The paper is aimed to identify SNP in Sarcandra glabra and Chloranthus spicatus, and authenticate S. glabra from Ch. spicatus and the mixture by using PCR amplification of specific alleles. SNPs in the ITS sequences of S. glabra and Ch. spicatus were found by ClustulX 2. 1 program and Bioedit software. Primers for authentic S. glabra and Ch. spicatus was designed according to the SNP site, and ITS sequence universal primers plus to the authentic primer to construct a multi-PCR reaction system, and then optimized the PCR reaction system. Five hundred and eighty band special for S. glabra and 470 bp band special for Ch. spicatus were found by using multi-PCR reaction. The multi-PCR reaction system could be applied to identify S. glabra and Ch. spicatus's leaves.
DNA, Plant ; analysis ; genetics ; DNA, Ribosomal ; genetics ; DNA, Ribosomal Spacer ; analysis ; genetics ; Magnoliopsida ; classification ; genetics ; Plant Leaves ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; RNA, Ribosomal ; genetics ; RNA, Ribosomal, 18S ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Species Specificity

OBJECTIVETo isolate and identify the anamorph of Shiraia bambusicola.

METHODFungus strains were isolated from mature ascospores and stroma. They were identified by means of morphological identification including colony and microscope characteristic, the molecular identification was done by 5.8S-ITS rDNA.

RESULTThe strains GZDXIFR-171 and GZDXIFR-181 were isolated and obtained with the separation of different methods, which had the same colony morphology. With the universal primers of the ITS1-5.8S rDNA-ITS2, the 5.8S-ITS rDNA sequence of GZDXIFR-171 and GZDXIFR-181 were obtained by the PCR amplification and sequencing. Compared with the published nucleotide sequence of 5.8S-ITS rDNA in NCBI (National Center for Biotechnology Information), GZDXIFR-171 and GZDXIFR-181 were highly identical with S. bambusicola.

CONCLUSIONThe isolated strains GZDXIFR-171 and GZDXIFR-181 were confirmed to be the true anamorph of S. bambusicola.

Ascomycota ; classification ; genetics ; isolation & purification ; DNA, Fungal ; genetics ; DNA, Ribosomal Spacer ; genetics ; Molecular Sequence Data ; Mycological Typing Techniques ; Phylogeny ; RNA, Ribosomal, 5.8S ; genetics

Molecular systematic techniques were applied to reveal the genetic diversity of medicinal plants and their related species in Salvia. The internal transcribed spacer (ITS) as well as 5.8S rDNA sequences of 27 samples of Salvia were amplified using PCR method and sequenced. Mega 3.1 was used to analyze the genetic diversity within genus. The complete sequences of ITS plus 5.8S rDNA are about 612-617 bp. A phylogenetic tree generated by Neighbor-Joining method partly supported the morphological classification within Salvia, but incompatible results were also obtained in the treatment of phylogenetic positions of some species such as Salvia trijuga, Salvia flava var. flava and Salvia flava var. megalentha. The ITS regions of present Salria species showed considerable variation between subgenera in contrast with the conservative 5.8S rDNA sequences. The native Salvia species might have a different origin from the foreign species. The phylogenetic positions of subgenera and sections inferred by ITS analysis were comparable with that of traditional classification, while the phylogeny within sections is still doubtful due to limited information in ITS sequence and need to be further proved by other evidence. ITS analysis in this study supports the rationality of using species from Drymosphace section as substitute drug resources of Dan shen, but also reveals significant genetic differences between high mountain Dan shen species such as Salvia przewalskii with traditional Dan shen origins.
Base Sequence ; DNA, Plant ; genetics ; DNA, Ribosomal ; genetics ; DNA, Ribosomal Spacer ; genetics ; Genetic Variation ; Phylogeny ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 5.8S ; genetics ; Salvia ; classification ; genetics ; Sequence Alignment ; Sequence Analysis, DNA

OBJECTIVETo study the ITS sequence variation of Pinellia ternata from different population in China, and it correlation to geographical distribution and morpha of the plant.

METHODThe rDNA ITS regions of various P. ternata were amplified and sequenced. And they were analyzed by means of the software of CLUSTRAL and MEGA.

RESULTComplete sequence of ITS and 5.8S rDNA of 16 different P. ternata population were obtained. The sequences of ITS1, 5.8S and ITS2 are 276,162 and 246 bp, respectively. ITS1 was more conservative than ITS2. Phylogenetic tree based on ITS1 and ITS2 sequences data was conducted by Neighbor-joining method.

CONCLUSIONRibosomal DNA ITS sequence analyses can be applied to the resource research of P. ternata.

