The first internal transcribed spacer(ITS1) of nuclear ribosomal DNA of three wild rice species and two subspecies of cultivated rice, which are distributed in China, was amplified using PCR technique and sequenced wi...The first internal transcribed spacer(ITS1) of nuclear ribosomal DNA of three wild rice species and two subspecies of cultivated rice, which are distributed in China, was amplified using PCR technique and sequenced with automated fluorescent sequencing. The sequences of ITS1 ranged from 193 bp to 218 bp in size and G/C content varied from 69.3% to 72.7%. In pairwise comparisons among the five taxa, sequence site divergence ranged from 1.5% to 10.6%. Phylogenetic analysis of ITS1 sequences using Wagner parsimony generated a single well resolved tree, which revealed that Oryza rufipogon was much more closely related to cultivated rice species than to the other two wild species. Oryza granulata was less closely related to either cultivated rice species or the other two wild species, and might be a unique and isolated taxon in the genus Oryza. The phylogenetic relationships of the three wild rice species and two cultivated rice subspecies inferred from ITS1 sequences is highly concordant with those based on the molecular evidence from isozyme, chloroplast DNA (cpDNA), mitochondrial DNA(mtDNA) and nuclear DNA (nDNA) of the genus Oryza .展开更多
The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA (including 5.8S rRNA) of 15 Rhododendron, species, representing most sections of the genus, one Ledum species and Cassiope fastigiata were sequenc...The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA (including 5.8S rRNA) of 15 Rhododendron, species, representing most sections of the genus, one Ledum species and Cassiope fastigiata were sequenced. Together with the ITS sequences of 13 selected Rhododendron species and Bejaria racemosa downloaded from GenBank, we explored the infrageneric and sectional relationships of this important North Temperate genus by employing maximum-parsimony analysis using PAUP software. C. fastigiata and B. racemosa were designated as outgroups. The ITS-based tree inferred that: (1) Rhododendron was a well-supported monophyletic group, while subg. Therorhodion was basal to the rest of the genus; (2) Ledum was a member of Rhododendron, and its close relationship with the lepidote rhododendron was confirmed; (3) the lepidote rhododendron plus Ledum formed a strongly-supported monophyletic clade which was sister to the rest of the elepidote rhododendron; (4) the elepidote rhododendron formed a weakly-supported clade within which the monophyly of subg. Hynwrianthes and subg. Tsutsusi were strongly supported, while subg. Pentanthera and subg. Azaleastrum were polyphyletic; and (5) the monophyly of sect. Choniastnini, (subg. Azaleastrum) was strongly-supported, while subg. Tsutsusi could be sister to a weakly-supported clade composed of two sampled species of sect. Azaleastrum (subg. Azaleastrum) together with R. sentibarbatum, of subg. Mumeazalea.展开更多
Pinus squamata X. W. Li, a recently-described species of the Pinaceae, is an extremely endangered pine with only 32 individuals in the wild. This species was thought to be intermediate between P. bungeana Zuec. ex End...Pinus squamata X. W. Li, a recently-described species of the Pinaceae, is an extremely endangered pine with only 32 individuals in the wild. This species was thought to be intermediate between P. bungeana Zuec. ex Endl. of subgen. Strobus and P. yunnanensis Franch. of subgen. Pinus by the original author. We made an effort to address the question of the phylogenetic relationship of this peculiar and important species in the context of the major groups of the genus Pinus by using sequences of the rbcL, matK,genes, rpl20-rps18 spacer, trnV intron of the chloroplast genome and the nuclear ribosomal ITS region. The results of the separated analysis and the combined analysis of the four cpDNA sequences and ITS sequence indicated that P. squamata was a stable member of subsect. Gerardianae and P. gerardiana Wall. was the closest species of P. squamata phylogenetically. As a result, the causes of the distribution pattern of subsect. Gerardianae were also discussed.展开更多
Presented is design concept for key parameters o f the reverse conducting gate commutated thyristor (RC-GCT),such as the thickness and concentration of n-base region and the transparent anode region,and the wi dth o...Presented is design concept for key parameters o f the reverse conducting gate commutated thyristor (RC-GCT),such as the thickness and concentration of n-base region and the transparent anode region,and the wi dth of separation region between asymmetric GCT and PIN diode.A structure model of the RC-GCT is set up based on the design concept and its characteristics are analyzed.The simulation results show the design concept is reasonable.展开更多
The microstructure, phase composition and cold shut defect of thick titanium alloy electron beam welded joint were studied. The results showed that the microstructure of weld zone was composed of α′ phase; the heat ...The microstructure, phase composition and cold shut defect of thick titanium alloy electron beam welded joint were studied. The results showed that the microstructure of weld zone was composed of α′ phase; the heat affected zone was divided into fine-grained zone and coarse-grained zone, the microstructure of fine-grained zone was primary α phase + β phase + equiaxed α phase, and the microstructure of coarse-grained zone was primary α phase + acicular α′ phase; the microstructure of base metal zone basically consisted of primary α phase, and a small amount of residual β phase sprinkled. The forming. reason of cold shut was analyzed, and the precaution of cold shut was proposed.展开更多
