Turkish Isolates Research Articles

Comparative proteomic and genomic analysis of Lactococcus garvieae and Lactococcus petauri isolates from Turkey

Piscine lactococcosis, a bacterial disease caused by Lactococcus garvieae, L. petauri and L. formosensis, induces mass mortalities in multiple farmed fish species globally. Following the identification of L. petauri as one of the emerging etiological agents of piscine lactococcosis, it was revealed that the majority of isolates previously recognized as L. garvieae actually belong to L. petauri. In this study, a total of 134 Lactococcus sp. isolates, including L. garvieae (n = 40) and L. petauri (n = 94) were analyzed. Differential identification of L. garvieae and L. petauri isolates was performed through basic sucrose fermentation and end-point PCR analysis using species-specific primer sequences. Comparative analysis of capsule formation in both species was conducted for the first time using transmission electron microscopy, whole-cell protein profile via 1D SDS-PAGE, antigenic protein characterization using Western blot analysis, and serological profile description by slide and microagglutination assays. Moreover, comprehensive genomic comparisons were made, deciphering overall genomic relatedness indices, virulence genes, and antimicrobial resistance patterns among these closely related species.Analysis showed microcapsule formation in some isolates of L. garvieae and L. petauri for the first time. While a high similarity rate was observed in the whole-cell protein profiles of these species, significant differences were identified among antigenic proteins. Variations between species were observed in slide agglutination and quantitative agglutination tests of L. garvieae and L. petauri field isolates. Capsule-deficient L. garvieae reference and field strain antisera did not agglutinate with each other; however, L. garvieae field strain antisera agglutinated with all L. garvieae field strains and L. petauri strains. These results indicated distinct serological characteristics of capsule-deficient L. garvieae isolates from Turkey compared to previously reported serological classifications.Genomic analyses revealed that none of the investigated L. garvieae and L. petauri isolates harbored the capsule gene cluster. In addition, Turkish isolates of L. garvieae and L. petauri clustered separately from isolates of other geographical origins but together with each other. This study aims to address the problem of lactococcosis, commonly referred to as “Summer Mortality Outbreak and Vaccine-Preventable Diseases,” in a wide variety of cultured fish species through the selected L. garvieae and L. petauri isolates as potential vaccine strains.

Insecticidal effect of two Turkish entomopathogenic nematode isolates against Tribolium castaneum (Herbst) larvae and adults at different conditions

Red flour beetle, Tribolium castaneum (Herbst.), is the most common pest of stored grains and products. Aerial and contact pesticides are used to control this pest. However, the red flour beetle has also developed significant resistance to phosphine, the main fumigant currently in use worldwide. The adverse effects of pesticides on non-target organisms, groundwater contamination, residues on food crops, and the emergence of insect resistance to chemicals have compelled the industry to prioritize the development of alternative control measures. Entomopathogenic nematodes find application in various agricultural systems. The insecticidal effect of two isolates of Steinernema feltiae Filipjev (Nematoda: Steinernematidae) and Heterorhabditis bacteriophora Poinar (Nematoda: Heterorhabditidae) against the different stages of T. castaneum. The entomopathogenic nematode isolates were examined at four concentrations (0, 250, 500, and 1000 IJs/ml) with a distilled water at 15 and 25 ± 1 °C temperatures. Results revealed that both S. feltiae (A-16) caused with 92.33% mortality rate while H. bacteriophora (O-7) caused with 76.77% mortality in the last instar larvae of the red flour beetle while these two species caused 75.38% and 66.85% mortality in adults, at 25 °C. The effect of nematodes decreased as the temperature decreased. According to results, the mortality rate of T. castaneum larvae treated with S. feltiae (A-16) with (54.62%) was significantly higher than that of H. bacteriophora (O-7) with (32.13%) 96 h after inoculation at 15 °C. The data indicated that 1000 IJs/ml was the most effective concentration of tested EPN isolates for all larval and adult stages.In this study, our findings indicate that S. feltiae (A-16) and H. bacteriophora (O-7) can be used as effective biological control agents against both the larval and adult stages of the red flour beetle. The results demonstrate that EPNs are promising candidates for biocontrol of T. castaneum. This information is useful in designing future biological control programs.

