The phylogeny of Ptychostomum was first spacer (ITS) region of the nuclear ribosomal (nr) DNA DNA rps4 sequences. Maximum parsimony, maximum undertaken based on analysis of the internal transcribed and by combinin...The phylogeny of Ptychostomum was first spacer (ITS) region of the nuclear ribosomal (nr) DNA DNA rps4 sequences. Maximum parsimony, maximum undertaken based on analysis of the internal transcribed and by combining data from nrDNA ITS and chloroplast likelihood, and Bayesian analyses all support the conclusion that the reinstated genus Ptychostomum is not monophyletic. Ptychostomum funkii (Schwagr.) J. R. Spence (≡ Bryum funkii Schwaigr.) is placed within a clade containing the type species of Bryum, B. argenteum Hedw. The remaining members of Ptychostomum investigated in the present study constitute another well-supported clade. The results are congruent with previous molecular analyses. On the basis of phylogenetic evidence, we agree with transferring B. amblyodon Mull. Hal. (≡ B. inclinatum (Brid.) Turton≡ Bryum archangelicum Bruch & Schimp.), Bryum lonchocaulon Mull. Hal., Bryum pallescens Schleich. ex Schwaigr., and Bryum pallens Sw. to Ptychostomum.展开更多
Sequence variation of the first internal transcribed spacer of ribosomal DNA ( ITS - 1 ) was examined and its application to the study of genetic variation was explored in four populations of farter' s scallop Chla...Sequence variation of the first internal transcribed spacer of ribosomal DNA ( ITS - 1 ) was examined and its application to the study of genetic variation was explored in four populations of farter' s scallop Chlamys farreri. ITS - 1 fragments, with a length of about 300 bp,of 78 individuals collected from Dalian, Qingdao, Yantai in China and Korea respectively were amplified via PCR, cloned and sequenced. Intra-genomic variation was examined by sequencing several clones of single individuals. Alignment and polymorphism analysis detected 44 haplotypes and 50 polymorphic sites which consist of 30 substitutions and 20 indels, indicating a high level of polymorphisms. Sequence analysis also showed a very low level of intra-individual variation. All these features validated the feasibility of application of ITS - 1 fragment to population analysis. Polymorphism analysis showed that the Korea sample has the richest genetic variation, followed by Yantai and Qingdao samples. AMOVA (analysis of molecular variance) showed that the majority (96.26%) of genetic variation was distributed within populations and 3.74% resulted from among populations, but with P 〈 0.05 ( = 0.042), indicating that the populations in this study have significant divergence. This output was basically concordant with the result arising from RAPD data and different from that from mitochondrial 16S rDNA sequence data. Discussion on this inconsistency was made accordingly.展开更多
Phylogenetic relationships of Arundinaria and related genera (Pleioblastus, Pseudosasa, Oligostachyum, Bashania, Clavinodum, etc.) were assessed by analyzing the sequences of the nrDNA internal transcribed spacer (ITS...Phylogenetic relationships of Arundinaria and related genera (Pleioblastus, Pseudosasa, Oligostachyum, Bashania, Clavinodum, etc.) were assessed by analyzing the sequences of the nrDNA internal transcribed spacer (ITS) and the cpDNA trnL-F intergenic spacer (IGS). Comparison with trnL-F IGS sequence, the ITS region provided the higher number of parsimony informative characters, and the interspecific variation of the ITS sequence was higher than that of the trnL-F IGS sequence.The tree obtained by combining both sets of data showed that the species sampled in Arundinaria and the related genera were monophyletic and divided into two clades. The relationships and positioning of all the taxa surveryed (including A. oleosa, A. hsienchuensis, A. chino, A. amara, A. yixingensis, A. amabilis, A. fortunei, A. pygmaea, A. gramineus, A. fargesii, A. faberi, A. hupehense, Pseudosasa japonica cv. Tsutsumiana, P. japonica and Brachystachyum densiflorum) were also discussed. The results from the sequences were broadly consistent with morphological characters, appearing all these taxa sampled belong to the genus of Arundinaria. The topologies of the trees generated from individual data and the combined data were similar.展开更多
Soil fungi are extremely important for maintaining soil health and plant production in agricultural systems.Currently,the effect of continuous cropping of sweet potato on soil fungal communities and physiochemical par...Soil fungi are extremely important for maintaining soil health and plant production in agricultural systems.Currently,the effect of continuous cropping of sweet potato on soil fungal communities and physiochemical parameters has not been well documented.In the present study,four sweet potato fields consecutively monocultured for 1,2,3,and 4 years were selected to investigate the effect of monoculture on soil fungal communities through Illumina MiSeq sequencing.Continuous cropping of sweet potatoes dramatically altered the fungal community composition,whereas fungal diversity was almost unchanged.Ascomycota and Basidiomycota were the most abundant phyla in all soil samples,accounting for 32.59%and 21.14%of the average relative abundance,respectively.The abundance of some potential pathogens,such as Ascobolus spp,specifically Ascobolus stercorarius,and some unknown fungi increased significantly as the sweet potato monoculture period increased,and their presence were highly positively correlated with disease incidence.In contrast,Basidiomycota,Bullera,Fusarium and Trichocladium most likely play roles as antagonists of sweet potato disease development,as their relative abundance decreased significantly over time and were negatively correlated with disease incidence.Redundancy and correlation analyses revealed that soil pH and organic carbon content were the most important factors driving these changes.Our findings provided a dynamic overview of the fungal community and presented a clear scope for screening beneficial fungi and pathogens of sweet potato.展开更多
