PLANT CELL REPORTS
·
2010
·Vol. 29
·期 8
·887-894
·
IF 2.301
Wang Cui-Ting,Liu Hua,Gao Xiao-Shu,Zhang Hong-Xia
A number of genes that function in the terpenoid indole alkaloids (TIAs) biosynthesis pathway have been identified in Catharanthus roseus . Except for the geraniol 10-hydroxylase (G10H) gene, which encodes a cytochrome P450 monooxygenase, several of these genes are up-regulated by ORCA3, a jasmonate-responsive APETALA2 (AP2)-domain transcript factor. In this study, the G10H gene was transformed independently, or co-transformed with ORCA3 into C. roseus , using Agrobacterium rhizogenes MSU440. Hairy root clones expressing the G10H gene alone, or both the G10H and ORCA3 genes, were obtained. Alkaloid accumulation level analyses showed that all transgenic clones accumulated more catharanthine, with the highest accumulation level in the transgenic clone OG12 (6.5-fold higher than that of the non-expression clone). Following treatment with ABA, accumulation of catharanthine reached 1.96 mg/g DW in the transgenic clone OG12. The expression levels of TIAs biosynthesis genes in transgenic and non-transgenic clones were also investigated.
研究领域:
植物学
·
代谢工程
·
药用植物研究
✓ 已验证
DOI
PubMed
高影响因子
Journal of Applied Research on Medicinal and Aromatic Plants
·
2024
·
IF 3.8
Yushu Sui,Liben Pan,Nan Yang,Xiaorui Guo,Zhonghua Tang
In the present study, developed a dependable HPLC technique to quantify asarinin and sesamin in Asarum heterotropoides Fr. Schmidt var. mandshuricum (Maxim.) Kitag. The optimal conditions for the ultrasonic-assisted extraction were determined by conducting single-factor experiment, Plackett-Burman design experiment, and Box-Behnken response surface method . Based on actual operation factors and analysis, the optimal extraction parameters were methanol volume fraction of 80% (v/v), liquid-to-material ratio of 10:1 (mL/g), ultrasonic time of 40 min, ultrasonic temperature of 51°C, and ultrasonic power of 300 W. When subjected to these conditions, the average asarinin and sesamin content were found to be 13.40 mg/g and 2.39 mg/g, respectively, with a relative error of 1.04% and 3.46%, which was consistent with the theoretical value. This study can offer a theoretical reference for the extraction of asarinin and sesamin in Asarum .
研究领域:
药学
·
分析化学
✓ 已验证
DOI
高影响因子
MOLECULES
·
2018
·Vol. 23
·期 12
·3276
·
IF 3.098
Chuxin Liang,Chang Chen,Pengfei Zhou,Lv Xu,Jianhua Zhu,Jincai Liang,Jiachen Zi,Rongmin Yu
This study reported the inducing effect ofAspergillus flavusfungal elicitor on biosynthesis of terpenoid indole alkaloids (TIAs) inCatharanthus roseuscambial meristematic cells (CMCs) and its inducing mechanism. According to the results determined by HPLC and HPLC-MS/MS, the optimal condition of theA. flavuselicitor was as follows: after suspension culture ofC. roseusCMCs for 6 day, 25 mg/LA. flavusmycelium elicitor were added, and the CMC suspensions were further cultured for another 48 h. In this condition, the contents of vindoline, catharanthine, and ajmaline were 1.45-, 3.29-, and 2.14-times as high as those of the control group, respectively. Transcriptome analysis showed thatD4H,G10H,GES,IRS,LAMT,SGD,STR,TDC, andORCA3were involved in the regulation of this induction process. The results of qRT-PCR indicated that the increasing accumulations of vindoline, catharanthine, and ajmaline inC. roseusCMCs were correlated with the increasing expression of the above genes. Therefore,A. flavusfungal elicitor could enhance the TIA production ofC. roseusCMCs, which might be used as an alternative biotechnological resource for obtaining bioactive alkaloids.Keywords:Catharanthus roseus;cambial meristematic cells;Aspergillus flavus;terpenoid indole alkaloids;biosynthesis
研究领域:
植物生物技术
·
植物化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Pharmacognosy Magazine
·
2017
·
IF 1.069
Zhou Pengfei,Zhu Jianhua,Yu Rongmin,Zi Jiachen
