Molecular Phylogenetic Studies Research Articles

Molecular Identification of Genes Resistant to Antibiotics in Staphylococcus aureus Isolated from Sub-clinical Animals Affected by Mastitis

Background and Aim: Mastitis is a prevalent and costly disease that significantly impacts the dairy industry, affecting both sheep's health and milk quality. In order to determine the bacterial causes of subclinical mastitis in Iraq and its susceptibility to antibiotics, the study aims to molecularly detect resistant Staphylococcus aureus isolated from sheep infected with mastitis. Methods: Molecular assay and phylogenetic study to detect Staph aureus isolates by using 16 Sr RNA and resistance virulence genes (aac-aph, tetk) by the conventional method of PCR. Two hundred samples were collected from the subclinical mastitis of infected ewes, Samples were reserved on an icebox and transported to the laboratories. Milk samples were cultured on Blood Agar and Mannitol Salt Agar (7.5%) plates. The culture plates were then incubated at 37°C for 24 hours. Results: The PCR assay revealed amplification of the 16S rRNA gene in all 20 isolates, amplification of the aac-aph gene in 19 isolates, and the tetK gene in all 20 isolates. The PCR products of 20 positive samples for (16 Sr RNA) target genes were sequenced, evaluated, and dropped on the Genbank-NCBI under the accession number, which became a reference to Iraq and the world. According to the current study, Staphylococcus aureus is commonly found in milk. To combat drug resistance, antimicrobial drug use should be normalized, and antimicrobial resistance surveillance should be conducted regularly. Conclusion: PCR-product of 8 positive samples for 16sr RNA target gene were sequenced, analyzed, and deposited on the Genbank-NCBI under the accession number and became a reference to Iraq and the Middle East and the world.

Molecular Identification of Genes Resistant to Antibiotics in Staphylococcus aureus Isolated from Sub-clinical Animals Affected by Mastitis

Background and Aim: Mastitis is a prevalent and costly disease that significantly impacts the dairy industry, affecting both sheep's health and milk quality. In order to determine the bacterial causes of subclinical mastitis in Iraq and its susceptibility to antibiotics, the study aims to molecularly detect resistant Staphylococcus aureus isolated from sheep infected with mastitis. Methods: Molecular assay and phylogenetic study to detect Staph aureus isolates by using 16 Sr RNA and resistance virulence genes (aac-aph, tetk) by the conventional method of PCR. Two hundred samples were collected from the subclinical mastitis of infected ewes, Samples were reserved on an icebox and transported to the laboratories. Milk samples were cultured on Blood Agar and Mannitol Salt Agar (7.5%) plates. The culture plates were then incubated at 37°C for 24 hours. Results: The PCR assay revealed amplification of the 16S rRNA gene in all 20 isolates, amplification of the aac-aph gene in 19 isolates, and the tetK gene in all 20 isolates. The PCR products of 20 positive samples for (16 Sr RNA) target genes were sequenced, evaluated, and dropped on the Genbank-NCBI under the accession number, which became a reference to Iraq and the world. According to the current study, Staphylococcus aureus is commonly found in milk. To combat drug resistance, antimicrobial drug use should be normalized, and antimicrobial resistance surveillance should be conducted regularly. Conclusion: PCR-product of 8 positive samples for 16sr RNA target gene were sequenced, analyzed, and deposited on the Genbank-NCBI under the accession number and became a reference to Iraq and the Middle East and the world.

Cryptanthawhippleae (Boraginaceae), a new serpentine-adapted species endemic to northern California, U.S.A.

Cryptanthawhippleae D.A.York & M.G.Simpson (Boraginaceae) is described as new. This species is currently known to occur in serpentine barrens in the Shasta-Trinity National Forest of Siskiyou County, California, with one outlier population in possible serpentine of Lake County, California. The new species is most similar to Cryptanthagrandiflora and to C.milobakeri, these three likely each others' closest relatives. All three have a relatively large corolla limb width and similar smooth, lance-ovate to ovate, marginally rounded, acuminate and abaxially transversely flattened nutlets. Cryptanthawhippleae differs from C.grandiflora in having a short, as opposed to a tall, stem height; bifurcate as opposed to trifurcate primary axis cymules; and typically 2-3 nutlets per fruit, as opposed to usually one nutlet per fruit. Cryptanthawhippleae differs from C.milobakeri also in having a short, versus tall, stem height; appressed-strigose and spreading-hispid stem vestiture, as opposed to strigose only or strigose and hirsute; calyx trichomes with two distinct vestiture types, these marginally appressed hirsute and medially hispid, as opposed to calyx trichomes of one type, dense, appressed to ascending, whitish sericeous; and 2-3 nutlets per fruit, as opposed to one nutlet per fruit. Cryptanthawhippleae is relatively rare and joins seven other Cryptantha species that are found on serpentine, either obligately or facultatively. Current molecular phylogenetic studies support the mostly convergent evolution of serpentine adaptation in Cryptantha, but additional studies are needed.

