Nucleosides-based identification model for Fritillariae Cirrhosae Bulbus / 中国中药杂志

A high performance liquid chromatography( HPLC) method was established for the fast,and precise determination of ten nucleosides in Fritillariae Cirrhosae Bulbus and its counterfeits. Then multivariate statistical analyses,such as clustering analysis,principal component analysis( PCA),and Fisher' s linear discriminant analysis( LDA),were conducted to establish a discriminant function model for an integrated analysis. The results indicated that data acquisition time of a single sample was shortened within 16 min by the HPLC method. In the range of 5-1 000 mg·kg~(-1),the mass concentrations of all nucleosides exhibited good linear relationships with the corresponding peak areas( R2> 0. 999). The spiked recoveries were in the range of 93. 83%-108. 9% with RSDs of0. 12%-1. 3%( n = 5). The limit of quantitation( LOQ) was 0. 98-4. 13 mg·kg~(-1). As revealed by the clustering analysis,Fritillariae Cirrhosae Bulbus and the counterfeits could be discriminated into two clusters based on the content of nucleosides. Fisher's LDA could achieve this discrimination,while PCA dimension reduction failed. The accuracy of the discriminant function model established on the screened characteristic indicators reached 97. 5%. The present study proposed a new identification method of Fritillariae Cirrhosae Bulbus with one-dimensional indicators,which is simple,accurate,and reliable. It can provide a scientific basis for further optimizing the identification techniques for Fritillariae Cirrhosae Bulbus and inspiration for quality control strategy development of Chinese medicinal materials.

A rapid identification of authenticity and specifications of Chinese medicine Fritillariae Cirrhosae Bulbus based on E-eye technology / 中国中药杂志

The quality of traditional Chinese medicine tablets is correlated with clinical efficacy and drug safety, and plays a great role in promoting the development of traditional Chinese medicine. However, the existing traditional artificial identification and modern instrument detection in terms of accuracy and timeliness have both advantages and disadvantages. Therefore, how to quickly and accurately identify the quality of traditional Chinese medicine tablets has become a high-profile issue. The purpose of this paper is to explore the feasibility of the application of electronic eye technology in the study of rapid identification of traditional Chinese medicine quality. A total of 80 batches of samples were collected and tested by Fritillariae Cirrhosae Bulbus for traditional empirical identification(M_1) and modern pharmacopeia(M_2). The optical data was collected from electronic eyes, and the chemical metrology was used to establish suitable discrimination models(M_3). Four authenticity and commodity specification models, namely identification analysis(DA), minimum bidirectional support vector machine(LS-SVM), partial minimum two-multiplier analysis(PLS-DA), main component analysis identification analysis(PCA-DA), were established, respectively. The accuracies of the authenticity identification models were 82.5%, 90.0%, 96.2% and 93.8%, while the accuracies of the commodity specification identification models were 89.3%, 96.0%, 90.7% and 97.3%, respectively. The models were well judged, the authenticity identification was based on the final identification model of PLS-DA, and the commodity specification was based on the final identification model of PCA-DA. There was no significant difference between its accuracy and M_1, and the time of determination was much shorter than M_2(P<0.01). Therefore, electronic-eye technology could be used for the rapid identification of the quality of Fritillariae Cirrhosae Bulbus.

Breeding and variety characteristics of a new variety of Fritillaria thunbergii "Zhebei 3" / 中国中药杂志

A new variety "Zhebei 3(Zhejiao Pharmaceutical 2018002)" was selected and bred from multi seeded Fritillaria thunbergii mutants by systematic breeding method. From 2012 to 2016, the traits assessment, disease resistance appraisal, plot ratios and regional trials of the variety were continuously carried out. The results showed that "Zhebei 3" emerged early and had late seedlings. The average growth period was about 100 days, which was 6 days and 12 days higher than the "Zhebei 1" and "Zhebei 2". The average yield was 5 095.5 kg·hm~(-2), which was 14.42% and 17.71% higher than of the control respectively. The average proliferation rate of bulbs was 261.2%, which was 37.46% and 31.58% higher than that of the control, respectively. The propagation coefficient of bulbs was about 1∶2.6, and the total amount of peimine and peiminine was 0.172 2%, which was 4.49% and 29.47% higher than the control, respectively. The identification of disease resistance showed that it was resistance to bulb stem(soft) rot, better than the control. "Zhebei 3" has stable characters, high yield, good quality, strong disease resistance, and moderate propagation coefficient which is suitable for planting in Zhejiang province.

