Molecular and metabolic traits of some Egyptian species of Cassia L. and Senna Mill (Fabaceae-Caesalpinioideae).

The genus Cassia and Senna have been classified under subfamily Caesalpinioideae of family Fabaceae (Leguminosae) of order Fabales. There is a scarce taxonomical studies of the genus Cassia and Senna inhabiting Egyptian environments, thus, the main objective of the current was to revise and authenticate the phylogenetic relationship between studied taxa of the species of the genera Cassia and Senna in Egypt using the recent tools of ITS barcoding, RAPD analysis and metabolic profiling, in comparing to the traditional taxonomical features. From the cluster analysis of the traditional 27 morphological characters, the studied taxa were categorized into two major clades with an average taxonomic distance of 4.3. The clade I include Cassia fistula, C. renigera, C. javanica L subsp. nodosa and C. roughiia that belongs to series Obolospermae, and C. grandis that belongs to series Grandes. The clade (II) includes Senna surattensis and S. alata at taxonomic level 3.6. The taxonomical description of the studied taxa was confirmed from the molecular analysis of ITS sequences and RAPD analysis. The ITS sequences of the tested plants species C. fistula L, C. grandis MD4, C. javanica subsp. nodosa MD7, C. roxburghii MD5, C. renigera MD5 were deposited at genbank with accession numbers MW367973, MZ960447, MW386305, MW326753 and MW32685, respectively. While, the ITS sequences of the S. surrattensis and S. alata were deposited into genbank accession # MD14 MW367670 and MD20 MW412635, respectively. Thus, from the molecular analysis, two clades were clearly separated into Clade I of Cassia and Clade II of Senna. The cluster I represented by C. fistula, C. renigera, C. roxburghii, and C. javanica sub nodosa, and the cluster II represented by S. alata and S. surattensis. From the PCA of RAPD, a clearly discrimination between the two Taxa was observed revealing the characteristic grouping of Cassia and Senna. The species Senna alata and Senna surattensis were grouped together, but the species of C. renigera, C. javanica, C. roxburghii and C. grandis was grouped on a distinct group. The separation of Cassia and Senna species into two clusters verify the segregation of the genus Cassia L. senso lato into two distinct genera namely Senna P. and Cassia L. The morphological, molecular traits of the studied plants were authenticated from the metabolic profiling by GC-MS analysis. Among the 23 identified metabolites, four compounds namely hexadecanoic acid, methyl ester, 9-Octadecenoic acid (Z)-ethyl ester and Vitamin E were detected with fluctuated concentrations, among C. fistula, C. grandis, C. javanica subsp. nodosa and C. roxburghii. Conclusively, the traditional morphological features, molecular barcoding using ITS sequences, RAPD analysis and metabolic traits by GC-MS analysis, authenticates the taxonomical diversity of the genus Cassia and Senna.

In depth search of the Sequence Read Archive database reveals global distribution of the emerging pathogenic fungus Scedosporium aurantiacum.

Scedosporium species are emerging opportunistic fungal pathogens causing various infections mainly in immunocompromised patients, but also in immunocompetent individuals, following traumatic injuries. Clinical manifestations range from local infections, such as subcutaneous mycetoma or bone and joint infections, to pulmonary colonization and severe disseminated diseases. They are commonly found in soil and other environmental sources. To date S. aurantiacum has been reported only from a handful of countries. To identify the worldwide distribution of this species we screened publicly available sequencing data from fungal metabarcoding studies in the Sequence Read Archive (SRA) of The National Centre for Biotechnology Information (NCBI) by multiple BLAST searches. S. aurantiacum was found in 26 countries and two islands, throughout every climatic region. This distribution is like that of other Scedosporium species. Several new environmental sources of S. aurantiacum including human and bovine milk, chicken and canine gut, freshwater, and feces of the giant white-tailed rat (Uromys caudimaculatus) were identified. This study demonstrated that raw sequence data stored in the SRA database can be repurposed using a big data analysis approach to answer biological questions of interest. LAY SUMMARY: To understand the distribution and natural habitat of S. aurantiacum, species-specific DNA sequences were searched in the SRA database. Our large-scale data analysis illustrates that S. aurantiacum is more widely distributed than previously thought and new environmental sources were identified.

Screening saikosaponin d (SSd)-producing endophytic fungi from Bupleurum scorzonerifolium Willd.