Redescription of Rhabditis maupasi Seurat in Maupas, 1919 (Rhabditida: Rhabditidae) from northwest Iran

Summary Rhabditis maupasi is re-described and reviewed based on morphological, morphometric, and molecular (D2-D3 expansion segment of the rDNA 28S) analysis collected from northwest Iran. The species is characterised by having a 1.1-1.9 mm long body, lateral fields with three incisures, the lip region offset by a depression, 12-15 μm wide, with six globular lips, the stoma rhabditoid, 14-20 μm long, with distinct cheilo-, gymno- and stegostom; cheilostom slightly cuticularised, gymno-promesostegostom a short tube with well-cuticularised walls, metastegostom with glottoid apparatus bearing small teeth, stomatal or pharyngeal collar covering 40-60% of stoma; pharynx well developed, the pharyngeal corpus consisting of a more or less cylindrical procorpus and swollen metacorpus (median bulb), basal bulb ovoid to pyriform, with grinder and narrow haustrulum, excretory pore at basal bulb level; vulva a transverse slit with protruding lips, the female tail conical-elongated (65-75 μm, c = 16-26, c′ = 2.0-2.5), the male tail conoid, ventrally arcuate (29-34 μm, c = 22-41, c′ = 0.7-1.0); spicules free, with pointed end and dagger-shaped, 61-72 μm long, bursa pseudopeloderan, and with eight pairs of genital papillae plus the phasmid, and gubernaculum 34-36 μm long. Furthermore, its relationship with four closely related species of the genus is discussed. A molecular phylogenetic study using a partial sequence of large subunit ribosomal DNA fragments revealed that R. maupasi forms a clade with R. terricola, recorded previously from Iran.

Prevalence and identification of cyathocotylid trematodes infecting African catfish in Egypt.

The trematode family Cyathocotylidae infects various hosts worldwide, including birds, mammals, reptiles, and fish. However, the lack of molecular data from adult worms hinders phylogenetic, epidemiological, and host association studies. This study aims to identify the common cyathocotylid trematodes infecting African catfish in Egypt using morphological and molecular evidence. Out of 142Clarias gariepinus, 123 fish (86.6%) harbored cyathocotylid metacercariae, with a mean metacercarial intensity of 201 ± 38.5/g. Cyathocotylid metacercariae prevalence gradually rose as host size increased. Although there was no significant difference between groups, larger fish had a higher mean metacercarial intensity. The prevalence and intensity were unrelated to the fish gender. Histopathological examination of metacercariae-infected catfish revealed varying degrees of degenerative changes, including intermuscular edema leading to muscle fiber dispersion and atrophy, involving 11% to over 81% of muscle sections. We identified three cyathocotylid metacercariae and eight cyathocotylid adult species from experimental infection using morphometric and molecular data, including internal transcribed spacer (ITS) and/or mitochondrial cytochrome c oxidase subunit 1 (cox1) sequences. We determined the phylogenetic position of these cyathocotylid samples.The ITS sequence analysis linked the isolated Cyathocotylidae sp. 1 and 2 metacercariae to Prohemistomum vivax adults. Mesostephanus appendiculatoides and Paracoenogonimus ovatus were reported for the first time in Egypt. These findings may provide valuable genetic data for future molecular epidemiological and phylogenetic studies of cyathocotylid trematodes.