Studies on alkaloid constituents of Fritillaria yuminensis / 中国中药杂志

Twelve alkaloids were isolated from the bulbs of Fritillaria yuminensis by column chromatography over silica gel, ODS, and Sephadex LH-20, as well as RP-HPLC. Their structures were identified mainly by NMR and MS analyses as yubeinine(1), imperialine(2), delavinone(3), tortifoline(4), hupehenizioiside(5), imperialine-β-D-glucoside(6), kuroyurinidine(7), pengbeisine A(8), walujewine A(9), peimisine-3-O-β-D-glucopyranoside(10), solanidine-3-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside(11), and solanidine-3-O-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside(12). Compounds 4-12 were obtained from F. yuminensis for the first time.

Correlation between rhizospheric microorganisms distribution and alkaloid content of Fritillaria taipaiensis / 中国中药杂志

In order to reveal the relationship between the amount of soil microorganisms and the quality of Fritillaria taipaiensis, both cultivated and wild F. taipaiensis were collected from Chongqing, Wuxi at different stages of their growth as objects of the research. The mycorrhizal infection rate and colonization intensity, peimisine and total alkaloid content in bulbs, the amount of microorganisms and biomass carbon content in rhizospheric soil were all determined using common methods. The results showed that the typical arbuscular-vesicle roots were formed after the AM fungi infected the F. taipaiensis roots which were collected from different origins. The mycorrhizal infection rates were ranged from 78.74% to 98.68% and the colonization intensities were ranged from 13.29% to 37.06%. The rhizospheric microorganisms of F. taipaiensis showed abundant resources. The distribution rule of them in the rhizospheric soil was as follows: the amount of bacteria>the amount of actinomycetes>the amount of fungi. The rhizospheric bacteria, decomposition inorganic phosphorus bacteria, decomposition organic phosphorus bacteria, actinomycetes amount and the total number of microbes increased first and then decreased with the increase of years, while decomposition potassium bacteria showed decreasing trend and fungi showed gradual increasing trend. The soil microbial flora content in the soil changed from "bacterial type" with a high fertility to "fungal type" with a low fertility. The mass fraction of peimisine and total alkaloid content increased first and then decreased with the increase of over the years, the same trend of culturable rhizosphere soil bacteria and actinomycetes indicated that the growth years affected the quality of soil and medicinal materials on different levels. Therefore, the diversity of microbial communities in rhizosphere soil reduced with the increase of years leading to the continuous cropping obstacles and the destruction of medicinal quality of F. taipaiensis.

Cloning and Characterization of the Gene Encoding 3-hydroxy-3- Methylglutaryl-coenzyme A (HMG-CoA) Reductase from Fritillaria Cirrhosa D. Don

ABSTRACT The enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR; EC1.1.1.34) catalyzes the first committed step of isoprenoids biosynthesis in Mevalonate (MVA) pathway. Here we report for the first time the cloning and characterization of a full-length cDNA encoding HMGR from Fritillaria cirrhosa (FcHMGR), a bulbous medicinal plant. The full-length cDNA of FcHMGR was 2072 base pair (bp), containing a 1680-bp open reading frame. Bioinformatical analyses revealed that FcHMGR had HMG CoA-binding domains and two NADPH binding domains, which are required for HMGR activity. Quantitative real-time PCR (qRT-PCR) analysis revealed that FcHMGR expressed high in mature bulbs. A truncated version of FcHMGR protein lacking the N-terminal 249-bp GC rich area was expressed in Escherichia coli. The crude cell lysate containing the recombinant protein showed a better HMGR activity than the control and the relative enzyme activity was calculated to be 1.62 U/mg. The cloning, characterization and functional analysis of FcHMGR gene allowed us to further understand the role of FcHMGR involved in steroidal alkaloid biosynthetic pathway in F. cirrhosa at the molecular level.

Study on accumulation law of peimine in different Fritillaria thunbergii cultivar / 中国中药杂志

In order to study the accumulation of Fritillaria thunbergii cultivar, peimine content in Xiaye, Kuanye, Duozi and Xiaosanzi bulbs of different sizes and parts was determined by HPLC-ELSE. The results indicated that the peimine content varied significantly with the cultivar type, the size and part of bulb. The distribution laws of peimine were as follow: Xiaosanzi > Duozi > Xiaye > Kuanye, small-size bulb > big-size bulb, core bud > scale. The peimine yield per plant in Duozi was the highest.

