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Two species of Pratylenchoides recovered from the grasslands in Sabalan region and one species recovered from natural habitats of Tehran are illustrated based on morphological, morphometric and molecular characters. The first species, P. crenicauda is characterized mainly by its lip region with three-four annuli, lateral field with four incisures areolated throughout the length and having rod shaped sperm cells. It is further distinguished by the positions of the pharyngeal glands nuclei. P. magnicauda was found in Tehran and its morphological characters and phylogenetic relations with other species are discussed. The Iranian populations of P. variabilis are characterized by three lip annuli, stylet 20-22 µm long, four and six incisures in lateral field, rounded sperm and one of the pharyngeal glands nuclei located posterior to pharyngo-intestinal valve. The phylogenetic tree inferred from the partial sequences of D2-D3 segment of 28S rDNA revealed the three sequenced species are separate from each other and form a clade with high (1.00) Bayesian posterior probability (BPP) in Bayesian inference (BI) and 86% bootstrap support value (BS) in maximum likelihood (ML) analyses with other two sequenced species of the genus for this genomic region.

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Radula as the main feeding organ in gastropods not only functions in obtaining food, but also considered as an important organ in classification and phylogenic studies. In this research, the radula morphology of the two species, Conomurex persicus (Strombidae) and Mauritia grayana(Cypraeidae) from BandareLengeh and the Larak Island were analyzed. The collected samples were fixed in 70% alcohol, and radulae were isolated and processed in the laboratory, then photographed with a scanning electron microscope (SEM). Radula in these two species is of taenioglossan type with 7 teeth per row, but structure of radula teeth in two species are different from each other. Due to various type of feeding habits in C. expersicus and M. grayana, they require radulae with structure to suit their needs. On other hand, any radula structure can also imply special dietary habit, so it can be used to understand the feeding habit of a particular gastropod species.

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The chloroplast gene matK, formerly known as ORF509, has been identified as one of the most rapidly evolving genes at the nucleotide and corresponding amino acid levels. This gene is located in the large single-copy region of the chloroplast genome, and placed between the 5’ and the 3’ exons of trnK (tRNA-lysine) within a group II intron. The matK RNA and protein levels are affected by light and developmental stages, suggesting functional roles for this putative maturase that affect in photosynthesis indirectly. The matK has been considered as one of the most useful genes for resolving phylogenetic and evolutionary relationships at a range of taxonomic levels, from closely related species to the generic, familial, and even supra-familial levels among land plants, especially Angiosperms. The matK as a DNA barcode for land plants showed high levels of discrimination among angiosperm species that can be used single or in combination with other genes.

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Entomopathogenic nematodes (EPNs) are commonly used as biological agents for control of insect pests. This study was carried out to identify EPNs, determine dominant and frequent species in Kurdistan province, Iran and characterize their symbiotic bacteria. Identification of EPNs was performed based on morphological and morphometrical characters and also rRNA-ITS gene sequences. Two EPNs, Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae) and Steinernema feltiae (Rhabditida: Steinernematidae) were identified. Out of totally 150 soil samples collected mainly from mid-southern parts of the province, 60% were positive for EPNs. Heterorhabditis bacteriphora showed the highest frequency in this region, remarkably 59.3% of soil samples contained this species. Incidences of H. bacteriphora in grasslands, woodlands and alfalfa fields were 57.3, 14.6 and 28.0 percent, respectively. Steinernema feltiae was found only in alfalfa fields with 0.66% frequency. Bacterial symbionts of H. bacteriophora and S. feltiae were also identified based on biochemical characters and recA gene sequencing. In this research, two species of Photorhabdus were isolated from H. bacteriophora namely P. luminescens subsp. kayaii and P. temperata subsp. thracensis. Xenorhabdus bovienii was identified from infective juveniles of S. feltiae.

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There are very few reports discussing the higher level phylogeny of the chewing lice (Phthiraptera) and their roles in the evaluation of these insects. So, the aim of this work is to study the phylogeny of genera of the suborder Ischnocera in Saudi Arabia using morphological, ecological and molecular criteria trying to answer a very important question about ecological factors that affecting their evolution. The results show high degree of similarity between phylograms produced by morphological and ecological criteria and that produced by genes sequencing data of Cytochrome Oxidase Subunit I (COI) and elongation factor-1 alpha (EF-1). The rooted trees showed several clusters for head chewing lice and this indicated that the phenomenon of parasitizing birds’ heads evolved several times in the clade of chewing louse. This is also indicating that the position on host played a very important role in shaping the diversity of these insects. The unrooted tree which produced by analysis of COI and (EF-1) also supports the previous conclusion but added to it the impact of host habitat on such diversity. The evolutionary distance between marine birds’ chewing lice was very small and all of them appear in single line or group which far from that of terrestrial taxa of the chewing lice. Both the host habitat and the feeding position of chewing lice on their hosts played a very important role in the way by which this group of insects evolved.