Saikosaponin d (SSd) is an important bioactive compound of traditional Chinese medicinal plant Bupleurum scorzonerifolium Willd. and exhibits many effects, such as anti-tumor, anti-inflammation and immunomodulatory. Since endophytic fungi possess the natural capacity to produce the similar secondary metabolite to that of their host plants, they are promising as alternative sources of plant bioactive natural products. In this study, in order to search for SSd-producing strains, endophytes were isolated from B. scorzonerifolium and were authenticated by the ITS sequence and the translation elongation factor-1alpha gene (TEF-1α) sequence analysis. The profile of metabolites present in the crude exacts was carried out by ultra performance liquid chromatography time-of-flight mass spectrometry (UPLC/Q-TOF-MS) analysis. The results showed that two strains, CHS2 and CHS3 from B. scorzonerifolium could produce SSd by UPLC/Q-TOF-MS analysis, and the amount of SSd produced by strain CHS2 and CHS3 were about 2.17 and 2.40 µg/mL, respectively. CHS2 and CHS3 showed a close phylogenetic relationship to Fusarium oxysporum and Fusarium acuminatum, respectively. According to our concern, no endophytic fungi capable of producing SSd from B. scorzonerifolium have been found before. Our clear intention was to isolate and identify these endophytic fungi that produce important active secondary metabolites, and then study the strains that produce this compound on a large scale through fermentation or even genetic study, to provide a feasible and more convenient way for the production of SSd.

Biosystematic studies of some Egyptian species of Cestrum (Solanaceae).

Cestrum is the second largest genus of family Solanaceae, after Solanum, distributed in warm to subtropical regions. Species of genus Cestrum are one of the most ethnopharmacological relevant plants, for their broad biological and pharmacological properties. There is a scarcity to taxonomical studies and identification of these plants in Egypt, thus, the objective of this study was to implement various morphological features, chemical markers and molecular tools to emphasize the taxonomical features of the different Cestrum species. Morphologically, the epidermal cells of C. diurnum, C. elegans and C. parqui were irregular with sinuate anticlinal wall patterns for both surfaces, while, C. nocturnum has anticlinal walls, sinuolate with polygonal to irregular epidermal cells on the abaxial surface. The species of Cestrum have hypostomatic leaves, except C. parqui that has amphistomatic leaves. The experimented species of Cestrum have Anomocytic and anisocytic stomata, while, C. elegans has a diacytic stomata. The morphologically identified Cestrum spp were molecular confirmed based on their ITS sequences, the sequences of C. diurnum, C. nocturnum, C. elegans and C. parqui were deposited on genbank with accession # MT742788.1, MT749390.1, MW091481.1 and MW023744.1, respectively. From the SCOT analyses, the four species of Cestrum were grouped into 2 clusters (I, II), cluster I contains C. elegans, C. nocturnum and C. parqui, while cluster II contains only C. diurnum with 100% polymorphism for all primers. From the GC-MS profile, the C. diurnum exhibited a diverse metabolic paradigm, ensuring their richness with different metabolites comparing to other experimented Cestrum species. Among the total resolved metabolites, 15-methyltricyclo 6.5.2-pentadeca-1,3,5,7,9, 11,13-heptene was the highly incident compound in C. elegans (35.89%) followed by C. parqui (21.81%) and C. diurnum (11.28%), while it absent on C. nocturnum. The compound, 2,2',6,6'-tetra-tert-butyl-4,4'-methylenediphenol was highly detected in C. elegans and C. dirunum with minor amounts in the other Cestrum species. Cypermethrin and 3-butynyl-2,2,5-trimethyl-1,3-dioxane-5-methanol were pivotally reported in C. nocturnum. Taken together, from molecular and metabolic markers, C. diurnum, C. parqui and C. elegans have higher proximity unlike to C. nocturnum.

Molecular identification and phylogenetic analysis of fungal pathogens isolated from diseased fish in Xinjiang, China.