Differences of fungal diversity and structure in rhizosphere of Fritillaria thunbergii from different provenances / 中国中药杂志

To explore the mechanism of soil microbial ecology, the differences of fungal diversities in rhizosphere of different provenances of Fritillaria thunbergii were analyzed. The diversities and compositions of rhizo-fungi of the samples were analyzed by using DGGE and 454 pyrosequencing. DGGE results showed the Shannon index of Ninbo provenance planted in Ninbo was the highest one. And its dominant fungi were Ascomycota, Deuteromycota and Zygomycota. Except the same fungi, every provenance planted in Ninbo had its own special ones. From the 454 pyrosequencing, the fungal diversity in Panan producing was the highest which was similar with DGGE result. Among the ten phylum detected in its rhizosoil, Fungi_incertae_sedis, Ascomycota, Mucoromycotina, Basidiomycota and Chytridiomycota almost amounted to 90% of the whole community. The fungal types and amounts in Panan were more than those in Ninbo indicating the differences between producing areas and the advantage of macro genome sequencing. There were 10 phyla, 29 families, 28 genus and 159 species of fungi in Panan provenance, 6 phyla, 20 families, 19 genus, 136 species in Ninbo provenance, 8 phyla, 37 families, 47 genus, 289 species in Nantong provenance and 7 phyla, 25 families, 24 genus, 102 species in the bulk soil. Some genus such as Dothidea, Capnobotryella and Conidiobolus were only existed in Nantong provenance, while Pyrenochae- ta, Glomus and Pseudonectria were only in Panan provenance, which implied these species could grew because F. thunbergii influenced the existence of fungi. Experiments of provenance and producing area of F. thunbergii showed that the fungal diversity of indigenous provenance was higher than that of exotic provenance and each provenance had unique fungal species in the rhizosphere, which indicated that the diversity and structure was shaped cooperatively by the species and soil type. These fungal species are interacted with the soil-rhizhosphere-microbe microecological system, which in turn influence the growth of F. thunbergii.

Pollination biology of Fritillaria delavayi / 中国中药杂志

In this study, the processes of pollination ecology of Fritillaria delavayi were investigated to document its reproductive characteristics. Some individuals of F. delavayi could produce seeds under bagging without emasculation (11%), but the rate was significantly lower than that of the natural control (87%). It is suggesting that pollination of F. delavayi largely depends on pollen vectors. Bombus sushikini was the only effective pollinator of F. delavayi and the visitation frequency was 0.003 time xXflower(-1) x min(-1). Flowering of F. delavayi in whole population lasted for 35 d and single flower for 11 d. Pollen viability and stigma receptivity lasted for 9 d and were relatively long compared with other Fritillaria genus plants. Consequently, bumblebee pollination and long floral longevity seem to be important for reproductive assurance of F. delavayi in harsh alpine environments.

Phylogenetic analysis for Fritillaria hupehensis: evidence from ITS, rpl16 and matK sequences / 中国中药杂志

The systematic position of Fritillaria hupehensis has been in dispute. Phylogentic analyses were conducted on sequences of ITS, rpl16, matK sequences for species of F. hupehensis and allies. Lilium davidii was designed as outgroup. The analyses were performed using MP and ML methods. Conclusions could be achieved as follow. The topologies of MP and ML are consistent. The samples of F. hepehensis from different places form a supported clade with a strong bootstrap. And then form a strongly supported clade with F. anhuiensis, F. monantha. The results suggests that although F. hupehensis has a closet relation with the two ones, it exists some difference.