Genomic analysis of Brucella isolates from animals and humans, Türkiye, 2010 to 2020.

BackgroundBrucellosis is a bacterial zoonosis causing severe illness in humans and animals and leading to economic losses in the livestock production in Türkiye and other endemic countries.AimWe aimed at investigating genomic differences of Brucella isolates from animals and humans in Türkiye.MethodsWe used whole genome sequencing (WGS) to assess the genetic diversity of Brucella isolates from 41 provinces in Türkiye and compared with isolates from other countries. We applied allele-based typing and core genome single nucleotide polymorphism (cgSNP) determination.ResultsOf the 106 Turkish Brucella isolates included, 57 were B. abortus and 49 were B. melitensis. One B. melitensis and two B. abortus isolates were identified as vaccine strains. Most (n = 55) B. abortus isolates clustered in three major branches, with no spatial discernible pattern. Of the B. melitensis isolates, 48 were assigned to the Eastern Mediterranean lineage with no discernible patterns between host species, location and sampling date. The Turkish isolates clustered with isolates from neighbouring countries such as Greece and Syria, but some also with isolates from human patients in European countries, like Germany, Norway and Sweden, suggesting that the source may be travel-related.ConclusionSeveral B. melitensis and B. abortus lineages are circulating in Türkiye. To decrease the prevalence and prevent brucellosis in animals and humans, stricter control measures are needed, particularly in areas where humans and animals have close contact. Furthermore, illegal transportation of animals across borders should be more closely controlled and regulated.

Prevalence and molecular analysis of some important viruses in honey bee colonies in Türkiye: the status of multiple infections.

In this study, seven bee viruses of significant importance for bee health in Türkiye were investigated using one-step RT-PCR. For this purpose, larvae from 1183 hives and adult bees from 1196 hives were sampled from 400 apiaries in 40 provinces. The prevalence of viral infections in hives was as follows: acute bee paralysis virus (ABPV), 6.4%; black queen cell virus (BQCV), 77%; chronic bee paralysis virus (CBPV), 3.2%; deformed wing virus (DWV), 63.8%; Israel acute bee paralysis virus (IAPV), 7%; Kashmir bee virus (KBV), 2.7%; sacbrood virus (SBV), 49.7%. Moreover, 50 different combinations of viral infections were identified in the hives. While dual infections (36.1%) were the most common in hives, triple infections with BQCV, DWV, and SBV were found to have the highest prevalence (22.1%). At least one viral infection was detected in all of the apiaries tested. Phylogenetic analysis showed that the isolates from this study generally exhibited the highest similarity to previously reported Turkish isolates. When similarity ratios and the locations and types of amino acid mutations were analyzed, it was observed that the isolates from our study exhibited high similarity to isolates from various countries, including China, the United Kingdom, Syria, and Germany.

Effectiveness of four Turkish entomopathogenic nematode isolates against Bactrocera oleae (Diptera: Tephritidae) pupae at different temperatures

Türkiye, is the country of origin of the olive tree and is among the world’s leading olive producers. Most of the olive production in Türkiye is concentrated in Izmir, Aydın, Çanakkale, Balıkesir, Muğla and Bursa Provinces. Bactrocera oleae (Diptera: Tephritidae), being one of the main pests of olive trees, is an important problem in olive cultivation in Türkiye. The efficacy of four Turkish populations of entomopathogenic nematodes against B. oleae pupae was determined under laboratory conditions. They were Steinernema feltiae/12, S. carpocapsae/1133, Heterorhabditis bacteriophora/70, H. bacteriophora/91, which were collected in different regions of Türkiye during our earlier studies. They were tested against the pest by using a single dose at 5 different temperatures (10, 15, 20, 25 and 30 °C). The highest infection rate was observed forH. bacteriophora/91 isolate at 25 °C, and the lowest infection rate was observed for S. feltiae/12 isolate at 10 °C.

An Evaluation of the Lineage of Brucella Isolates in Turkey by a Whole-Genome Single-Nucleotide Polymorphism Analysis.

