ITS DNA Research Articles

Plastome evolution in the genus Sium (Apiaceae, Oenantheae) inferred from phylogenomic and comparative analyses

BackgroundSium L. (Apiaceae) is a small genus distributed primarily in Eurasia, with one species also occurring in North America. Recently, its circumscription has been revised to include 10 species, however, the phylogenetic relationships within its two inclusive clades were poorly supported or collapsed in previous studies based on nuclear ribosomal DNA ITS or cpDNA sequences. To identify molecular markers suitable for future intraspecific phylogeographic and population genetic studies, and to evaluate the efficacy of plastome in resolving the phylogenetic relationships of the genus, the complete chloroplast (cp) genomes of six Sium species were sequenced.ResultsThe Sium plastomes exhibited typical quadripartite structures of Apiaceae and most other higher plant plastid DNAs, and were relatively conserved in their size (153,029–155,006 bp), gene arrangement and content (with 114 unique genes). A total of 61–67 SSRs, along with 12 highly divergent regions (trnQ, trnG-atpA, trnE-trnT, rps4-trnT, accD-psbI, rpl16, ycf1-ndhF, ndhF-rpl32, rpl32-trnL, ndhE-ndhG, ycf1a and ycf1b) were discovered in the plastomes. No significant IR length variation was detected showing that plastome evolution was conserved within this genus. Phylogenomic analysis based on whole chloroplast genome sequences produced a highly resolved phylogenetic tree, in which the monophyly of Sium, as well as the sister relationship of its two inclusive clades were strongly supported.ConclusionsThe plastome sequences could greatly improve phylogenetic resolution, and will provide genomic resources and potential markers useful for future studies of the genus.

Effects of company and season on blood fluke (Cardicola spp.) infection in ranched Southern Bluefin Tuna: preliminary evidence infection has a negative effect on fish growth.

Aporocotylid blood flukes Cardicola forsteri and C. orientalis are an ongoing health concern for Southern Bluefin Tuna (SBT), Thunnus maccoyii, ranched in Australia. Therapeutic application of praziquantel (PZQ) has reduced SBT mortalities, however PZQ is not a residual treatment therefore reinfection can occur after the single treatment application. This study documents the epidemiology of Cardicola spp. infection in ranched SBT post treatment over three ranching seasons (2018, 2019 and 2021). Infection prevalence (percentage of SBT affected) and intensity (parasite load) was determined by adult fluke counts from heart, egg counts from gill filaments and the use of specific quantitative polymerase chain reaction (qPCR) for detection of C. forsteri and C. orientalis ITS-2 DNA in SBT hearts and gills. SBT Condition Index decreased as intensity of Cardicola spp. DNA in SBT gills increased, suggesting blood fluke infection had a negative effect on SBT growth (Spearman's r = -0.2426, d.f. = 138, p = 0.0041). Prevalence and intensity of infection indicated PZQ remained highly effective at controlling Cardicola spp. infection in ranched SBT, 10 years after PZQ administration began in this industry. Company A had the highest prevalence and intensity of Cardicola spp. infection in 2018, and Company G had the highest in 2019. No consistent pattern was seen in 2021. Overall, intensity of infection did not increase as ranching duration increased post treatment. Results from this study improve our knowledge of the biology of blood flukes and helps the SBT industry to modify or design new blood fluke management strategies to reduce health risks and improve performance of SBT.