Base Sequence ; China ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Ecosystem ; Molecular Sequence Data ; Phylogeny ; Pinellia ; genetics ; Plant Tubers ; genetics ; Plants, Medicinal ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Sequence Analysis, DNA

OBJECTIVETo find the patterns of the rDNA ITS sequence variation of Schisandra sphenanthera and S. viridis, and to establish the molecular biological method for the identification of Fructus Schisandrae Sphenantherae and the fruits of S. viridis.

METHODPCR products were sequenced directly and the sequences were analyzed with PAUP 4.0b10. NJ systematic tree was obtained with neighbor-joining method.

RESULTThe Complete ITS sequence of S. sphenanthera was 691-692 bp, of which there were 282 bp of ITS1 and 246-247 bp of ITS2. The complete sequence of S. viridis was 694-695 bp, consisting of 285-286 bp of ITS1 and 246-247 bp of ITS2. There were three informative sites in ITS1 regions for the two species. In the NJ tree with Kadsura anamosma and K. coccinea as outgroups, five different populations of S. viridis were the monophyletic group with the bootstrap value of 68%. These populations included one from Tianmushan, Zhejiang province, three populations from Jigongshan, Henan Province and the other two populations of S. viridis cited the sequences from GeneBank (registration numbers are AF263438 and AF163703 respectively).

CONCLUSIONThe rDNA internal transcribed spacer is a good marker to distinguish the Fructus Schisandrae Sphenantherae from the fruits of S. viridis.

Base Sequence ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; Fruit ; genetics ; Molecular Sequence Data ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Schisandra ; classification ; genetics ; Sequence Analysis, DNA

OBJECTIVETo explore a new method for identification Astragali Radix from its adulterants by using ITS sequence.

METHODThirteen samples of the different Astragali Radix materials and 6 samples of the adulterants of the roots of Hedysarum polybotrys, Medicago sativa and Althaea rosea were collected. ITS sequence was amplified by PCR and sequenced unidirectionally. The interspecific K-2-P distances of Astragali Radix and its adulterants were calculated, and NJ tree and UPGMA tree were constructed by MEGA 4.

RESULTITS sequences were obtained from 19 samples respectively, there were Astragali Radix 646-650 bp, H. polybotrys 664 bp, Medicago sativa 659 bp, Althaea rosea 728 bp, which were registered in the GenBank. Phylogeny trees reconstruction using NJ and UPGMA analysis based on ITS nucleotide sequences can effectively distinguish Astragali Radix from adulterants.

CONCLUSIONITS sequence can be used to identify Astragali Radix from its adulterants successfully and is an efficient molecular marker for authentication of Astragali Radix and its adulterants.

Althaea ; classification ; genetics ; Astragalus membranaceus ; classification ; genetics ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal ; chemistry ; genetics ; DNA, Ribosomal Spacer ; genetics ; Fabaceae ; classification ; genetics ; Medicago sativa ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Roots ; genetics ; RNA, Ribosomal ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Sequence Analysis, DNA ; Species Specificity

OBJECTIVETo provide DNA molecular marker for identification of Nelumbo nucifera by exploring the differences of nrDNA-ITS sequence of N. nucifera originated from different habitats.

METHODTo compare nrDNA-ITS base sequence using specific PCR-ITS.

RESULTThe completed sequence of ITS and 5.8 S rDNA, and the partial sequences of 18S rDNA and 26S rDNA, totally 750 bp, from N. nucifera were obtained. The differences among N. nucifera from different habitats and from different cultivars were found.

CONCLUSIONThe method can be used to identify N. nucifera among different species and to distinguish their fakes. It provided the basis for identifying N. nucifera from different geographical regions by comparison of their ITS sequences.

Base Sequence ; China ; DNA, Plant ; chemistry ; genetics ; metabolism ; DNA, Ribosomal Spacer ; classification ; genetics ; Deoxyribonuclease EcoRI ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Drug Contamination ; prevention & control ; Geography ; Nelumbo ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; RNA, Ribosomal ; genetics ; RNA, Ribosomal, 18S ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Sequence Analysis, DNA ; Species Specificity

OBJECTIVETo screen out fungus strains with acetylcholinesterase inhibitory activity from Huperzia serrata.

METHODEndophytic fungi fermentation products from 59 H. serrata strains were stained with acetylcholinesterase hydrolyzed alpha-naphthaleneacetic ethyl ester and fast blue B salt, and screened for acetylcholinesterase inhibitory activity with thin-layer chromatography-bioautography. Target strains were classified and identified through the sequence analysis on 18s rDNA and 5.8s rDNA combined with morphological characteristics.

