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Background@#In 2021, 5981 of cancer new cases was registered in Mongolian population. Among those cases, liver cancer was commonly registered with a prevalence of 32.7%. Studies on anticancer agents with no-adverse effects and good-preventive efficacy against cancer have been attracted more attention from the researchers in the field of pharmaceutical sciences. Scutellaria baicalensis Georgi, Saussurrea amara.L, Chiazospermum erectum Berh, and Carthamus tinctorius.L are well recognized as effective agent against liver diseases. Using these raw materials, researchers have been invented a traditional prescription and named as Hepaclin-4. In this study, we aimed to investigate the qualitative study of raw materials and some biologically active sub- stances in the compounds.@*Purpose@#To study the qualitative study of raw materials for Hepaclin-4 prescription@*Materials and methods@#Some qualitative properties of raw materials for Hepaclin-4 prescription, including appearance, minerals, some organic compounds, total ash, water-soluble substances and fungi, were investigated according to Mongolian pharmacopeia and total flavonoid was detected by thin layer chromatography.@*Results@#No changes were observed on the appearance of raw materials, and minerals and organic compounds weren’t detected in the prescription. No contamination with fungi and insects were identified. The moist in the raw materials were 5.9 to 8.1%, total ash was 4.7 to 13.3% and the water-soluble substances were detected 33.8 to 42.9%. Number of aerobic bacteria, fungi and E.coli, Salmonella species were detected in normal range, indicating that the prescription was matched with the requirement of pharmacopeia. According to the thin layer chromatography study of the raw materials, a yellow spot on the chromatogram were identified and same as quercetin (Rf=0.9-0.98) and rutin ((Rf=0.18-0.23)) as standard compounds, which indicated that the spot which indicated that the spot was flavonoids in the prescription.@*Conclusions@#These results showed that the appearance, moist, minerals, organic compound, water-soluble substances, ash and biologically active substances of the raw materials for Hepaclin-4 prescription was corresponded with the requirements of pharmacopeia, and flavonoid was detected in raw materials of Hepaclin-4.
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OBJECTIVE@#Flavonoids are the bioactive compounds in safflower (Carthamus tinctorius), in which chalcone synthase (CHS) is the first limiting enzyme. However, it is unclear that which chalcone synthase genes (CHSs) are participated in flavonoids biosynthesis in C. tinctorius. In this study, the CHSs in the molecular characterization and enzyme activities were investigated.@*METHODS@#Putative chalcone biosynthase genes were screened by the full-length transcriptome sequences data in C. tinctorius. Chalcone biosynthase genes in C. tinctorius (CtCHSs) were cloned from cDNA of flowers of C. tinctorius. The cloned gene sequences were analyzed by bioinformatics, and their expression patterns were analyzed by real-time PCR (RT-PCR). The protein of CtCHS in the development of flowers was detected by polyclonal antibody Western blot. A recombinant vector of CtCHS was constructed. The CtCHS recombinant protein was induced and purified to detect the enzyme reaction (catalyzing the reaction of p-coumaryl-CoA and malonyl-CoA to produce naringin chalcone). The reaction product was detected by HPLC and LC-MS.@*RESULTS@#Two full-length CtCHS genes were successfully cloned from the flowers of safflower (CtCHS1 and CtCHS3), with gene lengths of 1525 bp and 1358 bp, respectively. RT-PCR analysis showed that both genes were highly expressed in the flowers, but the expression of CtCHS1 was higher than that of CtCHS3 at each developmental stage of the flowers. WB analysis showed that only CtCHS1 protein could be detected at each developmental stage of the flowers. HPLC and LC-MS analyses showed that CtCHS1 could catalyze the conversion of p-coumaryl-CoA and malonyl-CoA substrates to naringin chalcone.@*CONCLUSION@#CtCHS1 is involved in the biosynthesis of naringin chalcone in safflower.