Scalp microbiome of healthy women wearing hijab compared to those not wearing hijab: a cross-sectional study

Use of hijab can influence the scalp’s condition, including its microbiome. To date there has been no study comparing scalp microbiome in women wearing hijab to that in women not wearing hijab. This was a cross-sectional study conducted from August 2019 to April 2021. Healthy women aged 18 years old or older who had not undergone menopause were recruited. Those in the hijab group should wear hijab minimum 8 h a day for at least 5 years. After wash-out period, the sample was collected from the subject’s scalp. Next Generation Sequencing (NGS) was performed with primer V3-V4 region of 16S rRNA and ITS1 DNA for bacteria and fungi, respectively. Alpha diversity and beta diversity were identified, along with functional analysis. Actinobacteria and Ascomycota were the most dominant phyla on the scalp. S. capitis was more prominent in the hijab group while S. cohnii was more prominent in non-hijab group. Additionally, M. restricta was more common in hijab group while M. globosa was more common in non-hijab group. This study emphasizes the difference of scalp microbiome in women wearing hijab compared to women not wearing hijab, which indicated that women wearing hijab are more prone to seborrheic dermatitis.

Sorption and Biodestruction of Microcystin-LR by <i>Penicillium verrucosum</i> CP4 Strain Isolated from the Bottom Sediments of Sestroretsky Razliv Lake

The strain of fungus СР4 capable of degrading microcystin – LR (MC-LR) was isolated from the bottom sediments of Sestroretsky Razliv Lake. Based on DNA ITS sequencing and morphological analysis, the CP4 strain was identified as Penicillium verrucosum. The decrease in the content of MC-LR during the cultivation of strain СР4 from 0.64 μg/mL to 0.31 μg/mL occurs mainly due to biodegradation and, to a lesser extent, due to the sorption of the toxin by fungal cells. The method of biotesting (Daphnia magna) showed a decrease in the toxicity of the culture liquid in the process of MC-LR biodegradation by the strain СР4. The obtained results allow us to consider Penicillium verrucosum СР4 as a promising strain for mycoremediation of water bodies contaminated with microcystins.

DBALM: A novel method for identifying ornamental flowering plants based on DNA barcodes–leaf morphology

AbstractWhereas the presence of flowers on ornamental flowering plants is essential for their identification via traditional methods, ornamental flowering plants cannot be reliably identified in non‐flowering stages likewise. Here, DBALM (DNA Barcodes–Leaf Morphology), a new approach that combines DNA barcoding data with micromorphological features of the leaf epidermis and that is not limited by the flowering stage, was used to identify 16 evergreen rhododendron cultivars. First, the sequences of DNA barcodes, ITS, matK, psbA‐trnH, and rbcL, were obtained from the DNA of leaves. Phylogenetic analysis was conducted to clarify the groupings among all the samples based on the four markers. Then, microscopic features of the leaf epidermis were used to further distinguish individuals from the same clade. DNA barcoding permitted the 16 cultivars to be divided into eight groups. The microscopic features of the leaf epidermis permitted cultivars within the same clade to be distinguished. The matK + psbA‐trnH combination was the most effective barcode combination in this study. In addition, the new primer matK‐Rh_R was designed, and it increased the amplification rate of evergreen rhododendron cultivars to 100%. In sum, DBALM was capable of accurately identifying the 16 evergreen rhododendron cultivars using data collected from a single leaf in the vegetative growth stage. This method can greatly facilitate the identification and breeding of ornamental flowering plants.

Accidental cultivation of the European truffle Tuber brumale in North American truffle orchards.

Tuber brumale is a European edible truffle species that is often viewed as a contaminant in truffle orchards, as it visually resembles more valuable black truffles such as T. melanosporum, but differs in aroma and flavor and sells for a much lower price. Although T. brumale is not native to or intentionally cultivated in North America, it was reported to have been accidently introduced into British Columbia in 2014 and North Carolina in 2020. However, in winter of 2021, various truffle orchards in eastern North America produced truffles that differed from the anticipated harvest of T. melanosporum. Molecular analysis of these specimens confirmed T. brumale truffle fruiting bodies from ten orchards distributed across six eastern USA states. Phylogenetic analysis of nuclear ribosomal ITS and 28S DNA sequences indicated that all samples belong to the T. brumale A1 haplogroup, the genetic subgroup of T. brumale that is more common in western Europe. This pattern of widespread fruiting of T. brumale in North American truffle orchards is likely the result of T. brumale being introduced in the initial inoculation of trees used as hosts in T. melanosporum truffle cultivation. We review other examples of introduced non-target truffle species and strategies for limiting their impact on truffle cultivation.

