Morphological Identity Research Articles

First report of Needle Cast of Cathaya argyrophylla caused by Neofusicoccum parvum in China.

Cathaya argyrophylla Chun et Kuang is a first-grade protected tree in China with significant conservation value. In June 2022, needle cast was observed in approximately 27 to 34% of C. argyrophylla (n=200, covering about 1 ha) at Dashahe National Nature Reserve (28.89° N, 107.6° E) in Daozhen County, Guizhou, China. Initial symptoms were small red lesions (3 to 5 mm in diameter) on needles. Over time, these lesions expanded, turned necrotic, and led to needle cast. In addition, the diseased needles exhibit brown spots surrounded by a yellowish discoloured area measuring 0.5 mm2. The disease is most severe in young trees, with over 60% of the needles displaying discoloration in 15% of young trees (n=100). Forty infected conifer tissues were randomly selected, surface sterilized, and incubated on PDA in the dark at 26°C for 5d. Among the 40 samples, 39 exhibited fungi with similar morphology. Single-spore isolation method was used to obtain pure cultures for 9 isolates. The isolates were morphologically identical. On PDA medium, colonies were white with abundant aerial mycelium. After 14 days of growth under light, the colonies became greyish black with septate mycelium. Conidia were hyaline, thin-walled, smooth-surfaced, and ranged from ellipsoid to ovoid in shape, measuring 6.5 to 15.2 ×2.0 to 3.1 μm (n=50). Based on these morphological characteristics, the isolates were identified as belonging to Neofusicoccum spp. (Pavlic et al., 2009). For molecular identification, genomic DNA was extracted from 3 selected isolates. The internal transcribed spacer (ITS) region was amplified using primers ITS1/ITS4 (White et al., 1990). In addition, chitin synthase 1 (CS1) was amplified using primers CT-WK3-S/CT-WK3-A (Zimoch et al., 2003). The ITS (OR710949) and CS1 (OR714767) sequences of isolate JY1-1 were deposited in GenBank, exhibiting a homology of 99% to 100% (537/538, 191/192) with N. parvum MUCC211 (accession numbers EU301017 and EU339495). The reconstructed phylogenetic tree further substantiated the genetic relationship between isolate JY1-1 and N. parvum. Consequently, the isolate associated with needle cast on C. argyrophylla was identified as N. parvum. Isolates JY1-1, JY1-2, and JY1-3 were used to confirm Koch's postulates. The needles of 15 healthy 2-year-old C. argyrophylla saplings were inoculated with a conidial suspension of N. parvum (2.0 × 105 conidia/mL). As a control, the needles of C. argyrophylla saplings were treated with sterile water (n=10). The needles were covered with Ziplock bags, and maintained at humidity levels exceeding 90%. All treatments were placed in a greenhouse at 26℃. After 7 days of inoculation, the needles exhibited a change in color, with reddish-brown symptoms observed on day 30 (disease rate=100%). Furthermore, the lesion expanded, and needles were shed after 4 months. No symptoms were observed in the control group. Pathogens reisolated from all diseased plants exhibited morphological and ITS sequence identity to N. parvum (separation rate=100%), and no pathogens were isolated from control plants. Consistent results were obtained through two repeated experiments. While this pathogen is known to induce foliar disease in various plants (Sun et al., 2020; Shi et al., 2019), our study represents the first report Needle Cast of C. argyrophylla caused by N. parvum in China. These findings provide a foundation for developing effective strategies to monitor and manage C. argyrophylla.

Identification and Characterization of Myxobolus pronini (Cnidaria: Myxozoa) from Gibel Carp Carassius auratus gibelio and Goldfish C. auratus: New Fish Host, Infection Site, and Geographic Distribution in China