[Objective] The aim was to select suitable gene for Chlorella identification and to identify the oil-producing microalgae.[Method] Four candidate gene sequences,the nuclear genomic rDNA of the 18S rRNA gene,internal t...[Objective] The aim was to select suitable gene for Chlorella identification and to identify the oil-producing microalgae.[Method] Four candidate gene sequences,the nuclear genomic rDNA of the 18S rRNA gene,internal transcribed spacer(ITS),internal transcribed spacer Ⅱ(ITS Ⅱ)and the chloroplast rbcL gene,were selected for Chlorella molecular identification.Through these four candidate genes,the genetic variability and distinguish ability between intra-species and inter-species was analyzed to choose the right genes for identification of the high oil-content Chlorella.On this basis,application of these gene segments were classified and identified for five fresh-water isolated Chlorella,which oil-content is more than 30%.[Result] ITS gene was a suitable gene because of its high variation and short fragment length,meanwhile its genetic distance intra-species(0.439 6±0.135 9)was larger than inter-species(0.045 7±0.084 3).Its sequence length varied between different species whereas highly conserved in the same species.By the application of ITS sequences,respectively,five high oil-content stains were identified as one C.vulgaris,two strains of C.sorokiniana and two strains of algae Chlorella sp.[Conclusion] This study had provided reference for the establishment of identification gene pool of Chlorella.展开更多
Aim To investigate the plant origin and the identity of the red substance on the surface of Hong Dangshen, a unique medicinal material for diarrhea in Hong Kong. Methods The HPLC fingerprints and 5S rRNA gene spacer s...Aim To investigate the plant origin and the identity of the red substance on the surface of Hong Dangshen, a unique medicinal material for diarrhea in Hong Kong. Methods The HPLC fingerprints and 5S rRNA gene spacer sequences of Hong Dangshen were obtained and compared with those of genuine species of Radix Codonopsis. The X-ray diffraction spectrum of the red substance was analyzed and compared with that of Halloysitum fingerprints to the Codonopsis species and the highest similarity to Rubrum. Results Hong Dangshen showed very similar HPLC Codonopsis pilosula var. modesta in terms of 5S rRNA gene spacer sequences. The X-ray diffraction spectrum of the red substance was consistent with that of Halloysitum Rubrum. Conclusion The source plant of Hong Dangshen was suggested to be Codonopsis pilosula var. modesta, one of the genuine original plants of Radix Codonopsis (Dangshen) in the China Pharmacopoeia (2005 edition). The red substance on the surface of Hong Dangshen was indicated to be Halloysitum Rubrum, a traditional medicinal mineral for chronic diarrhea. Our data suggest that Hong Dangshen is derived from the roots of Codonopsis pilosula var. modesta which has been processed with Halloysitum Rubrum, and the name is suggested to be Radix Codonopsis Praeparata Halloysita Rubra.展开更多
Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientis...Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientists. In this paper, we sequenced the 18S rDNA genes, the internal transcribed spacer (ITS) regions and the rbcL genes in seven organisms and obtained 536-566 bp long ITS sequences, 1 377-I 407 bp long rbcL sequences and 1 718-1 761 bp long partial 18S rDNA sequences. The GC base pair content was highest in the ITS regions and lowest in the rbcL genes. The sequencing results showed that the three Ulvaprolifera (or U. pertusa) gene sequences from Qingdao and Nan'ao Island were identical. The ITS, 18S rDNA and rbcL genes in U. prolifera and U. pertusa from different sea areas in China were unchanged by geographic distance. U.flexuosa had the least evolutionary distance from U. californica in both the ITS regions (0.009) and the 18S rDNA (0.002). These data verified that Ulva and Enteromorpha are not separate genera.展开更多
An improved method of PCR in which the small segment of conchocelis is amplified directly without DNA extraction was used to amplify a RUBISCO intergenic spacer DNA fragment from nine species of red algal genus Porphy...An improved method of PCR in which the small segment of conchocelis is amplified directly without DNA extraction was used to amplify a RUBISCO intergenic spacer DNA fragment from nine species of red algal genus Porphyra(Bangiales,Rhodophyta),including Porphyra yezoensis(Jiangsu,China),P.haitanensis(Fujian,China),P.oligospermatangia(Qingdao,China),P.katadai(Qingdao,China),P.tenera(Qingdao,China),P.suborboculata(Fujian,China),P.pseudolinearis(Kogendo,Korea),P.linearis(Devon,England),and P.fallax(Seattle,USA).Standard PCR and the method developed here were both conducted using primers specific for the RUBISCO spacer region,after which the two PCR products were sequenced.The sequencing data of the amplicons obtained using both methods were identical,suggesting that the improved PCR method was functional.These findings indicate that the method developed here may be useful for the rapid identification of species of Porphyra in a germplasm bank.In addition,a phylogenetic tree was constructed using the RUBISCO spacer and partial rbcS sequence,and the results were in concordant with possible alternative phylogenies based on traditional morphological taxonomic characteristics,indicating that the RUBISCO spacer is a useful region for phylogenetic studies.展开更多