Identification of Egyptian Mangrove Species Based on DNA Barcoding

Aims: Mangroves are woody trees or shrubs that grow in the intertidal zone and are distributed along tropical and subtropical coasts. These plants are resilient to various environmental challenges; they are also one of the most efficient terrestrial and coastal ecosystem for carbon fixation and storage. In recent years, mangrove reforestation has attracted much attention as a strategy to reduce the effects of climate change. In Egypt, there are two types of mangroves, Rhizophora mucronata and Avicennia marina, between 30°N and 30°S of the equator. Mangrove management presents a difficult task, particularly when it comes to managing molecular mangroves for long-term sustainability. With the impact of human activity on mangrove ecosystems increasing each year, molecular research on mangrove correlates remains to be conducted. For this reason, using DNA barcoding technology to quickly identify species, mangrove ecosystems may be protected.
 Methodology: In this work, the two Egyptian mangrove species were assessed through morphological, cytological, and molecular approaches. Two universal DNA barcodes, rbcL and ITS, were examined to identify their efficacy for Egyptian mangrove species identification and phylogenetic reconstruction.
 Results: According to pairwise alignments, the rbcL region had the highest level of variability (73.2%), whereas the ITS region was the least variable (11.96%). The selected Egyptian mangrove species can potentially be distinguished by barcoding loci rbcL and ITS due to the existence of distinctive variable sites.

Entoloma species of subgenus Cyanula (Tricholomatinae, Basidiomycota) in Norway, with emphasis on habitat preferences and distribution

This is the third paper with results from the Norwegian Entoloma project 2015-2017. An overview is given of the species of Entolomasubgenus Cyanula in Norway. Cyanula includes, among other elements, many grassland fungi. This paper is part of a larger studyconcerning the European species of the subgenus Cyanula, including a multigene phylogeny that will be published in due course.Furthermore, a monograph with updated taxonomic revision is recently issued. This paper focuses mainly on the species recorded from Norway with help of ITS DNA barcoding, with reference to recent collections also from Northern Sweden, with extensive notes of their habitat requirements and distribution. Altogether 53 Cyanula species are presented here, with an updated circumscription and naming. More than half of these are reported new to Norway since the start of the Norwegian Entoloma project. Of these, 18 have been described as new during the last three years, largely based on Norwegian material. Four of these are known exclusively from Scandinavia.

Insights into Diversity, Distribution, and Systematics of Rust Genus Puccinia.

Puccinia, which comprises 4000 species, is the largest genus of rust fungi and one of the destructive plant pathogenic rust genera that are reported to infect both agricultural and nonagricultural plants with severe illnesses. The presence of bi-celled teliospores is one of the major features of these rust fungi that differentiated them from Uromyces, which is another largest genus of rust fungi. In the present study, an overview of the current knowledge on the general taxonomy and ecology of the rust genus Puccinia is presented. The status of the molecular identification of this genus along with updated species numbers and their current statuses in the 21st century are also presented, in addition to their threats to both agricultural and nonagricultural plants. Furthermore, a phylogenetic analysis based on ITS and LSU DNA sequence data available in GenBank and the published literature was performed to examine the intergeneric relationships of Puccinia. The obtained results revealed the worldwide distribution of Puccinia. Compared with other nations, a reasonable increase in research publications over the current century was demonstrated in Asian countries. The plant families Asteraceae and Poaceae were observed as the most infected in the 21st century. The phylogenetic studies of the LSU and ITS sequence data revealed the polyphyletic nature of Puccinia. In addition, the presences of too short, too lengthy, and incomplete sequences in the NCBI database demonstrate the need for extensive DNA-based analyses for a better understanding of the taxonomic placement of Puccinia.