Transposable Elements and Genome Size Variations in Plants

Although the number of protein-coding genes is not highly variable between plant taxa, the DNA content in their genomes is highly variable, by as much as 2,056-fold from a 1C amount of 0.0648 pg to 132.5 pg. The mean 1C-value in plants is 2.4 pg, and genome size expansion/contraction is lineage-specific in plant taxonomy. Transposable element fractions in plant genomes are also variable, as low as ~3% in small genomes and as high as ~85% in large genomes, indicating that genome size is a linear function of transposable element content. Of the 2 classes of transposable elements, the dynamics of class 1 long terminal repeat (LTR) retrotransposons is a major contributor to the 1C value differences among plants. The activity of LTR retrotransposons is under the control of epigenetic suppressing mechanisms. Also, genome-purging mechanisms have been adopted to counter-balance the genome size amplification. With a wealth of information on whole-genome sequences in plant genomes, it was revealed that several genome-purging mechanisms have been employed, depending on plant taxa. Two genera, Lilium and Fritillaria, are known to have large genomes in angiosperms. There were twice times of concerted genome size evolutions in the family Liliaceae during the divergence of the current genera in Liliaceae. In addition to the LTR retrotransposons, non-LTR retrotransposons and satellite DNAs contributed to the huge genomes in the two genera by possible failure of genome counter-balancing mechanisms.

Phytochemical and biological research of Fritillaria medicine resources / 中国天然药物

The genus Fritillaria is a botanical source for various pharmaceutically active components, which have been commonly used in traditional Chinese medicine for thousands of years. Increasing interest in Fritillaria medicinal resources has led to additional discoveries of steroidal alkaloids, saponins, terpenoids, glycosides and many other compounds in various Fritillaria species, and to investigations on their chemotaxonomy, molecular phylogeny and pharmacology. In continuation of studies on Fritillaria pharmacophylogeny, the phytochemistry, chemotaxonomy, molecular biology and phylogeny of Fritillaria and their relevance to drug efficacy is reviewed. Literature searching is used to characterize the global scientific effort in the flexible technologies being applied. The interrelationship within Chinese Bei Mu species and between Chinese species, and species distributed outside of China, is clarified by the molecular phylogenetic inferences based on nuclear and chloroplast DNA sequences. The incongruence between chemotaxonomy and molecular phylogeny is revealed and discussed. It is essential to study more species for both the sustainable utilization of Fritillaria medicinal resources and for finding novel compounds with potential clinical utility. Systems biology and omics technologies will play an increasingly important role in future pharmaceutical research involving the bioactive compounds of Fritillaria.

Rat intestinal absorption trait of peimine and peiminine in Thunberg fritillary bulb extract / 药学学报

To study the in situ intestinal absorption kinetics and compatibility influence of peimine and peiminine in rats, the absorption of peimine and peiminine in small intestine (duodenum, jejunum and ileum) and colon of rats was investigated using in situ single-pass perfusion method and the drug content was measured by HPLC-ELSD. Perfusion rate, pH, concentration of drug, gender and bile duct ligation can significantly affect the absorption of peimine and peiminine, the Ka, and Papp values in the condition of pH 6.8 and pH 7.4 had significant difference (P<0.01), as drug concentration irlcreased, the absorption parameters of peimine and peiminine decreased, Ka and Papp between low concentrations and middle concentrations was significant difference (P<0.01). Verapamil can not affect Ka and Papp of peimine and peiminine which are in the extract (P> 0.05). Bitter almonds and licorice can significantly reduce the absorption of peimine and peiminine with the usual dose (P<0.01), extracted separately and together had no significant difference on Ka and Papp (P> 0.05). Experimental results show that the absorption features of peimine and peiminine are basically the same, both of them could be absorbed at all segments of the intestine in rats and had no special absorption window, and with significant differences between male and female individuals. The absorption of peimine and peiminine complies with the active transport and facilitated diffusion in the general intestinal segments. Bitter almond and licorice can reduce the intestinal absorption rate ofpeimine and peiminine.

Fingerprint analysis of Fritillaria thunbergii using rapid resolution liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry / 中国中药杂志

A fingerprint method for quality assessment of Fritillaria thunbergii was developed by rapid resolution liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (RRLC-Q-TOF-MS). The separation was performed using Agilent Eclipse Plus C18 column (2.1 mm x 100 mm, 1.8 microm) by gradient elution with acetonitrile and 0.1% formic acid aqueous solution (containing 10 mmol x L(-1) ammonium formate) as the mobile phase. Q-TOF-MS was used to obtain the accurate mass for precursor and product ions. Under this chromatographic and MS condition, 12 batches of F. thunbergii and its adulterants (F. hupehensis and F. pallidiflora) were analyzed by RRLC-Q-TOF-MS. Fifteen steroidal alkaloids were identified from F. thunbergii, F. hupehensis and F. pallidiflora and nine were assigned as the common characteristic peaks for F. thunbergii. The RRLC-Q-TOF-MS fingerprint of F. thunbergii was different significantly with those of F. hupehensis and F. pallidiflora. The quality of 12 batches of F. thunbergii samples were finally evaluated by hierarchical clustering analysis (HCA) and principle component analysis (PCA). This convenient and high specific method could be used to identify and evaluate the quality of the F. thunbergii.