Myxozoans represent a significant group of parasitic pathogens affecting both wild and farmed fish. For accurate and comprehensive early detection, the composition of myxozoan pathogens in fish has consistently been a primary focus for parasitologists. In China, we undertook an investigation into myxozoan infection in fish from Shandong Province, China, successfully isolating a myxozoan species from gibel carp Carassius auratus gibelio Bloch, 1782 and goldfish C. auratus Linnaeus, 1758. In the present study, this myxozoan species was identified by detailed morphological and molecular analysis. This parasite is characterized by the presence of visible plasmodia in various organs (intestine, liver, gallbladder, and abdominal cavity) of gibel carp as well as the abdominal cavities of goldfish. In comparison to all documented myxozoan species, this myxozoan species exhibits morphological identity with Myxobolus pronini Liu, Batueva, Zhao, Zhang, Zhang, Li, Li, 2016, in terms of spore morphology. Molecular sequence analysis, including sequence similarity, variable site, and the secondary structure of SSU rRNA, indicates that the present myxozoan species closely resembles M. pronini. Furthermore, phylogenetic analyses revealed that all isolates collected from different organs and hosts in this study clustered with M. pronini, forming a distinct branch within the Carassius-infecting clade. Consequently, the present myxozoan species can be confidently identified as M. pronini. Compared with the previous reports on M. pronini, this study documents, for the first time, goldfish as a host, intestines and gallbladders as the sites of infection, and Shandong Province as the geographic distribution of this myxozoan species.

Biopriming of Maize with their endophyte Aspergillus fumigatus reinforces their resistance to salinity stress and improves their physiological traits

Zea mays L. (Maize) is one of the most crucial world’s crops, for their nutritional values, however, the water scarcity and consequent soil salinization are the major challenges that limit the growth and productivity of this plant, particularly in the semi-arid regions in Egypt. Recently, biopriming has been recognized as one of the most efficient natural-ecofriendly approaches to mitigate the abiotic salt stress on plants. The haploid (128) and triploid (368) seeds of maize were selected as model verities for assessing their resistance to salt stress and mitigating their effect by fungal-biopriming. Overall, the haploid and triploid plants viabilities were drastically affected by salt concentration, at 500 mM of NaCl. At 500 mM NaCl, the fresh weights of the triploid and haploid seedlings were reduced by ~ 5 and 6.1 folds, compared to the controls, ensuring slightly higher salt resistance of the triploid than haploid ones. The pattern of the endophytic fugal isolates was plausibly changed with the salt concentration for both plant types, Aspergillus fumigatus isolate was emerged with the higher NaCl concentration (400–500 mM), and their morphological identity was molecularly confirmed and deposited into Genbank with accession # PQ200673. The fungal bioprimed seeds of the haploid and triploid plants were irrigated with 400 mM NaCl. The fungal-bioprimed plants displayed a significant improvement on the shoot density, fibrous roots, root length, shoot length, and leaves numbers and areas of the stressed-plants by ~ 1.7 folds, compared to control, ensures the triggering of different salt resistance machineries in plants upon fungal biopriming. The total antioxidant enzymes activities “catalase, peroxidase, superoxide dismutase” of the salt-stressed bioprimed maize plants were increased by ~ 4.7–5.3%, compared to control, confirming the mitigating effect of the salinity stress on plants upon fungal biopriming. The chlorophyll and carotenoids contents were significantly increased of the salt stressed maize upon biopriming with A. fumigatus. The expression of the sod, apx2, nhx11, hkt1, H + -PPase, nced of the plant salt stressed was strongly increased in response to A. fumigatus biopriming, normalized to β-actin gene. The expression of apx2 was dramatically increased by about 30 and 43 folds, in response to fungal biopriming. The nhx1 was significantly up-regulated by 18.9 fold in response to fungal biopriming, compared to control.

Diversity of the ‘eerie’ glass squid genus Taonius Steenstrup, 1861 (Cephalopoda: Oegopsida) in the Pacific Ocean

ABSTRACT ‘Glass’ squids (family Cranchiidae Prosch, 1849) are diverse and widely distributed, but many cranchiid genera are in need of global taxonomic revision. Although two species of Taonius are generally recognised from the Pacific Ocean (T. belone and T. borealis), a recent review of material from this basin has revealed the presence of at least six species, including at least four new to science, described here as T. expolitus, T. notalia, T. robisoni, and T. tanuki. Morphological species identities are supported by several characters, including arm sucker count, and arm and manus sucker dentition. Proposed species delimitations are further supported by differences in mitochondrial DNA (cytochrome c oxidase subunit I). Although a full systematic review of the Cranchiidae is still needed tripling the known diversity of Taonius in the Pacific Ocean and clarifying identities of its locally occurring species represent an important step forward.