Langbiangia, a new genus of Gesneriaceae endemic to Langbiang Plateau, southern Vietnam and a taxonomic endeavor to achieve key targets of the post-2020 global biodiversity framework.

Situated in the southern end of the Annamite Mountain Range, Langbiang Plateau is a major biodiversity hotspot of southern Vietnam known for high species diversity and endemicity. To achieve effective conservation, parts of the plateau were designated as the Langbiang Biosphere Reserve, an UNESCO World Network aiming to improve relationships between inhabitants and their environments. Amongst the rich endemic flora of the plateau are three gesneriads ascribed to Primulina, a calciphilous genus with high species diversity in the vast limestone karsts stretching from southern China to northern Vietnam. However, a recent phylogenetic study questioned the generic placement of the Langbiang Primulina, corroborating with observations on the geographical distribution, habitat preference, and phyllotaxy of the three species. Based on phylogenetic analyses of nuclear ITS and plastid trnL-F DNA sequences of a comprehensive sampling covering nearly all genera of the Old World Gesneriaceae, we demonstrate that the three Langbiang Primulina species form a fully supported clade distantly related to other Primulina. As this clade is biogeographically, ecologically, morphologically, and phylogenetically distinct worthy of generic recognition, we propose to name it Langbiangia gen. nov. to highlight the rich and unique biodiversity of the Langbiang Plateau. By means of this taxonomic endeavor, we are hoping to raise the conservation awareness of this biodiversity heritage of southern Vietnam and promote the importance of Langbiang Biosphere Reserve that is crucial for achieving action-oriented global targets of the post-2020 global biodiversity framework (GBF) of the UN Convention on Biological Diversity (CBD)-effective conservation and management of at least 30% of biodiverse terrestrial, inland water, and costal and marine areas by 2030-that has been agreed at the COP15 in Montréal in December 2022.

Taxonomy and Phylogeny of Hyphomycetous Muriform Conidial Taxa from the Tibetan Plateau, China

During the investigation of lignicolous freshwater fungi in the Tibetan Plateau habitat, fifteen collections were isolated from submerged decaying wood. Fungal characteristics are commonly found as punctiform or powdery colonies with dark pigmented and muriform conidia. Multigene phylogenetic analyses of combined ITS, LSU, SSU and TEF DNA sequences showed that they belong to three families in Pleosporales. Among them, Paramonodictys dispersa, Pleopunctum megalosporum, Pl. multicellularum and Pl. rotundatum are established as new species. Paradictyoarthrinium hydei, Pleopunctum ellipsoideum and Pl. pseudoellipsoideum are reported as new records on the freshwater habitats in Tibetan Plateau, China. The morphological descriptions and illustrations of the new collections are provided.

Phylogenetical Position versus Pollination Syndromes: Floral Trichomes of Central American and Mexican Pinguicula.

Central American and Mexican Pinguicula species are characterized by enormous divergence in size and color of flowers and are pollinated by butterflies, flies, bees, and hummingbirds. It is known that floral trichomes are key characters in plant-pollinator interaction. The main aim of our study was to verify our hypothesis that the distribution and diversity of non-glandular and glandular trichomes are related to the pollinator syndromes rather than the phylogenetic relationships. The studied sample consisted of Central American and Mexican species. In our study, we relied on light microscopy and scanning electron microscopy with a phylogenetic perspective based on ITS DNA sequences. The flower morphology of species pollinated by butterflies and hummingbirds was similar in contrast to species pollinated by flies and bees. Species pollinated by butterflies and hummingbirds contained low diversity of non-glandular trichomes, which occurred mostly in the tube and basal part of the spur. Surprisingly, in P. esseriana and P. mesophytica, non-glandular trichomes also occurred at the base of lower lip petals. In the case of species pollinated by flies/bees, we observed a high variety of non-glandular trichomes, which occurred on the surface of corolla petals, in the tube, and at the entrance to the spur. Furthermore, we did not identify any non-glandular trichomes in the spur. The capitate glandular trichomes were of similar morphology in all examined species. There were minor differences in the shape of the trichome head, as well as the length and the number of stalk cells. The distribution and the diversity of non-glandular and glandular trichomes and pollinator syndromes were mapped onto a phylogenetic reconstruction of the genus. Most micromorphological characters appear to be associated more with floral adaptation to pollinators and less with phylogeny.