Karyotype and pollen morphology of main Fritillaria thunbergii cultivars / 中国中药杂志

<p><b>OBJECTIVE</b>To study the differences in pollen morphology and karyotype among main Fritillari thunbergii cultivars.</p><p><b>METHOD</b>Pollen morphologies of three F. thunbergii cultivars were examined by scanning electron microscopy (SEM), and the chromosome numbers and karyotypes were studied by applying traditional squash technique.</p><p><b>RESULT</b>The pollen shape of F. thunbergii (Xiaye) was ovoid, while those of the other F. thunbergii (Kuanye, Duozi) were oblong. There were significant differences in mesh ridge width, mesh size among three F. thunbergii cultivars. The karyotype formula ofthree cultivars were as follows: F. thunbergii (Xiaye) was 2n =2x =3m +1sm + 8st(2SAT) + 12t(4SAT), F. thunbergii (Kuanye) was 2n = 2x =2m + 2sm + 10st(2SAT) + 10t (2SAT), F. thunbergii (Duozi) was 2n =2x = 24 =2m +2sm +5st(2SAT) +15t(4SAT). The three cultivars of karyotype belonged to 3B; There were the heterozygosity of homologous chromosome in both F. thunbergii (Xiaye) and F. thunbergii (Duozi).</p><p><b>CONCLUSION</b>The genetic diversity of F. thunbergii is very rich, which could enhance the adaptability, and lay the foundations for new variety selection of F. thunbergii.</p>

Valuation on analgesic, expectorant and antitussive effects of compatible use of Aconiti radix cocta and Fritillaria cirrhosa or Fritillaria thunbergii / 中国中药杂志

<p><b>OBJECTIVE</b>To study the analgesic, expectorant and antitussive effects of the compatible use of Aconiti Radix Cocta and Fritillaria cirrhosa or F. thunbergii with different matching ratio or dose in mice.</p><p><b>METHOD</b>The two-factor, seven-level uniform design method was adopted to observe the analgesic, expectorant and antitussive effects of the oral administration with the two combined decoctions in rats, with frequency of body torsions induced by acetum, secretion of phenol red in tracheas and frequency of coughs as indexes. Significant matching proportions and doses were collected for verification.</p><p><b>RESULT</b>The effect on the frequency of body torsions: The combined decoctions could effectively reduce the frequency of body torsions. According to a regression analysis, Aconiti Radix Cocta and F. cirrhosa had the synergistic effect, which was maximized with a ratio of 1: 1. The 1: 1 combined decoction played the least role in reducing the frequency of body torsions with a total dose of more than 5 g x kg(-1). The effect on the secretion of phenol red in tracheas. The combined decoctions could effectively increase the secretion of phenol red in tracheas. According to a regression analysis, Aconiti Radix Cocta and F. thunbergii had the antagonism, which was maximized at the ratio of 1: 1, and minimized with a total dose of less than 10 g x kg(-1) and a ratio of 5: 1 between F. thunbergii and Aconiti Radix Cocta. The effect on the frequency of coughs. The combined decoctions could effectively reduce the frequency of coughs. According to a regression analysis, Aconiti Radix Cocta and F. cirrhosa had the antagonism, which was maximized at the ratio of more than 1: 5 and less than 10: 1. There was no interaction between Aconiti Radix Cocta and F. thunbergii. F. thunbergii could reduce the frequency of coughs, whereas Aconiti Radix Cocta showed no effect.</p><p><b>CONCLUSION</b>The compatible application of Aconiti Radix Cocta and F. cirrhosa could enhance the analgesic effect of Aconiti Radix Cocta and reduce the expectorant and antitussive effects of F. cirrhosa, which vary according to different matching ratio and dose. The compatible application of Aconiti Radix Cocta and F. thunbergii shows no effect on the antitussive effect of F. thunbergii. This study provides experimental basis for in-depth studies on the combined effect of Aconiti Radix Cocta and Fritillaria--two of eighteen incompatible pairs.</p>

Taxonomic Study of Amanita Subgenus Lepidella and Three Unrecorded Amanita Species in Korea