Characterization of Fusarium falciforme Inciting Wilt in Peace Lily (Spathiphyllum wallisii)

Severe wilt incidence with characteristic yellowing of older leaves was observed in Spathiphyllum plants of different commercial nurseries of Andhra Pradesh, India during 2021-23. Upon isolation on potato dextrose agar medium, the fungus displayed aerial greyish white mycelium on upper surface and with yellow pink pigmentation on the lower surface. Microscopic observation of the three types of conidia namely microconidia, macroconidia and chlamydospores confirmed the morphological identity of the pathogen as genus Fusarium. Multi-gene sequencing analysis, utilizing ITS, Tef-1α, RPB1, and RPB2 gene sequences, further confirmed the pathogen's identity at the species level as Fusarium falciforme. Pathogenicity assays in 30 days old Spathiphyllum plants demonstrated that yellowing of leaves was observed seven days after pathogen inoculation using a non-invasive method, followed by complete wilting of the plants within 15 days. In addition, host range analysis revealed that the isolated pathogen isolate from wilted Spathiphyllum plants could cause complete wilting within 12-15 days post-inoculation in other economically important solanaceous crops such as tomato, brinjal, chili and tobacco, further highlighting its economic significance. Studies evaluating different fungicides revealed that root zone application of carbendazim 50%WP at 1g/l and tebuconazole 25.9% EC at 1ml/l were effective in controlling wilt incidence, achieving 77.78% reduction as a prophylactic treatment and 66.67% reduction as a curative treatment. To the best of our knowledge, this is the first comprehensive study on Spathiphyllum wilt incidence and its management worldwide.

Molecular phylogeny of Boninia (Platyhelminthes: Polycladida), with description of a new species from the Pacific coasts of Panama.

Mesopsammic polyclad members in the family Boniniidae have attracted attention in terms of their evolutionary shifts of microhabitat and their unique morphology such as a pair of pointed tentacles extending from the anterolateral margins and prostatoid organs harbouring stylets. Here, we establish a new species of this family as Boninia panamensis sp. nov. from the Pacific coasts of Panama, based on its morphological characteristics of (i) four cerebral and 61-80 marginal eyespots, (ii) two prostatoid organs located anterior and posterior to the penis papilla, and (iii) two uterine canals departing from the anterior part of the Lang's vesicle. We also report Boninia cf. uru from Hawai'i, USA, based on its morphological identity with B. uru from Okinawa, Japan, along with their genetic distances for the partial cytochrome c oxidase subunit I (COI ) sequences, which were beyond the range of intraspecific differences observed in congeners in this study. Boninia oaxaquensis is also reported from Panama as a new locality for the species. Involving the above-mentioned three species sequenced herein, we reconstructed molecular phylogenetic trees of Boninia based on the four gene markers (18S rDNA, 28S rDNA, 16S rDNA and COI ). Our phylogenetic trees indicated the synapomorphy within the genus Boninia of the small numbers of stylets (2-4) and the connection route of the uterine canals to the Lang's vesicle. The results also showed a characteristic distribution pattern in which pairs of species in distinct lineages occurred sympatrically with different microhabitats, as observed in Boninia uru and Boninia yambarensis in Okinawa and B. panamensis sp. nov. and B. oaxaquensis in Panama. In addition, we discuss possible speciation pathways in this genus based on the tree topology. ZooBank: urn:lsid:zoobank.org:pub:D414BACD-C14A-4B34-8AF9-7ACBA28F46D0.

First blood-meal record of Simulium asakoae (Diptera: Simuliidae) in Malaysia, with notes on its distribution in Asia and status as a potential vector.

Simulium asakoae Takaoka and Davies has been confirmed to bite humans and has been incriminated as a vector of blood protozoan parasites of the genera Leucocytozoon and Trypanosoma, as well as an unknown filarial parasite in Thailand. However, its attraction to humans has remained uninvestigated in Malaysia. Recently, 27 black flies were collected in Pahang, Malaysia, of which 25 were captured in CO2-baited Malaise traps and two were collected from humans during trapping activity. All specimens were morphologically identified as S. asakoae. Cytochrome c oxidase I sequences of the two specimens caught on humans showed 100% similarity with those of S. asakoae in the NCBI GenBank, confirming their morphological identity. Blood-meal analysis using a HAL·HARTM kit did not show the presence of domestic or wild animal DNA. However, human DNA was amplified from one engorged fly in the cytochrome b gene amplification assay, providing the first evidence of blood-feeding by S. asakoae in Malaysia.

First Report of Powdery mildew caused by Golovinomyces ambrosiae on Bidens pilosa in Guangdong Province, China.