Harmful algal blooms often occurred in the East China Sea (ECS) and the German coastal waters of the North Sea but Karlodinium veneficum had not been taxonomically reported. Two strains of Karlodinium (LAMB090611 a...Harmful algal blooms often occurred in the East China Sea (ECS) and the German coastal waters of the North Sea but Karlodinium veneficum had not been taxonomically reported. Two strains of Karlodinium (LAMB090611 and LAMB010601) were isolated from the two areas in 2009. The morphological characteristics and molecular phylogeny of two strains are compared on the basis of observation of a light microscope, a scanning electron microscope (SEM), a laser scanning micro-scope (LSM) and an internal transcribed spacer (ITS) sequence data. The mean cell length of strain LAMB090611 is (14.2 ± 1.8) μm (range 11.1–18.7 μm) and mean width is (10.8 ± 1.5) μm (range 8.2– 14.7 μm). The mean cell length of strain LAMB010601 is (15.1 ± 1.2) μm (range 12.7–17.9 μm) and the mean width is (11.4 ± 1.1) μm (range 9.1–14.7 μm), respectively. The two strains are similar in morphological characteristics, including a straight apical groove, distinct ventral pore, sulcal extension, cingulum displacement, two or four irregular shaped chloroplasts within the cell and almost equal sized epicone and hypocone. The large and round nucleus is located at the center or at the hypocone of the cell. The sequence length of the ECS strain LAMB090611 and the German strain LAMB010601 is 640 and 646 bp, respectively. The GC content is 49%. The nucleotide similarity of the two strains is 98.1%. The sequence divergence is 0.003. Both strains are confirmed as Karlo-dinium veneficum (D. Ballantine) J. Larsen and this is the first taxonomic report from China and Germany coastal waters. The population dynamics of this toxic species in the ECS and German coastal waters needs to be investigated in the near future.展开更多
Genera Lamiophlomis and Paraphlomis were originally separated from genus Phlomis s.1. on the basis of particular morphological characteristics. However, their relationship was highly contentious, as evidenced by the l...Genera Lamiophlomis and Paraphlomis were originally separated from genus Phlomis s.1. on the basis of particular morphological characteristics. However, their relationship was highly contentious, as evidenced by the literature. In the present paper, the systematic positions of Lamiophlomis, Paraphlomis, and their related genera were assessed based on nuclear internal transcribed spacer (ITS) and chloroplast rpl16 and trnL-F sequence data using maximum parsimony (MP) and Bayesian methods. and outgroup were sampled. Analyses of both separate In total, 24 species representing six genera of the ingroup and combined sequence data were conducted to resolve the systematic relationships of these genera. The results reveal that Lamiophlomis is nested within Phlomis sect. Phlomoides and its generic status is not supported. With the inclusion ofLamiophlomis rotata in sect. Phlomoides, sections Phlomis and Phlomoides of Phlomis were resolved as monophyletic. Paraphlomis was supported as an independent genus. However, the resolution of its monophyly conflicted between MP and Bayesian analyses, suggesting the need for expended sampling and further evidence.展开更多
The phylogenetic origin of Beckmannia remains unknown. The genus has been placed within the Chlorideae, Aveneae (Agrostideae), Poeae, or treated as an isolate lineage, Beckmanniinae. In the present study, we used nu...The phylogenetic origin of Beckmannia remains unknown. The genus has been placed within the Chlorideae, Aveneae (Agrostideae), Poeae, or treated as an isolate lineage, Beckmanniinae. In the present study, we used nuclear internal transcribed spacer (ITS) and chloroplast trnL-F sequences to examine the phylogenetic relationship between Beckmannia and those genera that have assumed to be related. On the basis of the results of our studies, the following conclusions could be drawn: (i) Beckmannia and Alopecurus are sister groups with high support; and (ii) Beckmannia and Alopecurus are nested in the Poeae clade with high support. The results of our analysis suggest that Beckmannia should be placed in Poeae.展开更多
Filamentous Bangia,which are distributed extensively throughout the world,have simple and similar morphological characteristics.Scientists can classify these organisms using molecular markers in combination with morph...Filamentous Bangia,which are distributed extensively throughout the world,have simple and similar morphological characteristics.Scientists can classify these organisms using molecular markers in combination with morphology.We successfully sequenced the complete nuclear ribosomal DNA.approximately 13 kb in length,from a marine Bangia population.We further analyzed the small subunit ribosomal DNA gene(nrSSU) and the internal transcribed spacer(ITS) sequence regions along with nine other marine,and two freshwater Bangia samples from China.Pairwise distances of the nrSSU and 5.8S ribosomal DNA gene sequences show the marine samples grouping together with low divergences(0-0.003;0-0.006,respectively) from each other,but high divergences(0.123-0.126;0.198,respectively) from freshwater samples.An exception is the marine sample collected from Weihai,which shows high divergence from both other marine samples(0.063-0.065;0.129,respectively) and the freshwater samples(0.097;0.120,respectively).A maximum likelihood phylogenetic tree based on a combined SSU-ITS dataset with maximum likelihood method shows the samples divided into three clades,with the two marine sample clades containing Bangia spp.from North America,Europe,Asia,and Australia;and one freshwater clade,containing Bangia atropurpurea from North America and China.展开更多
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 spe...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.展开更多