The outbreaks of fungal diseases in cultured fish have been severe in recent years, which is harmful to the healthy and sustainable development of fish farming. In this study, an investigation was conducted for significant fungal infections of 12 species of fish in four regions in Xinjiang, China, to understand the distribution of local fish fungal pathogens. Twenty-six fungal strains with pathogenicity were isolated, and the challenge experiment showed that eight strains from Changji area had high infection rate to fish eggs. Based on internal transcribed spacer sequence data and molecular analysis, the 26 strains were classified into nine different species of six fungal genera. Phylogenetic analysis showed that all strains were divided into two clades, namely Cluster 1 (contains only the genus Mucor) and Cluster 2 (consists of five small branches), and the distribution of strains from the same region was scattered in two clusters. There is no strict host selectivity for these fungi to infect fish. Mucor sp. are the main fungal pathogen of fish in these four regions, whereas Hypophthalmichthys molitrix and Carassius auratus are two types of fish that were susceptible to pathogen. In addition, the environmental adaptability experiments showed that eight highly pathogenic strains have different adaptability to the environment, and their optimum temperature and pH were 25°C and 7.0, respectively, whereas the concentration of NaCl was negatively correlated with the growth of strains. Therefore, these results indicated that the coinfection of multiple fungal pathogens in a culture region should be considered in the future study.

A pilot study: a possible implication of Candida as an etiologically endogenous pathogen for oral lichen planus.

BACKGROUND: The aim of this study was to investigate the prevalence and genotypic profiles of Candida albicans in patients with oral lichen planus (OLP). MATERIALS AND METHODS: Positive rates and genotypic profiles of Candida albicans strains from OLP patients and healthy controls were analyzed. Random amplified polymorphic DNA and internal transcribed spacer of ribosome DNA polymerase chain reactions were used to sequence the DNA of these strains, and then their genetic similarity was measured using BLAST, UIV Band, and Vector NTI Suite Sequence Analyses Software. RESULTS: The prevalence of C. albicans strains detected from erosive-OLP, non-erosive OLP, and normal individuals was 18.87, 18.75, and 7.92%, respectively. Four different genotypes were revealed by the two methods. To be specific, type I was found only in the healthy subjects; type II a and II b were found in non-erosive OLP, and type III was identified in erosive OLP. Intragroup similarity coefficients, i.e. SAB were 100%, and inter-groups similarity coefficients, i.e. SAB were less than 30%. CONCLUSIONS: The genotypic results of C. albicans in OLP revealed an endogenous rather than exogenous infection of C. albicans. In addition, a possible pathogenic role of C. albicans in OLP, with the etiologic sense contributing to a more proper recognition on the pathogenesis, development, and progression of OLP, as well as some strategies for its diagnosis and treatment were identified.

[Isolation and identification of endophytic fungi from Huperzia serrata and their metabolites' inhibitory activities against acetylcholinesterase and anti-inflammatory activities].

A total of 27 endophytic fungal strains were isolated from Huperzia serrata,which were richly distributed in the stems and leaves while less distributed in roots. The 27 strains were identified by Internal Transcribed Spacer( ITS) r DNA molecular method and one of the strains belongs to Basidiomycota phylum,and other 26 stains belong to 26 species,9 general,6 families,5 orders,3 classes of Ascomycota Phylum. The dominant strains were Colletotrichum genus,belonging to Glomerellaceae family,Glomerellales order,Sordariomycetes class,Ascomycota Phylum,with the percentage of 48. 15%. The inhibitory activities of the crude extracts of 27 endophytic fungal strains against acetylcholinesterase( ACh E) and nitric oxide( NO) production were evaluated by Ellman's method and Griess method,respectively. Crude extracts of four fungi exhibited inhibitory activities against ACh E with an IC50 value of 42. 5-62. 4 mg·L~(-1),and some fungi's crude extracts were found to inhibit nitric oxide( NO) production in lipopolysaccharide( LPS)-activated RAW264. 7 macrophage cells with an IC50 value of 2. 2-51. 3 mg·L~(-1),which indicated that these fungi had potential anti-inflammatory activities.The chemical composition of the Et OAc extract of endophytic fungus HS21 was also analyzed by LCMS-IT-TOF. Seventeen compounds including six polyketides,four diphenyl ether derivatives and seven meroterpenoids were putatively identified.

Variation of rDNA Internal Transcribed Spacer Sequences in Rhizoctonia cerealis.