Phylogenomics reveal species limits and inter-relationships in the narrow-range endemic lycian salamanders

Salamanders of the genus Lyciasalamandra are represented by as many as 20 narrow-range endemic taxa inhabiting the Mediterranean coast of Turkey and a handful of Aegean Islands. Despite recent molecular phylogenetic studies, the genus is rife with uncertainty about the number of contained species and their phylogenetic relationships, both of which can interfere with needed conservation actions. To test species limits and infer interrelationships we generated as many as 113,176 RAD loci containing 229,427 single nucleotide polymorphisms (SNPs), for 110 specimens of Lyciasalamandra representing 19 of the 20 described taxa. Through a conservative species delimitation approach, we found support for eight species in the genus which broadly agree with currently described species-level diversity. We then use multiple coalescent-based species tree methods to resolve relationships in this relatively old, synchronous species radiation. We recommend synonymization of the largely over-split subspecific taxa, and the elevation of L. luschani finikensis to full species status as L. finikensis. Our hope is that this revised taxonomic framework provides a stable foundation for conservation management in these fragile, microendemic taxa.

Phylogenomics supports a single origin of terrestriality in isopods.

Terrestriality, the adaptation to life on land, is one of the key evolutionary transitions, occurring numerous times across the tree of life. Within Arthropoda, there have been several independent transitions: in hexapods, myriapods, arachnids and isopods. Isopoda is a morphologically diverse order within Crustacea, with species adapted to almost every environment on Earth. The order is divided into 11 suborders with the most speciose, Oniscidea, including terrestrial isopods such as woodlice and sea-slaters. Recent molecular phylogenetic studies have challenged traditional isopod morphological taxonomy, suggesting that several well-accepted suborders, including Oniscidea, may be non-monophyletic. This implies that terrestriality may have evolved multiple times. Current molecular hypotheses, however, are based on limited sequence data. Here, I collate available genome and transcriptome datasets for 36 isopods and four peracarid crustaceans from public sources, generate assemblies and use 970 single-copy orthologues to estimate isopod relationships and divergence times with molecular dating. The resulting phylogenetic analyses support monophyly of terrestrial isopods and suggest conflicting relationships based on nuclear ribosomal RNA sequences may be caused by long-branch attraction. Dating analyses suggest a Permo-Carboniferous origin of isopod terrestriality, much more recently than other terrestrial arthropods.

Disentangling conflicting molecular phylogenetic signals in nuclear and plastid DNA of the western Eurasian-Mediterranean grass genus Cynosurus and its relatives (Poaceae subtribes Cynosurinae and Parapholiinae)

The western Eurasian-Mediterranean grass genus Cynosurus, comprising about 11 species, is morphologically well delimited by the regular occurrence of conspicuous sterile spikelets distal to the fertile ones on the outer, abaxial side of the inflorescences. However, our molecular phylogenetic study using nuclear ribosomal DNA (ITS, ETS) and plastid DNA sequences (trnL–F, matK) has shown that the genus is not monophyletic in its current delimitation, but consists of three distinct lineages. These lineages were found to be closely related to a group of 6–7 genera taxonomically assigned to the subtribe Parapholiinae. These Parapholiinae genera were consistently monophyletic in our analyses, but the suggested relationships to the three lineages of Cynosurus varied depending on the particular DNA region examined. This was the case for both plastid and nuclear DNA, with cytonuclear discordance and ‘chloroplast capture’ indicating earlier hybridization. Interestingly, hybridization also proved to be the most likely explanation even with regard to the 18S–26S cistrons of the nuclear ribosomal DNA, where an exceptional evolutionary divergence between ITS and ETS was found. The results highlight and illustrate the important role of hybridization in the evolution of grasses. In terms of taxonomy, our findings argue against maintaining a polyphyletic genus Cynosurus s.l. but instead argue for dividing it into three monophyletic genera: Cynosurus s.s., Falona, which is reestablished here, and Ciliochloa, which is described as a new genus. In addition, it is proposed that the two subtribes Cynosurinae and Parapholiinae be combined into a single subtribe Cynosurinae, which is also monophyletic. The possible genetic background of the formation of sterile spikelets and the occasional occurrence of inflorescences with consistently fertile spikelets are discussed. New combinations are Ciliochloa effusa, C. effusa var. obliquata, C. effusa var. fertilis, C. elegans, C. gracilis, C. turcomanica and Falona colorata.