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UDP-glucose: flavonoid 3-O-glucosyltransferase (UF3GT) uses flavones, dihydroflavonol or anthocyanin as the acceptor and uridine 5′-diphosphate-sugar as the donor to catalyze the production of flavonoid 3-O-glycoside compounds. Based on sequence homology and transcriptome data, we screened and cloned a UF3GT gene named CtUF3GT (GenBank No. OM948976) from safflower. Biological information analysis demonstrate that CtUF3GT has highly conserved PSPG motif. The open reading frame of CtUF3GT is 1 446 bp, encoding 481 amino acids, with a presumed molecular weight of 52.36 kD and a theoretical isoelectric point of 5.33. Multiple sequence alignment indicate that CtUF3GT has a high homology with UF3GT from Asteraceae, and phylogenetic analysis showed that CtUF3GT clusters with functional identified UF3GTs from other species. The purified recombinant protein glucosylated kaempferol and quercetin to biosynthesis of kaempferol 3-O-glucoside and quercetin 3-O-glucoside, respectively. And CtUF3GT prefered to use kaempferol as substrate. qRT-PCR analysis showed that the UF3GT gene was most highly expressed in flowers, followed by leaves, with very low expression in bracts and stems, and no expression in roots. The expression of UF3GT gene showed a trend of increasing and then decreasing at different stages of flower development. The expression of CtUF3GT gene in safflower with different flower color was highly significant (P < 0.01) at S1, S2, S5, S6 and S7 stages of flower development, in which the expression of CtUF3GT in white safflower was 5.3 and 3.1 times higher than that in red safflower at S6 and S7 stages. This study lays the foundation for further exploring the role of CtUF3GT in the mechanism of safflower flavonoid secondary metabolite biosynthesis and accumulation.
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ABSTRACT: The safflower (Carthamus tinctoriusL.) has an uneven flowering and fruiting, which can cause problems in seed production and harvesting in regions with hot and humid climates. However, little is known about the optimal safflower harvest time. Therefore, this study evaluated the optimumtiming for seed harvest of three safflower genotypes (2106, S-325, and 7329).The experiment was a randomized complete block design with six replications. The harvest started 16 days after flowering (DAF) and ended at 52 DAF. Ten harvests were made in total. Seed water content, seeds fresh and dry matter, seed germination, and first germination counts were evaluated.Genotypes 2106 and 7329 had germination rates of 79% and 91%, respectively, at 34 and 38 DAF, while genotype S-325 had 90% germination at 37 DAF. Harvesting at 52 DAF combined with a rainy season impaired the germination of safflower seeds. The harvest time most suitable for safflower occurred between 34 and 42DAF, when the seeds have the seed water content between 26% and 33%.
RESUMO: O cártamo (Carthamus tinctoriusL.) apresenta flores e frutos irregulares, o que pode causar problemas na produção e colheita de sementes em regiões com clima quente e úmido. No entanto, pouco se sabe sobre o tempo ideal de colheita de cártamo. Portanto, o objetivo desteestudo foi avaliar o momento ideal para a colheita de sementes de três genótipos de cártamo(2106, S-325 e 7329). O experimento foi delineado em blocos casualizados, com seis repetições. A colheita começou 16 dias após o florescimento (DAF) e terminou aos 52 DAF. Dez colheitas foram feitas no total. Foram avaliados o teor de água das sementes, matéria fresca e seca das sementes, germinação das sementes e primeira contagem de germinação. Os genótipos 2106 e 7329 tiveram taxas de germinação de 79% e 91%, respectivamente, aos 34 e 38 dias após a floração (DAF), enquanto o genótipo S-325 teve 90% de germinação aos 37 DAF. A colheita aos 52 DAF combinada com uma estação chuvosa prejudicou a germinação das sementes de cártamo. A época de colheita mais adequada para o cártamo ocorreu entre 34 e 42 DAF, quando as sementes apresentam teor de água entre 26% e 33%.
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Objective: In order to obtain new glycosyltransferases with highly efficient catalysis, the glycosyltransferases from Carthamus tinctorius which contains diverse types of glycosides were mined. Methods: A new glycosyltransferase gene (UGT88B2) with full length was obtained by PCR and further transformed into Escherichia coli for heterologous expression. The catalytic activity of recombinant UGT88B2 was determined by HPLC-MSn. The structures of representative catalytic products were elucidated by MS and NMR. Results: UGT88B2 exhibited catalytic promiscuity and various patterns in glycosylation of flavonoids with high efficiency. Conclusion: A new glycosyltransferase named UGT88B2 was successfully mined and can be employed as enzymatic tools in glycosylation of flavonoids.