Bidens pilosa L., an annual herbaceous plant with a wide distribution, possesses novel medicinal properties. In January 2021, a powdery mildew disease outbreak was documented on B. pilosa plants located in the roadside areas in Shenzhen, Guangdong Province, China, with 60 to 80% disease incidence. Initial symptoms manifested as small, irregular white powdery patches, primarily on the adaxial surfaces of leaves. Subsequently, the colonies expanded, forming coalescent colonies that spread across the leaves, petioles, and stems, eventually leading to the distortion and senescence of leaves. Hyphae are hyaline, flexuous to straight, septate, with thin walls and a width ranging from 2 to 8 μm. Hyphal appressoria are nipple-shaped. Conidophores are erect or slightly flexuous, ranging from 80 to 150 µm in length and 12 to 18 µm in width (n = 30). Typically, these conidophores bear chains of 2 to 5 immature conidia, displaying a sinuate outline. Foot-cells, located at the base of conidophores, are cylindrical and erect, approximately 33 to 100 µm in length and 6 to 10 µm in width (n = 30). Conidia are hyaline, ellipsoid-ovoid to barrel-shaped, and lack fibrosin bodies. Primary conidia are ellipsoid-ovoid in shape, characterized by a rounded apex and a subtruncate base, 25 to 40 µm × 15 to 22 µm in width. Secondary conidia are barrel-shaped with truncate or subtruncate ends, 27 to 35 µm × 15 to 20 µm in width. Germ tubes exhibit a longitubus pattern and are prominently produced at the perihilar or apical region of the conidia. No chasmothecia were observed in the collected samples. In order to conduct a molecular-level identification, mycelium and conidia were collected from B. pilosa leaves. Genomic DNA was subsequently extracted from these samples. The internal transcribed spacer (ITS), intergenic spacer (IGS) and beta-tubulin (tub2) sequences were performed using primer pairs ITS1/ITS4, IGS-12a/NS1R, and tub2, respectively (Carbone and Kohn 1999; Scholin et al. 1994; White et al.,1990). A 568-bp ITS, a 366-bp IGS, and a 354-bp tub2 sequences (GenBank accession nos. OR647592, OR978282 and OR978283) were obtained. The ITS sequence exhibited over 99.6% similarity to Golovinomyces ambrosiae (MT929773) and G. cichoracearum (MH590731). The IGS sequence displayed 100% similarity to G. ambrosiae (MK383490) and G. ambrosiae (OK349420). The tub2 sequence displayed 100% similarity to G. ambrosiae (MW981257) and G. ambrosiae (MW981256). Phylogenetic analysis of IGS, ITS and tub2 also grouped obtained sequences within the G. ambrosiae complex. Based on the analysis of morphological characteristics and sequence identity, the pathogen was identified as G. ambrosiae. In order to satisfy Koch's postulates, an infected leaf was carefully pressed onto leaves of six healthy young B. pilosa plants, each grown in a separate pot. Additionally, a control group consisted of six non-inoculated plants. All plants were placed in a greenhouse: 25°C, 14/10-h light/dark photoperiod, and relative humidity 50%. After 10 days, the inoculated leaves exhibited powdery mildew colonies similar to those observed in the original infected plants. At 16 days, the inoculated leaves exhibited discoloration and premature leaf drop. The pathogenicity test was conducted twice. Microscopic observation and sequencing confirmed that isolated fungus was identical to the original pathogen. G. ambrosiae has previously been documented on B. pilosa in Fuzhou, Fujian Province, China (Mukhtar et al., 2022). However, to the best of our knowledge, this study represents the first report of powdery mildew caused by G. ambrosiae on B. pilosa in Shenzhen, Guangdong Province, China.

Zinc oxide nano-fertilizer differentially effect on morphological and physiological identity of redox-enzymes and biochemical attributes in wheat (Triticum aestivum L.)

