[Objective] The research aimed to establish the cell suspension culture system of Pinellia ternate.[Method] The petiole of Pinellia ternate was as the explant.The orthogonal test was used to study the influences of fo...[Objective] The research aimed to establish the cell suspension culture system of Pinellia ternate.[Method] The petiole of Pinellia ternate was as the explant.The orthogonal test was used to study the influences of four plant growth substances（2,4-D,NAA,picloram and KT）and their mixture ratios on the formation of loose callus.[Result] The induction effect of 2,4-D and picloram on the petiole callus of Pinellia ternate was the most significant.Then,the second ones were KT and NAA.The optimal medium which induced Pinellia ternate petiole to form the loose callus was MS＋0.5 mg/L of 2,4-D＋1.0 mg/L of NAA＋1.0 mg/L of picloram＋1.5 mg/L of KT.[Conclusion] The research laid the foundation for extracting the active ingredient from the cell suspension of Pinellia ternate and producing the artificial seed.展开更多
Total proteins extracted from Pinellia ternata with different types of leaves have been assayed by UV absorbance, blue native polyacrylamide gel electrophoresis （BN-PAGE） and sodium dodecyl sulfate-polyacrylamide ge...Total proteins extracted from Pinellia ternata with different types of leaves have been assayed by UV absorbance, blue native polyacrylamide gel electrophoresis （BN-PAGE） and sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS- PAGE）. We found that the total protein contents of P. ternata with the herbaceous peony and claw types of leaves were much higher than that with other types of leaves. P. ternata collected from Nanchuan County, except the ones with herbaceous peony leaves, displayed a high similarity with each other. Both UV absorbance and cluster analysis indicated that the protein content in P. ternata with the herbaceous peony leaves was less affected by habitats than that with other types of leaves. The results showed that it was necessary to increase the homogeneity of seeds for improving the protein content ofP. ternata.展开更多
The full-length cDNA of Pinellia ternata agglutinin (PTA) was cloned from inflorescences using RACE-PCR. Through comparative analysis of PTA gene (pta) and its deduced amino acid sequence with those of other Araceae s...The full-length cDNA of Pinellia ternata agglutinin (PTA) was cloned from inflorescences using RACE-PCR. Through comparative analysis of PTA gene (pta) and its deduced amino acid sequence with those of other Araceae species, pta was found to encode a precursor lectin with signal peptide and to have extensive homology with those of other Araceae species. PTA was a heterotetrameric mannose-binding lectin with three mannose-binding boxes like lectins from other Araceae and Amaryllidaceae species. Southern blot analysis of the genomic DNA revealed that pta belonged to a low-copy gene family. Northern blot analysis demonstrated that pta constitutively expressed in various plant tissues including root, leaf, stem and inflorescence. The pta cDNA sequence encoding for mature PTA protein was cloned into pET-32a plasmid and the resulting plasmid, pET-32a-PTA containing Trx-PTA fusion protein, was investigated for the expression in E. coli BL21. SDS-PAGE gel analysis showed that the Trx-PTA fusion protein was successfully expressed in E. coli BL21 when induced by IPTG. Artificial diet assay revealed that PTA fusion protein had significant levels of resistance against peach potato aphids when incorporated into artificial diet at 0.1% (w/v). The cloning of the pta gene will enable us to further test its effect in depth on aphids by transferring the gene into crop plants.展开更多
Pinellia ternata is an important medicinal plant,and its growth and development are easily threatened by high temperature.In this study,comprehensive research on physiological,cytological and transcriptional responses...Pinellia ternata is an important medicinal plant,and its growth and development are easily threatened by high temperature.In this study,comprehensive research on physiological,cytological and transcriptional responses to different levels of heat stress were conducted on a typical phenotype of P.ternata.First,P.ternata exhibited tolerance to the increased temperature,which was supported by normal growing leaves,as well as decreased and sustained photosynthetic parameters.Severe stress aggravated the damages,and P.ternata displayed an obvious leaf senescence phenotype,with significantly increased SOD and POD activities(46%and 213%).In addition,mesophyll cells were seriously damaged,chloroplast thylakoid was fuzzy,grana lamellae and stroma lamellae were obviously broken,and grana thylakoids were stacked,resulting in a dramatically declined photosynthetic rate(74.6%).Moreover,a total of 16808 genes were significantly differential expressed during this process,most of which were involved in photosynthesis,transmembrane transporter activity and plastid metabolism.The number of differentially expressed transcription factors in MYB and bHLH families was the largest,indicating that these genes might participate in heat stress response in P.ternata.These findings provide insight into the response to high temperature and facilitate the standardized cultivation of P.ternata.展开更多