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We determined a component-target-disease network for Carthamus tinctorius L. and the key compounds, identified by topological analysis, were related to vasculitis, coronary heart and cerebrovascular disease. Based on these compounds, the chromatographic fingerprint of Carthamus tinctorius L. was established. Firstly, 132 compounds were obtained from TCMID and TCMSP databases. Their targets were predicted in the PharmMapp and HemMapper databases. CardioGenBase, Therapeutic Target Database and DisGeNET databases were used to collect targets of vasculitis, coronary heart disease and cerebrovascular disease. The corresponding relationships between component and target protein were established by mapping. Finally, the "component-target-disease" network was built with Cytoscape software. The core network and key nodes were analyzed with the Cytohubba plug-in. The results showed that the 24 key compounds were alpha-tocopherol, adenosine, quinone chalcone pigments such as hydroxysafflor yellow A, safflower yellow, quercetin, kaempferol and other flavonoids, organic acids such as stearic acid, linolenic acid, coumaric acid and cinnamic acid. This resulting chromatographic fingerprint of Carthamus tinctorius L. showed good consistency, and the core chemical compounds obtained by topological analysis of the network of "component-target-disease", could be used as quality control markers. Our research provides a new approach for the identification of quality control indicators in Chinese medicinal materials.
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Objective: To optimize the preparation technology of phospholipid complex of Carthamus tinctorius (safflower) extract and investigate its permeability. Methods: On the basis of single factor experiment, the preparation process was optimized by using the response surface analysis method, taking the compound rate of phospholipid complex of safflower extract as the index. It was characterized by UV-vis absorption spectrum and infrared spectrum. The modified Franz diffusion cell was used to evaluate the membrane permeability of safflower extract and phospholipid complex of safflower extract with different drug-lipid ratios in vitro. Results: The optimum preparation technology of phospholipid complex of safflower extract was as follows: methanol was used as compound solvent, the concentration of safflower extract was 5.0 mg/mL, and the mass ratio of phospholipid to phospholipid was 1∶1, the reaction time was 1.5 h, and the reaction temperature was 55 ℃. The results of transmembrane experiment showed that the 24-hour cumulative permeability (Q24) of safflower extract phospholipid complex with drug-fat ratio of 2, 1, and 0.5 was (15.07 ± 1.24), (15.61 ± 0.92), (21.94 ± 1.54), and (21.05 ± 1.39) μg/cm2, respectively. Conclusion: The optimized preparation process is reasonable and feasible, and the phospholipid complex of safflower extract can obviously improve its membrane permeability.
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Objective: To establish a Logistic model for quality assessment of Carthamus tinctorius based on content determination and bioactivity. Methods: A method to determinate hydroxy safflor yellow A by using ultra-high performance liquid chromatography (UPLC) was proposed. The activity of anticoagulant was reflected by thrombin time (TT) and the anti-oxidant activity was expressed by scavenging ability of hydroxyl radical and DPPH radical. Later, a Logistic model grade evaluation of C. tinctorius was constructed based on correlation analysis between content and biological activity Logistic.Results: The constructed Logistic model had outstanding stability and high prediction accuracy of grades and it divides 19 batches of C. tinctorius into four grades. Conclusion: The Logistic model based on content determination and biological activity is applicable to assess quality of C. tinctorius slices and provides some reference for quality control.
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Objective To study the alkaloids from leaves of Carthamus tinctorius. Methods The alkaloids were isolated and purified by silica gel, MCI, Sephadex LH-20 column chromatographies, and semi-preparative HPLC, and their structures were elucidated by physical and spectroscopic analysis. Results A new β-carboline alkaloid, 4,9-dimethoxy-1-ethyl-β-carboline (1) along with one known analogue 4-methoxy-1-ethyl-β-carboline (2), were isolated from the leaves of C. tinctorius. Compounds 1 and 2 showed the cytotoxicity against HepG2 cell lines with IC50 values of (15.2 ± 0.58) μmol/L and (17.4 ± 0.33) μmol/L, respectively. Conclusion Compounds 1 and 2 are firstly obtained from Carthamus genus, and compound 1 is a new compound named carthine A. Both compounds 1 and 2 exhibited cytotoxicity against HepG2 cell lines.
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Objective: To clone a coding region sequence of AP2/ERF transcription factor family from Carthamus tinctorius, and construct a plant expression vector. Methods: A gene (CtERF1) of AP2/ERF family transcription factor was cloned by RT-PCR based on the sequence of C. tinctorius transcription sequencing, the phylogenetic tree was constructed by ClustalW 1.83 software, Spe I and Xba I restriction sites were introduced to construct over-expression vector pBASTA-CtERF1 containing 35S promoter. Results: CtERF1 gene had a functional domain of a typical AP2/ERF gene encoding 297 amino acids, and contained an AP2 region speculated to be located in cytoplasm and nucleus, which was ERF subprotein. Systematic evolution analysis showed that CtERF1 gene had some homology with other plant species, among which the relationship with Populus deltoides and Panax japonicus were the closest. The pBASTA-CtERF1 plant expression vector was constructed successfully by molecular biology. Conclusion: A CtERF1 gene of C. tinctorius AP2/ERF transcription factor family was cloned and the plant expression vector pBASTA-CtERF1 was constructed successfully.
