Identification and expression profiling of R2R3-MYB transcription factors in Erigeron breviscapus / 中国中药杂志

R2 R3-MYB transcription factors are ubiquitous in plants, playing a role in the regulation of plant growth, development, and secondary metabolism. In this paper, the R2 R3-MYB transcription factors were identified by bioinformatics analysis of the genomic data of Erigeron breviscapus, and their gene sequences, structures, physical and chemical properties were analyzed. The functions of R2 R3-MYB transcription factors were predicted by cluster analysis. Meanwhile, the expression patterns of R2 R3-MYB transcription factors in response to hormone treatments were analyzed. A total of 108 R2 R3-MYB transcription factors, named EbMYB1-EbMYB108, were identified from the genome of E. breviscapus. Most of the R2 R3-MYB genes carried 2-4 exons. The phylogenetic tree of MYBs in E. breviscapus and Arabidopsis thaliala was constructed, which classified 234 MYBs into 30 subfamilies. The MYBs in the five MYB subfamilies of A.thaliala were clustered into independent clades, and those in E. breviscapus were clustered into four clades. The transcriptome data showed that MYB genes were differentially expressed in different tissues of E. breviscapus and in response to the treatments with exogenous hormones such as ABA, SA, and GA for different time. The transcription of 13 R2 R3-MYB genes did not change significantly, and the expression patterns of some genes were up-regulated or down-regulated with the extension of hormone treatment time. This study provides a theoretical basis for revealing the mechanisms of R2 R3-MYB transcription factors in regulating the growth and development, stress(hormone) response, and active ingredient accumulation in E. breviscapus.

A semimicroquality evaluation method on Panax notoginseng and its application in analysis of continuous cropping obstacles research samples / 中国中药杂志

Panax notoginseng is a commonly used traditional Chinese medicine with blood activating effect while has continuous cropping obstacle problem in planting process. In present study, a semimicroextraction method with water-saturated n-butanol on 0.1 g notoginseng sample was established with good repeatability (RSD<2.5%) and 9.6%-20.6% higher extraction efficiency of seven saponins than the conventional method. A total of 16 characteristic peaks were identified by LC-MS-IT-TOF, including eight 20(S)-protopanaxatriol (PPT) type saponins and eight 20(S)-protopanaxadiol (PPD) type saponins. The established method was utilized to evaluate the quality of notoginseng samples cultivated by manual intervened methods to overcome continuous cropping obstacles.As a result, HPLC fingerprint similarity, content of Fa and ratio of notoginsenoside K and notoginsenoside Fa (N-K/Fa) were found out to be as valuatable markers of the quality of samples in continuous cropping obstacle research, of which N-K/Fa could also be applied to the analysis of notoginseng samples with different growth years.Notoginseng samples with continuous cropping obstacle had HPLC fingerprint similarity lower than 0.87, in consistent with normal sample, and had significant lower content of notoginsenoside Fa and significant higher N-K/Fa (2.35-4.74) than normal group (0.45-1.33). All samples in the first group with manual intervention showed high similarity with normal group (>0.87), similar content of common peaks and N-K/Fa (0.42-2.06). The content of notoginsenoside K in the second group with manual intervention was higher than normal group. All samples except two displayed similarity higher than 0.87 and possessed content of 16 saponins close to normal group. The result showed that notoginseng samples with continuous cropping obstacle had lower quality than normal sample. And manual intervened methods could improve their quality in different levels.The method established in this study was simple, fast and accurate, and the markers may provide new guides for quality control in continuous cropping obstacle research of notoginseng.

Effects of seed priming on physiology of seed germination and seeding growth of Marsdenia tenacissima under NaCl stress / 中国中药杂志

To offer the reference and method for salt damage in the cultivation of Marsdenia tenacissima, the seeds of M. tenacissima collected from Maguan city ( Yunnan province) were taken as the test materials to study the effects of different priming materials on improving germination and growth under high-level salt stress condition. Four different treatments, which were GA3, KNO3-KH2PO4, PEG-6000, NaCl, combined with ANOVA were applied to test the performance of germination energy, germination percentage, germination index, MDA, SOD, and CAT. The results showed that the seed germination was obviously inhibited under salt stress and the soaked seeds with different priming materials could alleviate the damage of salt stress. Under these treatments, the activities of SOD, CAT the content of soluble protein significantly increased. While the content of MDA significantly decreased. The maximum index was obtained when treated with 1.20% KNO3-KH2PO4, the germination percentage increased from 52.67% to 87.33% and the activity of SOD increased from 138.01 to 219.44 respectively. Comparing with the treatment of 1.20% KNO3-KH2PO4, the germination percentage of treating with 300 mg x L(-1) GA3 increased from 52.67% to 80.67%, while the activity of SOD increased from 138.01 to 444.61.