Objective:Pinellia Tuber,the dried tuber of Pinellia ternata,is widely used in Japanese Kampo medicines and traditional Chinese medicines.The unprocessed Pinellia Tuber is known to cause very strong acrid irritation a...Objective:Pinellia Tuber,the dried tuber of Pinellia ternata,is widely used in Japanese Kampo medicines and traditional Chinese medicines.The unprocessed Pinellia Tuber is known to cause very strong acrid irritation at oral and laryngopharynx mucosa.Recent studies have shown that the sharp needle-like crystals called raphides,that are composed of calcium oxalate and proteins,are the main causative substances of the irritation.Ginger,the rhizome of Zingiber officinale,has been used in the processing to reduce the acridity of Pinellia Tuber since before the sixth century,however,the mechanisms of reducing acridity have not been scientifically proved yet.Methods:We developed the raphides denaturation assay(RDA)to quantify the degree of denaturation in the raphides to cause irritation.By their lipophilic characters,the raphides could be extracted in petroleum ether(PE)layer from powdered Pinellia Tuber suspended in water,and the contents of the raphides in PE layer were measured by the absorbance.By this assay,we conducted the activity-guided fractionation from the boiling water extract of ginger to find the ingredients to denature the raphides.We also conducted the gustatory tests to detect the change of the irritation of the denatured raphides.Results:The treatment of powdered Pinellia Tuber suspension with ginger extract reduced the distribution of raphides in PE layer in RDA in a concentration-dependent manner.The activity-guided fractionation using RDA revealed that oxalic acid was the main active ingredient in ginger extract to denature the raphides of Pinellia Tuber.Oxalic acid reduced the lipophilicity of the raphides in the thermo-,time-,and concentration-dependent manners,and its activity was affected by pH.The treatment of powdered Pinellia Tuber suspension with oxalic acid significantly reduced its acrid irritation in gustatory test in human.Conclusions:We found that oxalic acid is the main active ingredient in ginger to reduce the acrid irritation of Pinellia Tuber.展开更多
Based on literature reviews and analysis of research reports on Pinellia ternata found locally and abroad in recent years,this article summarizes and arranges them.The research on Pinellia ternata mainly focuses on it...Based on literature reviews and analysis of research reports on Pinellia ternata found locally and abroad in recent years,this article summarizes and arranges them.The research on Pinellia ternata mainly focuses on its cultivation,tissue culture,and so on.There are only a few research on its active components and its regulation mechanism.The wild resources of Pinellia ternata are gradually decreasing,hence it is urgent to take effective measures to protect these wild resources as well as to establish germplasm resources bank and nursery.In order to meet the needs of the domestic market,it is necessary to investigate the distribution of wild Pinellia ternata resources,explore the best growing environment and conditions,artificially cultivate Pinellia ternata,as well as implement resource industrialization,sustainable development,and utilization.展开更多
[ Objective] This study aimed to investigate the variation rule of total RNA concentration during formation of microtubers of PineUia ternata induced by sucrose. [ Method ] Plantlets of P. ternata were used as experim...[ Objective] This study aimed to investigate the variation rule of total RNA concentration during formation of microtubers of PineUia ternata induced by sucrose. [ Method ] Plantlets of P. ternata were used as experimental materials. Total RNA concentration was determined with spectrophotometer, and the variation of total RNA was analyzed before and after formation of P. ternata microtubers treated with different concentrations of sucrose using DDRT-PCR technology. [ Result] Total RNA concentration of P. ternata treated with different concentrations of sucrose varied greatly, which showed an upward trend in the first 15 d, de- clined from 15~ d to 25th d, and slightly raised from 25＇h d to 30＇8 d. Microtubers formed under various sucrose treatments showed an increase in RNA concentration after induction, and it was assumed that the treatment of high concentrations of sucrose might have activated the expression of development-related＇ genes in P. ter- nata microtubers. [ Conclusion] This study provides the basis for revealing the related gene expression information and molecular mechanism during formation of P. ternata microtubers induced by sucrose.展开更多