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Objective: To clone CtWD40 transcription factors (TFs) and analyze its expression level in different tissues of Carthamus tinctorius and relationship with the content of hydroxysafflor yellow A (HSYA). Methods: CtWD40 gene was obtained by cloning the WD40 candidate gene from transcriptome of C. tinctorius as reference. Its conserved domain, three-dimensional structure and phylogeny analysis were analyzed by bioinformatics methods. The CtWD40 expression pattern was also analyzed by qRT-PCR method, in the same time, HSYA content in different petals were analyzed by HPLC. Results: Gene sequence of CtWD40 was obtained and eight conserved WD domains were found in CtWD40 gene sequences. Phylogenetic analysis revealed that CtWD40 had a closed homology with WD40 from composite plants. Conclusion: The expression of CtWD40 gene was first increased and then decreased in petal tissues of different flowering stages.Pearson coefficient revealed significant correlation between CtWD40 expression and HSYA content in C. tinctorius petal.
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Objective To explore the protective effects of Salvia miltiorrhiza and Carthamus tinctorius active ingredients in different compatibility on cerebral ischemia-reperfusion injury in rats. Methods Adult male Sprague-Dawley (SD) rats were selected to establish a cerebral ischemia-reperfusion (I/R) injury model by middle cerebral artery occlusion (MCAO) method. The rats were randomly divided into sham operation group, model group, and positive control group (Danhong Injection 2 mL/kg); And orthogonal test method L9(34) was adopted to compose nine compatibility groups from main active ingredients of S. miltiorrhiza and C. tinctorius (trashinol, salvianolic acid A, salvianolic acid B, and HYSA). Rats were tail iv administrated once daily for continuous 3 d. Score neurological deficit was evaluated after 3 d; qRT-PCR was used to detect mRNA expression of glucose regulated protein 78 (GRP-78), nuclear factor-κB (NF-κB), C/EBP homologous protein (CHOP), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6); HE staining was used to detect the pathological changes of cerebral cortex, and the expression of NF-κB p65 protein in cerebral cortex was detected by immunohistochemistry. Results Compared to the model group, Danhong Injection group and orthogonal compatibility nine groups significantly reduced the neurological deficit scores of rats with cerebral ischemia-reperfusion; increased the mRNA expression of GRP-78 in the cerebral cortex; decrease the mRNA expression of NF-κB, CHOP, TNF-α, and IL-6, and decrease the protein expression of NF-κB p65 in the cerebral cortex. The results also showed that the protective effects of the 4th group (danshensu 30 mg/kg, salvianolic acid A 2.5 mg/kg, salvianolic acid B 16 mg/kg, and HYSA 8 mg/kg), 6th group (trashinol 30 mg/kg, salvianolic acid A 10 mg/kg, salvianolic acid B 8 mg/kg, and HYSA 4 mg/kg) in the 9th group were more significant for cerebral ischemia reperfusion injury in rats. Conclusion The active components of S. miltiorrhiza and C. tinctorius can play a good protective role in endoplasmic reticulum stress and inflammation in rats with cerebral ischemia reperfusion injury.
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ABSTRACT Water deficit is one of the major stresses affecting plant growth and productivity worldwide. Plants induce various morphological, physiological, biochemical and molecular changes to adapt to the changing environment. Safflower (Carthamus tinctorius L.), a potential oil producer, is highly adaptable to various environmental conditions, such as lack of rainfall and temperatures. The objective of this work was to study the physiological and production characteristics of six safflower lines in response to water deficit followed by rehydration. The experiment was conducted in a protected environment and consisted of 30 days of water deficit followed by 18 days of rehydration. A differential response in terms of photosynthetic pigments, electrolyte leakage, water potential, relative water content, grain yield, oil content, oil yield and water use efficiency was observed in the six lines under water stress. Lines IMA 04, IMA 10, IMA 14 showed physiological characteristics of drought tolerance, with IMA 14 and IMA 16 being the most productive after water deficit. IMA 02 and IMA 21 lines displayed intermediate characteristics of drought tolerance. It was concluded that the lines responded differently to water deficit stress, showing considerable genetic variation and influence to the environment.