Background:Cryptococcal meningitis is a severe infectious disease associated with high morbidity and mortality.Rapidity and accuracy of diagnosis contribute to better prognosis,but readily available tools,such as micr...Background:Cryptococcal meningitis is a severe infectious disease associated with high morbidity and mortality.Rapidity and accuracy of diagnosis contribute to better prognosis,but readily available tools,such as microscopy,culture,and antigens do not perform well all the time.Our study attempted to diagnose and genotype cryptococcus in the cerebrospinal fluid(CSF)samples from patients with cryptococcal meningitis using the approach of metataxonomics of Internal Transcribed Spacer(ITS)amplicons.Methods:The CSF samples were collected from 11 clinically suspected cryptococcal meningitis patients and four non-infectious controls.Samples were recruited from the First Affiliated Hospital of Fujian Medical University Hospital,Fuzhou Fourth Hospital and the 476th Hospital of Chinese People's Liberation Army from December 2017 to December 2018.ITS1 ribosomal deoxyribonucleic acid(rDNA)genes of 15 whole samples were amplified by universal forward primer ITS1(CTTGGTCATTTAGAGGAAGTAA)and reverse primer ITS2(GCTGCGTTCTTCATCGATGC),sequenced by Illumina MiSeq Benchtop Sequencer.The results were confirmed by sanger sequencing of ITS1 region and partial CAP59 gene of microbial isolates from 11 meningitic samples.Pair-wise comparison between infectious group and control group was conducted through permutational multivariate analysis(PERMANOVA)in R software.Results:The 30,000 to 340,000 high-quality clean reads were obtained from each of the positively stained or cultured CSF samples and 8 to 60 reads from each control.The samples from 11 infected patients yielded detectable cryptococcal-specific ITS1 DNA with top abundance(from 95.90%to 99.97%),followed by many other fungal groups(each<1.41%).ITS genotype was defined in 11 CSF samples,corresponding to ITS type 1,and confirmed by Sanger sequencing.A statistically significant difference(r2=0.65869,P=0.0014)between infectious group and control group was observed.Conclusions:The metataxonomics of ITS amplicons facilitates the diagnosis and genotype of cryptococcus in CSF samples,which may provide a better diagnostic approach of cryptococcal infection.展开更多
A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning (...A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning (PSP). Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer (ITS) of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection lim- its of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction (PCR) method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum.展开更多
Dumasia taxonomy and classification have long been problematic.Species within this genus have few morphological differences and plants without flowers or fruits are difficult to accurately identify.In this study,we ev...Dumasia taxonomy and classification have long been problematic.Species within this genus have few morphological differences and plants without flowers or fruits are difficult to accurately identify.In this study,we evaluated the ability of six DNA barcoding sequences,one nuclear(ITS)and five chloroplast regions(trnH-psbA,matK,rbcL,trnL-trnF,psbB-psbF),to efficiently identify Dumasia species.Most single markers or their combinations identify obvious barcoding gaps between intraspecific and interspecific genetic variation.Most combined analyses including ITS showed good species resolution and identification efficiency.We therefore suggest that ITS alone or a combination of ITS with any cpDNA marker are most suitable for DNA barcoding of Dumasia.The phylogenetic analyses clearly indicated that Dumasia yunnanensis is not monophyletic and is separated as two independent branches,which may result from cryptic differentiation.Our results demonstrate that molecular data can deepen the comprehension of taxonomy of Dumasia and provide an efficient approach for identification of the species.展开更多
Although intestinal fungi play important roles in host health and disease,the composition and diversity of fungal communities remain poorly reported in fish.In this study,fungi in the fore-,mid-,and hindintestine of t...Although intestinal fungi play important roles in host health and disease,the composition and diversity of fungal communities remain poorly reported in fish.In this study,fungi in the fore-,mid-,and hindintestine of tilapia(Oreochromis mossambicus)and bighead carp(Aristichthys nobilis)from Hongchaojiang Reservoir in Guangxi,China were investigated by ITS sequencing.Based on this,we obtained 1763478 high-quality tags,which clustered into 1089 operational taxonomic units(OTUs).In total,404 OTUs were annotated,of which 310,68,and 26 belonged to Ascomycota,Basidiomycota,and other,respectively.Results show significant differences in the community composition of intestinal fungi between tilapia and bighead carp but not within their different intestinal segments.Furthermore,154 of the 404 annotated OTUs were considered reliable and were classified into three trophic modes and nine guilds.The three trophic modes consisted of 108 OTUs of saprotrophic fungi,41 OTUs of pathotrophic fungi,and five OTUs of symbiotrophic fungi.The top three most abundant OTUs overall(i.e.,Otu000002,Scopulariopsis acremonium;Otu000018,Alternaria palandui;Otu000034,Aureobasidium pullulans)showed lower abundance in the hind-intestinal segments of bighead carp,suggesting uneven distribution of these fungi in this species.In addition,saprotrophic and pathotrophic fungi were markedly decreased in the hindintestine.It is indicated that the fungal community was not only related to host species specificity but also to the respective physiological functions of different intestinal segments.These findings provide valuable information on the composition,structure,and potential function of the intestinal fungi community associated with different intestinal segments in tilapia and bighead carp under natural conditions,thus highlighting the importance of fungi as an integral part of the inte stinal microbiota in maintaining host health.展开更多