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The nucleotide sequences of 28S nuclear rDNA were determined for 34 powdery mil-dew taxa mostly collected from Iran in order to infer the phylogenetic relationships of these fungi. Total DNA was isolated from cleistothecia or mycelia using the chelex method. About a 650 nucleotide length of the 5´end of the 28S rDNA was amplified twice by the PCR using a nested primer set, PM3, TW14 and NLP2. Direct sequencing of the PCR product was done in an Applied Biosystems 373A sequencer. The results showed that powdery mildew taxa are divided into five groups, which were distinguished by their morphology. Members of Erysiphe section Erysiphe, Microsphaera and Uncinula clustered together. E. sect. Galeopsidis and E. sect. Golovinomyces were seperated from E. sect. Ery-siphe and formed the Euoidium without fibrosin bodies group. Leveillula and Phyllactinia showed a close evolutionary relationship and clustered together. The genera Cystotheca, Podosphaera, Sawadaea, and Sphaerotheca formed a monophyletic group (fibrosin body lineage) with 98% bootstrap support. These fungi are well characterized by the presence of fibrosin bodies in their conidia. Blumeria graminis, which is characterized by some unique morphological characters, clustered with fibrosin body lineage with a low boot-strap value. This result shows that B. graminis is not closely related to the Erysiphe spe-cies. The nucleotide divergence between the genera analyzed in this study ranged from 0.50 to 14.10%. The lowest nucleotide divergence was found between Microsphaera and E. sect. Erysiphe (0.50–4.50%). Podosphaera and Sphaerotheca showed a low level of diver-gence, too (2.30-2.60%), which suggests a close relationship between these two genera.

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In this study, 32 plant samples with leaf spot symptoms were collected from Chaharmahal and Bakhtiari province, Iran, in the spring and summer of 2018. Isolation and purification of 26 fungal isolates were performed on 2% water agar and potato dextrose agar to identify the causal agents. Fungal species were identified according to morphological characteristics and molecular data obtained from glyceraldehyde 3-phosphate dehydrogenase (gpdh) gene sequences. In this research, 11 species belonging to four genera of hyphomycetous fungi, including Alternaria cantlous, A. consortialis, A. multiformis, Bipolaris sorokiniana, B. zeicola, Curvularia spicifera, C. nicotiae, C. inaequalis, Stemphylium beticola, S. symphyti, and S. vesicarium are introduced. Among them, three species, including C. nicotiae from Salvia officinalis L., S. beticola from Plantago major L., and S. symphyti from Mentha pulegium L., are reported as new records for mycobiota of Iran. All collected plant species are reported as new hosts (matrix nova) for the identified fungal taxa.

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Some saprophytic fungi were isolated during fungi isolation causing black spot or scab-like symptoms (genus Venturia) from Iran. One saprophytic isolate from apple leaf was identified as a member of the family Phaeosphaericeae based on morphological characteristics of pseudothecia, asci, and ascospores on oatmeal agar (OA). Mature pseudothecia were produced on OA after one-month incubation at 24 °C and continuous dark conditions; however, the pseudothecia produced on PDA were immature. Pseudothecia were dark brown and globose in shape. Numerous bitunicate asci formed in a broad hymenium. Asci were clavate, apically rounded, short pedicellate, with eight overlapping biseriate fusiform ascospores. Ascospores with the smooth wall, straight or slightly curved and slightly constricted at the second septum. Phylogenetic analysis based on sequence data of ITS and LSU regions of ribosomal DNA confirmed the morphological identification and specified the isolate as Neosetophoma poaceicola. The N. poaceicola is the first report from Iran and apple leaf in the world.
 