Subject(s)
Stress, Physiological/physiology , Water/physiology , Carthamus tinctorius/physiology , Water/metabolism , Principal Component Analysis , Carthamus tinctorius/growth & development , Carthamus tinctorius/metabolism , Droughts , Fluid TherapyABSTRACT
Objective To establish an HPLC method for the simultaneous determination of hydroxysafflor yellow A, rutin, quercetin, and kaempferol in Carthamus tinctorius L..Methods The Agilent ZORBAX SB-C18 column (4.6 mm × 250 mm, 5 μm) was used with diode array detection. The mobile phase was 0.5% phosphoric acid (A) and methanol (B) to separate the four components by gradient elution at 30℃; the flow rate was 0.8 mL/min; the injection Volume was 10 μL; the detection wavelength was at 360 nm.Results The linear ranges of hydroxysafflor yellow A, rutin, quercetin, and kaempferol were 0.0776–0.7760 μg (r=0.9999), 0.0460–0.4600 μg (r=0.9996), 0.0064–0.0640 μg (r=0.9999) and 0.0128–0.1280 μg (r=0.9997), respectively. The average recoveries of four components were 99.68% with RSD=2.29% (n=6), 99.78% with RSD=1.52% (n=6), 102.03% with RSD=1.02% (n=6), 97.03% with RSD=2.05% (n=6), respectively.Conclusion The method is convenient, accurate, sensitive, stable and repeatable, which can be a method for quality control of Carthamus tinctorius L..
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Objective: To clone the full-length cDNA sequence of DHDPS gene encoding the key enzyme DHDPS in lysing biosynthesis pathway of Carthamus tinctorius, and to construct the plant expression vector. Methods: The dihydrodipicolinate synthase (DHDPS) gene fragment was acquired according to the sequence of transcriptome in C. tinctorius, and the full-length cDNA sequence of CtDHDPS gene from C. tinctorius seeds was cloned by RT-PCR and RACE technologies. The pBasta-DHDPS plant expression vector was constructed using traditional molecular cloning and recombination technique. Results: Bioinformatics analysis showed that the full-length cDNA of CtDHDPS was 1309 bp, open reading frame was 954 bp, encoding a polypeptide of 317 amino acids, the theoretical isoelectric point of the coded protein was 5.93, and the molecular weight was about 34 750.79. The plant expression vector pBasta-DHDPS was successfully constructed by traditional molecular cloning and recombinant technique. Conclusion: The full-length sequence of CtDHDPS gene is obtained and the plant expression vector is successfully constructed, which lays a foundation for the further study on the mechanism of CtDHDPS in regulation of essential amino acid metabolism.
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Objective To clone the MYB transcription factor gene in safflower, the sequence information and gene expression were analyzed to preliminarily identify the MYB transcription factor gene involved in regulation of flavonoid biosynthesis. Methods All the sequences of MYB transcription factor gene that was reported to involved in regulation of flavonoid biosynthesis were analyzed. The degenerate primers were designed to clone the core sequence. The full length of MYB transcription factor genes were cloned by RACE. The bioinformatics were used to analyze the sequences. The expression of MYB transcription factor genes in different tissues and different developmental stages of flower were analyzed by semi-quantitative PCR. Results Three candidate MYB transcription factors were cloned, named as CtFRMYB1, CtFRMYB2 and CtFRMYB3. The full lengths of the sequence were 1 223 bp, 1 080 bp and 1 348 bp, respectively. And the molecular weight were 17 878.15, 28 766.45 and 27 987.89, respectively. All three transcription factors have DNA binding domain and belong to the MYB family. Homologous analysis showed that CtFRMYB1 and CtFRMYB2 were closely related to AtMYB12. Expression analysis showed that CtFRMYB1 and CtFRMYB2 were only expressed in flowers, and the expression level was higher in flowering stage 3. Conclusion Three candidate MYB transcription factor genes involved in regulation of flavonoid biosynthesis (CtFRMYB1, CtFRMYB2 and CtFRMYB30) were successfully cloned. Two MYB transcription factor genes (CtFRMYB1 and CtFRMYB2) were identified by bioinformatics and expression analysis. These result laid a foundation for the molecular mechanism analysis of flavonoid biosynthesis.