Brucellosis is a disease caused by the Brucella (B.) species. It is a zoonotic disease that affects farm animals and causes economic losses in many countries worldwide. Brucella has the ability to persist in the environment and infect the host at low doses. Thus, it is more important to trace brucellosis outbreaks, identify their sources of infection, and interrupt their transmission. Some countries already have initial data, but most of these data are based on a Multiple-Locus Variable-Number Tandem-Repeat Analysis (MLVA), which is completely unsuitable for studying the Brucella genome. Since brucellosis is an endemic disease in Turkey, this study aimed to examine the genome of Turkish Brucella isolates collected between 2018 and 2020, except for one isolate, which was from 2012. A total of 28 strains of B. melitensis (n = 15) and B. abortus (n = 13) were analyzed using a core-genome single-nucleotide polymorphism (cgSNP) analysis. A potential connection between the Turkish isolates and entries from Sweden, Israel, Syria, Austria, and India for B. melitensis was detected. For B. abortus, there may be potential associations with entries from China. This explains the tight ties found between Brucella strains from neighboring countries and isolates from Turkey. Therefore, it is recommended that strict measures be taken and the possible effects of uncontrolled animal introduction are emphasized.

Phylogenomic characterization of Flavobacterium psychrophilum isolates retrieved from Turkish rainbow trout farms.

Flavobacterium psychrophilum, a devastating fish pathogen, is responsible for bacterial cold-water disease (BCWD), also known as rainbow trout fry syndrome. F. psychrophilum is the main causative agent of outbreaks in rainbow trout farms, especially at early live stages. In the present study, we aimed to characterize F. psychrophilum Turkish isolates. Eighteen isolates were retrieved from BCWD outbreaks between 2014 and 2021. Invitro phenotypic characterization showed gelatin and casein hydrolysis capacities and invitro adhesion for all isolates, whereas elastinolytic activity was present for 16 of 18 isolates. We used complete genome sequencing to infer MLST-type, serotype and phylogenetic reconstruction. Strikingly, one strain isolated from Coruh trout (FP-369) belongs to ST393, a previously undescribed ST, and is phylogenetically distant from the other isolates. However, all strains retrieved from rainbow trout belong to the well-characterized clonal complex CC-ST10, 12 of 17 were tightly connected in a single cluster. Several serotypes (Types -1, -2 and -3) were represented among isolates, but no correlation was observed with geographic origins. This analysis suggests a regional dissemination of an epidemic, disease-producing bacterial population. This study provides a basis for epidemiological surveillance of isolates circulating in Turkey and phenotypic data for future molecular studies of virulence traits of this important fish pathogen.

Global Spread, Genetic Differentiation, and Selection of Barley Spot Form Net Blotch Isolates.

Spot form net blotch, caused by Pyrenophora teres f. maculata, is a significant necrotrophic disease of barley that spread worldwide in the twentieth century. Genetic relationships were analyzed to determine the diversity, survival, and dispersal of a diverse collection of 346 isolates from Australia, Southern Africa, North America, Asia Minor, and Europe. The results, based on genome-wide DArTseq data, indicated that isolates from Turkey were the most differentiated with regional sub-structuring, together with individuals closely related to geographically distant genotypes. Elsewhere, population subdivision related to country of origin was evident, although low levels of admixturing was found that may represent rare genotypes or migration from unsampled populations. Canadian isolates were the next most diverged, and Australian and South African the most closely related. With the exception of Turkish isolates, multiple independent Cyp51A mutation events (which confer insensitivity to demethylation inhibitor fungicides) between countries and within regions was evident, with strong selection for a transposable element insertion at the 3' end of the promoter and counterselection elsewhere. Individuals from Western Australia shared genomic regions and Cyp51A haplotypes with South African isolates, suggesting a recent common origin. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Turkish isolates of alfalfa mosaic virus belong to a distinct lineage among global population

