To clarify the occurrence and distribution of broad bean wilt virus 2(BBWV2) on Rehmannia glutinosa, this study collected 87 R. glutinosa samples with typical symptoms of viral disease such as chlorosis and crumple from Wenxian county and Wuzhi county in Jiaozuo city, Henan province and Qiaocheng district in Bozhou city, Anhui province. The BBWV2 CP target band was amplified from 37 R. glutinosa samples by RT-PCR technology. The total detection rate reached 42.5%, among which 43.0% was detected in samples from Henan province. The detection rate in samples from Anhui province was 37.5%. 37 BBWV2 CP sequences were obtained by cloning and sequencing of BBWV2 positive samples(data has been submitted to GenBank, accession numbers: PP407959-PP407995), and the sequence analysis of these CP sequences with 91 other BBWV2 isolates in GenBank showed a high genetic diversity with a consistency rate of 70.8%-100%. Meanwhile, phylogenetic analysis showed that BBWV2 could be divided into three groups according to CP sequences, among which the BBWV2 in R. glutinosa isolates obtained in this study were all located in group 3. This study identified the differences in the occurrence, distribution, and genetic diversity of BBWV2 in R. glutinosa from Henan province and Anhui province and provided a theoretical basis for the prevention and control of BBWV2.
Rehmannia/virology* ; Phylogeny ; Plant Diseases/virology* ; China ; Molecular Sequence Data ; Fabavirus/classification*

The differences in chemical quality characteristics between Xinjiang Rehmannia glutinosa and R. glutinosa were analyzed to provide a theoretical basis for the introduction and quality control of R. glutinosa. In this study, the high performance liquid chromatography(HPLC) fingerprints of 6 batches of Xinjiang R. glutinosa and 10 batches of R. glutinosa samples were established. The content of iridoid glycosides, phenylethanoid glycosides, monosaccharides, oligosaccharides, and polysaccharides in Xinjiang R. glutinosa and R. glutinosa was determined by high performance liquid chromatography-diode array detection(HPLC-DAD), high performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD), and ultraviolet-visible spectroscopy(UV-Vis). The determination results were analyzed with by chemical pattern recognition and entropy weight TOPSIS method. The results showed that there were 19 common peaks in the HPLC fingerprints of the 16 batches of R. glutinosa, and catalpol, aucubin, rehmannioside D, rehmannioside A, hydroxytyrosol, leonuride, salidroside, cistanoside A, and verbascoside were identified. Hierarchical cluster analysis(HCA) and principal component analysis(PCA) showed that Qinyang R. glutinosa, Mengzhou R. glutinosa, and Xinjiang R. glutinosa were grouped into three different categories, and eight common components causing the chemical quality difference between Xinjiang R. glutinosa and R. glutinosa in Mengzhou and Qinyang of Henan province were screened out by orthogonal partial least squares discriminant analysis(OPLS-DA). The results of content determination showed that there were glucose, sucrose, raffinose, stachyose, polysaccharides, and nine glycosides in Xinjiang R. glutinosa and R. glutinosa samples, and the content of catalpol, rehmannioside A, leonuride, cistanoside A, verbascoside, sucrose, and glucose was significantly different between Xinjiang R. glutinosa and R. glutinosa. The analysis with entropy weight TOPSIS method showed that the comprehensive quality of R. glutinosa in Mengzhou and Qinyang of Henan province was better than that of Xinjiang R. glutinosa. In conclusion, the types of main chemical components of R. glutinosa and Xinjiang R. glutinosa were the same, but their content was different. The chemical quality of R. glutinosa was better than Xinjiang R. glutinosa, and other components in R. glutinosa from two producing areas and their effects need further study.
Rehmannia/classification* ; Drugs, Chinese Herbal/chemistry* ; Chromatography, High Pressure Liquid/methods* ; Quality Control

To clone and analyze a member of the Auxin/indole-3-acetic acid (Aux/IAA) gene family, RgIAA1, from Rehmannia glutinosa. The transcriptional EST database of R. glutinosa was used to clone the new Aux/IAA gene by cDNA probe of AtIAA14. Bioinformatics was applied to analyze the sequence characteristics of RgIAA1 protein and construct phylogenetiC trees. Quantitative RT-PCR has been applied to detect the transcription level of RgIAA1 in seven tissues as well as in leaves under three stresses. The results showed that, the cDNA sequence of RgIAA1 contains 903 bp was obtained. The open reading frame (ORF) of RgIAA1 was 681 bp encoding 226 amino acids, which has typical structural domains and characteristic sequence of Aux/IAA family proteins. RgIAA1 showed the highest expression level in unfolded leaf, followed by the stem. And the expression of RglAA1 was quickly decreased with leaf growing up. The transcription level increased under continuous cropping conditions while it reduced both in salinity and waterlogging stresses. RgIAA1, an Aux/IAA gene from R. glutinosa has been obtained for the first time, which can lay the foundation for further studies about its molecular function in development and responses to stress.
Amino Acid Sequence ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Indoleacetic Acids ; metabolism ; Molecular Sequence Data ; Organ Specificity ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; Rehmannia ; classification ; genetics ; physiology ; Stress, Physiological ; genetics