Fifty-four single protoplast isolates (SPIs) were regenerated from three Rhizoctonia cerealis strains. A total of 169 rDNA-ITS regions were cloned and sequenced from these 54 SPIs. Variations in the ITS1 and ITS2 regions that flank the 5.8S gene were found within clones from the same strain, as well as within clones from the same SPI. These include variations in GC content and ITS length, and single-nucleotide polymorphisms (SNPs). The different strains and SPIs GC contents range from 40.25 to 41.74% and from 42.40 to 45.02%, in the ITS1 and ITS2 regions, respectively. All SNPs occur in the ITS1 and ITS2 regions, with 3-6 and 4-6 polymorphic sites in each region, respectively, in the different strains. SNP variation is relatively stable within the same strain. For example, the 89 ITS sequences generated from isolate WK-207, regardless of SPI or clone, predominantly cluster into two separate clades on a phylogenetic tree, suggesting that nuclei genetic heterogeneity is related to ITS variation in R. cerealis. Although rDNA-ITS sequences from the three strains and different SPIs are somewhat variable, all of our ITS sequences cluster together in anastomosis subgroup AG-DI during phylogenetic analysis. The ITS variation we observed does not negatively influence R. cerealis anastomosis group or subgroup classification.

Terpenoids and Phenylpropanoids in Ligularia duciformis, L. kongkalingensis, L. nelumbifolia, and L. limprichtii.

The diversity in root chemicals and evolutionally neutral DNA regions in the complex of Ligularia duciformis, L. kongkalingensis, and L. nelumbifolia (the d/k/n complex) was studied using eight samples collected in central and northern Sichuan Province of China. Cacalol (14) and epicacalone (15), rearranged eremophilanes, were isolated from the complex for the first time. Two new phenylpropanoids were also obtained. Seven of the eight samples produced phenylpropanoids and the other produced lupeol alone. Two of the seven samples also produced furanoeremophilanes or their derivatives and one produced oplopanes. The geographical distribution of the sesquiterpene-producing populations suggests that the production of sesquiterpenes evolved independently in separate regions. L. limprichtii collected in northern Sichuan was also analyzed and its chemical composition and the sequence of internal transcribed spacers (ITSs) in the ribosomal RNA gene cluster were found to be similar to that in the d/k/n complex and L. yunnanensis, which are morphologically similar.

Performance of the Real Fungus-ID kit based on multiplex RT-PCR assay for the rapid detection and identification of Trichophyton spp. and Microsporum spp. in clinical specimens with suspected dermatophyte infection.

AIMS: The aim of this study was to evaluate the performance of a commercially available multiplex RT-PCR assay for the rapid detection and identification of dermatophytes directly from clinical samples and cultures. METHODS AND RESULTS: The multiplex RT-PCR assay was used to evaluate 118 clinical isolates from various specimen types and a total of 140 known specimens were compared with both conventional methods, commercially available PCR-REBA, and ITS sequence analysis. In this study, multiplex RT-PCR assay yield significantly more positive results than culture (91·9 vs 39·5%) and conventional methods including KOH microscopy (91·9 vs 71·3%). Although the results among the multiplex RT-PCR, PCR-REBA and ITS sequence analysis were concordant (100%) in 118 clinical isolates, concordant results between multiplex RT-PCR assay and culture were at 66% (78/118). The overall positive rates for the PCR-REBA, multiplex RT-PCR assay and ITS sequence analysis were 98·8, 91·9, and 52·9% respectively. In addition, the concordance rate of multiplex RT-PCR assay and the PCR-REBA assay was 93% (95% confidence interval (CI), 89·9-96·1, P < 0·0001), 93·7% (95% CI, 90·5-96·4, P < 0·0001) sensitivity, 100% (95% CI, 80·0-100, P < 0·0001) specificity, and 99·6% positive and 81% negative predictive values, respectively. Among the 258 samples, the most frequently identified dermatophyte species were Trichophyton rubrum (n = 199, 77·1%) and Trichophyton mentagrophytes (n = 28, 10·9%). CONCLUSIONS: The entire multiplex RT-PCR procedure takes about 3 h, while results from culture can take up to 2-3 weeks. The use of the multiplex RT-PCR molecular diagnostic assay was rapid and reliable for detecting pathogen infections. SIGNIFICANCE AND IMPACT OF THE STUDY: Even though the use of molecular diagnostic technology is more expensive than conventional methods, the clinical and economic benefit of saving time relative to expense remains to be elucidated. Therefore, the multiplex RT-PCR assay may provide the essential information to accelerate therapeutic decisions for earlier and adequate antibiotic treatment in the acute phase of fungal pathogen infections.