Phylogenetic analysis based on the ITS, matK and rbcL DNA barcodes and comparison of chemical contents of twelve Paeonia taxa in Türkiye

Twelve taxa of herbaceous Paeonia species were recorded in Türkiye. All definitions were performed morphologically and/or anatomically and there is no study based on DNA barcode sequences. Three barcode regions were sequenced to determine the phylogenetic relationships of Turkish Paeonia taxa. The chemical comparison of roots was also investigated. The taxons were collected between May and June 2021 from nine cities. Leaf materials were used for DNA isolation and ITS, matK and rbcL regions were amplified and sequenced. There was no difference among taxa in terms of rbcL sequences. But the ITS and matK regions distinguished 12 taxa and structured them in two groups. ITS region distinguished P. peregrina, P. arietina, and P. tenuifolia from other taxa, while matK region distinguished P. arietina and P. witmanniana from other taxa. Both barcode sequences actually showed that the registration of P. mascula subsp. arasicola was actually 100% similar to P. arietina. ITS was the most polymorphic region (n = 54) followed by matK (n = 9). These sequences could successfully discriminate Paoenia species from each other and diploid P. tenuifolia. The methanolic root (100 gr) extracts were examined for total phenolic and flavonoid content, and antioxidant activities. Significant variation was found for polyphenolic content, and antioxidant properties (TPC from 204.23 to 2343.89mg, TFC from 7.73 to 66.16mg, and FRAP from 523.81 to 4338.62mg). SC50 values of ABTS and DPPH were ranged from 115.08 to 1115.52μg/ml and 73.83 to 963.59μg/ml, respectively. It was concluded that 11 of 12 taxa had differences in terms of ITS and matK sequences and these region must be used for the correct identification of Turkish Paeonia.

RETRACTED: Clinical evolution of equine leishmaniasis with self-limiting cutaneous disease caused by Leishmania infantum in northeastern Brazil: A case report

Leishmania infantum infections have long been described in humans and dogs worldwide, but characterization of equine cases remains scarce. We describe the clinical evolution of a natural L. infantum infection to contribute to the diagnostic knowledge and epidemiology of equine leishmaniasis (EL). An auction-acquired four-year-old Mangalarga Marchador mare from Pernambuco state, presented a few subcutaneous nodules on the head and neck upon arrival at the purchaser's stud at Bahia state, in November of 2019. They progressed to multiple ulcerated and non-ulcerated nodules and spread to both right limbs in seven weeks. Hematology revealed anemia, lymphocytosis, monocytosis, and elevated plasma fibrinogen. Histopathology of the biopsied nodules identified a granulomatous dermatitis with macrophages containing Leishmania amastigotes. PCR detected Leishmania in skin lesions, but not in blood or spleen aspirate samples; ITS1 PCR-RFLP and DNA sequencing confirmed L. infantum species. A topical antiseptic and insect-repellent therapy and a monthly follow-up were established. All lesions improved progressively, without specific anti-Leishmania treatment, and 14months later there was a consistent resolution. This first description of EL by L. infantum in an endemic area is relevant to emphasize the need for epidemiological studies, and to enhance clinicians' awareness for differential diagnosis.

Plant DNA barcode library for native flowering plants in the arid region of northwestern China.