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 ...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.展开更多
During an exploration in the virgin coniferous forest of Jammu and Kashmir,a little known taxon,Clitocella popinalis(Basidiomycota,Entolomataceae)was gathered along with other macrofungi.Based on morphological and mol...During an exploration in the virgin coniferous forest of Jammu and Kashmir,a little known taxon,Clitocella popinalis(Basidiomycota,Entolomataceae)was gathered along with other macrofungi.Based on morphological and molecular(nrITS sequence)data,detailed taxonomic information related to this species is described and illustrated for the first time from India.Morphologically and genetically allied taxa are also compared with this species.展开更多
Molecular markers were used to identify and assess cultivars of Laminaria Lamx. and to delineate their phylogenetic relationships. Random amplified polymorphic DNA (RAPD) analysis was used for detection. After screeni...Molecular markers were used to identify and assess cultivars of Laminaria Lamx. and to delineate their phylogenetic relationships. Random amplified polymorphic DNA (RAPD) analysis was used for detection. After screening, 11 primers were selected and they yielded 133 bands in all, of which approximately 99.2% were polymorphic. The genetic distances between gametophytes ranged from 0.412 to 0.956. Two clusters were formed with the unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on the simple matching coefficient. All cultivars of Laminaria japonica Aresch. used for breeding in China fell into one cluster. L. japonica from Japan, L. saccharina (L.) Lam., and L. angustata Kjellm. formed the other cluster and showed higher genetic variation than L. japonica from China. Nuclear ribosomal DNA (rDNA) sequences, including internal transcribed spacers (ITS 1 and ITS2) were studied and aligned. The nucleotides of the sequences ranged from 634 to 668, with a total of 692 positions including ITS1, ITS2, and the 5.8S coding region. The phylogenetic tree obtained by the neighbor-joining method favored, to some extent, the results revealed by RAPD analysis. The present study indicates that RAPD and ITS analyses could be used to identify and assess Laminaria germplasm and to distinguish some species and, even intraspecies, in Laminaria.展开更多
Using the basic GenBank local alignment search tool program(BLAST)to identify fungi collected in a recently protected beech forest at Montricher(Switzerland),the number of ITS sequences associated to the wrong taxon n...Using the basic GenBank local alignment search tool program(BLAST)to identify fungi collected in a recently protected beech forest at Montricher(Switzerland),the number of ITS sequences associated to the wrong taxon name appears to be around 30%,even higher than previously estimated.Such results rely on the in-depth re-examination of BLAST results for the most interesting species that were collected,viz.first records for Switzerland,rare or patrimonial species and problematic species(when BLAST top scores were equally high for different species),all belonging to Agaricomycotina.This paper dissects for the first time a number of sequence-based identifications,thereby showing in every detail—particularly to the user community of taxonomic information—why sequence-based identification in the context of a fungal inventory can easily go wrong.Our first conclusion is that in-depth examination of BLAST results is too time consuming to be considered as a routine approach for future inventories:we spent two months on verification of approx.20 identifications.Apart from the fact that poor taxon coverage in public depositories remains the principal impediment for successful species identification,it can be deplored that even very recent fungal sequence deposits in GenBank involve an uncomfortably high number of misidentifications or errors with associated metadata.While checking the original publications associated with top score sequences for the few examples that were here re-examined,a positive consequence is that we uncovered over 80 type sequences that were not annotated as types in GenBank.Advantages and pitfalls of sequence-based identification are discussed,particularly in the light of undertaking fungal inventories.Recommendations are made to avoid or reduce some of the major problems with sequence-based identification.Nevertheless,the prospects for a more reliable sequence-based identification of fungi remain quite dim,unless authors are ready to check and update the metadata associated with previously deposited sequences in their publications.展开更多
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,th...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.展开更多
基金supported by the National Natural Science Foundation of China(grantno.30670152)the National Infrastructure of Natural Resources for Science and Technology(grant no.2005DKA21403)the Natural Science Foundation of Hebei Province,China(no.C2008000158)
文摘The phylogeny of Ptychostomum was first spacer (ITS) region of the nuclear ribosomal (nr) DNA DNA rps4 sequences. Maximum parsimony, maximum undertaken based on analysis of the internal transcribed and by combining data from nrDNA ITS and chloroplast likelihood, and Bayesian analyses all support the conclusion that the reinstated genus Ptychostomum is not monophyletic. Ptychostomum funkii (Schwagr.) J. R. Spence (≡ Bryum funkii Schwaigr.) is placed within a clade containing the type species of Bryum, B. argenteum Hedw. The remaining members of Ptychostomum investigated in the present study constitute another well-supported clade. The results are congruent with previous molecular analyses. On the basis of phylogenetic evidence, we agree with transferring B. amblyodon Mull. Hal. (≡ B. inclinatum (Brid.) Turton≡ Bryum archangelicum Bruch & Schimp.), Bryum lonchocaulon Mull. Hal., Bryum pallescens Schleich. ex Schwaigr., and Bryum pallens Sw. to Ptychostomum.