The Intra-familial Relationships of Pentaphylacaceae s.l. as Revealed by DNA Sequence Analysis.

During the past three decades, molecular taxonomy has made considerable changes in the systematic delimitations of several families in the order Ericales which were formed earlier based on morphology. For instance, the Pentaphylacaceae s.l. has been treated differently by both modern and traditional taxonomists. Modern molecular taxonomists constituted this family by combining the traditionally defined Pentaphylacaceae s.s. (Pentaphylax), Sladeniaceae s.s. (Sladenia), the subfamily Ternstroemioideae with 11 genera of Theaceae s.l. and the genus Ficalhoa. There are also treatments placing the genus Pentaphylax with Ternstroemioideae in Pentaphylacaceae and Ficalhoa with Sladenia in Sladeniaceae. Because most of these genera are poorly studied, investigations on all aspects are important to understand the phylogeny to settle the issues surrounding the treatment of the 14 genera in this family. In the present study, DNA sequences of nrITS and trnL-F genes from species of 11 genera from these 14 genera were generated and analyzed together with sequences from other closely related members of Ericales. The results suggested existence of four distinct lineages viz., Sladenia, Pentaphylax, and tribes Frezierieae (9 genera) and Ternstroemieae (2 genera). Further, it demonstrated that within the biggest lineage, Frezierieae, the Visnea remained sister to the clades Adinandra+Cleyera, Euryodendron+Symplococarpon, Freziera, and finally, Eurya. Based on the evidence, it can be concluded that Sladeniaceae and Pentaphylacaceae are very close to each other and the proposal of merging them into a mega family Pentaphylacaceae s.l. with four tribes, i.e., Sladenieae, Pentaphylaceae, Ternstroemieae, and Freziereae should be considered seriously.

Disseminated Histoplasmosis presenting with panniculitis and macrophage activation syndrome in a patient undergoing immunosuppressive therapy for Mixed Connective Tissue Disease (MCTD).

Histoplasmosis, a fungal infection caused by Histoplasma capsulatum, is endemic in many parts of the world. However, this is not common in Japan. We herein present a unique case of military histoplasmosis in a 45-year-old female with mixed connective tissue disease (MCTD) who was receiving immunosuppressive therapy. The histological findings coupled with molecular confirmation led to final a diagnosis. This case emphasizes the diagnostic challenges associated with histoplasmosis in immunocompromised patients and underscores the importance of considering it in the differential diagnosis of any atypical presentation in rheumatic patients.

Conserved DNA sequence analysis reveals the phylogeography and evolutionary events of Akebia trifoliata in the region across the eastern edge of the Tibetan Plateau and subtropical China.

BACKGROUND: The eastern edge of the Qinghai‒Tibet Plateau (QTP) and subtropical China have various regions where plant species originate and thrive, but these regions have been the focus of very few integrative studies. Here, we elucidated the phylogeographic structure of a continuous and widespread Akebia trifoliata population across these two regions. RESULTS: Sixty-one populations consisting of 391 genotypes were examined to assess population diversity and structure via network distribution analysis, maximum likelihood phylogenetic tree reconstruction, divergence time estimation, demographic history inference, and ancestral area reconstruction of both conserved internal transcribed spacer (ITS) and chloroplast (rps16) DNA sequences. The results showed that the ITS region was more variable than the rps16 region and could be suitable for studying intraspecific phylogeography. The A. trifoliata population displayed high genetic diversity, genetic differentiation and obvious phylogeographical structure, possibly originating on the eastern QTP, expanding during the last glacial-interglacial cycle, diverging in the early Pleistocene and middle Pleistocene, and extensively migrating thereafter. The migration route from west to east along rivers could be largely responsible for the long-distance dispersal of this species, while three main refuges (Qinba Mountains, Nanling Mountains and Yunnan-Guizhou Plateau) with multiple ice shelters facilitated its wide distribution. CONCLUSIONS: Our results suggested that the from west to east long migration accompanying with the minor short reciprocal migration in the south-north direction, and the three main refuges (the Qinba Mountains, Nanling Mountains and Yunnan-Guizhou Plateau) contributed to the extant geographical distribution of A. trifoliata. In addition, this finding also strongly reduced the discrepancy between glacial contraction and postglacial expansion and the in situ survival hypothesis by simultaneously considering the existence of many similar climate-related ecological niches and migration influences.