Amanita Pers. is a well-known monophyletic mushroom genus with a broad distribution. However, the diversity of Korean Amanita species has been underestimated, and most taxonomic studies conducted in Korea have only investigated their morphological characteristics. This approach is frequently insufficient for correct identification in fungal classification; therefore, we constructed a phylogeny of Amanita subgen. Lepidella in order to understand the phylogenetic placements of 16 Amanita specimens collected in Korea in 2012. The phylogeny constructed using the sequence data of the internal transcribed spacers and the partial large subunit of ribosomal RNA identified nine Amanita species (A. citrina, A. excelsa var. spissa, A. flavipes, A. fritillaria, A. oberwinklerana, A. pallidorosea, A. rubescens, A. subjunquillea, and A. volvata); of these, A. fritillaria, A. oberwinklerana, and A. pallidorosea are new to Korea.

Clinical study on effect of compound granule prescription of thunberg fritillary bulb in relieving post-chemotherapy bone marrow suppression in RAL patients / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of compound granule prescription of thunberg fritillary bulb in relieving the post-chemotherapy bone marrow depression in refractory acute leukemia (RAL) patients.</p><p><b>METHOD</b>Two hundred and thirty eight RAL patients collected from 7 third-grade class-A hospitals were randomly divided into the treatment group and the control group. Both groups were treated with conventional chemotherapy. They were administered with compound granule prescription of thunberg fritillary bulb or placebo three days before chemotherapy for consecutively 14 days. A standardized chemotherapy course was a treatment cycle. The changes in peripheral hemogram of two groups were detected before and after chemotherapy.</p><p><b>RESULT</b>After the treatment, the white blood cell counts of the two groups were significantly different (P < 0.05), but there was no statistical significance in qualitative comparison. Before and after the treatment, the difference of the white cell counts of the two groups detected had significant statistics (P < 0.05). The white cell counts of both groups declined after chemotherapy, but the treatment group decreased more significantly than the control group. Before and after the treatment, there were no statistical significances in quantitative and qualitative comparisons both in HGB and PLT. After the treatment, HGB and PLT contents of both groups declined, but there was no statistical difference between the two groups.</p><p><b>CONCLUSION</b>Compound granule prescription of thunberg fritillary bulb can relieve the bone marrow suppression in RAL patients caused by the chemotherapy, which is mainly reflected by the slowdown of reduction in white blood cells.</p>

Study on water-soluble constituents of Fritillaria unibracteata by HPLC fingerprints / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a HPLC fingerprint of water-soluble constituents of Fritillaria unibracteata.</p><p><b>METHOD</b>Zorbax SB Aq C18 chromatographic column (4. 6 mm x 250 mm, 5 microm) was adopted for gradient elute with the mobile phase consisting of methanol and water. The flow rate was 1.0 mL x min(-1); the detection wavelength was 260 nm, and the temperature of sample manager was set at 25 degrees C. Similarity Evaluation System for Chromatographic Fingerprint of traditional Chinese medicine (version 2.0) published by the State Pharmacopeia Committee of China was adopted for the fingerprint analysis on the 11 batches of F. unibracteata herbs.</p><p><b>RESULT</b>The 11 batches of F. unibracteata herbs had 14 common peaks, nine of which were identified with good separating degrees. The similarities of the 11 batches were more than 0. 970, with good quality homogeneity.</p><p><b>CONCLUSION</b>The method is so accurate, highly reproducible and stable that it is suitable for the comprehensive quality evaluation of F. unibracteata herbs.</p>

Comparison of different methods for isolating total RNA from bulblet of Fritillaria anhuiensis / 中国中药杂志

<p><b>OBJECTIVE</b>To optimize a simple and effective method for total RNA extraction from bulblet of Fritillaria anhuiensis.</p><p><b>METHOD</b>Four methods, i. e. guanidine isothiocyanate, bentonite, modified SDS/phenol and the RNAiso plus, were used to extract total RNA from bulblet of F. anhuiensis. Then the results of the extraction were compared and analyzed by electrophoresis detection and RT-PCR verification.</p><p><b>RESULT</b>The total RNA extracted by bentonite method were clear and no dispersion, the integrity of the RNA was well, and there was no obvious contamination with DNA and other impurities, was suitable for RT-PCR test.</p><p><b>CONCLUSION</b>The bentonite method is quick, economic, and efficient for total RNA extraction from bulblet of F. anhuiensis.</p>