基金This work was financially supported by the"863"Project of China under contract No.2002AA626020the National Nalural Science Foundation of China under contract No.30570242.
文摘Sequence variation of the first internal transcribed spacer of ribosomal DNA ( ITS - 1 ) was examined and its application to the study of genetic variation was explored in four populations of farter' s scallop Chlamys farreri. ITS - 1 fragments, with a length of about 300 bp,of 78 individuals collected from Dalian, Qingdao, Yantai in China and Korea respectively were amplified via PCR, cloned and sequenced. Intra-genomic variation was examined by sequencing several clones of single individuals. Alignment and polymorphism analysis detected 44 haplotypes and 50 polymorphic sites which consist of 30 substitutions and 20 indels, indicating a high level of polymorphisms. Sequence analysis also showed a very low level of intra-individual variation. All these features validated the feasibility of application of ITS - 1 fragment to population analysis. Polymorphism analysis showed that the Korea sample has the richest genetic variation, followed by Yantai and Qingdao samples. AMOVA (analysis of molecular variance) showed that the majority (96.26%) of genetic variation was distributed within populations and 3.74% resulted from among populations, but with P 〈 0.05 ( = 0.042), indicating that the populations in this study have significant divergence. This output was basically concordant with the result arising from RAPD data and different from that from mitochondrial 16S rDNA sequence data. Discussion on this inconsistency was made accordingly.
文摘Phylogenetic relationships of Arundinaria and related genera (Pleioblastus, Pseudosasa, Oligostachyum, Bashania, Clavinodum, etc.) were assessed by analyzing the sequences of the nrDNA internal transcribed spacer (ITS) and the cpDNA trnL-F intergenic spacer (IGS). Comparison with trnL-F IGS sequence, the ITS region provided the higher number of parsimony informative characters, and the interspecific variation of the ITS sequence was higher than that of the trnL-F IGS sequence.The tree obtained by combining both sets of data showed that the species sampled in Arundinaria and the related genera were monophyletic and divided into two clades. The relationships and positioning of all the taxa surveryed (including A. oleosa, A. hsienchuensis, A. chino, A. amara, A. yixingensis, A. amabilis, A. fortunei, A. pygmaea, A. gramineus, A. fargesii, A. faberi, A. hupehense, Pseudosasa japonica cv. Tsutsumiana, P. japonica and Brachystachyum densiflorum) were also discussed. The results from the sequences were broadly consistent with morphological characters, appearing all these taxa sampled belong to the genus of Arundinaria. The topologies of the trees generated from individual data and the combined data were similar.
基金supported by Key laboratory of Degraded and Unused Land Consolidation Engineering,the Ministry of Land and Resources(SXDJ2018-06)National Natural Science Foundation of China(Nos.41501271 and 41601339)+1 种基金China Agriculture Research System(No.CARS-10-B10)Support Plan on Youth Innovation Science and Technology for Higher Education of Shandong Province(2019KJD014).
文摘Soil fungi are extremely important for maintaining soil health and plant production in agricultural systems.Currently,the effect of continuous cropping of sweet potato on soil fungal communities and physiochemical parameters has not been well documented.In the present study,four sweet potato fields consecutively monocultured for 1,2,3,and 4 years were selected to investigate the effect of monoculture on soil fungal communities through Illumina MiSeq sequencing.Continuous cropping of sweet potatoes dramatically altered the fungal community composition,whereas fungal diversity was almost unchanged.Ascomycota and Basidiomycota were the most abundant phyla in all soil samples,accounting for 32.59%and 21.14%of the average relative abundance,respectively.The abundance of some potential pathogens,such as Ascobolus spp,specifically Ascobolus stercorarius,and some unknown fungi increased significantly as the sweet potato monoculture period increased,and their presence were highly positively correlated with disease incidence.In contrast,Basidiomycota,Bullera,Fusarium and Trichocladium most likely play roles as antagonists of sweet potato disease development,as their relative abundance decreased significantly over time and were negatively correlated with disease incidence.Redundancy and correlation analyses revealed that soil pH and organic carbon content were the most important factors driving these changes.Our findings provided a dynamic overview of the fungal community and presented a clear scope for screening beneficial fungi and pathogens of sweet potato.