Abstract The internal transcribed spacer regions (ITS1 and ITS2) and 5.8S rRNA genes were sequenced in six populations of four Aspidisca species, namely A. leptaspis, A. orthopogon, A. rnagna and A. aculeata. Phylog...Abstract The internal transcribed spacer regions (ITS1 and ITS2) and 5.8S rRNA genes were sequenced in six populations of four Aspidisca species, namely A. leptaspis, A. orthopogon, A. rnagna and A. aculeata. Phylogenetic trees were constructed by means of Bayesian inference (BI), Maximum Parsimony (MP), Neighbor-Joining (NJ), and Maximum Likelihood (ML) to assess the inter- and intra-species relationships within the genus Aspidisca. All trees show similar topologies with stable supports and indicate that: (1) four well known groups, i.e., Oligotrichia, Stichotrichia, Choreotrichia and Hypotrichia, are distinctly outlined within the class Spirotrichea, and all are monophyletic other than Hypotrichia; (2) members of Aspidisca can be distinguished well, based on the ITSI-5.SS-ITS2 region sequences, and A. leptaspis and A. magna shared a closer relationship than other species; (3) Aspidisca and Euplotes branch early in the subclass Hypotrichia. To compare the phylogenetic relationships based on different genes, SSU rRNA trees were also constructed with nearly the same species inclusion, which revealed different topologies of inter-species, inter-genera and inter-subclasses.展开更多
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...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.展开更多
We collected nine Enteromorpha specimens from the coast of Yantai and evaluated their diversity based on analyses of their ITS(internal transcribed spacer) and 5S rDNA NTS(non-transcribed spacer) sequences.The ITS seq...We collected nine Enteromorpha specimens from the coast of Yantai and evaluated their diversity based on analyses of their ITS(internal transcribed spacer) and 5S rDNA NTS(non-transcribed spacer) sequences.The ITS sequences showed slight nucleotide divergences between Enteromorpha linza and Enteromorpha prolifera.In contrast,multiple highly variable regions were found in the ITS region of Enteromorpha flexuosa.In general,there were more variable sites in the NTS region than in the ITS region in the three species.The variations in 5S rDNA NTS sequences indicated that the molecular diversity of Enteromorpha from the coast of Yantai is very high.However,a phylogenetic tree constructed using 5S rDNA NTS sequence data indicated that genetic differences were not directly related to geographical distribution.展开更多
The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fuj...The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fujian coast of China were introduced from Vietnam, the Philippines and Indonesia. Combined with morphological characteristics, all Kappaphycus and Eucheuma cultivated strains were identified by internal transcribed spacer (ITS) sequences. The phylogenetic tree was constructed using neighbor-joining and maximum likelihood methods. The results indicate that different ITS sequence lengths occurred in the different genera and species. An obvious difference in morphology could be found in the protuberance shape between Kappaphycus and Eucheuma. The protuberance in Eucheuma was thorn-like and in Kappaphycus was wartlike or papillate. Their ITS sequence lengths differed significantly in nucleotide variation rates up to 58.55%-63.90%. All nucleotide variations occurred in the ITS1 and ITS2 regions except for five nucleotide transversions in the 5.8S rDNA region. In addition, the difference was at the branches among congeneric species. Kappaphycus sp. had branches with small buds, while K. alvarezii did not have such a feature. The nucleotide variation rates varied from 7.02% to 7.48% among species; within the same species of the clades it was <1.20%. Eucheumatoideae algae cultivated in China consisted of three clades, K. alvarezii, Kappaphycus sp., and E. denticulatum. The results indicate that ITS sequence analysis was an effective way for identification of interspecies and intraspecies phylogenetic relationships and might provide a clue for molecular identification of algal Eucheumatoideae.展开更多
Batrachospermum arcuatum Kylin is typically dioecious,although some monoecious specimens have been collected from four locations in North China.In this study,B.arcuatum populations,including monoecious and dioecious t...Batrachospermum arcuatum Kylin is typically dioecious,although some monoecious specimens have been collected from four locations in North China.In this study,B.arcuatum populations,including monoecious and dioecious thalli,were collected from seven stream segments.The nuclear DNA internal transcribed spacer(ITS)region was sequenced for the seven populations,and a phylogenetic tree was constructed using Bayesian inference to assess intraspecifi c relationships.A haplotype network was also created.The ITS region in B.arcuatum from North China comprised 822–853 bp,with 10 haplotypes detected from the seven locations.The results of this study support the inclusion of monoecious individuals in the species B.arcuatum.展开更多
Microsatellites or simple sequence repeats(SSR) function widely and locate dependently in genome. However, their characteristics are often ignored due to the lack of genomic sequences of most species. Kelp(Saccharina ...Microsatellites or simple sequence repeats(SSR) function widely and locate dependently in genome. However, their characteristics are often ignored due to the lack of genomic sequences of most species. Kelp(Saccharina japonica), a brown macroalga, is extensively cultured in China. In this study, the genome of S. japonica was surveyed using an Illumina sequencing platform, and its microsatellites were characterized. The preliminarily assembled genome was 469.4 Mb in size, with a scaffold N_(50) of 20529 bp. Among the 128370 identified microsatellites, 90671, 25726 and 11973 were found in intergenic regions, introns and exons, averaging 339.3, 178.8 and 205.4 microsatellites per Mb, respectively. These microsatellites distributed unevenly in S. japonica genome. Mononucleotide motifs were the most abundant in the genome, while trinucleotide ones were the most prevalent in exons. The microsatellite abundance decreased significantly with the increase of motif repeat numbers, and the microsatellites with a small number of repeats accounted for a higher proportion of the exons than those of the intergenic regions and introns. C/G-rich motifs were more common in exons than in intergenic regions and introns. These characteristics of microsatellites in S. japonica genome may associate with their functions, and ultimately their adaptation and evolution. Among the 120140 pairs of designed microsatellite primers, approximately 75% were predicted to be able to amplify S. japonica DNA. These microsatellite markers will be extremely useful for the genetic breeding and population evolution studies of kelp.展开更多