Complete RNA3 sequences of 15 alfalfa mosaic virus (AMV) isolates sampled from Afyonkarahisar, Ankara, Konya, Tokat, Kayseri, and Nevşehir Provinces of Türkiye were obtained using RT-PCR with two primer pairs designed in this study. Molecular and population analyses were performed on them together with 63 global isolates which full genome sequences available in NCBI GenBank. Phylogenetic analysis based on movement protein (MP) and coat protein (CP) regions showed unrelated isolates clustering between the constructed trees, and the new Turkish isolates were dispersed in different phylogroups. Particularly, three new Turkish potato isolates: B6, K1, and K2 occupied a divergent MP lineage, together with one alfalfa isolate from China: ZM1 NX3. S, η, and k parameters of genetic diversity confirmed that CP were more conserved than MP sequences. The 15 new Turkish isolates shared 95.4–100% nucleotide and 94–100% amino acid, and 92.8–100% nucleotide and 93.3–100% amino acid identities among themselves at MP and CP regions, respectively. All tested populations were found to be under very strict purifying constraints (dN/dS ratios of 0.061–0.219), and had undergone recent expansion, probably due to new mutations in their genome, according to diversity and neutrality tests. Fixation index (FST) values obtained by pairwise comparisons of phylogroups were all above 0.33, suggested that the isolates clustering has been done convincingly.

Genetic diversity and phylogenetic relationships of Streptococcus thermophilus isolates from traditional Turkish yogurt: multilocus sequence typing (MLST).

Yogurt, a globally consumed fermented dairy product, is recognized for its taste and potential health benefits attributed to probiotic bacteria, particularly Streptococcus thermophilus. In this study, we employed Multilocus Sequence Typing (MLST) to investigate the genetic diversity and phylogenetic relationships of 13 S. thermophilus isolates from traditional Turkish yogurt samples. We also assessed potential correlations between genetic traits and geographic origins. The isolates were identified as S. thermophilus using VITEK® MALDI-TOF MS, ribotyping, and 16S rRNA analysis methods. MLST analysis revealed 13 different sequence types (STs), with seven new STs for Turkey. The most prevalent STs were ST/83 (n = 3), ST/135 (n = 2), and ST/134 (n = 2). eBURST analysis showed that these isolates mainly were singletons (n = 7) defined as sequence types (STs) that cannot be assigned to any group and differ at two or more alleles from every other ST in the sample. This information suggests that the isolates under study were genetically distinct from the other isolates in the dataset, highlighting their unique genetic profiles within the population. Genetic diversity analysis of ten housekeeping genes revealed polymorphism, with some genes showing higher allelic variation than others. Tajima's D values suggested that selection pressures differed among these genes, with some being more conserved, likely due to their vital functions. Phylogenetic analysis revealed distinct genetic diversity between Turkish isolates and European and Asian counterparts. These findings demonstrate the genetic diversity of S. thermophilus isolates in Turkish yogurt and highlight their unique evolutionary patterns. This research contributes to our understanding of local microbial diversity associated with yogurt production in Turkey and holds the potential for identifyic strains with enhanced functional attributes.

First Report of Natural Infection of Watermelon mosaic virus (WMV) Infecting Bottle Gourd and Snake Melon

Cucurbitaceous crops, one of the main crops of agriculture, are sensitive to many plant viruses. In August 2019, virus-like symptoms were observed on some cucurbit plants grown in private home gardens in Antalya and Denizli provinces (Turkey). A total of 53 leaf samples were sampled from plants with the most symptoms (melon (Cucumis melo L.), watermelon (Citrullus lanatus L.), bottle gourd (Lagenaria siceraria (Molina) Standl.), and snake melon (Cucumis melo var. flexuosus) and tested by Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR) against possible watermelon mosaic potyvirus (WMV) infection. The coat protein gene (CP) specific primer sets amplified a gene product of nearly 820 bp fragment from symptomatic plants. WMV infections were detected in 31 individual cucurbit plants, including 11 melons, 8 watermelons, 7 snake melons and 5 bottle gourds. The presence of viral infection was found only in ornamental squash plants in Antalya province and in all cucurbits sampled in Denizli province. To better comprehend the molecular characteristics of virus isolates, the amplified viral DNA fragments were cloned in a proper prokaryotic plasmid, sequenced by Next Generation Sequencing (NGS) and recorded to GenBank. Bioinformatic analyses using the Basic Local Alignment Search Tool (BLAST) showed that the identified CP gene sequences exhibited significant nucleotide homogeneity, supported by a high nucleotide similarity index with that of other isolates around the world. In addition, Turkish isolates isolated from Antalya and Denizli regions showed approximately 94% nucleotide similarity among themselves. For phylogenetic inference, WMV sequences were subjected to multiple alignments with isolates from different geographic origins of the same viruses. Molecular phylogeny showed that all WMV isolates are closely related to other world WMV isolates at variable rates. WMV is wide host range viruses in cucurbit crops, however, this work is the first scientific report of WMV isolates detected in bottle gourd and snake melon from the South and West Regions of Turkey all over the world.WMV are broad spectrum viruses in cucurbit crops. This work is the first scientific report of Watermelon mosaic potyvirus isolates detected in bottle gourd and snake melon from the South-West Region of Turkey in all over the world.