In this paper, Pinellia ternata lectin (PTL) purified from 95% saturated ammonium sulfate precipitation by applying mannose-Sepharose 4B affinity column chromatography was first described. The mannose-Sepharose 4B aff...In this paper, Pinellia ternata lectin (PTL) purified from 95% saturated ammonium sulfate precipitation by applying mannose-Sepharose 4B affinity column chromatography was first described. The mannose-Sepharose 4B affinity adsorption gel has already been designed and prepared before the experiments according the combinative characterization between lectin and mannose. Mass spectrometry analysis result shows that PTL is a glycoprotein with the molecular weight of 12.165 kD. It was conclude that PTL had strong agglutination effects on mouse red blood cells, and the minimum reaction concentration was 25 μg/ml according the Hemagglutination. Results of automatic amino acid analysis indicated that PTL mainly contained 15 varieties of amino acids, of which the minimum content was cysteine and aspartate was the maximum. Cell experiment results suggested that PTL of low concentrations (0.004 mg/ml, 0.02 mg/ml and 0.1 mg/ml) promoted HeLa cell proliferation, but the effect weakened with the concentrations and treated-time increasing. However, the HeLa cells pro- liferation was intensively inhibited by higher PTL concentrations (0.5 mg/ml and 1 mg/ml), and the effect increased in a dose and time dependent manner.展开更多
As its tuberous alkaloids induce valuable pharmacological effects,Pinellia ternata has considerable clinical value.However,its production currently fails to meet its demand.In vitro microtuber culture,combined with sa...As its tuberous alkaloids induce valuable pharmacological effects,Pinellia ternata has considerable clinical value.However,its production currently fails to meet its demand.In vitro microtuber culture,combined with salicylic acid(SA)elicitation,may provide an effective alternative to traditional field production.In this study,an in vitro P.ternata microtuber induction system was developed and used to evaluate SA-induced elicitation of alkaloid accumulation.The quality of in vitro microtubers was assessed by total alkaloid measurement,a cytotoxicity assay,and transcriptomic analysis.With or without SA treatment,P.ternata microtuber induction was achieved within 60 d using petiole-derived explants,with a microtuber proliferation rate of approximately 17 microtubers per explant.The total alkaloid content of in vitro microtubers elicited with 100μM SA was equivalent to that of field-grown tubers,while those not treated with SA contained lower alkaloid content.The cytotoxicity assay showed preliminary cytotoxic effects of SA-treated microtubers against the breast cancer cell line SUM159,comparable to field-grown tubers.Transcriptomic analysis revealed many differentially expressed genes(DEGs)in SA-treated in vitro microtubers.Six and twelve DEGs were annotated to the tropane,piperidine,and pyridine alkaloid pathway and the isoquinoline alkaloid pathway,respectively.RT-qPCR confirmed that the genes encoding spermidine synthase(c64642_g1),hyoscyamine(6S)-dioxygenase(c62620_g1),catechol oxidase(c61704_g3),monoamine oxidase(c65996_g3),and aspartate transaminase(c71069_g1)were significantly induced by SA.This study advances the production of P.ternata microtubers without field cultivation and provides considerable genetic information regarding SA-promoted alkaloid accumulation in P.ternata.展开更多
Objective: To investigate the toxicity difference between raw and processed Pinelliae Rhizoma(Banxia in Chinese, BX), the rhizoma of Pinellia ternata, from the view of chemical composition.Methods: Sixteen samples of ...Objective: To investigate the toxicity difference between raw and processed Pinelliae Rhizoma(Banxia in Chinese, BX), the rhizoma of Pinellia ternata, from the view of chemical composition.Methods: Sixteen samples of raw and processed BX were prepared and analyzed by UPLC/Q-TOF-MS/MS.The discrimination(chemical marker) between the two group was investigated by principal component analysis(PCA) and T-test analysis. According to the accurate charge-to-mass ratio, MS/MS fragments, and comparison of corresponding data with the reference or database, the chemical markers were identified preliminarily.Results: Liquiritin, liquiritigenin, and lysophosphatidylcholine(LPC) were identified as the characteristic markers. The reducing of LPC in processed BX was one of the main reasons for detoxification because LPC could induce the inflammatory response;Liquiritin and liquiritigenin showed the anti-inflammatory effect and reduced liver injury, therefore the appearance of them in processed BX was an another reason for detoxification.Conclusion: An approach to explain the mechanisms of reducing the toxicity in medicinal plants by processing was proposed. Moreover, the chemical markers of toxicity could be used to differentiate the raw material from processed herbs for the quality control and safety application in clinical practice.展开更多
