de 31 días; para Saquisilí se mantuvieron con una humedad relativa promedio de 75%, y una temperatura de 13,9°C con un ciclo total de 33 días y en Machachi se mantuvo una temperatura alrededor de 17°C y una humedad relativa de 74% con un ciclo total de 34 días.  Los datos registrados en todo el proceso fueron: número de huevos depositados, tiempo de desarrollo de cada estado ninfal y duración de la etapa adulto. Las observaciones se realizaron durante los meses de octubre, noviembre y diciembre, en donde se registraron los datos de los primeros instares de B. cockerelli hasta su última etapa, logrando determinar que la localidad Saquisilí tuvo un desarrollo oval rápido promedio de 2 a 6 días y la localidad más longeva en etapa adulta de B. cockerelli fue Salache, la misma que además obtuvo el mejor desarrollo en el ciclo biológico total de la especie.   

SummaryThe Candidate Phyla Radiation (CPR) is a very large group of bacteria with no pure culture representatives, first discovered by metagenomic analyses. Within the CPR, candidate phylum Parcubacteria (previously referred to as OD1) within the candidate superphylum Patescibacteria is prevalent in anoxic sediments and groundwater. Previously, we had identified a specific member of the Parcubacteria (referred to as DGGOD1a) as an important member of a methanogenic benzene-degrading consortium. Phylogenetic analyses herein place DGGOD1a within theCandidateclade Nealsonbacteria. Because of its persistence over many years, we hypothesized thatCa. Nealsonbacteria DGGOD1a must serve an important role in sustaining anaerobic benzene metabolism in the consortium. To try to identify its growth substrate, we amended the culture with a variety of defined compounds (pyruvate, acetate, hydrogen, DNA, phospholipid), as well as crude culture lysate and three subfractions thereof. We observed the greatest (10 fold) increase in the absolute abundance ofCa. Nealsonbacteria DGGOD1a only when the consortium was amended with crude cell lysate. These results implicateCa. Nealsonbacteria in biomass recycling. Fluorescent in situ hybridization and cryogenic transmission electron microscope images revealed thatCa. Nealsonbacteria DGGOD1a cells were attached to larger archaealMethanothrixcells. This apparent epibiont lifestyle was supported by metabolic predictions from a manually curated complete genome. This is one of the first examples of bacterial-archaeal episymbiosis and may be a feature of otherCa. Nealsonbacteria found in anoxic environments.

Anaerobic methanotrophic (ANME) archaea can drive anaerobic oxidation of methane (AOM) using solid iron or manganese oxides as the electron acceptors, hypothetically via direct extracellular electron transfer (EET). This study investigated the response of Candidatus “Methanoperedens nitroreducens TS” (type strain), an ANME archaeon previously characterized to perform nitrate-dependent AOM, to an Fe(III)-amended condition over a prolonged period. Simultaneous consumption of methane and production of dissolved Fe(II) were observed for more than 500 days in the presence of Ca. “M. nitroreducens TS,” indicating that this archaeon can carry out Fe(III)-dependent AOM for a long period. Ca. “M. nitroreducens TS” possesses multiple multiheme c-type cytochromes (MHCs), suggesting that it may have the capability to reduce Fe(III) via EET. Intriguingly, most of these MHCs are orthologous to those identified in Candidatus “Methanoperedens ferrireducens,” an Fe(III)-reducing ANME archaeon. In contrast, the population of Ca. “M. nitroreducens TS” declined and was eventually replaced by Ca. “M. ferrireducens,” implying niche differentiation between these two ANME archaea in the environment.

Anaerobic methanotrophic (ANME) archaea have recently been reported to be capable of using insoluble extracellular electron acceptors via extracellular electron transfer (EET). In this study, we investigated EET by a microbial community dominated by “Candidatus Methanoperedens” archaea at the anode of a bioelectrochemical system (BES) poised at 0 V vs. standard hydrogen electrode (SHE), in this way measuring current as a direct proxy of EET by this community. After inoculation of the BES, the maximum current density was 274 mA m–2 (stable current up to 39 mA m–2). Concomitant conversion of 13CH4 into 13CO2 demonstrated that current production was methane-dependent, with 38% of the current attributed directly to methane supply. Based on the current production and methane uptake in a closed system, the Coulombic efficiency was about 17%. Polarization curves demonstrated that the current was limited by microbial activity at potentials above 0 V. The metatranscriptome of the inoculum was mined for the expression of c-type cytochromes potentially used for EET, which led to the identification of several multiheme c-type cytochrome-encoding genes among the most abundant transcripts in “Ca. Methanoperedens.” Our study provides strong indications of EET in ANME archaea and describes a system in which ANME-mediated EET can be investigated under laboratory conditions, which provides new research opportunities for mechanistic studies and possibly the generation of axenic ANME cultures.

Amaç: Bu çalışmada Elma Çoklu Sürgün Fitoplazma Hastalığı (EÇSF) (Candidatus Pyhtoplasma mali)’nın biyolojik mücadelesinde endofitik fungusların etkinliğinin belirlenmesi amaçlanmıştır. Böylece pratikte kimyasal mücadele olanağı olmayan hastalık etmenine karşı farklı kontrol stratejileri geliştirilebilme olanağının sağlanması hedeflenmiştir. Yöntem ve Bulgular: EÇSF ve endofit fungus izolasyonları için toplam 50 farklı bahçe gezilerek fitoplazma testleri için 360 ağaçtan, endofit funguslar için ise hem sağlıklı hem de fitoplazma ile enfekteli 198 ağaçtan bitki örneği toplanmıştır. Fitoplazma açısından testlenen 360 örnekten 29 tanesi ‘Ca. P. mali’ ile enfekteli bulunmuştur. ‘Ca. P. mali’ ile enfekteli ve sağlıklı elma ağaçlarından endofit fungus türleri izole edilmiş, bu fungusların morfolojik ve moleküler tanısı ile birlikte patojenisite çalışmaları yapılmıştır. Sağlıklı elma ağaçlarından en yaygın olarak izole edilen endofit funguslardan Epicoccum nigrum, Aureobasidium pullulans, Ulocladium consortiale’nin EÇSF hastalığına karşı etkinlikleri incelenmiştir. Her bir fungus izolatının EÇSF hastalık yoğunluğunu farklı düzeylerde etkilediği, en yüksek etkinin A. pullulans’a ait olduğu ve EÇSF hastalık yoğunluğunu uygulama sonrası %34,18 etki değeri ile azalttığı belirlenmiştir. Genel Yorum: EÇSF hastalığının etkili bir kimyasal mücadelesinin bulunmaması ve etmenin karantina listesinde yer alması nedeniyle olası biyolojik mücadele uygulamaları oldukça önemlidir. EÇSF ile enfekteli ağaçlara inokule edilen 3 farklı endofit fungus türünden Aureobasidium pullulans’ın fitoplazma yoğunluğunu diğer 2 etmene göre daha fazla azaltmasından dolayı A. pullulans hastalıkla mücadelede kullanılabilecek biyolojik mücadele ajanı olabilir. Çalışmanın Önemi ve Etkisi: Bu çalışma ile biyolojik mücadele ajanı olan endofit fungusların elmalarda EÇSF hastalığı üzerine etkileri Türkiye’de ilk kez araştırılmış ve hastalığın kontrol stratejilerinin oluşturulmasında önemli ön bulgular elde edilmiştir.