Characterisation of the complete chloroplast genome of Solanum tuberosum cv. White Lady

AbstractPotato (Solanum tuberosum) is considered worldwide as one of the most important non-cereal food crops. As a result of its adaptability and worldwide production area, potato displays a vast phenotypical variability as well as genomic diversity. Chloroplast genomes have long been a core issue in plant molecular evolution and phylogenetic studies, and have an important role in revealing photosynthetic mechanisms, metabolic regulations and the adaptive evolution of plants. We sequenced the complete chloroplast genome of the Hungarian cultivar White Lady, which is 155 549 base pairs (bp) in length and is characterised by the typical quadripartite structure composed of a large- and small single-copy region (85 991 bp and 18 374 bp, respectively) interspersed by two identical inverted repeats (25 592 bp). The genome consists of 127 genes of which 82 are protein-coding, eight are ribosomal RNAs and 37 are transfer RNAs. The overall gene content and distribution of the genes on the White Lady chloroplast was the same as found in other potato chloroplasts. The alignment of S. tuberosum chloroplast genome sequences resulted in a highly resolved tree, with 10 out of the 13 nodes recovered having bootstrap values over 90%. By comparing the White Lady chloroplast genome with available S. tuberosum sequences we found that gene content and synteny are highly conserved. The new chloroplast sequence can support further studies of genetic diversity, resource conservation, evolution and applied agricultural research. The new sequence can support further potato genetic diversity and evolutionary studies, resource conservation, and also applied agricultural research.

Molecular evidence of hepatozoonosis in tigers of Vidarbha region of Maharashtra State of India

BackgroundHepatozoonosis has been reported in many species around the world. Few incidences have been reported in various species of wild felids. Tigers are endangered large cats and are protected under the Wildlife Protection Act, 1972 under Schedule I. The study was carried out to estimate the positivity rate of hepatozoonosis in tigers of the Vidarbha region of Maharashtra, India.MethodsBlood (n = 21) or tissue samples (n = 5) were collected from 26 wild captured / zoo-born or dead tigers during the quarantine period/post-mortem examination. Blood smear examination along with Polymerase Chain Reaction (PCR) studies were conducted for the detection of hepatozoonosis. All the amplicons from the positive samples were purified and sequenced, and the sequences were subjected to nBLAST analysis to detect the species of Hepatozoon. The sequences were deposited into public domain database of National Center for Biotechnology Information (NCBI) and accession numbers were allotted. A phylogenetic study was undertaken to understand the evolutionary lineage of the pathogen. Tissue distribution studies were carried out on tissue samples received during post mortem. A clinical case in a tiger cub was managed and sub-clinical cases were monitored for relapse. Age-wise, sex-wise, region-wise and captive time-wise positivity rate was estimated. The data was analyzed using statistical tools.ResultsA total of 12 tigers were found positive for H. felis during the screening. A clinical case was diagnosed and successfully treated. The age group of 0–3 years reported a positivity rate of 66.66%, and all the cases found positive were reported between the age group of 0–7 years. Males reported a positivity rate of 58.33 per cent, while females reported 35.71%. Taboba and Andhari Tiger Reserve of the state had a positivity rate of 52.94 per cent. However, the statistical analysis for blood parameters and positivity rate by ‘t’ test and Chi-squared test were found to be non-significant.ConclusionsAn overall positivity rate of 46.15% indicates the wide distribution of hepatozoonosis among wild tigers of the Vidarbha region of Maharashtra, India, which is strategically important considering the gene flow and migration of tigers. Hepatozoonosis can progress to clinical outcomes in young animals and require veterinary intervention. Molecular tools and phylogenetic studies can supplement important data on circulating species of Hepatozoon in the field. Further studies on the clinical management and epidemiology of the infection in wild felids will comprehend the cause of wildlife conservation.

Heracleum munzurense (Apiaceae), a new species from East Anatolia, Turkey and taxonomic position of Tetrataenium lasiopetalum

A new species, Heracleum munzurense (Apiaceae), is described from East Anatolia, Turkey. It grows in damp places formation of the Munzur Mountains in the district of Pülümür (Tunceli Province). The taxonomic affinities of the new species were determined by analyzing morphological data and through a molecular phylogenetic study of nrITS/ETS. The new species is morphologically and genetically related to H. apiifolium and H. pastinaca, but differs mainly in stems, leaves, pedicels, rays and petals pubescence, petals size, fruit hairs, and size features. The anatomical structure of the fruit is given. The diagnostic morphological characters of the new species are discussed. In this study, the geographical distribution of the new species and the related taxa H. apiifolium, H. lasiopetalum, and H. pastinaca is mapped. In addition, the habitat, ecology, phenology, and conservation status of the new species are also provided. Tetrataenium lasiopetalum is also closely related to this group of species. Common morphological features and results of molecular phylogenetic study suggest to transfer this species back to the genus Heracleum.