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The spiders of the genus Trichopelma Simon, 1888 present in Cuba, are revised. Currently, the genus Trichopelma comprises 22 known species distributed in the Caribbean, and Central and the upper region of South America. Cuba currently hosts five valid species and, in this study, the descriptions of seven new species distributed throughout the island are presented: T. baracoense sp. nov. (♂♀, Guantanamo prov.), T. cheguevarai sp. nov. (♂, Ciego de Ávila prov.), T. citma sp. nov. (♀, Granma prov.), T. fidelcastroi sp. nov. (♂♀, Holguín prov.), T. granmense sp. nov. (♂♀, Granma prov.), T. rudloffi sp. nov. (♂♀, Holguín prov.) and T. soroense sp. nov. (♂♀, Artemisa prov.). Based on morphological characters, a cladistic analysis was performed, revealing the phylogenetic position of the new species compared to the species previously described. Based on this phylogeny, morphological characters and close proximity in distribution, T. banksia Özdikmen & Demir, 2012 syn. n., is proposed as a junior synonym of T. cubanum (Simon, 1903). The genus Thalerommata Ausserer, 1871 is reported from Cuba for the first time, with the description of T. anae sp. nov. (♂, Sancti Spíritus prov.).

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Microgastrines are diverse group of endoparasitoid wasps attacking caterpillars (Lepidoptera). Despite their importance in biological control, there is still no consensus concerning the phylogeny relationships among taxa. Although previous phylogenetic analyses have advanced the overall understanding of phylogenetic relationships of Microgastrinae, the small numbers of sampled taxa have led to disagreement in taxonomic assignments. In the present study, we performed a molecular genetic survey using both mitochondrial and nuclear data, increasing the taxons' sampling, to clarify the generic relationships and improve the inferences of the taxonomic status within Microgastrinae. We reconstructed a phylogenomic tree of Microgastrinae with sequences that exist up till now, from fifty-five genera for COI and thirty genera for 28S rDNA, both new and from previous studies. Several species and genera have been sequenced for the first time. In this study, we identified some of the closest phylogenetic relatives of Microgastrinae genera by analyzing DNA sequences from the mitochondrial COI and 28S rDNA. Most clades of the current findings correspond to the latest morphological classification of Microgastrinae. New clades and several well-supported clades, conform to the most previously recorded clades and provide an increased understanding of the Microgastrinae evolution. Based on molecular examination, Pholetesor psedocircumscriptus Abdoli, 2019 is synonymized with Pholetesor circumscriptus (Nees, 1834).

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Chitinases are essential enzymes in crustaceans that play an important role in the molting cycle and digestion of chitin. Based on the present study, the chitinase encoding cDNA of Penaeus mergueinsis with a length of 1440 bp containing 467 amino acids was sequenced by PCR and then its phylogenetic and bioinformatics analysis was performed. The new sequence was registered in the gene bank with the accessition number MT250539 and the molecular weight of the protein resulting from this sequence was predicted to be 51.84 KDa and the theoretical isoelectric point of 4.79. Comparison of amino acid sequences among penaeid chitinases showed the highest identification (about 97 to 92%) with P. mondon chi-3, F. chinensis, P. vannamei and P. japonicus chi-3, respectively. Phylogenetic studies showed that chitinase in the present study belongs to group 3 chitinases. Revealed protein pattern analyzes showed that chitinase from P. mergueinsis contained the catalytic domain Glyco-18 at position 2-347, a chitin-binding site of pritrophin A at position 403-456, a disulfide bridge formed by two cysteines at position 436-421 is a chitin-binding domain type 2, active site (¹¹7FDGLDMDWE¹²⁵), a proline / threonine-rich region at positions 376-412, and a putative N-glycosylation site at position 427-424 (NTSG). The present study shows that the P. mergueinsis sequence contains active chitinase motifs similar to previously sequenced chitinases, and in the case of cloning, expression and purification probably has functional and structural features similar to the enzymes of the above species.

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To identify the endophytic fungi of the caper plant, samples were collected from the healthy leaves, stems, and roots from Bushehr and Qom provinces during the summer and early autumn of 2022. Isolation and purification of fungal isolates were performed on potato dextrose agar and 2% water agar culture media, respectively. A total of 102 fungal isolates were obtained. Morphological characteristics and molecular data of the ITS region and TUB gene were used to identify the fungal species. Based on the results, Microascus cirrosus and M. terreus were identified as new endophytic fungi of caper for the first time in the world, and M. terreus was introduced as a new record for the funga of Iran.


 

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Fungal endophytes are defined as microorganisms with the ability to colonize plants asymptomatically throughout or at least a significant part of their life cycle, thereby establishing a plant-fungal association. In the present study, 44 plant samples, including healthy and symptomless fruit, leaf and branch samples, were collected from Eucalyptus trees located in Tehran, Qom, Alborz, Esfahan, and Mazandaran provinces to isolate and identify the endophytic fungi. Among 170 fungal isolates from collected plant samples, two species were new based on morphological and molecular phylogeny of the ITS rDNA for the Funga of Iran, including Phaeophleospora eucalypticola and Pseudosydowia eucalypti. Furthermore, both species are reported for the first time as endophytic fungi of eucalyptus trees in the world.