The aim of current study was to prepared zinc oxide nanofertilzers by ecofriendly friendly, economically feasible, free of chemical contamination and safe for biological use. The study focused on crude extract of Withania coagulans as reducing agent for the green synthesis of ZnO nano-particles. Biosynthesized ZnO NPs were characterized by UV–Vis spectroscopy, XRD, FTIR and GC–MS analysis. However, zinc oxide as green Nano fertilizer was used to analyze responses induced by different doses of ZnO NPs [0, 25, 50,100, 200 mg/l and Zn acetate (100 mg/l)] in Triticum aestivum (wheat). The stimulatory and inhibitory effects of foliar application of ZnO NPs were studied on wheat (Triticum aestivum) with aspect of biomass accumulation, morphological attributes, biochemical parameters and anatomical modifications. Wheat plant showed significant (p < 0.01) enhancement of growth parameters upon exposure to ZnO NPs at specific concentrations. In addition, wheat plant showed significant increase in biochemical attributes, chlorophyll content, carotenoids, carbohydrate and protein contents. Antioxidant enzyme (POD, SOD, CAT) and total flavonoid content also confirmed nurturing impact on wheat plant. Increased stem, leaf and root anatomical parameters, all showed ZnO NPs mitigating capacity when applied to wheat. According to the current research, ZnO NPs application on wheat might be used to increase growth, yield, and Zn biofortification in wheat plants.

Advancements and current research in orofacial tissue regeneration

The orofacial system, an intricate assembly of diverse tissues that underpin the unique biologic and morphological identity of an individual, presents a formidable challenge in the realm of tissue regeneration within oral and maxillofacial surgery. This review conducts a retrospective appraisal of advancements in the field of orofacial tissue regeneration, elucidating the current research landscape while critically addressing the persisting challenges. It underscores the pivotal roles of orofacial mesenchymal stem cells in orchestrating regenerative processes, offering an insightful outlook on future developments. The objective is to demarcate innovative therapeutic avenues and clinical implications by fostering a comprehensive understanding of this domain among dental practitioners. As such, it aspires to serve as a valuable reference for prospective investigations and to elevate the knowledge base pertaining to orofacial tissue regeneration.

Combining biometrics and genetics to distinguish two subspecies of the Great Cormorant Phalacrocorax carbo carbo and P. c. sinensis at an inland lake in southeast Norway

Cover photo: Adult Great Cormorant of the subspecies Phalacrocorax carbo sinensis in breeding plumage. Photo: Frode Falkenberg. Great Cormorants Phalacrocorax carbo are now regularly seen in inland watercourses in southeast Norway, after the P. c. sinensis subspecies first established breeding colonies in coastal south Norway in 1996. Although both the sinensis and carbo subspecies occur, the ratio between them is unknown. We tested the accuracy of subspecific identification based on biometrics and genetic analyses in 75 Great Cormorants that drowned in fishing nets in a lake in southeast Norway during the period 2009-2020. Primarily based on the gular pouch angle (GPA), we classified 40 individuals to the carbo subspecies and 35 individuals to the sinensis subspecies. Eight of the carbo individuals and 14 of the sinensis individuals were within the overlapping range of GPA and were therefore classified to subspecies by supplementary measurements (bill depth minimum and bill length). Genetic analyses were based on seven polymorphic microsatellite markers. Assuming one panmictic population, we performed Structure analyses to separate the genotypes into two assumed genetic clusters. The two clusters, carbo and sinensis, could only be separated when adding information about morphological subspecies identities for the carbo, sinensis and carbo/sinensis groups. The sinensis and the carbo/sinensis groups belonged almost entirely to one genetic cluster, whereas the carbo group consisted of individuals with varying proportions of mixed ancestry. Furthermore, we found significant genetic differentiation between the carbo and sinensis groups, and between the carbo and the carbo/sinensis groups, but no significant differentiation between the sinensis and the carbo/sinensis groups. Our results suggest that gene flow is more common from sinensis into carbo than vice-versa, and that the use of GPA &lt; 73° has limitations for the identification of Great Cormorant subspecies in areas where both forms occur. We conclude that the numbers of carbo vs. sinensis individuals in our sample were 32 and 43, respectively.

The -eci Syncretism in Korean: Implications for the Theory of v and Voice

Abstract This paper investigates the syncretism exhibited by the Korean verbal suffix -eci. In addition to its widely known appearance in the passive construction, -eci can also be used to derive verbs expressing potentiality. In this paper, I show that two independently motivated theoretical tools — (i) the articulated verbal structure with root, verbalizer, and Voice; and (ii) the assumption that morphological identity signifies the morpheme's realization of an identical syntactic head — accurately explain the passive-potential syncretism in Korean. Specifically, I argue that -eci realizes a syntactic head that the passive and potential structures have in common: vGO, the verbalizer marking the eventuality of ‘change’. I attribute the systematic morpho-syntactic and semantic contrasts between passives and potentials to the (non)existence of VoicePASS, the projection introducing an implicit external argument. The analysis successfully captures the properties of the other constructions formed upon -eci — namely, derived change-of-state and lexical inchoative predicates.