Different from ribosomal genes, which contain highly conserved sequences that are detected in all organisms, the intergenic spacer of rDNA (IGS) appears to be the most rapidly-evolving spacer region. For this reason w...Different from ribosomal genes, which contain highly conserved sequences that are detected in all organisms, the intergenic spacer of rDNA (IGS) appears to be the most rapidly-evolving spacer region. For this reason we tested this region for phylogenetic studies. This report focuses on the study of IGS sequences of isolates belonging to Trichoderma section (T. viride, T. koningii, T. hamatum, T. erinaceus, T. asperellum) and Pachybasium section (T. harzianum, T. crassum, T. fasciculatum, T. oblongisporum, T. virens). Using the primer pair 28STD and CNS1, the Fast Start Taq DNA Polymerase (Roche), and a three temperature PCR protocol, products ranging from ca 1900 to 2400 bp were obtained from all tested isolates. The PCR product of 16 Trichoderma spp. isolates was cloned into a pGEM-TEeasy Vector (Promega) and sequenced. Based on a BLAST search we can conclude that the PCR product represents the whole IGS region. Multiple alignments of IGS sequences revealed two portions with different homology level. Portion A (ca 1660 bp) is the portion that contains 3’ end of 28S gene and is the more variable, while portion B (ca 830 bp), that contains the 3’ end of IGS region and the 5’ end of 18S gene, is the less variable. Comparing all sequences in region A 705 identical pairs occur out of 1704 total nucleotides (41.4%), while in region B identical pairs were 723 out of 832 total nucleotides (86.9%) . Sequence comparison of the two regions at intraspecific level (where it was possible) showed higher variability in region A (0.17%-6.8%) than in region B (0.0%-1.0%) . At interspecific level, performing all possible comparisons, the variability of region A (19.5%-52.7%) and B (0.8%- 16.9%). were significantly higher. Comparing sequences of species belonging to Trichoderma section variability of the two regions appears reduced if compared with that obtained from comparisons of species belonging to Pachybasium section. On the basis of sequence alignment, phylogenetic trees were obtained either with entire IGS, with region A, and with region B. Results of this analysis revealed that all isolates belonging to Trichoderma section grouped separately from isolates belonging to Pachybasium section. IGS region allowed us to group species according to their taxonomic position. The topology of the tree did not change substantially, varying in genetic distance only. Performing a GenBank search sequences representing the final portion of the IGS region of other fungal species were found, and we carried out a multiple alignment using also our sequences of Trichoderma spp. and Diaporthe helianthi. The phylogeny inferred from sequence alignment matched the generally accepted morphology-based classification and was identical to other molecular schemes at high taxonomic level. Data analysis was useful in establishing a broad-scale phylogeny of Ascomycota and was also useful in sorting them into statistically-supported clades. The tree showed that Trichoderma occurred in a well-supported terminal subclade of a larger clade that also contained other genera belonging to Hypocreales order. Sequence analysis of the Trichoderma spp. IGS region allowed us to design a specific PCR primer that was successfully used to amplify region A. The new reverse primer LCR2, that recognize all Trichoderma isolates, was identified in region B and confirmed for its specificity on the DNA of fungi belonging to other Ascomycota genera. Results obtained showed that IGS region seems to be an interesting and versatile tool for phylogenetic analysis, for resolving some taxonomic problems and for constructing specific primer useful for different purposes.展开更多
基金The project supported by the Grant from Presidentof Chinese Academy of Sciences
文摘The first internal transcribed spacer(ITS1) of nuclear ribosomal DNA of three wild rice species and two subspecies of cultivated rice, which are distributed in China, was amplified using PCR technique and sequenced with automated fluorescent sequencing. The sequences of ITS1 ranged from 193 bp to 218 bp in size and G/C content varied from 69.3% to 72.7%. In pairwise comparisons among the five taxa, sequence site divergence ranged from 1.5% to 10.6%. Phylogenetic analysis of ITS1 sequences using Wagner parsimony generated a single well resolved tree, which revealed that Oryza rufipogon was much more closely related to cultivated rice species than to the other two wild species. Oryza granulata was less closely related to either cultivated rice species or the other two wild species, and might be a unique and isolated taxon in the genus Oryza. The phylogenetic relationships of the three wild rice species and two cultivated rice subspecies inferred from ITS1 sequences is highly concordant with those based on the molecular evidence from isozyme, chloroplast DNA (cpDNA), mitochondrial DNA(mtDNA) and nuclear DNA (nDNA) of the genus Oryza .