FTIR fingerprint spectrograms of traditional Chinese medicine Marsdenia tenacissima / 中国中药杂志

In this paper, Fourier transform infrared spectroscopy fingerprint analysis of Marsdenia tenacissima samples was used to develop a reliable method of tracing the geographical origins. Forty-eight samples from four provinces of China were analyzed by FTIR. We analyzed and characterized the fingerprints in both the full spectrum peaks and characteristic peaks, then the principal component analysis and the cluster analysis were carried out. The results of fingerprint analysis, correlation analysis, principal component analysis and cluster analysis can identify the geographic origins correctly, which verified and supplemented each other; the identification results and the actual location showed a high degree of consistency, namely the lower the space distance, the greater the similarity of different samples. These results revealed the obvious superiority and practical value in comparison to the more tedious and time-consuming wet chemistry method normally used. Using appropriate metrology methods can trace the geographical source correctly. The M. tenacissima materials from the region of Maguan should be considered as genuine medicinal materials taking into account the good quality.

Wild resources survey of Marsdenia tenacissima in Honghe, Yunnan / 中国中药杂志

To ascertain current situation of wild Marsdenia tenacissima resources in Honghe, Yunnan province, the distribution, habitat characteristic and resources reserves of M. tenacissima were surveyed based on interviews and investigation. The results showed that M. tenacissima was found in 7 counties such as Jinping, Mengzi etc, and distributed mainly on the mountainsides from 800 m to 1 200 m. And distribution was affected by many factors, such as light, heat, topography, soil, and vegetation. M. tenacissima grew well in distribution areas. M. tenacissima had averagely a weight of 2.8 kg per plant. Resources reserve of M. tenacissima in Honghe was estimated to 1 300 tons by now but it reduced rapidly in resent years, the wild resources reserve may not meet demand of market. Resources protection and wildlife tending would be conducted to deal with increasing medication requirements.

SSR information in Erigeron breviscapus transcriptome and polymorphism analysis / 中国中药杂志

<p><b>OBJECTIVE</b>The SSR information in the transcriptome of Erigeron breviscapus was analyzed in this study, in order to further develop new functional genes SSR markers laid a solid foundation.</p><p><b>METHOD</b>SSR loci were searched in all of 52,060 unigenes by using est_timmer. Perl program and SSR primers were designed by Primer3. Furthermore, 36 pairs of primers were randomly selected for the polymorphism analysis on 13 Erigeron breviscapus plants collected from different places.</p><p><b>RESULT</b>A total of 3639 SSRs were found in the transcriptome of Erigeron breviscapus, distributed in 3260 unigenes with the distribution frequency of 6.99%. Di-nucleotide repeat was the main type, account for as much as 34.41% of all SSRs, followed by mono-nucleotide (31.41%) and tri-nucleotide repeat motif (28.08%). The di-nucleotide repeat motifs of AT/AT and AC/GT were the predominant repeat types (28.71%). The tri-nucleotide repeat motifs of AAT/AT was the predominant repeat types (7.94%). For validation the availability of those SSR primers, we randomly selected 36 pairs of primers for PCR amplification. Among them, 34 pair primers (94.44%) produced clear and reproductive bands, 19 pair primers showed polymorphism (52.78%), and 13 Erigeron breviscapus plants were divided into 2 groups.</p><p><b>CONCLUSION</b>There are numerous SSRs in Erigeron breviscapus transcriptome with high frequency and various types, this will provide abundant candidate molecular markers for genetic diversity study and genetic map in this plant.</p>

Research on situation and countermeasure of Erigeron breviscapus plant production / 中国中药杂志

For more than ten years, Erigeron breviscapus has been used for the treatment of cardiovascular and cerebrovascular diseases, it experienced the procedure from wild species to the cultivars, and the quality of drug was rapidly improved. In order to further promote the development of E. breviscapus planting industry, this paper analyzes the development status and existing problems of E. breviscapus planting. Some measures would be come forward, such as strengthen the government's policy support and industrial regulate, accelerate the industry standard and technological innovation, expand brand effect of E. breviscapus of Yunnan, so that the industry has the health and sustainable development.

