[Objectives]To establish an acute yunaconitine poisoning rat model with a single oral administration and to determine the contents of yunaconitine in rat tissues by UPLC-MS/MS method,then investigate the distribution ...[Objectives]To establish an acute yunaconitine poisoning rat model with a single oral administration and to determine the contents of yunaconitine in rat tissues by UPLC-MS/MS method,then investigate the distribution of yunaconitine in rats. [Methods]The rats were randomly divided into three groups and were intragastrically administered a single dose of 2. 2,1. 1,and 0. 7 mg/kg of yunaconitine,respectively.The rats were killed 2 h later,the stomach tissue,intestine tissue,liver tissue,pancreas tissue,kidney tissue,lung tissue,spleen tissue,heart tissue,bladder tissue,testis tissue,brain tissue and heart blood samples were collected. The contents of yunaconitine in the biological materials were determined by UPLC-MS/MS method after the biological samples extracted by liquid-liquid extraction. [Results] A rat model of the yunaconitine poisoning was made with a single dose of 1. 1 mg/kg,the concentrations of yunaconitine shown in the organs with the following order: stomach,small intestine,liver,pancreas,kidney,lung,spleen,heart,bladder,testis,heart blood and brain. [Conclusions]Yunaconitine was widely distributed in rats,especially the levels in the stomach,small intestine and liver were the highest. The conclusion is expected to provide a basis for the selection of test materials for the poisoning of Aconitum vilmorinianum Kom.展开更多
Diterpenoid alkaloids(DAs) have been often utilized in clinical practice due to their analgesic and anti-infammatory properties. Natural DAs are prevalent in the family Ranunculaceae, notably in the Aconitum genus. Ne...Diterpenoid alkaloids(DAs) have been often utilized in clinical practice due to their analgesic and anti-infammatory properties. Natural DAs are prevalent in the family Ranunculaceae, notably in the Aconitum genus. Nevertheless, the evolutionary origin of the biosynthesis pathway responsible for DA production remains unknown.In this study, we successfully assembled a highquality, pseudochromosome-level genome of the DA-rich species Aconitum vilmorinianum(A.vilmorinianum)(5.76 Gb). An A. vilmorinianumspecific whole-genome duplication event was discovered using comparative genomic analysis,which may aid in the evolution of the DA biosynthesis pathway. We identified several genes involved in DA biosynthesis via integrated genomic, transcriptomic, and metabolomic analyses. These genes included enzymes encoding target ent-kaurene oxidases and aminotransferases, which facilitated the activation of diterpenes and insertion of nitrogen atoms into diterpene skeletons, thereby mediating the transformation of diterpenes into DAs. The divergence periods of these genes in A. vilmorinianum were further assessed, and it was shown that two major types of genes were involved in the establishment of the DA biosynthesis pathway. Our integrated analysis offers fresh insights into the evolutionary origin of DAs in A.vilmorinianum as well as suggestions for engineering the biosynthetic pathways to obtain desired DAs.展开更多
Aconitum vilmorinianum,a well-known traditional Chinese herb,is recently being threatened by overexploitation and environment disturbance.This study was conducted to provide propagation methods through in vitro germi...Aconitum vilmorinianum,a well-known traditional Chinese herb,is recently being threatened by overexploitation and environment disturbance.This study was conducted to provide propagation methods through in vitro germination and explant cultivation.Germination was stimulated up to 66.00%on Murashige and Skoog(MS)medium containing 2.0 mg L^(−1)6-benzylaminopurine(BAP),0.1 mg L^(−1)1-napthaleneacetic acid(NAA),and 30 g L^(−1)sucrose.Three bacteria(Pantoea agglomerans,Erwinia persicina,and Pseudomonas tolaasii)would be responsible for consistent contamination during germination.The latter two were effectively eradicated after disinfected.The influence of explant types and hormone combinations on direct and indirect organogenesis was evaluated in the present work.The frequency of shoot induction from axillary bud explants was 100%on the MS fortified with 2.0 mg L^(−1)BAP and 0.3 mg L^(−1)NAA.Shoots multiplication was optimized on MS medium supplemented with 0.1 mg L^(−1)thidiazuron(TDZ)and 0.1 mg L^(−1)NAA.High callus induction percentage(96.67%)was obtained from stem segments on MS medium with 2.0 mg L^(−1)2,4-D,then successfully regenerated into shoots on MS medium in the presence of 0.1 mg L^(−1)TDZ and 0.2 mg L^(−1)NAA.The present work could be useful for the utilization and conservation of this valuable species.展开更多
基金Supported by Scientific and Technological Planning Project of Yunnan Province(2013FB118)Innovation Team Project of Kunming Medical University(CXTD201604)Scientific Research Foundation of Yunnan Provincial Department of Education(2017zDX161)
文摘[Objectives]To establish an acute yunaconitine poisoning rat model with a single oral administration and to determine the contents of yunaconitine in rat tissues by UPLC-MS/MS method,then investigate the distribution of yunaconitine in rats. [Methods]The rats were randomly divided into three groups and were intragastrically administered a single dose of 2. 2,1. 1,and 0. 7 mg/kg of yunaconitine,respectively.The rats were killed 2 h later,the stomach tissue,intestine tissue,liver tissue,pancreas tissue,kidney tissue,lung tissue,spleen tissue,heart tissue,bladder tissue,testis tissue,brain tissue and heart blood samples were collected. The contents of yunaconitine in the biological materials were determined by UPLC-MS/MS method after the biological samples extracted by liquid-liquid extraction. [Results] A rat model of the yunaconitine poisoning was made with a single dose of 1. 1 mg/kg,the concentrations of yunaconitine shown in the organs with the following order: stomach,small intestine,liver,pancreas,kidney,lung,spleen,heart,bladder,testis,heart blood and brain. [Conclusions]Yunaconitine was widely distributed in rats,especially the levels in the stomach,small intestine and liver were the highest. The conclusion is expected to provide a basis for the selection of test materials for the poisoning of Aconitum vilmorinianum Kom.