 

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During a survey in the spring of 2024, noticeable severe leaf yellow spots, yellowing, fruit necrosis, and chlorotic ring spots symptoms were observed in Cucumber plants cultivated near Isfahan City, central Iran, to assess Orthotospovirus infection. Total genomic RNA was individually extracted from the leaves of 5 symptomatic and one asymptomatic leaf sample. The extracted RNA samples were subsequently subjected to RT-PCR using an Orthotospovirus-universal primer pair. An amplicon of the expected size was obtained with all diseased samples, and a BLAST search of its nucleotide sequences exhibited a high level of nucleotide identity with several MYSP isolates. This is the first report of the melon yellow spot virus (MYSV) Orthotospovirus meloflavi occurrence in Iran.



 

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Hop stunt viroid (HSVd) isolates have been reported as the causal agent of citrus cachexia in Mazandaran Province and recently shown to be associated with yellow corky vein disease of sweet orange and split bark disorder of sweet lime in the Fars Province, Iran. In the present work isolation and partial characterization of viroids from citrus trees affected by gummy stem blight is reported from Kohgiluyeh–Boyerahmad (KB) Province of Iran. Fifteen samples of citrus trees from Dehdasht area (KB Province) showing bark necrosis, gum exudation and die-back as well as seven citrus symptompless trees from the same area were tested for the prevalence of viroids, through Reverse Transcription Polymerase Chain Reaction (RT-PCR) followed by sequencing of PCR products. They were also tested for Citrus tristeza virus through Double-Antibody Sandwich Enzyme Linked Immunosorbent Assay (DAS-ELISA). Two variants of HSVd which differed from GenBank isolates in nucleotide sequence and two variants of Citrus Bent Leaf Viroid (CBLVd) were identified in any of the symptomatic samples. Moreover, a Citrus Exocortis Viroid (CEVd) was found only in symptomatic sweet lime. An HSVd isolate from KB (HSVd-bn₁) was selected and used for comparison with a number of HSVd variants from Iran (Fars and Mazandaran Provinces) and the related accessions from GenBank. On the basis of nucleotide sequence and secondary structure analysis, HSVd-bn1 and HSVd-bn2 belong to non-cachexia variants of HSVd and have about 95% similarity to Citrus gummy bark viroid, a sub-species of HSVd. CTV was not detected in the diseased plants. It is yet to be determined whether bark necrosis of sweet lime and of sweet orange plants is caused solely by the associated viroid(s) or other factors are involved as well.

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Theplant pathogenic Phytophthora drechsleri is morphologically similar to some other non-papillate Phytophthora spp., especially P. melonis, and it is difficult to discriminate these convergent taxa. It seems that the Iranian putative P. drechsleri isolates from different cucurbit species have been generally misidentified and their characteristics do not match with P. drechsleri. In order to compare these two groups, authentic P. drechsleri isolates and isolates from different cucurbits were assessed for morphological, physiological (cultural, temperature relations, mating type), and molecular traits. Multiple gene genealogy analysis were performed on regions of nuclear (ITS, β-tubulin, translation elongation factor 1α, elicitin) and mitochondrial (cytochrome c oxidase subunit I) gene sequences. Congruence was observed in different phylogenetic data sets. The present study demonstrated that putative P. drechsleri isolates from cucurbits and pistachio trees were a distinct species and belonged to P. melonis. Data showed that P. melonis was a homogenous species and there were no considerable molecular intraspecific variations between isolates from cucurbits and isolates from other hosts. Design of a molecular species-specific identification tool for P. melonis isolates is under investigation. 

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Cyclotides are small disulfide-rich proteins that have the unusual feature of a cyclic backbone. Cyclotides have a range of interesting biological activities and are found in a variety of tropical plants from the Rubiaceae, Violaceae, Cucurbitaceae and Fabaceae families. We have cloned and characterized cyclotides in Viola modesta, a Viola species native to western Asia, which was collected from the Kurdistan Province of Iran. Fifteen cyclotide sequences were obtained using homology based PCR strategy. Sequence analysis showed that 14 of them had continued open reading frames and showed high level of similarity to cyclotide genes from other species of the Violaceae. After analyzing the full endoplasmic reticulum signals of V. modesta cyclotides, two conserved sequences, AAFALPA and ATAFALP, were detected. Analysis of isolated cyclotide sequences showed that they all belonged to bracelet family and were separated into two subclasses. Phylogenetic analysis of cyclotide genes from V. modesta and other Viola species revealed that most V. modesta genes showed close relationship with their homologs from the Violaceae, while the V. modesta genes formed two separate clades. Transcription analysis by semi-quantitative RT-PCR revealed that Vmcyc1 and Vmcyc7 were differentially expressed in all tested tissues including roots, stems, leaves, flowers, seeds, peduncles, and capsules with the highest transcript level in the capsules.