Tokat İli Asma Alanında Bazı Asma Virüslerinin Moleküler Olarak Belirlenmesi ve Grapevine Virus A'nın Filogenetik Analizi

Grapevine (Vitis spp.) is one of the major fruit crop with high socioeconomic importance for Turkey. In vineyards, many harmful organism, especially virus infections, weaken the plant and lead to decreases in yield and quality, so it takes the lead in quarantine and certification. This study was carried out to determine some viral agents that cause yield loss in vines produced in Tokat, where viticulture is very important. Samples were collected from young leaves and one-year-old shoots of grapevines showing virus symptoms from some vineyard areas in Tokat Center and its districts. Collected 189 grapevine samples were subjected to the RT-PCR test, which is a molecular method using virus-specific primers, to detect the presence of Grapevine pinot gris virus (GPGV), Grapevine virus A (GVA), Strawberry latent ringspot virus (SLRSV). Out of a total of 189 plant samples, 80 (42.32%) of GVA, 3 (1.58%) of GPGV were detected and SLRSV (0%) was not detected. More than one virus was found in 2 (1.05%) of 189 tested samples. It was determined that the most common virus was GVA, the least detected virus was GPGV in plant samples collected from Tokat Center and its districts. Bidirectional sequence analysis of RT-PCR products of GVA-infected isolates were performed and phylogenetic analyzes were done by comparing them with reference isolates after they were aligned with the MEGAX computer program. Based on phylogenetic analysis studies, GVA showed differential branching with isolates registered in GenBank and isolates obtained in the study. GVA-infected isolates showed similarity with reference isolates at rates of 92-94%. In this study, molecular analysis of Turkish GVA isolates was performed. This molecular information is important as it will shed light on future studies.

The association of Lactococcus petauri with lactococcosis is older than expected

Lactococcosis is a globally prevalent infectious disease that has a significant economic and sanitary impact on the rainbow trout industry. Lactococcus garvieae has traditionally been considered the only species implicated in the etiology of this disease, but Lactococcus petauri, a new species, has recently been implicated as another etiological agent. Both species cannot be distinguished by routine methods commonly used in diagnostic laboratories, resulting in their misidentification. In the present study, the identification of 48 isolates initially identified as L. garvieae was studied by determining their in-silico DNA–DNA hybridization (dDDH) and average nucleotide identity (ANI) values using pairwise comparisons of their whole genome sequences and the genomes of the type strains of L. garvieae and L. petauri. The genome sequences of 37 isolates from countries in which lactococcosis can be considered endemic (Spain, Italy, Türkiye, and Greece) were obtained in this study, and the genomes of 11 isolates were retrieved from the GenBank database. Isolates from Italy, Singapore, Japan, South Korea, India, one Turkish isolate from 2013 and two Spanish isolates recovered in 1992 and 1996 were confirmed as L. garvieae. The remaining isolates from Spain and Türkiye, as well as those from Portugal, Israel, USA, and Greece were identified as L. petauri. Overall, 60.4% of isolates previously identified as L. garvieae were found to be L. petauri. These results confirm the implication of both species in the etiology of lactococcosis and suggest that L. petauri plays a significant role in the epidemiology of this disease. Some of the isolates identified as L. petauri in the present study were isolated three decades ago, indicating that its association with lactococcosis is older than might be expected from the recent descriptions. The commercial Rapid ID32 Strep system was unable to discriminate between L. garvieae and L. petauri. However, both species exhibited some biochemical differences that might serve as phenotypic markers for their presumptive recognition. Consequently, isolates that hydrolyze hippurate and produce acid from sucrose and tagatose could be presumptively recognized as L. petauri, while those that fail these tests could be identified as L. garvieae. The results of this work indicate that great attention should be given to L. petauri in the epidemiology of lactococcosis.