By means of the two-compartment system. PTL-channels, the first lectin channels formed on planar lipid bilayers by Pinellia ternata lectin (PTL) have been studied. The results show (i) PTL-channels are voltage-indepen...By means of the two-compartment system. PTL-channels, the first lectin channels formed on planar lipid bilayers by Pinellia ternata lectin (PTL) have been studied. The results show (i) PTL-channels are voltage-independent and have apparent subunits; (ii) in 50 mmol/L KC1 and 25 mmol/L BaCl2 solutions, a single channel has unit conductance of 21 pS and 42 pS, respectively; (iii) the channel exhibits a slightly higher permeability to divalent than monovalent cation (PBa/PK=4.1), and (iv) the selectivity among divalent or monovalent cations is poor. The cation selectivity sequence for the channel will follow PBa(7. 0)-Psr (6. 4)~PMg(6. 4)>PK(1. 7)>PN.(1. 0)~PLi(l. 0).Moreover, these data also give explanation to the facilitatory action of PTL on the release of acetyl-choline from motor nerve terminals.展开更多
Pinellia ternata lectin (PTL) extracted from the fresh juice of rhizome of pinellia ternata used as a traditional Chinese medicine facilitated the quantal release of acetylcholine (ACh) in the mouse motor nerve termin...Pinellia ternata lectin (PTL) extracted from the fresh juice of rhizome of pinellia ternata used as a traditional Chinese medicine facilitated the quantal release of acetylcholine (ACh) in the mouse motor nerve terminals and formed cation channels in artificial lipid bilayer. Here we report the action of PTL on presynaptic membrane currents of motor nerve terminals.The experiments were performed on the intercostal nerve triangularis sterni muscle preparations. By means of the perineurial recording, the effects of PTL on the sodium current in the preterminal part , three potassium currents and two calcium currents generated from the nerve terminals were investigated. The results show that PTL increases voltage-dependent fast Ca2+ current (ICa,f), Na+ current (INa) and Ca2+-acti-vated K+ current (IK,Ca) without action on either the voltage-dependent fast K+ current (IK,f) or the slow K+ current (IK,S). These effects are irreversible, but can be reversed by mannan, the specific binding sugar for PTL.The total action of PTL can be considered to raise [Ca2+], by promoting Ca2+ influx, and then facilitate the neurotransmitter release. To our knowledge, this is the first report about the influence of lectin on presynaptic membrane currents at mammalian motor nerve terminals.展开更多
基金Supported by National Natural Science Fund Item(30973963)Anhui Natural Science Fund Item(090413252)+1 种基金Natural Science ResearchKey Project of Anhui Provincial University(KJ2009A160)"Key Labora-tory Project"of Plant Resource and Biology(KLPRB200914)~~
文摘[Objective] The research aimed to establish the cell suspension culture system of Pinellia ternate.[Method] The petiole of Pinellia ternate was as the explant.The orthogonal test was used to study the influences of four plant growth substances（2,4-D,NAA,picloram and KT）and their mixture ratios on the formation of loose callus.[Result] The induction effect of 2,4-D and picloram on the petiole callus of Pinellia ternate was the most significant.Then,the second ones were KT and NAA.The optimal medium which induced Pinellia ternate petiole to form the loose callus was MS＋0.5 mg/L of 2,4-D＋1.0 mg/L of NAA＋1.0 mg/L of picloram＋1.5 mg/L of KT.[Conclusion] The research laid the foundation for extracting the active ingredient from the cell suspension of Pinellia ternate and producing the artificial seed.
基金Natural Science Foundation of Chongqing (Grant No. 2006BB5403)
文摘Total proteins extracted from Pinellia ternata with different types of leaves have been assayed by UV absorbance, blue native polyacrylamide gel electrophoresis （BN-PAGE） and sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS- PAGE）. We found that the total protein contents of P. ternata with the herbaceous peony and claw types of leaves were much higher than that with other types of leaves. P. ternata collected from Nanchuan County, except the ones with herbaceous peony leaves, displayed a high similarity with each other. Both UV absorbance and cluster analysis indicated that the protein content in P. ternata with the herbaceous peony leaves was less affected by habitats than that with other types of leaves. The results showed that it was necessary to increase the homogeneity of seeds for improving the protein content ofP. ternata.