Identification and in vivo functional analysis of furanocoumarin-responsive cytochrome P450s in a Rutaceae-feeding Papilio butterfly.

The Order Lepidoptera contains nearly 160,000 described species and most of them are specialist herbivores that use restricted plant species as hosts. Speciation that originated from host shift is one of the important factors for the diversification of Lepidoptera. Because plants prepare secondary metabolites for defense against herbivores, with varying profiles of the components among different plant taxa, the specialist herbivores need to be adapted to the toxic substances unique to their host plants. Swallowtail butterflies of the genus Papilio consist of over 200 species. Approximately 80% of them utilize Rutaceae plants, and among the remaining species, a specific subgroup uses phylogenetically distant Apiaceae plants as larval hosts. Rutaceae and Apiaceae commonly contain toxic secondary metabolites, furanocoumarins, and molecular phylogenetic studies support the concept that Apiaceae feeders were derived from Rutaceae feeders. Molecular mechanisms underlying furanocoumarin tolerance in Papilio butterflies have been investigated almost exclusively in an Apiaceae feeder by an in vitro assay. In contrast, there is little information regarding the Rutaceae feeders. Here, we focused on a Rutaceae feeder, Papilio xuthus, and identified two furanocoumarin-responsive cytochrome P450-6B (CYP6B) genes, of which one was an ortholog of a furanocoumarin-metabolizing enzyme identified in the Apiaceae-feeding Papilio while the other was previously unreported. We further conducted in vivo functional analysis using the CRISPR/Cas9 system, revealing a contribution of these CYP6Bs to furanocoumarin tolerance of P. xuthus larvae. Our findings suggest that co-option of furanocoumarin-metabolizing CYP6B enzymes at least partially contributed to the host shift from Rutaceae to Apiaceae in Papilio butterflies.

Clarification of the distribution, conchological diversity, and taxonomic status of the Iberian snail Iberus globulosus C.R. Boettger, 1913 (Gastropoda: Stylommatophora: Helicidae)

The taxonomic status of Iberus globulosus has been unsatisfactorily resolved since its original description, but in 2008 a molecular phylogenetic study of the genus Iberus suggested that it could be a valid species. However, limited information on both its geographic distribution and its conchological variability has made it difficult to properly delineate the species. Our exhaustive sampling throughout the potential area of its geographic distribution and the morphological study herein, together with pre-existing genetic evidence, allow us to confirm the validity of I. globulosus. Consequently, we improve the characterization of I. globulosus by redescribing it. Additionally, we provide information on the potential for hybridization among I. globulosus and geographically and phylogenetically close congeneric species.

Species of the Sections Hedysarum and Multicaulia of the Genus Hedysarum (Fabaceae): Taxonomy, Distribution, Chromosomes, Genomes, and Phylogeny.

The genus Hedysarum L. (Fabaceae) includes about 200 species of annual and perennial herbs distributed in Asia, Europe, North Africa, and North America. Many species of this genus are valuable medicinal, melliferous, and forage resources. In this review, we consider the taxonomic history of the genus Hedysarum, the chromosomal organization of the species from the sections Hedysarum and Multicaulia, as well as phylogenetic relationships between these sections. According to morphological, genetic, and phylogenetic data, the genus Hedysarum is divided into three main sections: Hedysarum (= syn. Gamotion), Multicaulia, and Stracheya. In species of this genus, two basic chromosome numbers, x = 7 (section Hedysarum) and x = 8 (sections Multicaulia and Stracheya), were determined. The systematic positions of some species within the sections are still uncertain due to their morphological similarities. The patterns of distribution of molecular chromosomal markers (45S rDNA, 5S rDNA, and different satellite DNAs) in karyotypes of various Hedysarum species made it possible to determine their ploidy status and also specify genomic relationships within the sections Hedysarum and Multicaulia. Recent molecular phylogenetic studies clarified significantly the taxonomy and evolutionary development of the genus Hedysarum.