DNA barcoding is a well-established tool for rapid species identification and biodiversity monitoring. A reliable and traceable DNA barcode reference library with extensive coverage is necessary but unavailable for many geographical regions. The arid region in northwestern China, a vast area of about 2.5 million km2 , is ecologically fragile and often overlooked in biodiversity studies. In particular, DNA barcode data from the arid region in China are lacking. We develop and evaluate the efficacy of an extensive DNA barcode library for native flowering plants in the arid region of northwestern China. Plant specimens were collected, identified and vouchered for this purpose. The database utilized four DNA barcode markers, namely rbcL, matK, ITS and ITS2, for 1816 accessions (representing 890 species from 385 genera and 72 families), and consisted of 5196 barcode sequences. Individual barcodes varied in resolution rates: species- and genus-level rates for rbcL, matK, ITS and ITS2 were 79.9%-51.1%/76.1%, 79.9%-67.2%/88.9%, 85.0%-72.0%/88.2% and 81.0%-67.4%/84.9%, respectively. The three-barcode combination of rbcL + matK + ITS (RMI) revealed a higher species- and genus-level resolution (75.5%/92.1%, respectively). A total of 110 plastomes were newly generated as super-barcodes to increase species resolution for seven species-rich genera, namely Astragalus, Caragana, Lactuca, Lappula, Lepidium, Silene and Zygophyllum. Plastomes revealed higher species resolution compared to standard DNA barcodes and their combination. We suggest future databases include super-barcodes, especially for species-rich and complex genera. The plant DNA barcode library in the current study provides a valuable resource for future biological investigations in the arid regions of China.

Notes on four species of Russula subgenus Heterophyllidiae (Russulaceae, Russulales) from southern China.

Heterophyllidiae, one of the main subgenus of Russula (Russulaceae, Russulales), is both ecologically and economically important. Although many studies have focused on subgenus Heterophyllidiae in China, the diversity, taxonomy, and molecular phylogeny still remained incompletely understood. In the present study, two new species, R. discoidea and R. niveopicta, and two known taxa, R. xanthovirens and R. subatropurpurea, were described based on morphology and molecular phylogenetic analyses of ITS and 28S DNA sequences with new collections of subgenus Heterophyllidiae from southern China. Both morphological and phylogenetic analyses consistently confirmed that R. niveopicta and R. xanthovirens belong to the subsect. Virescentinae, R. discoidea and R. subatropurpurea come under subsect. Heterophyllae, and R. prasina is synonymized with R. xanthovirens.

Fungal Community Composition and Function Associated with Loose Smokeless Tobacco Products.

Smokeless tobacco products (STPs) contain several microbial communities which are responsible for the formation of carcinogens, like tobacco-specific nitrosamine (TSNAs). A majority of STPs are sold in loose/unpackaged form which can be loaded with a diverse microbial population. Here, the fungal population and mycotoxins level of three popular Indian loose STPs, Dohra, Mainpuri Kapoori (MK), and loose leaf-chewing tobacco (LCT) was examined using metagenomic sequencing of ITS1 DNA segment of the fungal genome and LC-MS/MS, respectively. We observed that Ascomycota was the most abundant phylum and Sterigmatomyces and Pichia were the predominant fungal genera in loose STPs. MK displayed the highest α-diversity being enriched with pathogenic fungi Apiotrichum, Aspergillus, Candida, Fusarium, Trichosporon, and Wallemia. Further, FUNGuild analysis revealed an abundance of saprotrophs in MK, while pathogen-saprotroph-symbiotroph were abundant in Dohra and LCT. The level of a fungal toxin (ochratoxins A) was high in the MK product. This study caution that loose STPs harbor various harmful fungi that can infect their users and deliver fungal toxins or disrupt the oral microbiome of SLT users which can contribute to several oral pathologies.