Cloning and bioinformation analysis of flavone synthase II gene of Erigeron breviscapus / 中国中药杂志

<p><b>OBJECTIVE</b>Erigeron breviscapus is a medicinal plant with the most developmental potential in Yunnan province, which is belongs to Erigeron genus of Compositae family. Scutellarin, the main active component of Erigeron breviscapus is one of flavone 7-O-glucuronide derivatives, its biosynthesis pathway is still not clear.</p><p><b>METHOD</b>Full length cDNA encoding flavone syhthase II gene in E. breviscapus was cloned in this study using R-PCR, 3'-RACE and 5'-RACE.</p><p><b>RESULT</b>The opening reading frame of FS II cDNA of E. breviscapus is 1 557 bp long and encoding 518 amino acids, designed as EbFS II, which is highly homologous with FS II of Compositae species, like Callistephus chinensis, Cynara cardunculus var. scolymus, Gerbera hybrida, Dahlia pinnata and Lobelia erinus.</p><p><b>CONCLUSION</b>Phylogenetic analysis showed that EbFS II might has the function of directly converting flavanone to flavone.</p>

Simultaneous determination of chlorogenic acid, scutellarin, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid in different parts of Erigeron breviscapus by high-performance liquid chromatography / 中国中药杂志

<p><b>OBJECTIVE</b>A high-performance liquid chromatographic (HPLC) method was developed for simultaneous determination of chlorogenic acid, scutellarin, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid in different parts of Erigerontis Herba.</p><p><b>METHOD</b>The four constituents were measured on an Agilent Zorbax SB-C18 column (4.6 mm x 450 mm, 5 microm) with a gradient elution of acetonitrile (A) -0.3% phosphoric acid solution (B) (0-10 min, 12%-15% A, 10-32 min, 15% A, 32-33 min, 15%-20% A, 33-50 min, 20%-22% A) at wavelength of 335 nm and 327 nm, and a flow rate of 1.0 mL x min(-1) and the column temperature was 30 degrees C.</p><p><b>RESULT</b>Linearity of each standard was established in the concentration range of 0.050 1-1.002 microg for chlorogenic acid, 0.165 9-3.318 microg for chlorogenic acid, 0.049 7-0.994 microg for 3,5-dicaffeoylquinic acid, 0.048 7-0.974 p.g for 4,5-dicaffeoylquinic acid respectively, with correlation coefficient r > 0.999 6. Average recoveries (n = 6) of 4 compounds were 98.53% with a RSD of 0.94%, 99.68% with a RSD of 0.49%, 98.78% with a RSD of 1.1%, 99.06% with a RSD of 0.81%, respectively.</p><p><b>CONCLUSION</b>The developed method is simple, accurate, and precise, it can be used for the quantitative analysis of Erigeron breviscapus.</p>

Relationship between expression of chalcone synthase gene (CHS) and scutellarin content in Erigeron breviscapus / 中国中药杂志

<p><b>OBJECTIVE</b>Scutellarin from Erigeron breviscapus is a flavonoid with remarkable pharmacological activity, whose route of biosynthesis is still fully clear. Chalcone synthase (CHS) is the key enzyme regulating flavonoids biosynthesis, and the aim of this study is to explain the relationship between patterns of the gene expression and scutellarin content through studying CHS gene expression patterns combined with scutellarin content in various parts of E. breviscapus.</p><p><b>METHOD</b>Through RT-PCR and RACE, the full length of CHS was cloned and analyzed by fluorescent quantitative PCR. The scutellarin content in tissues was analyzed by HPLC.</p><p><b>RESULT</b>The full-length gene sequence was 1 270 bp, encoding 405 amino acids. Software analysis found that the DNA sequence was 80% similarity with Compositae plant homeo-box gene. Fluorescence quantitative analysis showed that CHS had the highest expression level in leaves, far higher than that in root, stem and flower. HPLC analysis showed that the scutellarin was the highest in leaves, followed by the flowers and stems, scutellarin was not detected in root.</p><p><b>CONCLUSION</b>Correlation analysis showed that CHS expression amount and scutellarin content in different parts of E. breviscapus is positive correlation (r = 0.761, P < 0.05), it suggests that CHS gene expression level has important effect on biosynthesis of scutellarin.</p>

