First record of Biecheleria cincta (Dinophyceae) from Chinese coasts, with morphological and molecular characterization of the strains

The presence ofBiecheleria cincta （=Woloszynskia cincta） in the Chinese coasts is reported for the first time. In scanning electron microscope, three to five series of vesicles and an elongated apical vesicle （EAV） were visible in the epicone, and both the hypocone and the cingulum had three series of vesicles each. Thin sections revealed that B. cincta possesses stalked pyrenoids and an unusual eyespot consisting of a stack of cistemae with brick-like materials （type E）, thus supporting its transfer from Woloszynskia to Biecheleria. Spiny cysts formed spontaneously in culture, with an encystment rate of around 20%. Both large subunit ribosomal DNA （LSU rDNA） and internal transcribed spacer region （ITS） sequences in 12 strains from the Chinese coasts were determined. Phylogenetic analyses based on LSU rDNA and ITS sequences using Bayesian inference and maximum likelihood revealed two distinct ribotypes （referred to as ribotype A and B） in B. cincta. ITS region pairwise distances within B. eincta ranged from 0.024 to 0.072, suggesting the existence of a complex of cryptic species.

Molecular phylogenetic analyses based on the complete plastid genomes and nuclear sequences reveal Daphne(Thymelaeaceae)to be non-monophyletic as current circumscription

The diverse members of the genus Daphne are prized for their fragrant flowers.Despite being promising ornamental plants in many countries,genetic information of Daphne is scarce.In this study,the plastomes of four species and one variety of Daphne were sequenced and analyzed.The plastomes were typical and contained a pair of inverted repeat(IR)regions that separated the large single-copy(LSC)region from the small single-copy(SSC)region.With a length ranging from 132,869 bp(D.genkwa)to 174,773 bp(D.championii),106 to 141 genes were predicted.Comparative plastome analysis of the newly sequenced plastomes with four publicly available Daphne plastomes identified an expansion of the IRs,sequence variations,and mutational hotspots.Phylogenetic analyses indicated that the genus Daphne in its current circumscription is polyphyletic.Daphne genkwa was nested within the genus Wikstroemia,while D.championii was well resolved as sister to Edgeworthia.These findings concurred with results from our study that used nuclear ribosomal internal transcribed spacer sequence data.The conflicts on the molecular placement of D.championii and D.genkwa and the present taxonomic classification in Daphne suggest that a new intergeneric classification system of Daphneae warrants consideration.

Molecular identification of some wild medicinal macrofungi from Northern Iran

In last decades,macrofungi have attracted increasing attention because of their valuable nutritional and medicinal properties.In this study,a total of 180 macrofungal samples were collected from forests in Mazandaran province,Iran.The dominant orders were Polyporales(51%)and Agaricales(35%).Pure mycelial cultures were successfully obtained from 91 collected samples.Regarding morphological data,47 isolates were selected for molecular identification based on internal transcribed spacer region(ITS)sequence analysis.The results showed that the 38 macrofungal isolates were belonging to 22 species,19 genera,10 families and 5 orders.Most of the macrofungi(47%)were identified as Trametes species and Ganoderma species.Three isolates identified as Hohenbuehelia species,Polyporellus brumalis and Ceriporia lacerata were records as a new to the Iran fungal flora.This study increases the knowledge on Iranian macrofungal diversity and facilitates future genetic and biotechnological investigations on these macrofungi.

Relationship between DNA Barcoding and Chemical Classification of Salvia Medicinal Herbs

Objective To make the identification of medicinal herbs in Salvia L.quickly and accurately.Methods In this work,DNA barcoding and chemical fingerprint were compared for the identification of herbs in Salvia L.First,the nucleotide sequences of the internal transcribed spacer region two amplified from 48 medicinal plants in Salvia L.,and three other groups of medicinal plants in Lamiaceae were sequenced.A molecular phylogeny was constructed using the minimum evolution and maximum parsimony methods according to their sequence diversity.Second,the water-solution bioactive components and lipid soluble components were tested by HPLC.Then a chemical phylogeny was built using HPLC fingerprint data.Comparing the molecular and chemical phylogenetic trees revealed many similarities.Results DNA barcoding was sequencing based and could therefore provide more accurate results within a shorter time especially in large-scale studies.Conclusion The results show that ITS2 region is a novel DNA barcode for the authentication of the species in Salvia L.This is the first work to show the relationship between DNA barcoding and chemical components.

Environmental barcoding of the ectomycorrhizal fungal genus Cortinarius

Understanding the role of ectomycorrhizal fungi in plant communities is hampered by a lack of knowledge about fungal diversity.DNA barcoding of the ectomycorrhizal fungal genus Cortinarius was used to compare fungal diversity in soil from four plant communities:(i)Nothofagus forest(where Cortinarius is common and diverse),(ii)Kunzea forest(where Cortinarius is present but with low diversity),(iii)a Pinus radiata plantation(Cortinarius is not thought to be present)and(iv)a sub-Antarctic island(where known ectomycorrhizal hosts are absent).PCR primers specific for the ITS region of Cortinarius species were developed.Specificity was tested in vitro and in silico against DNA from basidiocarps of Cortinarius and non-Cortinarius species.The primers were tested for their ability to amplify Cortinarius DNA in soil from forests of the three ectomycorrhizal forest communities and a range of soils from the ectomycorrhiza-free subantarctic Campbell Island.High diversity of Cortinarius was associated with soil of all three ectomycorrhizal communities,despite Cortinarius being previously unrecorded from Pinus.Soil from all three communities share some ectomycorrhizal fungi(including fungi shared between native and exotic hosts),having implications for community succession,introduction of exotic fungi and biodiversity assessment.No Cortinarius was detected from Campbell Island samples.The validated molecular protocol assessed species diversity in a rapid and cost effective way.Baseline biodiversity assessment based on DNA barcoding is more effective at detecting diversity than traditional methods,but requires careful consideration of the difference between ectomycorrhizal fungal diversity in soil versus root-tips.