基金supported by funding from the CAS “Pioneer Hundred Talents” Program and Strategic Priority Research Program of the Chinese Academy of Sciences (XDB31000000) to Y.C.National Natural Science Foundation of China (Nos. 31960082, 32270395, and 31960096)Yunnan Fundamental Research Projects (No. 202101AS070021) to D.Z。
文摘Diterpenoid alkaloids(DAs) have been often utilized in clinical practice due to their analgesic and anti-infammatory properties. Natural DAs are prevalent in the family Ranunculaceae, notably in the Aconitum genus. Nevertheless, the evolutionary origin of the biosynthesis pathway responsible for DA production remains unknown.In this study, we successfully assembled a highquality, pseudochromosome-level genome of the DA-rich species Aconitum vilmorinianum(A.vilmorinianum)(5.76 Gb). An A. vilmorinianumspecific whole-genome duplication event was discovered using comparative genomic analysis,which may aid in the evolution of the DA biosynthesis pathway. We identified several genes involved in DA biosynthesis via integrated genomic, transcriptomic, and metabolomic analyses. These genes included enzymes encoding target ent-kaurene oxidases and aminotransferases, which facilitated the activation of diterpenes and insertion of nitrogen atoms into diterpene skeletons, thereby mediating the transformation of diterpenes into DAs. The divergence periods of these genes in A. vilmorinianum were further assessed, and it was shown that two major types of genes were involved in the establishment of the DA biosynthesis pathway. Our integrated analysis offers fresh insights into the evolutionary origin of DAs in A.vilmorinianum as well as suggestions for engineering the biosynthetic pathways to obtain desired DAs.
基金Yunnan Fundamental Research Projects(Nos.202101AS070021,202101AT070265)the National Natural Science Foundation of China(Grant Nos.31960082,32160249)the Scientific Research Fund Project of Yunnan Education Department(No.2021J0002).
文摘Aconitum vilmorinianum,a well-known traditional Chinese herb,is recently being threatened by overexploitation and environment disturbance.This study was conducted to provide propagation methods through in vitro germination and explant cultivation.Germination was stimulated up to 66.00%on Murashige and Skoog(MS)medium containing 2.0 mg L^(−1)6-benzylaminopurine(BAP),0.1 mg L^(−1)1-napthaleneacetic acid(NAA),and 30 g L^(−1)sucrose.Three bacteria(Pantoea agglomerans,Erwinia persicina,and Pseudomonas tolaasii)would be responsible for consistent contamination during germination.The latter two were effectively eradicated after disinfected.The influence of explant types and hormone combinations on direct and indirect organogenesis was evaluated in the present work.The frequency of shoot induction from axillary bud explants was 100%on the MS fortified with 2.0 mg L^(−1)BAP and 0.3 mg L^(−1)NAA.Shoots multiplication was optimized on MS medium supplemented with 0.1 mg L^(−1)thidiazuron(TDZ)and 0.1 mg L^(−1)NAA.High callus induction percentage(96.67%)was obtained from stem segments on MS medium with 2.0 mg L^(−1)2,4-D,then successfully regenerated into shoots on MS medium in the presence of 0.1 mg L^(−1)TDZ and 0.2 mg L^(−1)NAA.The present work could be useful for the utilization and conservation of this valuable species.