文摘The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA (including 5.8S rRNA) of 15 Rhododendron, species, representing most sections of the genus, one Ledum species and Cassiope fastigiata were sequenced. Together with the ITS sequences of 13 selected Rhododendron species and Bejaria racemosa downloaded from GenBank, we explored the infrageneric and sectional relationships of this important North Temperate genus by employing maximum-parsimony analysis using PAUP software. C. fastigiata and B. racemosa were designated as outgroups. The ITS-based tree inferred that: (1) Rhododendron was a well-supported monophyletic group, while subg. Therorhodion was basal to the rest of the genus; (2) Ledum was a member of Rhododendron, and its close relationship with the lepidote rhododendron was confirmed; (3) the lepidote rhododendron plus Ledum formed a strongly-supported monophyletic clade which was sister to the rest of the elepidote rhododendron; (4) the elepidote rhododendron formed a weakly-supported clade within which the monophyly of subg. Hynwrianthes and subg. Tsutsusi were strongly supported, while subg. Pentanthera and subg. Azaleastrum were polyphyletic; and (5) the monophyly of sect. Choniastnini, (subg. Azaleastrum) was strongly-supported, while subg. Tsutsusi could be sister to a weakly-supported clade composed of two sampled species of sect. Azaleastrum (subg. Azaleastrum) together with R. sentibarbatum, of subg. Mumeazalea.
文摘Pinus squamata X. W. Li, a recently-described species of the Pinaceae, is an extremely endangered pine with only 32 individuals in the wild. This species was thought to be intermediate between P. bungeana Zuec. ex Endl. of subgen. Strobus and P. yunnanensis Franch. of subgen. Pinus by the original author. We made an effort to address the question of the phylogenetic relationship of this peculiar and important species in the context of the major groups of the genus Pinus by using sequences of the rbcL, matK,genes, rpl20-rps18 spacer, trnV intron of the chloroplast genome and the nuclear ribosomal ITS region. The results of the separated analysis and the combined analysis of the four cpDNA sequences and ITS sequence indicated that P. squamata was a stable member of subsect. Gerardianae and P. gerardiana Wall. was the closest species of P. squamata phylogenetically. As a result, the causes of the distribution pattern of subsect. Gerardianae were also discussed.
文摘Presented is design concept for key parameters o f the reverse conducting gate commutated thyristor (RC-GCT),such as the thickness and concentration of n-base region and the transparent anode region,and the wi dth of separation region between asymmetric GCT and PIN diode.A structure model of the RC-GCT is set up based on the design concept and its characteristics are analyzed.The simulation results show the design concept is reasonable.
基金Project (2010CB731704) supported by the National Basic Research Program of China
文摘The microstructure, phase composition and cold shut defect of thick titanium alloy electron beam welded joint were studied. The results showed that the microstructure of weld zone was composed of α′ phase; the heat affected zone was divided into fine-grained zone and coarse-grained zone, the microstructure of fine-grained zone was primary α phase + β phase + equiaxed α phase, and the microstructure of coarse-grained zone was primary α phase + acicular α′ phase; the microstructure of base metal zone basically consisted of primary α phase, and a small amount of residual β phase sprinkled. The forming. reason of cold shut was analyzed, and the precaution of cold shut was proposed.