Bean Common Mosaic Potyvirus (BCMV) Characterized from Bean (Phaseolus vulgaris L.) Crops Affected by Mosaic Disease in Denizli Province, Türkiye

Bean (Phaseolus vulgaris L.) is one of the world's oldest crops with both financial and nutritional importance. Bean common mosaic potyvirus (BCMV) is one of the prevalent viral agents that affect beans across the globe. Determining the presence of the agent in the relevant region is critical for minimizing crop losses by implementing appropriate preventive and control measures. In this study, 73 bean leaf samples were collected from bean-growing areas in the Denizli province of Türkiye in 2022. The samples were screened for the presence of viral agents using Polymerase Chain Reaction (PCR) with specific primers targeting the polypeptide gene. 26 of the bean samples were found to be positive for BCMV. The coat protein gene sequences of two randomly selected positive isolates were sequenced and deposited in the GenBank with accession numbers OQ910196 and OQ910197. The nucleotide sequences of isolates were found to have high similarity with those of isolates identified in various regions of the world. Phylogenetic analysis indicated that these isolates from Denizli, Türkiye were closely related to other Turkish isolates. However, since some Turkish isolates in the cluster associated with the Denizli isolates were identified as belonging to the US-5 or NL-6 strain, the current sequences may be related to these strains. Further research is necessary to identify the exact strain of the Denizli isolates, which could be achieved through the use of a strain differentiation set.

Acute bee paralysis virus field isolates from apiaries suffering colony losses in Türkiye

Multiple viral infections are one of the main drivers of honey bee colony losses and threaten the sustainability of colony health. The Acute bee paralysis virus (ABPV) is one of the common honey bee viruses that can induce overt and/or covert infections in bee colonies. Globally more knowledge is needed about its effects leading to colony losses. ABPV-related pathology includes paralytic, trembling bees, unable to fly and, in some cases, ABPV causes the death of the bee colonies. The molecular phylogenetic of ABPV isolates obtained from apiaries suffering colony losses and/or with CCD-like complaints in Türkiye was performed in this study. The isolates’ structural and non-structural gene regions were analyzed phylogenetically and compared according to sampling years and apiaries. According to the results, the nucleotide identity of the RNA-dependent RNA polymerase (RdRp) gene in Turkish ABPV field isolates is more conserved than the capsid gene. The evaluation of molecular differentiation among the ABPV field isolates can be used for determining the origin and relatedness of ABPV strains and periodic monitoring of ABPV infections in apiaries.

Characterization of Turkish isolates of Pseudocercospora griseola the causal agent of angular leaf spot of common beans

Characterization of Turkish isolates of Pseudocercospora griseola the causal agent of angular leaf spot of common beans

First molecular characterization of plum pox virus strains in stone fruits of Tuscany (Central Italy)

Plum pox virus (PPV) a potyvirus in the family Potyviridae, is the etiologic agent of sharka, the worldwide most important and detrimental viral disease of stone fruits (genus Prunus). Although PPV was identified in Italy already in the 1970s and it has been largely reported in almost all Italian regions, a broad investigation and genetic characterization of PPV isolates was lacking in most of them, including Tuscany (Central Italy). To address this knowledge gap, in 2020 and 2021, field surveys were carried out in 80 commercial Tuscany orchards and a total of 690 leaf samples were collected from different Prunus species. PPV was identified in 25 and 12 peach and plum samples, respectively (more than 5% of tested samples), whereas no positive samples were reported in apricot. Eighteen of the 37 PPV positive samples showed mixed infections with other viruses and viroids, mostly Prunus necrotic ringspot virus in plum and peach latent mosaic viroid in peach. Molecular typing of PPV infected samples generated NIb/CP amplicons corresponding to PPV-Marcus (M) or PPV-Recombinant (Rec) strains. Furthermore, starting from the identification of eight PPV nucleotide sequences (among which five and two new PPV-M and PPV-Rec isolates, respectively), this study firstly identified the PPV-Mb subgroup in Italy, which was even prevalent than PPV-Ma. Finally, PPV-Rec isolates resulted phylogenetically close to Italian and Turkish isolates previously detected. Overall, the results here presented represent an important step to fill knowledge gaps about PPV in Tuscany, and we believe it may encourage other similar research to achieve more accurate data on PPV populations at both national and international levels.