RESULTFungus strain LQ2F01 from H. serrata showed positive color reaction in the screening for acetylcholinesterase inhibitory activity. The sequence analysis on 18s rDNA and 5.8s rDNA combined with morphological characteristics showed the strain LQ2F01 belonged to Acremonium.

CONCLUSIONEndophytic Fungi LQ2F01 from H. serrata shows identical acetylcholinesterase inhibitory activity with the host plant, which is of great significance to the development of natural medicines and the studies on the relationship between the endophytic gungi and the host plant.

Acetylcholinesterase ; metabolism ; Acremonium ; genetics ; metabolism ; Cholinesterase Inhibitors ; isolation & purification ; metabolism ; Chromatography, Thin Layer ; DNA, Fungal ; chemistry ; genetics ; DNA, Ribosomal ; chemistry ; genetics ; Diazonium Compounds ; metabolism ; Fungi ; classification ; genetics ; metabolism ; Huperzia ; microbiology ; Hydrolysis ; Naphthaleneacetic Acids ; metabolism ; Phylogeny ; RNA, Ribosomal, 18S ; classification ; genetics ; RNA, Ribosomal, 5.8S ; classification ; genetics ; Sequence Analysis, DNA

The endophytic fungi with plant growth promoting effects were screened by co-culture of each endophytic fungus and seedlings of Dendrobium officinale. Anatomical features of the inoculated roots were studied by paraffin sectioning. Morphological characteristics and rDNA ITS1-5. 8S-ITS2 sequences were applied for the taxonomy of endophytic fungi. The results showed that 8 strains inoculated to D. officinale seedlings greatly enhanced plant height, stem diameter, new roots number and biomass. According to the anatomical features of the inoculated roots, each fungus could infect the velamina of seedlings. The hyphae or pelotons were existed in the exodermis passage cells and cortex cells. The effective fungi could not infect the endodermis and vascular bundle sheath, but which was exception for other fungi with harmful to seedlings. Combined with classic morphologic classification, 2 effective strains were identified which were subjected to Pestalotiopsis and Eurotium. Six species of fungi without conidiophore belonged to Pyrenochaeta, Coprinellus, Pholiota, Alternaria, Helotiales, which were identified by sequencing the PCR-amplified rDNA ITS1-5. 8S-ITS2 regions. The co-culture technology of effective endophytic fungi and plant can apply to cultivate the seedlings of D. officinale. It is feasible to shorten growth cycle of D. officinale and increase the resource of Chinese herbs.
Biomass ; DNA, Fungal ; genetics ; DNA, Ribosomal ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; cytology ; growth & development ; microbiology ; Endophytes ; classification ; genetics ; physiology ; Fungi ; classification ; genetics ; physiology ; Host-Pathogen Interactions ; Hyphae ; physiology ; Plant Roots ; cytology ; growth & development ; microbiology ; Plants, Medicinal ; growth & development ; microbiology ; RNA, Ribosomal, 5.8S ; genetics ; Seedlings ; cytology ; growth & development ; microbiology ; Species Specificity

The 2 principal species of hookworms infecting humans are Necator americanus and Ancylostoma duodenale. Case studies on zoonotic hookworm infections with Ancylostoma ceylanicum and/or Ancylostoma caninum are known mainly from Asian countries. Of these 2 zoonotic species, only A. ceylanicum can develop to adulthood in humans. In the present study, we report a molecular-based survey of human hookworm infections present in southern and northeastern Thailand. Thirty larval hookworm samples were obtained from fecal agar plate cultures of 10 patients in northeastren Thailand and 20 in southern Thailand. Partial ITS1, 5.8S, and ITS2 regions of the ribosomal DNA genes were amplified using PCR. The amplicons were sequenced, aligned, and compared with other hookworm sequences in GenBank database. The results showed that, in Thailand, N. americanus is more prevalent than Ancylostoma spp. and is found in both study areas. Sporadic cases of A. ceylanicum and A. duodenale infection were seen in northeastern Thailand.
Ancylostoma/classification/genetics/*isolation & purification ; Ancylostomiasis/*epidemiology ; Animals ; Cluster Analysis ; DNA, Ribosomal Spacer/chemistry/genetics ; Feces/parasitology ; Humans ; Molecular Sequence Data ; Necator americanus/classification/genetics/*isolation & purification ; Necatoriasis/*epidemiology ; Phylogeny ; Polymerase Chain Reaction ; Prevalence ; RNA, Ribosomal, 5.8S/genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Thailand/epidemiology