基金Supported by Key Project of Knowledge Innovation Project of Chinese Academy of Sciences(KGCX2-YW-374-3)Scientific and Technological Project of Shandong Province(2008GG20007002)~~
文摘[Objective] The aim was to select suitable gene for Chlorella identification and to identify the oil-producing microalgae.[Method] Four candidate gene sequences,the nuclear genomic rDNA of the 18S rRNA gene,internal transcribed spacer(ITS),internal transcribed spacer Ⅱ(ITS Ⅱ)and the chloroplast rbcL gene,were selected for Chlorella molecular identification.Through these four candidate genes,the genetic variability and distinguish ability between intra-species and inter-species was analyzed to choose the right genes for identification of the high oil-content Chlorella.On this basis,application of these gene segments were classified and identified for five fresh-water isolated Chlorella,which oil-content is more than 30%.[Result] ITS gene was a suitable gene because of its high variation and short fragment length,meanwhile its genetic distance intra-species(0.439 6±0.135 9)was larger than inter-species(0.045 7±0.084 3).Its sequence length varied between different species whereas highly conserved in the same species.By the application of ITS sequences,respectively,five high oil-content stains were identified as one C.vulgaris,two strains of C.sorokiniana and two strains of algae Chlorella sp.[Conclusion] This study had provided reference for the establishment of identification gene pool of Chlorella.
基金Hong Kong Jockey Club Charities Trust (ProjectNo. HKJCICM-2-02R).
文摘Aim To investigate the plant origin and the identity of the red substance on the surface of Hong Dangshen, a unique medicinal material for diarrhea in Hong Kong. Methods The HPLC fingerprints and 5S rRNA gene spacer sequences of Hong Dangshen were obtained and compared with those of genuine species of Radix Codonopsis. The X-ray diffraction spectrum of the red substance was analyzed and compared with that of Halloysitum fingerprints to the Codonopsis species and the highest similarity to Rubrum. Results Hong Dangshen showed very similar HPLC Codonopsis pilosula var. modesta in terms of 5S rRNA gene spacer sequences. The X-ray diffraction spectrum of the red substance was consistent with that of Halloysitum Rubrum. Conclusion The source plant of Hong Dangshen was suggested to be Codonopsis pilosula var. modesta, one of the genuine original plants of Radix Codonopsis (Dangshen) in the China Pharmacopoeia (2005 edition). The red substance on the surface of Hong Dangshen was indicated to be Halloysitum Rubrum, a traditional medicinal mineral for chronic diarrhea. Our data suggest that Hong Dangshen is derived from the roots of Codonopsis pilosula var. modesta which has been processed with Halloysitum Rubrum, and the name is suggested to be Radix Codonopsis Praeparata Halloysita Rubra.
基金Supported by the National Natural Science Foundation of China (No.30570125)the Key Construction Laboratory of Marine Biotechnology of Jiangsu Province (No. 2010HS03)
文摘Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientists. In this paper, we sequenced the 18S rDNA genes, the internal transcribed spacer (ITS) regions and the rbcL genes in seven organisms and obtained 536-566 bp long ITS sequences, 1 377-I 407 bp long rbcL sequences and 1 718-1 761 bp long partial 18S rDNA sequences. The GC base pair content was highest in the ITS regions and lowest in the rbcL genes. The sequencing results showed that the three Ulvaprolifera (or U. pertusa) gene sequences from Qingdao and Nan'ao Island were identical. The ITS, 18S rDNA and rbcL genes in U. prolifera and U. pertusa from different sea areas in China were unchanged by geographic distance. U.flexuosa had the least evolutionary distance from U. californica in both the ITS regions (0.009) and the 18S rDNA (0.002). These data verified that Ulva and Enteromorpha are not separate genera.
基金Supported by the National High Technology Research and Development Program of China (863 Program)(No 2006AA10A402)Project for Supporting National Development (No 2006BAD09A04)+2 种基金the National Natural Science Foundation of China (Nos U0633006,40476059)the Natural Science Foundation of Qingdao (No 05-2-p-2)the Knowledge Innovation Program of the Chinese Academy of Sciences (No KZCX2-211)
文摘An improved method of PCR in which the small segment of conchocelis is amplified directly without DNA extraction was used to amplify a RUBISCO intergenic spacer DNA fragment from nine species of red algal genus Porphyra(Bangiales,Rhodophyta),including Porphyra yezoensis(Jiangsu,China),P.haitanensis(Fujian,China),P.oligospermatangia(Qingdao,China),P.katadai(Qingdao,China),P.tenera(Qingdao,China),P.suborboculata(Fujian,China),P.pseudolinearis(Kogendo,Korea),P.linearis(Devon,England),and P.fallax(Seattle,USA).Standard PCR and the method developed here were both conducted using primers specific for the RUBISCO spacer region,after which the two PCR products were sequenced.The sequencing data of the amplicons obtained using both methods were identical,suggesting that the improved PCR method was functional.These findings indicate that the method developed here may be useful for the rapid identification of species of Porphyra in a germplasm bank.In addition,a phylogenetic tree was constructed using the RUBISCO spacer and partial rbcS sequence,and the results were in concordant with possible alternative phylogenies based on traditional morphological taxonomic characteristics,indicating that the RUBISCO spacer is a useful region for phylogenetic studies.