基金The National High-Tech Project of China under contract No.2007AA09Z110Chinese Ecology and Oceanography of Harmful Algal Blooms Project Ⅱ under contract No.2010CB428702+3 种基金the National Science Foundation Project under contract No.41176141Foundation of State Oceanic Administration of China under contract Nos 2009123 and 20100310the Basic Research Fund of the Second Institute of Oceanography,the State Oceanic Administration of China under contract No.SZ0824Joint Project between Germany and China supported by International Bureau of Federal Ministry for Education and Research under contract No.CHN06/009
文摘Harmful algal blooms often occurred in the East China Sea (ECS) and the German coastal waters of the North Sea but Karlodinium veneficum had not been taxonomically reported. Two strains of Karlodinium (LAMB090611 and LAMB010601) were isolated from the two areas in 2009. The morphological characteristics and molecular phylogeny of two strains are compared on the basis of observation of a light microscope, a scanning electron microscope (SEM), a laser scanning micro-scope (LSM) and an internal transcribed spacer (ITS) sequence data. The mean cell length of strain LAMB090611 is (14.2 ± 1.8) μm (range 11.1–18.7 μm) and mean width is (10.8 ± 1.5) μm (range 8.2– 14.7 μm). The mean cell length of strain LAMB010601 is (15.1 ± 1.2) μm (range 12.7–17.9 μm) and the mean width is (11.4 ± 1.1) μm (range 9.1–14.7 μm), respectively. The two strains are similar in morphological characteristics, including a straight apical groove, distinct ventral pore, sulcal extension, cingulum displacement, two or four irregular shaped chloroplasts within the cell and almost equal sized epicone and hypocone. The large and round nucleus is located at the center or at the hypocone of the cell. The sequence length of the ECS strain LAMB090611 and the German strain LAMB010601 is 640 and 646 bp, respectively. The GC content is 49%. The nucleotide similarity of the two strains is 98.1%. The sequence divergence is 0.003. Both strains are confirmed as Karlo-dinium veneficum (D. Ballantine) J. Larsen and this is the first taxonomic report from China and Germany coastal waters. The population dynamics of this toxic species in the ECS and German coastal waters needs to be investigated in the near future.
基金supported by the Natural Science Foundation of Yunnan Province,as well as by a grant from the Technology Foundation and the National Key Project for Basic Research(973no.2007CB411600).
文摘Genera Lamiophlomis and Paraphlomis were originally separated from genus Phlomis s.1. on the basis of particular morphological characteristics. However, their relationship was highly contentious, as evidenced by the literature. In the present paper, the systematic positions of Lamiophlomis, Paraphlomis, and their related genera were assessed based on nuclear internal transcribed spacer (ITS) and chloroplast rpl16 and trnL-F sequence data using maximum parsimony (MP) and Bayesian methods. and outgroup were sampled. Analyses of both separate In total, 24 species representing six genera of the ingroup and combined sequence data were conducted to resolve the systematic relationships of these genera. The results reveal that Lamiophlomis is nested within Phlomis sect. Phlomoides and its generic status is not supported. With the inclusion ofLamiophlomis rotata in sect. Phlomoides, sections Phlomis and Phlomoides of Phlomis were resolved as monophyletic. Paraphlomis was supported as an independent genus. However, the resolution of its monophyly conflicted between MP and Bayesian analyses, suggesting the need for expended sampling and further evidence.
基金supported b ythe National Natural Science Foundation of China(30270107)the Project Sponsored by the Scientific Research Foundation for the Doctors,Weifang Medical University
文摘The phylogenetic origin of Beckmannia remains unknown. The genus has been placed within the Chlorideae, Aveneae (Agrostideae), Poeae, or treated as an isolate lineage, Beckmanniinae. In the present study, we used nuclear internal transcribed spacer (ITS) and chloroplast trnL-F sequences to examine the phylogenetic relationship between Beckmannia and those genera that have assumed to be related. On the basis of the results of our studies, the following conclusions could be drawn: (i) Beckmannia and Alopecurus are sister groups with high support; and (ii) Beckmannia and Alopecurus are nested in the Poeae clade with high support. The results of our analysis suggest that Beckmannia should be placed in Poeae.
基金Supported by the National Natural Science Foundation of China(Nos.31270256,41276134)the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A406-6)
文摘Filamentous Bangia,which are distributed extensively throughout the world,have simple and similar morphological characteristics.Scientists can classify these organisms using molecular markers in combination with morphology.We successfully sequenced the complete nuclear ribosomal DNA.approximately 13 kb in length,from a marine Bangia population.We further analyzed the small subunit ribosomal DNA gene(nrSSU) and the internal transcribed spacer(ITS) sequence regions along with nine other marine,and two freshwater Bangia samples from China.Pairwise distances of the nrSSU and 5.8S ribosomal DNA gene sequences show the marine samples grouping together with low divergences(0-0.003;0-0.006,respectively) from each other,but high divergences(0.123-0.126;0.198,respectively) from freshwater samples.An exception is the marine sample collected from Weihai,which shows high divergence from both other marine samples(0.063-0.065;0.129,respectively) and the freshwater samples(0.097;0.120,respectively).A maximum likelihood phylogenetic tree based on a combined SSU-ITS dataset with maximum likelihood method shows the samples divided into three clades,with the two marine sample clades containing Bangia spp.from North America,Europe,Asia,and Australia;and one freshwater clade,containing Bangia atropurpurea from North America and China.