A morpho-phylogenetic update on ixodid ticks infesting cattle and buffalos in Vietnam, with three new species to the fauna and a checklist of all species indigenous to the country

BackgroundSoutheast Asia is regarded as a hotspot for the diversity of ixodid ticks. In this geographical region, Vietnam extends through both temperate and tropical climate zones and therefore has a broad range of tick habitats. However, molecular-phylogenetic studies on ixodid tick species have not been reported from this country.MethodsIn this study, 1788 ixodid ticks were collected from cattle, buffalos and a dog at 10 locations in three provinces of northern Vietnam. Tick species were identified morphologically, and representative specimens were molecularly analyzed based on the cytochrome c oxidase subunit I (cox1) and 16S rRNA genes. Fifty-nine tick species that are indigenous in Vietnam were also reviewed in the context of their typical hosts in the region.ResultsMost ticks removed from cattle and buffalos were identified as Rhipicephalus microplus, including all developmental stages. Larvae and nymphs were found between January and July but adults until December. Further species identified from cattle were Rhipicephalus linnaei, Rhipicephalus haemaphysaloides, Amblyomma integrum and Haemaphysalis cornigera. Interestingly, the latter three species were represented only by adults, collected in one province: Son La. The dog was infested with nymphs and adults of R. linnaei in July. Phylogenetically, R. microplus from Vietnam belonged to clade A of this species, and R. haemaphysaloides clustered separately from ticks identified under this name in China, Taiwan and Pakistan. Amblyomma integrum from Vietnam belonged to the phylogenetic group of haplotypes of an Amblyomma sp. reported from Myanmar. The separate clustering of H. cornigera from Haemaphysalis shimoga received moderate support.ConclusionsThree tick species (R. linnaei, A. integrum and H. cornigera) are reported here for the first time in Vietnam, thus increasing the number of indigenous tick species to 62. Clade A of R. microplus and at least R. linnaei from the group of Rhipicephalus sanguineus sensu lato occur in the country. There is multiple phylogenetic evidence that different species might exist among the ticks that are reported under the name R. haemaphysaloides in South and East Asia. This is the first report of A. integrum in Southeastern Asia.Graphical

Exploring the interspecific relationships among Dendrobium species via electrochemical fingerprinting techniques

This study investigated the interspecific relationships among 29 Dendrobium species using electrochemical fingerprinting techniques. Differential pulse voltammetry was employed to record species-specific electrochemical profiles in both phosphate buffer solution (PBS) and acetic acid buffer solution (ABS). Distinct voltammetric patterns were observed for each species, with ABS yielding higher peak currents compared to PBS. On average, peak currents in ABS were 37.2 % ± 5.8 % higher than in PBS across all species. For example, D. nobile showed a peak current of 12.3 μA in ABS compared to 8.9 μA in PBS, a 38.2 % increase. Normalized two-dimensional plots of the electrochemical fingerprints served as unique signatures for species differentiation. Cluster analysis of the electrochemical data revealed phylogenetic relationships largely consistent with previous molecular studies, while also highlighting novel groupings. For instance, D. parishii and D. pulchellum formed separate branches, indicating distant relationships with other species. Common species such as D. officinale, D. nobile, D. primulinum, and D. loddigesii clustered together, confirming their close relationships. Interestingly, species from Sect. Dendrobium showed mixed clustering with species from four other sections, reflecting the genus's complex evolutionary history. While some inconsistencies with previous molecular phylogenetic studies were noted, such as the distant relationship between D. parishii and D. anosmum, the overall congruence between electrochemical and molecular data validates the potential of this approach. This study demonstrates that electrochemical fingerprinting, combined with appropriate data analysis techniques, can provide valuable insights into the taxonomic relationships and chemical diversity of Dendrobium species, offering a complementary tool to traditional morphological and molecular methods for plant systematics and conservation efforts.

A new cloud forest tree Lychnodiscus bali (Sapindaceae), Critically Endangered from the Bali-Ngemba Forest Reserve, NW Region, Cameroon