基金Supported by the National Natural Science Foundation of China(No. 41006098)a joint grant from the Center of Excellence in Biodiversity,King Saud Universitythe 111 project (No.B08049)
文摘Abstract The internal transcribed spacer regions (ITS1 and ITS2) and 5.8S rRNA genes were sequenced in six populations of four Aspidisca species, namely A. leptaspis, A. orthopogon, A. rnagna and A. aculeata. Phylogenetic trees were constructed by means of Bayesian inference (BI), Maximum Parsimony (MP), Neighbor-Joining (NJ), and Maximum Likelihood (ML) to assess the inter- and intra-species relationships within the genus Aspidisca. All trees show similar topologies with stable supports and indicate that: (1) four well known groups, i.e., Oligotrichia, Stichotrichia, Choreotrichia and Hypotrichia, are distinctly outlined within the class Spirotrichea, and all are monophyletic other than Hypotrichia; (2) members of Aspidisca can be distinguished well, based on the ITSI-5.SS-ITS2 region sequences, and A. leptaspis and A. magna shared a closer relationship than other species; (3) Aspidisca and Euplotes branch early in the subclass Hypotrichia. To compare the phylogenetic relationships based on different genes, SSU rRNA trees were also constructed with nearly the same species inclusion, which revealed different topologies of inter-species, inter-genera and inter-subclasses.
基金Supported by the National Scientific-Basic Special Fund(No.2009FY210400)
文摘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.
基金Supported by the National Key Technology Research and Development Program(No.2008BA49B01)the Key Innovation Program of Chinese Academy of Sciences(No.KZCX2-YW-209)
文摘We collected nine Enteromorpha specimens from the coast of Yantai and evaluated their diversity based on analyses of their ITS(internal transcribed spacer) and 5S rDNA NTS(non-transcribed spacer) sequences.The ITS sequences showed slight nucleotide divergences between Enteromorpha linza and Enteromorpha prolifera.In contrast,multiple highly variable regions were found in the ITS region of Enteromorpha flexuosa.In general,there were more variable sites in the NTS region than in the ITS region in the three species.The variations in 5S rDNA NTS sequences indicated that the molecular diversity of Enteromorpha from the coast of Yantai is very high.However,a phylogenetic tree constructed using 5S rDNA NTS sequence data indicated that genetic differences were not directly related to geographical distribution.
基金Supported by the Shanghai Pujiang Talent Plan Project(No.05PJ14086)Shanghai Excellent Academic Leaders Project(No.08XD14037)Shanghai Hydrobiology Key Disciplines Funded Projects(No.S30701)
文摘The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fujian coast of China were introduced from Vietnam, the Philippines and Indonesia. Combined with morphological characteristics, all Kappaphycus and Eucheuma cultivated strains were identified by internal transcribed spacer (ITS) sequences. The phylogenetic tree was constructed using neighbor-joining and maximum likelihood methods. The results indicate that different ITS sequence lengths occurred in the different genera and species. An obvious difference in morphology could be found in the protuberance shape between Kappaphycus and Eucheuma. The protuberance in Eucheuma was thorn-like and in Kappaphycus was wartlike or papillate. Their ITS sequence lengths differed significantly in nucleotide variation rates up to 58.55%-63.90%. All nucleotide variations occurred in the ITS1 and ITS2 regions except for five nucleotide transversions in the 5.8S rDNA region. In addition, the difference was at the branches among congeneric species. Kappaphycus sp. had branches with small buds, while K. alvarezii did not have such a feature. The nucleotide variation rates varied from 7.02% to 7.48% among species; within the same species of the clades it was <1.20%. Eucheumatoideae algae cultivated in China consisted of three clades, K. alvarezii, Kappaphycus sp., and E. denticulatum. The results indicate that ITS sequence analysis was an effective way for identification of interspecies and intraspecies phylogenetic relationships and might provide a clue for molecular identification of algal Eucheumatoideae.
基金Supported by the National Natural Science Foundation of China(Nos.30970187,31200164)
文摘Batrachospermum arcuatum Kylin is typically dioecious,although some monoecious specimens have been collected from four locations in North China.In this study,B.arcuatum populations,including monoecious and dioecious thalli,were collected from seven stream segments.The nuclear DNA internal transcribed spacer(ITS)region was sequenced for the seven populations,and a phylogenetic tree was constructed using Bayesian inference to assess intraspecifi c relationships.A haplotype network was also created.The ITS region in B.arcuatum from North China comprised 822–853 bp,with 10 haplotypes detected from the seven locations.The results of this study support the inclusion of monoecious individuals in the species B.arcuatum.