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Three species belonging to three genera of the family Tylenchidae, namely, Cephalenchus leptus, Eutylenchus excretorius, and Lelenchus leptosoma were discovered as first reports for Iran's nematode fauna. They were characterized by morphological, morphometric and molecular phylogenetic studies. The two latter genera are new for Iran. Iranian population of Cephalenchus leptus is characterized by lateral field with six lines in females, stylet length of 16-20 µm and tail length of 153-290 µm with pointed tip. The recovered population of Eutylenchus excretorius is characterized by 848-1,038 µm long females’ body, stylet length of 20.0-21.5 µm, having advulval flaps and absence of male. The Iranian population of Lelenchus leptosoma is defined by its narrow slender body, flattened lip region, sinuous amphidial opening and absence of lateral field. The phylogenetic relationships of the three recovered species were studied using the partial sequences of 28S rDNA D2/D3 segment and revealed the genera Cephalenchus and Eutylenchus forming a monophyletic clade, while, Lelenchus was placed inside the clade of currently sequenced species of Malenchus in Bayesian tree.

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Low Molecular Weight Glutenin Subunits (LMW-GS), encoded by GLU-3 loci located on the short arm of homologous chromosomes of wheat, play an important role in the bread making quality. Some of the most important gens for quality are located on the D genome, which are interesting in wheat breeding programs. In addition to the bread wheat (Triticum​​​​​​​ aestivum), some species of Aegilops including Ae. cylindrica, Ae. tauschii, Ae. crassa, Ae. juvenalis, and Ae. vavilovi carry D genome. In this study, the phylogenetic relationship among Aegilops species with D genome and bread wheat has been studied based on the sequence of low molecular weight glutenins loci. The results indicated a great diversity for these loci. Presence of several numbers of common protein bands among species suggested a close relationship and high genetic flow among species. Three primers for the LMW-GS proteins were able to reveal the relationship between the species. The results showed a close relationship among bread wheat (T. astivum) and Ae. tauschii species. Ae. crassa species is more distant from bread wheat. Also, the results indicated a close relationship between the Ae. cylindrica, Ae. juvenalis, and Ae. vavilovi. A great diversity of LMW-GS in wild relatives and close relationship between these species and wheat suggest them as a potential source of genes for wheat breeding programs.
 

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 To investigate the infection of carnations (­Dianthus caryophillus) to common viruses, a total of 130 samples showing suspected viral disease symptoms were collected from 10 commercial greenhouses located in Mahallat City, Markazi Province. Total DNA and RNA were extracted and amplified by PCR and RT-PCR methods, respectively, using generic primers for the genera Potyvirus and Orthotospovirus and also specific primers of Carnation mottle virus (CarMV), Carnation necrotic fleck virus (CNFV), Carnation etched ring virus (CERV), Carnation latent virus (CLV), Carnation vein mottle virus (CVMV) and Cucumber mosaic virus (CMV). Results revealed that the estimated rates of infection in the collected samples for different viruses in decreasing order were for CarMV, CNFV, CERV, CMV and CVMV, while CLV and species of genus Orthotospovirus were not detected. Mixed viral infections were also investigated and among all of the 130 collected samples, 3.1% showed mixed infections with CarMV, CNFV, CVMV, CERV and CMV, 32% with CarMV, CNFV, and CERV, 20% with CarMV and CNFV, and 9% with CarMV and CERV. For phylogenetical assays, NIb-CP fragment of the CVMV genome was amplified, cloned, and sequenced. Phylogenetic trees of two isolates of this virus revealed that the two Iranian isolates of CVMV (IBR and RCM) were phylogenetically divergent. Mechanical inoculation of CVMV on 16 different indicator plant species showed specific symptoms on six of the indicators that were confirmed by RT-PCR. To the best of our knowledge, this is the first report of CMV and first partial genome sequencing of CVMV from a carnation plant in Iran.