К вопросу о синтаксическом статусе субстантивов в русских предложениях тождества

A multi aspect study of bisubstantive sentences of characterizing semantics is necessary to solve complex problems of syntactic analysis. The grammatical specificity of bisubstantial sentences lies in the morphological identity of the predicative minimum. The study has theoretical and practical tasks: identify the structural, semantic and communicative organization of sentences, propose methodological recommendations for syntactic analysis of sentences. The material of the study is modern literature. Analytical and descriptive methods are used to analyze the linguistic material. The problems of teaching syntactic analysis of sentences are identified through experimental methods. The main conclusions of the study: syntactic analysis of bisubstantial sentences should consist of traditional structural and semantic parameters, roles and functions in the semantic structure and communicative organization of a sentence, take into account context, stylistic and pragmatic factors. Syntactic phenomena should be studied from the functional position at all stages and levels of teaching syntax of the modern Russian language.

Structural insights into the recognition of the A/T-rich motif in target gene promoters by the LMX1a homeobox domain.

LIM homeodomain transcription factor 1-alpha (LMX1a) is a neuronal lineage-specific transcription activator that plays an essential role during the development of midbrain dopaminergic (mDA) neurons. LMX1a induces the expression of multiple key genes, which ultimately determine the morphology, physiology, and functional identity of mDA neurons. This function of LMX1a is dependent on its homeobox domain. Here, we determined the structures of the LMX1a homeobox domain in complex with the promoter sequences of the Wnt family member 1 (WNT1) or paired like homeodomain 3 (Pitx3) gene, respectively. The complex structures revealed that the LMX1a homeobox domain employed its α3 helix and an N-terminal loop to achieve specific target recognition. The N-terminal loop (loop1) interacted with the minor groove of the double-stranded DNA (dsDNA), whereas the third α-helix (α3) was tightly packed into the major groove of the dsDNA. Structure-based mutations in the α3 helix of the homeobox domain significantly reduced the binding affinity of LMX1a to dsDNA. Moreover, we identified a nonsyndromic hearing loss (NSHL)-related mutation, R199, which yielded a more flexible loop and disturbed the recognition in the minor groove of dsDNA, consistent with the molecular dynamics (MD) simulations. Furthermore, overexpression of Lmx1a promoted the differentiation of SH-SY5Y cells and upregulated the transcription of WNT1 and PITX3 genes. Hence, our work provides a detailed elucidation of the specific recognition between the LMX1a homeobox domain and its specific dsDNA targets, which represents valuable information for future investigations of the functional pathways that are controlled by LMX1a during mDA neuron development.

Molecular insights versus morphological traits: rethinking identification of the closely related Angiostrongylus cantonensis and Angiostrongylus malaysiensis

BackgroundThe closely related Angiostrongylus cantonensis and Angiostrongylus malaysiensis have been reported to coexist in Thailand and share similar hosts and life cycles. Recently, in an angiostrongyliasis outbreak in Thailand, both A. cantonensis and A. malaysiensis were found in the cerebrospinal fluid of affected patients. Morphological similarities, overlapping distribution, shared hosts and habitats, and the close genetics of the two Angiostrongylus species can complicate accurate species identification. Addressing these challenges, this study aims to evaluate whether a correlation between the morphological and genetic identities of A. cantonensis and A. malaysiensis can improve species identification accuracy.MethodsAngiostrongylus spp. specimens from five zoogeographical regions in Thailand were subjected to morphological and molecular identification using the mitochondrial cytochrome b gene and the nuclear internal transcribed spacer 2 region (ITS2). The morphological characters for males and females were then validated using the species identity obtained from the nuclear ITS2 region.ResultsThe results revealed that morphological misidentifications between these two closely related species are common due to overlapping morphological characters. Although certain male traits such as body length and width aided species differentiation, female traits were found to be less reliable. Furthermore, hybrid forms (8.2%) were revealed through the ITS2 results, which can further complicate morphological identification. Mito-nuclear discordance was also present in 1.9% of the Angiostrongylus specimens from Thailand, suggesting a complex historical interbreeding between the species.ConclusionsBased on our findings, we suggest that nuclear ITS2 is a reliable marker for species identification of A. cantonensis and A. malaysiensis, especially in regions where both species coexist. Additionally, the scope and consequences of hybridization between the two closely related Angiostrongylus species should be further investigated in Thailand.Graphical

First Report of Charcoal Rot Caused by Macrophomina phaseolina on Dry Bean (Phaseolus vulgaris L.) in Western Canada.