基金financially supported by National High Technology Research and Development Program(863 Program)of China(2012AA10A406)National Science and Technology Supporting Program of China(2012 BAD55G01)+1 种基金Municipal Science and Technology Research and Development Project of Yantai(2013LGS002)Shandong Province Higher Educational Science and Technology Program(J15LE14)
文摘Microsatellites or simple sequence repeats(SSR) function widely and locate dependently in genome. However, their characteristics are often ignored due to the lack of genomic sequences of most species. Kelp(Saccharina japonica), a brown macroalga, is extensively cultured in China. In this study, the genome of S. japonica was surveyed using an Illumina sequencing platform, and its microsatellites were characterized. The preliminarily assembled genome was 469.4 Mb in size, with a scaffold N_(50) of 20529 bp. Among the 128370 identified microsatellites, 90671, 25726 and 11973 were found in intergenic regions, introns and exons, averaging 339.3, 178.8 and 205.4 microsatellites per Mb, respectively. These microsatellites distributed unevenly in S. japonica genome. Mononucleotide motifs were the most abundant in the genome, while trinucleotide ones were the most prevalent in exons. The microsatellite abundance decreased significantly with the increase of motif repeat numbers, and the microsatellites with a small number of repeats accounted for a higher proportion of the exons than those of the intergenic regions and introns. C/G-rich motifs were more common in exons than in intergenic regions and introns. These characteristics of microsatellites in S. japonica genome may associate with their functions, and ultimately their adaptation and evolution. Among the 120140 pairs of designed microsatellite primers, approximately 75% were predicted to be able to amplify S. japonica DNA. These microsatellite markers will be extremely useful for the genetic breeding and population evolution studies of kelp.
文摘Different from ribosomal genes, which contain highly conserved sequences that are detected in all organisms, the intergenic spacer of rDNA (IGS) appears to be the most rapidly-evolving spacer region. For this reason we tested this region for phylogenetic studies. This report focuses on the study of IGS sequences of isolates belonging to Trichoderma section (T. viride, T. koningii, T. hamatum, T. erinaceus, T. asperellum) and Pachybasium section (T. harzianum, T. crassum, T. fasciculatum, T. oblongisporum, T. virens). Using the primer pair 28STD and CNS1, the Fast Start Taq DNA Polymerase (Roche), and a three temperature PCR protocol, products ranging from ca 1900 to 2400 bp were obtained from all tested isolates. The PCR product of 16 Trichoderma spp. isolates was cloned into a pGEM-TEeasy Vector (Promega) and sequenced. Based on a BLAST search we can conclude that the PCR product represents the whole IGS region. Multiple alignments of IGS sequences revealed two portions with different homology level. Portion A (ca 1660 bp) is the portion that contains 3’ end of 28S gene and is the more variable, while portion B (ca 830 bp), that contains the 3’ end of IGS region and the 5’ end of 18S gene, is the less variable. Comparing all sequences in region A 705 identical pairs occur out of 1704 total nucleotides (41.4%), while in region B identical pairs were 723 out of 832 total nucleotides (86.9%) . Sequence comparison of the two regions at intraspecific level (where it was possible) showed higher variability in region A (0.17%-6.8%) than in region B (0.0%-1.0%) . At interspecific level, performing all possible comparisons, the variability of region A (19.5%-52.7%) and B (0.8%- 16.9%). were significantly higher. Comparing sequences of species belonging to Trichoderma section variability of the two regions appears reduced if compared with that obtained from comparisons of species belonging to Pachybasium section. On the basis of sequence alignment, phylogenetic trees were obtained either with entire IGS, with region A, and with region B. Results of this analysis revealed that all isolates belonging to Trichoderma section grouped separately from isolates belonging to Pachybasium section. IGS region allowed us to group species according to their taxonomic position. The topology of the tree did not change substantially, varying in genetic distance only. Performing a GenBank search sequences representing the final portion of the IGS region of other fungal species were found, and we carried out a multiple alignment using also our sequences of Trichoderma spp. and Diaporthe helianthi. The phylogeny inferred from sequence alignment matched the generally accepted morphology-based classification and was identical to other molecular schemes at high taxonomic level. Data analysis was useful in establishing a broad-scale phylogeny of Ascomycota and was also useful in sorting them into statistically-supported clades. The tree showed that Trichoderma occurred in a well-supported terminal subclade of a larger clade that also contained other genera belonging to Hypocreales order. Sequence analysis of the Trichoderma spp. IGS region allowed us to design a specific PCR primer that was successfully used to amplify region A. The new reverse primer LCR2, that recognize all Trichoderma isolates, was identified in region B and confirmed for its specificity on the DNA of fungi belonging to other Ascomycota genera. Results obtained showed that IGS region seems to be an interesting and versatile tool for phylogenetic analysis, for resolving some taxonomic problems and for constructing specific primer useful for different purposes.