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Objective: To clone bZIP20 (basic region/leucine zipper motif) gene from Carthamus tinctorius, analyze the expression level in different plant tissues, and construct the plant expression vector. Methods: The bZIP20 gene was cloned by RT-PCR techniques, and the protein characteristics were analyzed by bioinformatics, and phylogenetic tree was constructed. The expression of bZIP20 gene in different tissues and the roots after inoculated by Fusarium oxysporum were analyzed using real time-PCR, and the plant expression vector pBASTA-bZIP20 was constructed. Results: The ORF sequence of bZIP20 gene was 981 bp, encoded a protein of 326 amino acids (GenBank: KT692605). Sequence alignment and phylogenetic tree analyses showed that bZIP20 had 85.41% and 83.99% of consistency with bZIP of Sesamum indicum and Camellia assamica. Real-time PCR results showed significant differences, the highest expression level of bZIP20 gene was detected in flower, and was highest in the bud period, bZIP20 gene was significantly increased in root tissue inoculated with F. oxysporum. The plant expression vector pBASTA-bZIP20 was obtained. Conclusion: The bZIP20 gene of safflower is successfully cloned, and the expression is analyzed. The plant expression vector pBASTA-bZIP20 is constructed.
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Objective: Plant expression vector of chalcone isomerase in safflower was built and its function was verified by over- expression of CHI in Arabidopsis thaliana. Methods: CHI isolated from safflower in our previous study was introduced by BamH I and EcoR I restriction sites and constructed into the over-expression vector pBASTA-CHI containing 35S promoter, transformed into A. thaliana by Flora-dip method. T2 plants of transgenic A. thaliana were detected by PCR and content of total flavones. Results: Plant expression vector containing the safflower CHI gene was built and over expressed in A. thaliana successfully. T2 plants of transgenic A. thaliana were obtained. Conclusion: PCR of transgenic T2 in A. thaliana is detected that CHI gene of safflower has been integrated into the A. thaliana genome, flavone content is determined in leaves, and the results show that the flavone in transgenetic A. thaliana is higher than that in wild type, the highest strain increases by nearly 2.3 times.
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Objective: To investigate the correlation of water and different volume fractions of ethanol boiling point and saturated steam pressure when using the vacuum extraction technology, and to compare the impact on saturation steam pressure of the addition of different medicine materials (Andrographis paniculata, Carthamus tinctorius, Angelica dahurica, Uncaria rhynchophylla, Scutellaria baicalensis, Salvia miltiorrhiza, Citrus reticulata) or the addition of materials with different particle sizes. Methods: To obtain the theoretical value and measured value of saturated steam pressure under different boiling points by the combined method of theoretical calculation, experiment measurement, and statistical analysis Results: The regression equation for the saturated steam pressure (P) with the volume fraction of ethanol solvent (V) and the boiling point (T) was P = 76.467 1+0.035 2 T2+1.201 0 TV-30.749 4 V2-3.123 9 T-14.966 7 V. The results showed that measured values of the saturated steam pressure of pure solvent, water and different concentration of ethanol, was less than the theoretical value. After adding herbs, the measured value of saturated steam pressure was higher than the pure solvent. The saturated steam pressure would reduce exactly as the extraction temperature reaches a certain temperature after the herbs were added. Conclusion: The addition and crush of herbs have influence on the saturated vapor pressure of solvent with small range.
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Objective: To study the establishment of cell suspension culture system and build a platform for exploring the functional genes in the biosynthesis pathway for Carthamus tinctorius. Methods: Using the cotyledon of C. tinctorius as explants for callus induction, callus with good quality was selected for suspension cultivation. The factors influencing the establishment of C. tinctorius cell suspension culture system were investigated, such as hormone, inoculum amount, pH, and cell activity. The effect of MeJA at different concentration on suspension cell growth rate was studied. Chemical compounds in suspension cell system were preliminary identified by UPLC-Q-TOF/MS. Results: The optimal medium for the cell suspension culture was MS + 1.0 mg/LTDZ + 0.1 mg/L NAA, the pH was 5.5-6.0, and inoculum amount was 0.02 g/mL. MeJA at different concentration had effects on suspension cell growth rate: 50 μmol/L MeJA increased cell growth rate, 500 μmol/L MeJA depressed cell growth rate greatly. Thirteen potential chemical compounds in suspension cell system were preliminary identified. Conclusion: The cell suspension culture system of C. tinctorius is established, and MeJA with optimal concentration has effects on suspension cell growth rate.