基金supported by the Fundamental Research Funds for the Central Universities(33000-31611215)the Guangzhou Science and Technology Program(201903010076)the National Natural Science Foundation of China(31760048).
文摘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.
文摘Background:Cryptococcal meningitis is a severe infectious disease associated with high morbidity and mortality.Rapidity and accuracy of diagnosis contribute to better prognosis,but readily available tools,such as microscopy,culture,and antigens do not perform well all the time.Our study attempted to diagnose and genotype cryptococcus in the cerebrospinal fluid(CSF)samples from patients with cryptococcal meningitis using the approach of metataxonomics of Internal Transcribed Spacer(ITS)amplicons.Methods:The CSF samples were collected from 11 clinically suspected cryptococcal meningitis patients and four non-infectious controls.Samples were recruited from the First Affiliated Hospital of Fujian Medical University Hospital,Fuzhou Fourth Hospital and the 476th Hospital of Chinese People's Liberation Army from December 2017 to December 2018.ITS1 ribosomal deoxyribonucleic acid(rDNA)genes of 15 whole samples were amplified by universal forward primer ITS1(CTTGGTCATTTAGAGGAAGTAA)and reverse primer ITS2(GCTGCGTTCTTCATCGATGC),sequenced by Illumina MiSeq Benchtop Sequencer.The results were confirmed by sanger sequencing of ITS1 region and partial CAP59 gene of microbial isolates from 11 meningitic samples.Pair-wise comparison between infectious group and control group was conducted through permutational multivariate analysis(PERMANOVA)in R software.Results:The 30,000 to 340,000 high-quality clean reads were obtained from each of the positively stained or cultured CSF samples and 8 to 60 reads from each control.The samples from 11 infected patients yielded detectable cryptococcal-specific ITS1 DNA with top abundance(from 95.90%to 99.97%),followed by many other fungal groups(each<1.41%).ITS genotype was defined in 11 CSF samples,corresponding to ITS type 1,and confirmed by Sanger sequencing.A statistically significant difference(r2=0.65869,P=0.0014)between infectious group and control group was observed.Conclusions:The metataxonomics of ITS amplicons facilitates the diagnosis and genotype of cryptococcus in CSF samples,which may provide a better diagnostic approach of cryptococcal infection.
基金The Science and Technology Commission of Shanghai Municipality under contract Nos 06235810108DZ1980802 and 10JC1418600a special research fund for the national non-profit institutes (East China Sea Fisheries Research Institute) under contract Nos 2007M22 and 2007Z01
文摘A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning (PSP). Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer (ITS) of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection lim- its of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction (PCR) method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum.
基金We thank Dr.Zhi-qiang Lu and Mr.Yi Fu for help during the field survey.We are grateful to Dr.Ovidiu Paun for very helpful comments on earlier drafts of this manuscript.We thank Dr.Shu-feng Li for the distributional map,as well as Dr.Bing Liu,Dr.Ren-bin Zhu,and Mr.Yi Fu for their photos of some Dumasia species.The first author thanks Dr.Wen-bin Yu,Dr.Pei-liang Liu,Dr.Xue-li Zhao,and Dr.Zhu-qiu Song for their help during the writing process.Additional thanks go to Dr.Richard T.Corlett,Raymond Porter and Mr Yuan-qiong Zhang for polishing this work.The authors would also like to express gratitude to two anonymous reviewers for their valuable comments on the manuscript.This work was financially supported by the Second Tibetan Plateau Scientific Expedition and Research(STEP)program(2019QZKK0502)National Natural Science Foundation of China(NSFC 41861008)the 135 Karst‘breakthrough’project Grant 2017XTBG-T03.
文摘Dumasia taxonomy and classification have long been problematic.Species within this genus have few morphological differences and plants without flowers or fruits are difficult to accurately identify.In this study,we evaluated the ability of six DNA barcoding sequences,one nuclear(ITS)and five chloroplast regions(trnH-psbA,matK,rbcL,trnL-trnF,psbB-psbF),to efficiently identify Dumasia species.Most single markers or their combinations identify obvious barcoding gaps between intraspecific and interspecific genetic variation.Most combined analyses including ITS showed good species resolution and identification efficiency.We therefore suggest that ITS alone or a combination of ITS with any cpDNA marker are most suitable for DNA barcoding of Dumasia.The phylogenetic analyses clearly indicated that Dumasia yunnanensis is not monophyletic and is separated as two independent branches,which may result from cryptic differentiation.Our results demonstrate that molecular data can deepen the comprehension of taxonomy of Dumasia and provide an efficient approach for identification of the species.