Dematiaceous fungal keratitis caused by Cladophialophora boppii - A case report.

Purpose: To report a rare case of dematiaceous fungal keratitis caused by Cladophialophora boppii (C. boppii) in an immunocompromised patient. Observations: An 83-year-old male with chronic renal failure was referred to the Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan due to persistent corneal epithelial defects (PEDs) in his left eye. Initial examination revealed decreased central corneal sensitivity and decreased tear secretion in that eye, both thought to be associated with herpetic keratitis. Permanent punctal-plug surgery combined with therapeutic soft contact lens wear was performed to treat the PED, which initially healed, yet recurred. Follow-up examination revealed a 1.0-mm-diameter black lesion consistent with the PED site, which subsequently increased in size, so treatment with miconazole solution eye drops, natamycin ophthalmic ointment, and systemic itraconazole was initially performed. Since the region of the lesion had progressed to corneal perforation, corneal transplantation surgery under general anesthesia was scheduled, yet the patient refused to undergo surgery. Mycological testing via DNA sequencing of the internal transcribed spacer of ribosomal DNA regions revealed that the isolate or pathogen was C. boppii. Mycotic keratitis caused by C. boppii was found to be resistant to antifungal drugs. Conclusion and importance: This is a rare case of fungal keratitis caused by C. boppii in an elderly immunocompromised patient.

Gentianamopanshanensis (Gentianaceae), a new species from Yunnan, southwest China.

Gentianamopanshanensis, a new species of the family Gentianaceae is here described and illustrated. This species is presently known only from the Mopanshan Mountain, Yunnan Province, southwest China. Phylogenetic analysis based on ITS sequence data has shown that this new species is a member of the series Fimbriatae of the section Chondrophyllae. Morphologically, it mostly resembles G.mairei and G.panthaica, but differs clearly from the latter two species in the shape and size of the leaves, and the characters of the corolla throat and plicae.

Polymerase chain reaction-based methods for the rapid identification of Amanita exitialis.

Amanita exitialis, a deadly mushroom found in eastern Asia, causes the highest death rates among all poisonous mushrooms in China. The aim of the present study was to develop an efficient, accurate, and user-friendly PCR-based method for identifying A. exitialis that could facilitate the prevention, diagnosis, and treatment of associated food poisoning. A. exitialis-specific primers and probes were designed based on the internal transcribed spacer region variations of 27 mushroom species. Specificity was confirmed using conventional and real-time PCR for 23 non-target mushroom species, including morphologically similar and closely related species. Compared to conventional PCR, real-time PCR was more sensitive (detectable DNA concentration: 1.36 × 10-2 ng/µL vs. 1.36 × 10-3) and efficient (analysis time: 1 h vs. 40 min). Furthermore, the real-time PCR results could be immediately visualized using amplification curve analysis. The results present two robust PCR-based methods for A. exitialis identification that can facilitate food safety.

Sileneophioglossa (Caryophyllaceae, Sileneae), a new species from southwest China.

Sileneophioglossa Huan C. Wang & Feng Yang, a new species of Caryophyllaceae, is here described and illustrated based on morphological and molecular evidence. The new species was found in Sichuan and Yunnan provinces, southwest China. Phylogenetic analysis based on ITS sequences showed this new species belongs to section Cucubaloides. Morphologically, it resembles S.phoenicodonta and S.viscidula, which were also found in the southwest China, but clearly differs from the latter two species by having 5-7 mm long calyces with sparsely hirtellous and short glandular hairs, white petals, linear limbs and lobes, and absent or oblong-linear coronal scales. A distribution map and a table with morphological diagnostic characters of new species and its closest relatives are provided, as well as a preliminary conservation assessment of S.ophioglossa under the IUCN criteria.

Synotisjinpingensis (Asteraceae, Senecioneae), a new species with white ray florets from southeastern Yunnan, China.

Synotisjinpingensis (Asteraceae, Senecioneae), a new species from Jinping county in southeastern Yunnan province, China, is described and illustrated. This species is distinguished by having white ray florets in the genus Synotis, in which only species with yellow ray florets have been hitherto known. In habit and leaf shape S.jinpingensis is most closely similar to S.duclouxii, a species occurring in southwestern Guizhou, southern Sichuan and northeastern Yunnan, China, but differs, in addition to the color of ray florets, by having fewer lateral veins of leaves, obviously longer bracts of calyculus, and larger phyllaries. The membership of the new species within Synotis is strongly corroborated by evidence from floral micromorphology and phylogenetic analyses based on ITS sequence data. Color photographs of living plants, a distribution map, and provisional IUCN status of S.jinpingensis are provided.