Background: Enzyme inhibitors have been used for the clarification of biosynthesis of natural products. Catharanthus roseus cambial meristematic cell (CMC) culture has been established and proved to be a better monoterpeneindole alkaloid (MIA) producer than C. roseus dedifferentiated cell (DDC) culture. However, little is known about the inter-relationship of the MIA-biosynthetic genes with respect to their transcription. Objective: To clarify effects of alteration of one gene transcription on transcript levels of another genes in MIA-biosynthetic pathway, and how the accumulation of MIAs in CMCs are influenced by the alteration of their biosynthetic gene transcript levels. Materials and Methods: 3-Hydroxy-3-methylglutaryl-CoA reductase (HMGR) inhibitor lovastatin and 1-deoxy-D-xylulose 5-phosphate synthase (DXS) inhibitor clomazone were fed to C. roseus CMC cultures. The contents of MIAs were qualified by High Performance Liquid Chromatography and the transcript levels of the relevant genes were measured by qRT-PCR. Results: Lovastatin improved the accumulation of MIAs via increasing the transcription of their biosynthetic genes encoding DXS1, tryptonphan decarboxylase (TDC), loganic acid methyltransferase (LAMT), strictosidine synthase (STR), desacetoxyvindoline-4-hydroxylase (D4H) and ORCA3 (a jasmonate-responsive transcriptional regulator), whereas clomazone reduced the contents of MIAs and the mRNA levels of the corresponding genes. Conclusion: The biosynthesis of MIAs in C. roseus is is manipulated via a complex mechanism, the knowledge of which paves the way for rationally tuning metabolic flux to improve MIA production in C. roseus CMCs.
研究领域:
植物生物技术
·
代谢工程
·
植物化学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
Journal of Pharmaceutical Analysis
·
2020
·
IF 2.673
Fanrong Sun,Ligai Bai,Mingxue Li,Changqing Yu,Haiyan Liu,Xiaoqiang Qiao,Hongyuan Yan
In this study, a functionalized covalent-organic framework (COF) was first synthesized using porphyrin as the fabrication unit and showed an edge-curled, petal-like and well-ordered structure. The synthesized COF was then introduced to prepare porous organic polymer monolithic materials (POPMs). Two composite POPM/COF monolithic materials with rod shapes, referred to as sorbent A and sorbent B, were prepared in stainless steel tubes using different monomers. Sorbents A and B exhibited relatively uniform porous structures and enhanced specific surface areas of 153.14 m 2 /g and 80.01 m 2 /g, respectively. The prepared composite monoliths were used as in-tube solid-phase extraction (SPE) sorbents combined with HPLC for the on-line extraction and quantitative analytical systems. Indole alkaloids (from Catharanthus roseus G. Don and Uncaria rhynchophylla (Miq.) Miq. Ex Havil.) contained in mouse plasma were extracted and quantitatively analyzed using the online system. The two composite multifunctional monoliths showed excellent clean-up ability for complex biological matrices, as well as superior selectivity for target indole alkaloids. Method validation showed that the RSD values of the repeatability ( n =6) were ≤ 3.46%, and the accuracy expressed by the spiked recoveries was in the ranges of 99.38%–100.91% and 96.39%–103.50% for vinca alkaloids and Uncaria alkaloids, respectively. Furthermore, sorbents A and B exhibited strong reusability, with RSD values ≤ 5.32%, which were based on the peak area of the corresponding alkaloids with more than 100 injections. These results indicate that the composite POPM/COF rod-shaped monoliths are promising media as SPE sorbents for extracting trace compounds in complex biological samples.
研究领域:
材料科学
·
分析化学
·
药学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Natural Product Communications
·
2018
·
IF 0.809
Jianhua Zhu,Shuijie He,Pengfei Zhou,Jiachen Zi,Jincai Liang,Liyan Song,Rongmin Yu
Vallesiachotamine and isovallesiachotamine are pharmacologically active monoterpene indole alkaloids produced by Catharanthus roseus. However, the biosynthetic pathway for these two alkaloids has not been characterized. The results of our experiments demonstrated that catharanthine induced the biosynthesis of vallesiachotamine and isovallesiachotamine in C. roseus cambial meristematic cells (CMCs) in a time- and dosage-dependent manner. The highest yields of vallesiachotamine and isovallesiachotamine (i.e., 1.49 mg l−1 and 1.08 mg l−1, respectively) were observed in CMCs treated with 10 mg catharanthine l−1 for 12 h. The cells growth was inhibited by 20 mg catharanthine l−1 (i.e., 16.3% decrease in dry-cell weight). A quantitative reverse transcription polymerase chain reaction analysis revealed that catharanthine regulated the biosynthesis of vallesiachotamine and isovallesiachotamine by up-regulating the expression of specific genes, including STR (strictosidine synthase), SGD (strictosidine β-D-glucosidase) and ORCA3 (transcription factor octadecanoid-responsive Catharanthus AP2/ERF-domain 3). Finally, considering the up-regulated expression of STR and SGD was consistent with the accumulation of vallesiachotamine and isovallesiachotamine, a potential vallesiachotamine and isovallesiachotamine biosynthetic pathway was proposed.