SummaryWe describe and illustrate Lychnodiscus bali Cheek (Sapindaceae), a new species to science, from the Bali Ngemba Forest Reserve of NW Region Cameroon, the last major remnant of cloud forest in the Bamenda Highlands of Cameroon, recently evidenced as a Tropical Important Plant Area (TIPA or IPA). Confined on current evidence to upper submontane forest, the new species is threatened by expanding habitat clearance for farms and is assessed as Critically Endangered. A small tree, attaining 3 – 4 m height, it is the first new species to be added to this Guineo-Congolian tree genus in 50 years, the third recorded from Cameroon and takes the number of species in the genus to eight. It has the highest known altitudinal range (1700 – 1950 m alt.) of any species of the genus. We discuss its discovery in the context of other recently discovered and highly threatened or even extinct plant species in the Cameroon Highlands, and the importance of their conservation. The new species was previously identified as Lychnodiscus grandifolius Radlk., but differs in the shorter length of the distal leaflets (12 – 18 cm vs 22 – 39 cm long); in the abaxial leaf surface lacking glands (vs glands flat and conspicuous); in being sparsely hairy all over (vs glabrous except the main veins) and in its larger flowers, 8 – 11 mm long at anthesis (vs 5 – 7 mm long). We present an identification key of the species of Lychnodiscus and discuss their classification in the context of recent molecular phylogenetic and phylogenomic studies. The authors contend that Lychnodiscus Radlk., previously placed in Cupanieae by Radlkofer, should now be placed in the reconstituted Nephelieae in the revised 2021 intrafamilial classification of Buerki et al., probably close to the genera Aporrhiza Radlk. and Laccodiscus Radlk. However, until the genus is included in molecular studies this cannot be confirmed and a sister relationship remains speculative.

Chloroplast Genome Profiling and Phylogenetic Insights of the “Qixiadaxiangshui” Pear (Pyrus bretschneideri Rehd.1)

The “Qixiadaxiangshui” pear (Pyrus bretschneideri Rehd.1) is a highly valued cultivar known for its crisp texture, abundant juice, and rich aroma. In this study, we reported the first complete chloroplast genome sequence of the “Qixiadaxiangshui” pear, which is 159,885 bp in length with a GC content of 36.58%. The genome exhibits a typical circular quadripartite structure, comprising a large single-copy region (LSC), a small single-copy region (SSC), and a pair of inverted repeat regions (IRs). A total of 131 genes were identified, including 84 protein-coding genes, 8 rRNA genes, and 37 tRNA genes. We also identified 209 simple sequence repeats (SSRs) and several mutation hotspots, such as ndhC-trnM-CAU and trnR-UCU-atpA, which can be applied in molecular identification and phylogenetic studies of Pyrus. Comparative genomic analysis showed high conservation among ten pear cultivars. Phylogenetic analysis indicated that the “Qixiadaxiangshui” pear is closely related to germplasm Dangshansuli, Wonwhang, and Yali, suggesting a recent common ancestor. These findings provided valuable insights into the genetic diversity and evolutionary dynamics of the Pyrus species and contribute to the conservation and breeding of pear germplasm resources.

Taxonomic revision of the genus Punjuba (Leguminosae, Caesalpinioideae)

Punjuba is a Neotropical genus of the mimosoid clade (Leguminosae). The genus was recently restablished and is recognized by presenting shrubs and trees, with lax racemes or spiciform racemes, leaves with 1–3 pairs of pinnae and 2–7 pairs of leaflets in each pinna, thin and lateraly compressed legumes, curved to spiral, and seeds lacking pleurogram. Recent advances on legumes systematics have been producing great challenges on the circumscription of genera, especially on the mimosoid clade. Molecular phylogenetic studies confirm that Punjuba is monophyletic, and a revision of the genus and species circumscriptions are needed. Here we present a taxonomic revision of Punjuba, providing an identification key for all species currently recognized in the genus, morphological descriptions, and distribution maps. We recognize six species in the genus: P. callejasii, P. centiflora, P. killipii, P. lehmannii, P. racemiflora and P. foreroana, being the last one described here as a new species. Abarema josephi is proposed as a new synonym of P. lehmannii. Morphological characters including the inflorescence, indumentum, leaves and leaflets, besides quantitative characters, were applied as diagnostic to recognize the species.

A new species of Grandinenia Minato & Chen, 1984 (Gastropoda, Stylommatophora, Clausiliidae, Garnieriinae) from Guangxi, China

A molecular phylogenetic study was conducted on genus Grandinenia, based on COI and 16S sequences. A total of eight out of 26 species in the genus, as well as three unidentified species were sequenced. Phylogenetic results supported the monophyly of Grandinenia and the validity of all sampled species and subspecies. A new species, Grandinenia jiangjilini Chen, Lin, Wu & Ouyang, sp. nov., from Guangxi, southern China is identified and described, based on morphological comparison and molecular phylogeny. The discovery indicates that the inflated-fusiform shell is not isolated in genus Grandinenia and the species diversity of the genus still remains to be explored.