Multi-detection for paramphistomes using novel manually designed PAR-LAMP primers and its application in a lateral flow dipstick (LAMP-LFD) system

Loop-mediated isothermal amplification (LAMP) has been applied for the detection of various parasites, and its application in lateral flow dipstick (LFD) can improve the convenience of point-of-care diagnosis. A novel PAR-LAMP probe and primers were designed by manual selection from a region of low variation in the ITS-2 DNA sequence. Up to six species of rumen fluke were detected by LAMP and LAMP-LFD in this study. Target specificity and sensitivity were tested, revealing a high target specificity (accuracy) and a low limit of detection (sensitivity). Different target sensitivities of paramphistome were presented, including 5 pg for Gastrothylax crumenifer and Carmyerius sp.; 1 pg for Fischoederius elongatus, Orthocoelium parvipapillatum, and O. dicranocoelium; and 0.1 pg for Paramphistomum epiclitum. LAMP-LFD can detect a paramphistome egg even in contaminated in feces that was spiked with the egg under laboratory conditions. In addition, natural paramphistome infection in cattle from Surat Thani and Khon Kaen provinces, Thailand, was evaluated by detection of egg contamination in fecal specimens using PAR-LAMP primers. The PAR-LAMP detection result was also statistically evaluated by microscopic examination of feces. This study presents the application of novel manually designed primers in a LAMP-LFD system for improving performance in detection and diagnosis assays for paramphistomosis.

Polyploid hybrid speciation in theCalcarataspecies complex ofViolasectionMelanium(Violaceae): relating hybrid species to parent species distribution and ecology

AbstractTwo species of the Calcarata species complex of Viola section Melanium have been hypothesized to be of polyploid hybrid origin. To test this hypothesis, we used a modified restriction site associated (RAD) DNA-sequencing approach along with ITS and plastid DNA sequence data for reconstructing and dating the phylogeny of the group. For the detection of polyploid hybrid speciation and the parental lineages involved we used the four-taxon D-statistic metric (ABBA-BABA). Viola corsica (Elba, Corsica, Sardinia; calcifuge) and V. tineorum (Sicily; calcicole) were confirmed as polyploid hybrid species. They share V. cassinensis subsp. cassinensis (central Apennine; calcicole) as a parental lineage with V. bertolonii (Maritime Alps; serpentine) as the second parent of V. corsica and with V. nebrodensis (Sicily; calcicole) as the second parent of V. tineorum. The geographical distribution of the two hybrid species reflects the geographical distribution of the differential parent lineages. Substrate requirements of V. corsica are different, and those of V. tineorum are identical to those of their respective parents. We conclude that the differential distribution and ecology of V. corsica and V. tineorum can be traced back to properties of their respective parents.

Ganoderma multistipitatum sp. nov. from Chir pine tree (Pinus roxburghii Sarg.) in Pakistan

BackgroundGanoderma species are in state of flux in Pakistan as well as in the world. The diversity of this wood-inhabiting basidiomycete in Pakistan is still poorly known. A few species have been introduced from the country; even the country is rich with many Ganoderma species, which still need identification. MethodsGanoderma multistipitatum sp. nov. is described from Lahore, Punjab Pakistan by using the morpho-anatomical (via naked eye and compound microscope) characters and molecular data from the ITS regions of DNA. The specimens used in this study were collected from the Botanical Garden of Government College University Lahore. The G. multistipitatum sp. nov. is, to date, only known from Lahore on Chir pine tree. ResultsMorphologically, this species is characterized by laccate orange, brown basidiomata, maroon, brown multi stipitate, and anatomically by ellipsoid basidiospores (10.24–11.2 × 5.3–5.4 µm) and trimitic hyphal system. The G. multistipitatum sp. nov. is 21st species from the genus Ganoderma known from Pakistan. This new species is also described with photographs, line drawings and compared with nearby taxa of the phylogenetic tree constructed by software MEGA10. This species resembled with typical G. lucidum, but the ITS DNA sequences of this species were much far away from G. multistipitatum sp. nov. ones. ConclusionThe phylogeny based on ITS sequences supported this as a new distinct species, which finally compared with closely matrixed G. multipileum and related allies. The aim of this study was to introduce the new species from genus Ganoderma, which is a major contribution in biodiversity of the world.