In 2021, several dry bean (Phaseolus vulgaris L.) plants at the mid-seed-fill growth stage displaying wilting, chlorosis of the leaves and reduced vigor were collected near the Pembina - Emerson Border of Manitoba, Canada and North Dakota, USA. When symptomatic plants were examined, gray to dark brown discoloration was observed on the lower stem and the roots. Afterwards, brown to black discoloration was noticed on stem and root sections. Root and lower stem pieces (1 to 2 cm) from affected plants were surface sterilized with 70 % ethanol, followed by 1% NaOCl, rinsed twice in sterilized water, air dried on sterilized filter papers, and placed on potato dextrose agar (PDA) amended with 1 mg/mL of streptomycin sulfate. The PDA plates were incubated at 28°C with 12 h light/12 h dark for 10 days. The growing hyphae were transferred using the hyphal tip method to new PDA plates. Growing cultures were initially hyaline and turned from light gray to dark brown or black with age. Abundant dark and spherical to oblong shaped sclerotia with an average diameter of 97.9 µm (range: 66.8 to 143.5 µm, n =30) formed on the pure cultures 7 days after incubation. Additional pure culture was obtained through an isolation of a single microsclerotium followed by a single hyphal tip transfer. One isolate was identified as Macrophomina phaseolina based on morphological characteristics (Smith and Wyllie 1999). The morphological identity was confirmed by sequencing the rDNA internal transcribed spacer (ITS) region with universal primers ITS1/ITS4 (White et al. 1990) and calmodulin (CAL) and translation elongation factor-1 alpha (TEF-1α) genes with MpTefF/MpTefR, and MpCalF/MpCalR primer sets (Santos et al. 2020), respectively. The online resource Basic Local Alignment Search Tool (BLAST; https://www.ncbi.nlm.nih.gov/BLAST) confirmed the fungus identity as 100% M. phaseolina. The sequences of the original isolate BF21-25 were deposited in GenBank with accession numbers OQ615297 (ITS), OR357630 (CAL), and OR363106 (TEF-1α). To confirm pathogenicity, bioassays were conducted under controlled conditions. Four seeds of cultivar 'Etna' were sown per pot, and five pots were used for inoculated (approx. 4 × 105 microsclerotia/pot) and control (mock-inoculated with sterile PDA medium) treatments. For the inoculum, 20 g of macerated 10 to 14-day old M. phaseolina culture grown on PDA medium was applied to each pot using an inoculum layering technique. Pots were kept in the greenhouse with 28/17°C day/night, 13 h light/11 h dark cycle, and 70% relative humidity and watered weekly. Disease symptoms similar to those observed in the field were visible on all inoculated plants at the mid-seed-fill growth stage. Mock-inoculated control plants didn't show any symptoms. The experiment was repeated twice with similar results. The pathogen was re-isolated from infected plants to confirm Koch's postulates and identified as M. phaseolina based on the morphology and sequences of ITS, CAL and TEF-1α regions. To our knowledge, this is the first report of charcoal rot caused by M. phaseolina on dry bean in Western Canada.

New alien and invasive bamboo aphid species of the genus Takecallis (Hemiptera: Aphididae) recorded in Poland – morphological and molecular identity

The occurrence of three bamboo aphid species of the genus Takecallis was detected. T. arundicolens (Clarke) and T. arundinariae (Essig) were recorded for the first time in Poland, and new localities for T. nigroantennatus Wieczorek were found. Key diagnostic morphological characteristics to help distinguish these species and DNA barcoding to analyze individuals at the molecular level were provided.