The swollen root of Rehmannia glutinosa is used as one kind of important Chinese traditional medicine. The root of R. glutinosa usually swelled in rotational cropping but not in continuous cropping. The rhizosphere microorganisms of R. glutinosa under different farming condition were thought related to that. In this study, the endophytic fungi in the root of R. glutinosa growing in various soil conditions were isolated for the study of the relationship between the microorganisms and the root enlargement of their host plants. The dominant endophytes, Verticillium spp., Fusarium oxysporum, F. redolens and Ceratobasidium spp. were identified by morphological observation and 18S rDNA and ITS sequence analysis. The preliminary investigation showed that the excessive growth of Verticillium and Fusarium genus fungi is unfavorable for the R. glutinosa root swelling, but Ceratobasidium fungi has no effects on the root enlargement.
DNA, Fungal ; genetics ; DNA, Ribosomal ; genetics ; Fungi ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Plant Roots ; microbiology ; Rehmannia ; microbiology

OBJECTIVETo provide molecular evidences for phylogenetic analysis by studying ITS sequences of Rehmannia glutinosa from different areas.

METHODThe DNAs were extracted from leaves of R. glutinosa by means of CTAB method. The products of PCR amplification were cloned . The data were analyzed by MEGA4.0 software.

RESULTThe results showed that the size of the ITS of R. glutionsa tested was from 613 to 614 bp and the length variation was only 1 bp. The sequence of ITS1 was 224-225 bp, and G + C content varied from 60.4% to 63%. The sequence of ITS2 was 224-225 bp and G + C content varied from 57.1% to 65.3%. The sequence of 5. 8S rDNA was 164 bp, it's very conservative in these species. Phylogram tree based on ITS sequence data indicated that the kinship between Bejing No. 2 R. glutinosa and the others were far. There was obvious diversity within wild R. glutinosa varieties, while there was no different among cultivated R. glutinosa varieties. In cultivated R. glutinosa varieties, there was no diversity between R. glutinosa varieties from Henan and those from others provinces. In wild varieties, R. glutinosa from Shengnongshan and Qingtianhe of Henan province showed a closer systematic relationship with cultivated R. glutinosa from Shandong province, while there was no difference between wild R. glutinosa varieties and cultivated varieties from Henan and Shanxi provinces.

CONCLUSIONThe genetic relationship among R. glutinosa varieties was very close, there was no distinct systematic differentiation.

DNA, Plant ; analysis ; Genetic Variation ; Molecular Sequence Data ; Phylogeny ; Plant Leaves ; genetics ; Polymorphism, Single Nucleotide ; RNA, Ribosomal, 16S ; analysis ; Rehmannia ; classification ; genetics

OBJECTIVETo analyze the genetic diversity of wild Rehmannia glutinosa and evaluate and compare random amplified polymorphic DNA (RAPD) and inter sample sequence repeat (ISSR) for analysis of R. glutinosa accessions.

METHODTwo molecular markers, RAPD and ISSR were used for analyzing 55 wild R. glutinosa accessions.

RESULTAverage 16.00 and 19.08 bands were amplified by RAPD primers and ISSR primers respectively, and the percentage of polymorphic bands were 89.58% and 94.32% respectively; Fifty-five R. glutinosa accessions categorized into 7 clusters were identified by unweighted pair-group method, arithmetic average (UPGMA) method.

CONCLUSIONA high level of genetic diversity of wild Rehmannia glutinosa was displayed at DNA level, and genetic diversity coefficient of R. glutinosa from different production areas was 0.63-0.98, and ISSR marker can detect higher genetic diversity of R. glutinosa germplasms than RAPD marker.

Genetic Variation ; genetics ; Phylogeny ; Random Amplified Polymorphic DNA Technique ; methods ; Rehmannia ; classification ; genetics

OBJECTIVETo define the genetic relationship and other genetic characters of different varieties of Rehmannia glutinosa.

METHODRAPD reactions were conducted on total 54 samples from 16 varieties and 2 populations of R. glutinosa. The data were analyzed by Phylips, Popgene and Arlequin software.

RESULTThe majority rule consensus tree defined the 54 samples to 3 groups. Population genetics analysis and AMOVA showed that the genetic diversity among varieties/populations much greater than that within varieties/populations, but within group II the genetic diversity among varieties was similar to that within varieties.

CONCLUSIONDue to the long-time asexual propagation of R. glutinosa, some genetic differentiation has been accumulated and fixed within the species. It was shown that the genetic distance between wild population and cultivated varieties was greater than the genetic distance among cultivated varieties. The wild resource of the plant should be paid more attention and studies.

Analysis of Variance ; China ; DNA, Plant ; analysis ; genetics ; Ecosystem ; Genetic Variation ; Phylogeny ; Plants, Medicinal ; genetics ; growth & development ; Random Amplified Polymorphic DNA Technique ; Rehmannia ; classification ; genetics ; growth & development