Nematode-citrus plant interactions: host preference, damage rate and molecular characterization of Citrus root nematode Tylenchulus semipenetrans.

Citrus plants are host to several plant parasitic nematodes (PPNs), which are microscopic organisms. Among PPNs, the citrus root nematode, T. semipenetrans (Cobb 1913) (Tylenchida: Tylenchulidae), causes significant damage to citrus plantations worldwide. Understanding citrus nematode populations, precise identification, host preference among citrus species, and damage threshold are crucial to control T. semipenetrans. The minutiae of citrus plant-nematode interactions, nematode density and molecular nematode identification are not well understood. In this study, nematode species and density in citrus orchards, host specialization, molecular and morphological characteristics of nematodes were assessed. Molecular and morphological methods, host-nematode interactions, host (citrus species) preference, damage economic threshold (ET), and economic injury level (EIL) were determined using laboratory methods and field sampling. Citrus plantations in different provinces in the Mediterranean region of Turkey were investigated. Nematode species were identified molecularly and morphologically. ITS sequences revealed that samples were infected by citrus root nematode T. semipenetrans. The lowest nematode density was in C. reticulata in Mersin (53 2nd stage juveniles (J2s) 100 g-1 soil), while the highest density was from Hatay in C. sinensis (12173 J2s 100 g-1 soil). Highest citrus nematode population density was on roots of C. reticulata, followed by C. sinensis, C. limon, and C. paradisi. The citrus nematode is more common than was thought and population fluctuations change according to specific citrus species. Environmental conditions, host and ecological factors, such as temperature, soil pH, and soil nutrients, might influence nematode populations in citrus orchards. Investigating nematode density in diverse soil ecologies and the responses of different resistant/tolerant citrus species and cultivars to nematode populations is essential in future studies.

Distinguishing features of Lycium L. species (family Solanaceae) distributed in Egypt based on their anatomical, metabolic, molecular, and ecological characteristics.

Among the 70-80 species of the genus Lycium (family Solanaceae) disjunctly distributed around the world, only three are frequently distributed in different locations in Egypt. Due to the morphological similarities between these three species, there is a need for alternative tools to distinguish them. Thus, the objective of this study was to revise the taxonomic features of Lycium europaeum L., Lycium shawii Roem. & Schult., and Lycium schweinfurthii var. aschersonii (Dammer) Feinbrun in consideration of their anatomical, metabolic, molecular, and ecological characteristics. In addition to analysis of their anatomical and ecological features, DNA barcoding was performed for molecular characterization through internal transcribed spacer (ITS) sequencing and start codon targeted (SCoT) markers. Furthermore, metabolic profiling of the studied species was conducted based on gas chromatography-mass spectrometry (GC-MS). The observed anatomical features of the adaxial and abaxial epidermal layers, type of mesophyll, crystals, number of palisade and spongy layers, and the vascular system showed variations between the studied species. Beyond this, the anatomy of the leaves showed an isobilateral structure in the studied species, without distinct differences. Species were molecularly identified in terms of ITS sequences and SCoT markers. The ITS sequences were deposited in GenBank with accession numbers ON149839.1, OP597546.1, and ON521125.1 for L. europaeum L., L. shawii, and L. schweinfurthii var. aschersonii, respectively. The sequences showed variations in GC content between the studied species; this was 63.6% in L. europaeum, 61.53% in L. shawii, and 63.55% in L. schweinfurthii var. aschersonii. A total of 62 amplified fragments, including 44 polymorphic fragments with a ratio of 70.97%, were obtained in the SCoT analysis, as well as unique amplicons in L. europaeum L., shawii, and L. schweinfurthii var. aschersonii of 5, 11, and 4 fragments, respectively. Through GC-MS profiling, 38 compounds were identified with clear fluctuations in the extracts of each species. Of these, 23 were distinguishing chemicals that could help in chemical identification of the extracts of the studied species. The present study succeeds in identifying alternative clear and diverse characteristics that can be used to distinguish between L. europaeum, L. shawii, and L. schweinfurthii var. aschersonii.