Novel cellular systems unveil mucosal melanoma initiating cells and a role for PI3K/Akt/mTOR pathway in mucosal melanoma fitness

BackgroundMucosal Melanomas (MM) are highly aggressive neoplasms arising from mucosal melanocytes. Current treatments offer a limited survival benefit for patients with advanced MM; moreover, the lack of pre-clinical cellular systems has significantly limited the understanding of their immunobiology.MethodsFive novel cell lines were obtained from patient-derived biopsies of MM arising in the sino-nasal mucosa and designated as SN-MM1-5. The morphology, ultrastructure and melanocytic identity of SN-MM cell lines were validated by transmission electron microscopy and immunohistochemistry. Moreover, in vivo tumorigenicity of SN-MM1-5 was tested by subcutaneous injection in NOD/SCID mice. Molecular characterization of SN-MM cell lines was performed by a mass-spectrometry proteomic approach, and their sensitivity to PI3K chemical inhibitor LY294002 was validated by Akt activation, measured by pAkt(Ser473) and pAkt(Thr308) in immunoblots, and MTS assay.ResultsThis study reports the validation and functional characterization of five newly generated SN-MM cell lines. Compared to the normal counterpart, the proteomic profile of SN-MM is consistent with transformed melanocytes showing a heterogeneous degree of melanocytic differentiation and activation of cancer-related pathways. All SN-MM cell lines resulted tumorigenic in vivo and display recurrent structural variants according to aCGH analysis. Of relevance, the microscopic analysis of the corresponding xenotransplants allowed the identification of clusters of MITF-/CDH1-/CDH2 + /ZEB1 + /CD271 + cells, supporting the existence of melanoma-initiating cells also in MM, as confirmed in clinical samples. In vitro, SN-MM cell lines were sensitive to cisplatin, but not to temozolomide. Moreover, the proteomic analysis of SN-MM cell lines revealed that RICTOR, a subunit of mTORC2 complex, is the most significantly activated upstream regulator, suggesting a relevant role for the PI3K-Akt-mTOR pathway in these neoplasms. Consistently, phosphorylation of NDRG1 and Akt activation was observed in SN-MM, the latter being constitutive and sustained by PTEN loss in SN-MM2 and SN-MM3. The cell viability impairment induced by LY294002 confirmed a functional role for the PI3K-Akt-mTOR pathway in SN-MM cell lines.ConclusionsOverall, these novel and unique cellular systems represent relevant experimental tools for a better understanding of the biology of these neoplasms and, as an extension, to MM from other sites.

Estudos morfológicos e moleculares sobre um parasita nematoide, Syphacia obvelata, que infecta camundongos de laboratório (Mus musculus) na Arábia Saudita

ABSTRACT During screening laboratory mice (Mus musculus) at the Zoology Department of the College of Science, King Saud University, some tiny oxyurid nematodes were detected from the large intestine of these rodents. Worms were identified using morphological and morphometric description. Furthermore, DNA was extracted from worms and subjected to polymerase chain reaction to amplify 18S rDNA and ITS (ITS1, 5.8S and ITS2) regions. Worms detected from caecum and colon of mice were detected in 80% of mice investigated. Morphologically, worms showed elongated body with tapered anterior end and narrow posterior part with cuticular annulations. Male worms measured 0.71-1.12 (0.90) long and 0.01-0.12 (0.11) wide. Female worms measured 3.12-5.011 (4.30) long and 0.13-0.29 (0.17) wide. Esophagus followed by intestine which opens at the posterior end via anal opening in females and via cloacal opening in males. Males have a single spicule with a gubernaculum and an accessory hook. Females’ uteri were pack with eggs. Data from 18S rDNA revealed a sequence which was identical to Syphacia obvelata in NCBI GenBank. Similarly, sequences from ITS regions grouped with sequences from S. obvelata confirming the morphological identity of the worm. However, it showed 3 mutations at the ITS2 region from related sequences from S. obvelata at NCBI GenBank.

An inductive method for classifying building form in a city with implications for orientation

The utilization of deep learning for form analysis facilitates the classification of an extensive number of forms based on their morphological features. A critical consideration for implementing such analysis methods in architectural or urban forms is whether building orientation should be embedded within the data. Orientation functions as a form variable significantly influenced by environmental, social, and cultural contexts within a city. In contrast to other domains where forms are extrapolated in relation to their context, in the city, domain orientation uniquely characterizes building form. In this paper, we introduce a pipeline for constructing an extensive building form dataset and scrutinizing the morphological identity of building forms, with a particular focus on the implications of building orientation as a manifestation of urban locality. Through a case study situated in Montreal, we engage in a comparative analysis employing two distinct datasets—those with orientation-embedded forms and those with orientation-normalized forms. Our research aims to investigate the typo-morphological characteristics of the building forms of the city and to examine how building orientation contributes to the identification of these traits and mirrors urban locality.