Incidence and genetic diversity of raspberry bushy dwarf virus (RBDV) in Rubus spp. in Turkey

AbstractRaspberry bushy dwarf virus (RBDV), recently renamed to Idaeovirus rubi, is one of the most common viruses infecting Rubus species worldwide but there is still a limited number of genome sequences available in the GenBank database and the majority of the sequences include partial sequences of RNA‐1 and RNA‐2. The distribution and incidence of RBDV in main raspberry and blackberry growing provinces in Turkey were monitored during 2015–2019 and 537 Rubus spp. samples were tested by both DAS‐ELISA and RT‐PCR. Among the tested samples, 36 samples tested positive for RBDV by DAS‐ELISA and 67 samples by RT‐PCR. There was relatively low nucleotide diversity among the Turkish isolates. Turkish isolates shared 93%–97.7%, 84.3%–98.9%, and 85%–99.2% nucleotide sequence identities with available sequences in the GenBank, in partial RNA‐1, movement protein (MP) and coat protein (CP) genes, respectively. In the phylogenetic tree constructed for RNA‐1, MP, and CP sequences, all Turkish raspberry isolates were clustered in a distinct clade. However, the blackberry isolates showed considerable variation in nucleotide sequences and were placed in three distinct groups. The divergent blackberry isolates showed high variability in MP (84.5%–89.3%) and CP (85.5%–89.7%) regions and were placed in a distinct group. The rest of blackberry isolates clustered together with sweet cherry RBDV isolates adjacent to the grapevine clade or together with raspberry isolates. The comparative analysis conducted on three RNA segments of RBDV highlighted the high sequence diversity of Turkish RBDV isolates. This study also emphasizes the importance of regular monitoring of RBDV infections in Turkey, with special regard to those Rubus spp. and grapevine accessions employed in conservation and selection programmes. In particular, the presence of new RBDV genetic variants and infection of Rubus species must be taken into account to choose a correct detection protocol and management strategy.

Global Population Structure of Apple Mosaic Virus (ApMV, Genus Ilarvirus).

The gene sequence data for apple mosaic virus (ApMV) in NCBI GenBank were analyzed to determine the phylogeny and population structure of the virus at a global level. The phylogenies of the movement protein (MP) and coat protein (CP) genes, encoded by RNA3, were shown to be identical and consisted of three lineages but did not closely correlate with those of P1 and P2, suggesting the presence of recombinant isolates. Recombination Detection Program (RDP v.4.56) detected significant recombination signal in the P1 region of K75R1 (KY883318) and Apple (HE574162) and the P2 region of Apple (HE574163) and CITH GD (MN822138). Observation on several diversity parameters suggested that the isolates in group 3 had higher divergence among them, compared to isolates in groups 1 and 2. The neutrality tests assigned positive values to P1, indicating that only this region experiencing balanced or contracting selection. Comparisons of the three phylogroups demonstrated high Fixation index (FST) values and confirmed genetic separation and the lack of gene flow among them. Additionally, ±500 bp of partial MP + 'intergenic region' + partial CP coding regions of two Turkish isolates from apple and seven from hazelnut were sequenced and determined that their phylogenetic positions fell within group 1 and 3, respectively.

Genome characterization of a Turkish bovine rotavirus field isolate by shotgun metagenomics.

A bovine rotavirus (BRV) isolate from Kirsehir was isolated from feces of a neonatal calf with diarrhea, identified, and sequenced by shotgun sequencing. Its genotype constellation is G10-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3. The structural genes and the non-structural genes NSP1, NSP3, and NSP4 of the Kirsehir isolate were similar in sequence to those of BRVs identified in Turkey. However, VP2, NSP2, NSP4, and NSP5/6 showed similarity to those of rotaviruses from different animal hosts. These findings not only expand our current understanding of the diversity of rotaviruses but also contribute to our understanding of the evolution of rotaviruses at both the national and global levels and reinforce the significance of conducting further research on rotaviruses in Turkey.