Genetic diversity and group consistency of breeding strains of Erigeron breviscapus determined by AFLP marker / 中国中药杂志

<p><b>OBJECTIVE</b>To analyze the genetic diversity and breeding strains of the E. breviscapus germplasms, in order to provide theoretical information for Erigeron breviscapus breeding.</p><p><b>METHOD</b>The genetic diversity and genetic structure were assayed to six germplasm resource of E. breviscapus which collected from Yunnna with 11 pairs primers and AFLP molecular marker.</p><p><b>RESULT</b>Six hundred and four amplification bands among 636 DNA bands were from six accession of E. breviscapus, which are about 82.40% of total bands. The six germplasms could be divided into three group at the 0. 706 similarity coefficient level. The first category include QS-1, QS-2 and Dali, Shilin, Kunming population. The second category included wild population of Qiubei. The third category included several sample from different district. The mean genetic similarity coefficient of QS-1 and QS-2 was bigger, genetic similarity coefficient range was smaller, hereditary character was more stable. Molecular system clustering analysis showed that the geographical origin of the same part had relative polymerization phenomenon and its genetic relationship was close. Qiubei was a single group possibly relating to the specific genetic basis.</p><p><b>CONCLUSION</b>The analysis of genetic diversity of E. breviscapus by AFLP marker is reliable. The systematic E. breviscapus breeding is feasible.</p>

Research progress and strategy on genetic improvement of Erigeron breviscapus / 中国中药杂志

Cultivation research and the research progress of genetic improvement of Erigeron breviscapus were been described. Some messures would be come forward, Such as developed the genetic reasearch, germplasm resources and breeding of E. breviscapus. Also it must be reasearch the biological basis, seed-breeding technology and some critical cultivation technique of E. breviscapus.

Study on optimal harvest time of Erigeron breviscapus / 中国中药杂志

<p><b>OBJECTIVE</b>To ascertain the optimal harvest time of Erigeron breviscapus.</p><p><b>METHOD</b>The dry matter weight accumulation of different organs in growth process and contents of scutellarin and coffeic acid ester in whole plant of E. breviscapus were determined.</p><p><b>RESULT</b>The number of leaves per plant, the dried weight of single leaf and dry matter weight of whole plant and different organs reached the highest after seedling 130-140 d. The content of scutellarin gradually decreased with growth period, and sharply decreased after seedling 140 d. The content of coffeic acid ester varied irregularly with growth period.</p><p><b>CONCLUSION</b>The optimal harvest time of E. breviscapus is in early bloom period after seedling 130 d.</p>

RAPD analysis on genetic relationships of Erigeron breviscapus germplasm resources / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the genetic relationships of Erigeron breviscapus at the molecular biology level.</p><p><b>METHOD</b>Thirty seven germplasm resources of E. breviscapus which collected from Yunnan, Sichuang and Guizhou province in 2005 were analyzed by Random amplified polymorphic DNA (RAPD) and cluster analysis based on NTSYS2.</p><p><b>RESULT</b>A total of 10 primers were screened, and 107 bands were amplified, among which 94 (87.85%) bands were found to be polymorphic. Thirty seven germplasm resources of E. breviscapus were clustered into 3 groups at genetic distance 0.36, the I group include in 9 germplasm resources collected from Mile, Qiubei, Luxi, Gejiu, and Yanshan of south east of Yunnan province; the II group included 8 germplasm resources collected from Gucheng, and shangrila of north west of Yunnan province, and Mile and Qiubei of south east of Yunnan province; the III group included in 20 germplasm resources collected from the center of Yunnan province, north east of Yunnan province, Sichuan province, and Guizhou province.</p><p><b>CONCLUSION</b>There were abundant genetic diversity in the germplasm resources of E. breviscapus, and the genetic relationships are closely related to geographical distance where they were collected.</p>