研究领域:
植物学
·
药理学
·
植物生物技术
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2015
·Vol. 99
·期 17
·7035-7045
·
IF 3.337
Zhou Pengfei,Yang Jiazeng,Zhu Jianhua,He Shuijie,Zhang Wenjin,Yu Rongmin,Zi Jiachen,Song Liyan,Huang Xuesong
Long-term stable cell growth and production of vindoline, catharanthine, and ajmalicine of cambial meristematic cells (CMCs) from Catharanthus roseus were observed after 2 years of culture. C. roseus CMCs were treated with β-cyclodextrin (β-CD) and methyl jasmonate (MeJA) individually or in combination and were cultured both in conventional Erlenmeyer flasks (100, 250, and 500 mL) and in a 5-L stirred hybrid airlift bioreactor. CMCs of C. roseus cultured in the bioreactor showed higher yields of vindoline, catharanthine, and ajmalicine than those cultured in flasks. CMCs of C. roseus cultured in the bioreactor and treated with 10 mM β-CD and 150 μM MeJA gave the highest yields of vindoline (7.45 mg/L), catharanthine (1.76 mg/L), and ajmalicine (58.98 mg/L), concentrations that were 799, 654, and 426 % higher, respectively, than yields of CMCs cultured in 100-mL flasks without elicitors. Quantitative reverse transcription (RT)-PCR showed that β-CD and MeJA upregulated transcription levels of genes related to the biosynthesis of terpenoid indole alkaloids (TIAs). This is the first study to report that β-CD induced the generation of NO, which plays an important role in mediating the production of TIAs in C. roseus CMCs. These results suggest that β-CD and MeJA can enhance the production of TIAs in CMCs of C. roseus , and thus, CMCs of C. roseus have significant potential to be an industrial platform for production of bioactive alkaloids.
研究领域:
植物生物技术
·
植物化学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
PLANT CELL TISSUE AND ORGAN CULTURE
·
2025
·Vol. 162
·期 3
·1-12
·
IF 2.4
Li Haihua,Li Jiahe,Liu Ningjin,Li Huishan,Wang Jiarui,Zhang Rulan,Xue Jingyan,Kang Xiaotian,Guan Xinying,Lin Shiya,Zhang Ziyu,Kuang Jiaqi,Qian Chenlei,Zhou Pengfei
In Catharanthus roseus , the inherently low yield of bioactive alkaloids such as vindoline, catharanthine, ajmalicine, and vinblastine remains a major limitation to their broader therapeutic application, particularly in anticancer and antihypertensive treatments. To address this challenge, we developed a novel metabolic flux regulation strategy by combining the enzyme inhibitors lovastatin and naproxen with the abiotic elicitor copper(II)-chitosan (Cu-CS) complex. Compared to controls, treatment with 2 mg/L lovastatin significantly enhanced catharanthine, vindoline, ajmalicine, and vinblastine accumulation in C. roseus CMCs to 2.38, 0.63, 35.3, and 2.33 mg/L, respectively — increases of 48%, 121%, 71%, and 12%. Application of 8 mg/L naproxen further enhanced catharanthine, vindoline, and vinblastine levels to 3.78, 0.67, and 2.14 mg/L, representing increases of 151%, 135%, and 30%, respectively. The Cu-CS complex at 10 mg/L elevated catharanthine, vindoline, ajmalicine, and vinblastine to 2.13, 0.49, 32.11, and 2.46 mg/L, with corresponding increases of 49%, 63%, 55%, and 36%. Notably, the combined treatment of all three agents synergistically boosted catharanthine, vindoline, and vinblastine production to 5.94, 1.52, and 3.65 mg/L — representing 310%, 390%, and 98% increases over control levels. Quantitative RT-PCR analysis revealed that Cu-CS treatment upregulated the transcription of key genes and transcription factors involved in terpenoid indole alkaloid (TIA) biosynthesis. Overall, this integrated strategy effectively redirects metabolic flux toward enhanced TIA production in C. roseus CMCs, providing a practical and efficient approach for improving the yield of pharmaceutically important alkaloids in plant cell culture systems.
研究领域:
植物生物技术
·
次生代谢
·
医药生物技术
·
分子生物学
·
代谢工程
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）