基金the Guangxi Science and Technology Major Project(No.AA17204095-8)the Open Fund of Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Aquaculture(No.GXKEYLA2019-05)the China-ASEAN Maritime Cooperation Fund(No.CAMC-2018F)。
文摘Although intestinal fungi play important roles in host health and disease,the composition and diversity of fungal communities remain poorly reported in fish.In this study,fungi in the fore-,mid-,and hindintestine of tilapia(Oreochromis mossambicus)and bighead carp(Aristichthys nobilis)from Hongchaojiang Reservoir in Guangxi,China were investigated by ITS sequencing.Based on this,we obtained 1763478 high-quality tags,which clustered into 1089 operational taxonomic units(OTUs).In total,404 OTUs were annotated,of which 310,68,and 26 belonged to Ascomycota,Basidiomycota,and other,respectively.Results show significant differences in the community composition of intestinal fungi between tilapia and bighead carp but not within their different intestinal segments.Furthermore,154 of the 404 annotated OTUs were considered reliable and were classified into three trophic modes and nine guilds.The three trophic modes consisted of 108 OTUs of saprotrophic fungi,41 OTUs of pathotrophic fungi,and five OTUs of symbiotrophic fungi.The top three most abundant OTUs overall(i.e.,Otu000002,Scopulariopsis acremonium;Otu000018,Alternaria palandui;Otu000034,Aureobasidium pullulans)showed lower abundance in the hind-intestinal segments of bighead carp,suggesting uneven distribution of these fungi in this species.In addition,saprotrophic and pathotrophic fungi were markedly decreased in the hindintestine.It is indicated that the fungal community was not only related to host species specificity but also to the respective physiological functions of different intestinal segments.These findings provide valuable information on the composition,structure,and potential function of the intestinal fungi community associated with different intestinal segments in tilapia and bighead carp under natural conditions,thus highlighting the importance of fungi as an integral part of the inte stinal microbiota in maintaining host health.
基金financed by ACECR,Iran(code no 2283)granted to Dr.
文摘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.
文摘During an exploration in the virgin coniferous forest of Jammu and Kashmir,a little known taxon,Clitocella popinalis(Basidiomycota,Entolomataceae)was gathered along with other macrofungi.Based on morphological and molecular(nrITS sequence)data,detailed taxonomic information related to this species is described and illustrated for the first time from India.Morphologically and genetically allied taxa are also compared with this species.
文摘Molecular markers were used to identify and assess cultivars of Laminaria Lamx. and to delineate their phylogenetic relationships. Random amplified polymorphic DNA (RAPD) analysis was used for detection. After screening, 11 primers were selected and they yielded 133 bands in all, of which approximately 99.2% were polymorphic. The genetic distances between gametophytes ranged from 0.412 to 0.956. Two clusters were formed with the unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on the simple matching coefficient. All cultivars of Laminaria japonica Aresch. used for breeding in China fell into one cluster. L. japonica from Japan, L. saccharina (L.) Lam., and L. angustata Kjellm. formed the other cluster and showed higher genetic variation than L. japonica from China. Nuclear ribosomal DNA (rDNA) sequences, including internal transcribed spacers (ITS 1 and ITS2) were studied and aligned. The nucleotides of the sequences ranged from 634 to 668, with a total of 692 positions including ITS1, ITS2, and the 5.8S coding region. The phylogenetic tree obtained by the neighbor-joining method favored, to some extent, the results revealed by RAPD analysis. The present study indicates that RAPD and ITS analyses could be used to identify and assess Laminaria germplasm and to distinguish some species and, even intraspecies, in Laminaria.
文摘Using the basic GenBank local alignment search tool program(BLAST)to identify fungi collected in a recently protected beech forest at Montricher(Switzerland),the number of ITS sequences associated to the wrong taxon name appears to be around 30%,even higher than previously estimated.Such results rely on the in-depth re-examination of BLAST results for the most interesting species that were collected,viz.first records for Switzerland,rare or patrimonial species and problematic species(when BLAST top scores were equally high for different species),all belonging to Agaricomycotina.This paper dissects for the first time a number of sequence-based identifications,thereby showing in every detail—particularly to the user community of taxonomic information—why sequence-based identification in the context of a fungal inventory can easily go wrong.Our first conclusion is that in-depth examination of BLAST results is too time consuming to be considered as a routine approach for future inventories:we spent two months on verification of approx.20 identifications.Apart from the fact that poor taxon coverage in public depositories remains the principal impediment for successful species identification,it can be deplored that even very recent fungal sequence deposits in GenBank involve an uncomfortably high number of misidentifications or errors with associated metadata.While checking the original publications associated with top score sequences for the few examples that were here re-examined,a positive consequence is that we uncovered over 80 type sequences that were not annotated as types in GenBank.Advantages and pitfalls of sequence-based identification are discussed,particularly in the light of undertaking fungal inventories.Recommendations are made to avoid or reduce some of the major problems with sequence-based identification.Nevertheless,the prospects for a more reliable sequence-based identification of fungi remain quite dim,unless authors are ready to check and update the metadata associated with previously deposited sequences in their publications.
基金supported by the International Cooperation Program of Science and Technology (No. 2007DFA30990)the Special Founding for Healthy Field (No. 200802043) awarded by the Chinese Ministry of Science and Technologysupported by grants from the Hong Kong Research Grant Council (HKU 7526/06M) to C.L.
文摘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.