文摘The full-length cDNA of Pinellia ternata agglutinin (PTA) was cloned from inflorescences using RACE-PCR. Through comparative analysis of PTA gene (pta) and its deduced amino acid sequence with those of other Araceae species, pta was found to encode a precursor lectin with signal peptide and to have extensive homology with those of other Araceae species. PTA was a heterotetrameric mannose-binding lectin with three mannose-binding boxes like lectins from other Araceae and Amaryllidaceae species. Southern blot analysis of the genomic DNA revealed that pta belonged to a low-copy gene family. Northern blot analysis demonstrated that pta constitutively expressed in various plant tissues including root, leaf, stem and inflorescence. The pta cDNA sequence encoding for mature PTA protein was cloned into pET-32a plasmid and the resulting plasmid, pET-32a-PTA containing Trx-PTA fusion protein, was investigated for the expression in E. coli BL21. SDS-PAGE gel analysis showed that the Trx-PTA fusion protein was successfully expressed in E. coli BL21 when induced by IPTG. Artificial diet assay revealed that PTA fusion protein had significant levels of resistance against peach potato aphids when incorporated into artificial diet at 0.1% (w/v). The cloning of the pta gene will enable us to further test its effect in depth on aphids by transferring the gene into crop plants.
基金supported by the Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences(No.CI2021A04106)the Fundamental Research Funds for the Central Public Welfare Research Institutes of China(Nos.ZXKT21026 and ZZ15-YQ-033)the Major Special Project of Scientific and Technological Cooperation of Bijie City(2021-02).
文摘Pinellia ternata is an important medicinal plant,and its growth and development are easily threatened by high temperature.In this study,comprehensive research on physiological,cytological and transcriptional responses to different levels of heat stress were conducted on a typical phenotype of P.ternata.First,P.ternata exhibited tolerance to the increased temperature,which was supported by normal growing leaves,as well as decreased and sustained photosynthetic parameters.Severe stress aggravated the damages,and P.ternata displayed an obvious leaf senescence phenotype,with significantly increased SOD and POD activities(46%and 213%).In addition,mesophyll cells were seriously damaged,chloroplast thylakoid was fuzzy,grana lamellae and stroma lamellae were obviously broken,and grana thylakoids were stacked,resulting in a dramatically declined photosynthetic rate(74.6%).Moreover,a total of 16808 genes were significantly differential expressed during this process,most of which were involved in photosynthesis,transmembrane transporter activity and plastid metabolism.The number of differentially expressed transcription factors in MYB and bHLH families was the largest,indicating that these genes might participate in heat stress response in P.ternata.These findings provide insight into the response to high temperature and facilitate the standardized cultivation of P.ternata.
基金the grant from Oriental Medicine Research Foundation in 2020.
文摘Objective:Pinellia Tuber,the dried tuber of Pinellia ternata,is widely used in Japanese Kampo medicines and traditional Chinese medicines.The unprocessed Pinellia Tuber is known to cause very strong acrid irritation at oral and laryngopharynx mucosa.Recent studies have shown that the sharp needle-like crystals called raphides,that are composed of calcium oxalate and proteins,are the main causative substances of the irritation.Ginger,the rhizome of Zingiber officinale,has been used in the processing to reduce the acridity of Pinellia Tuber since before the sixth century,however,the mechanisms of reducing acridity have not been scientifically proved yet.Methods:We developed the raphides denaturation assay(RDA)to quantify the degree of denaturation in the raphides to cause irritation.By their lipophilic characters,the raphides could be extracted in petroleum ether(PE)layer from powdered Pinellia Tuber suspended in water,and the contents of the raphides in PE layer were measured by the absorbance.By this assay,we conducted the activity-guided fractionation from the boiling water extract of ginger to find the ingredients to denature the raphides.We also conducted the gustatory tests to detect the change of the irritation of the denatured raphides.Results:The treatment of powdered Pinellia Tuber suspension with ginger extract reduced the distribution of raphides in PE layer in RDA in a concentration-dependent manner.The activity-guided fractionation using RDA revealed that oxalic acid was the main active ingredient in ginger extract to denature the raphides of Pinellia Tuber.Oxalic acid reduced the lipophilicity of the raphides in the thermo-,time-,and concentration-dependent manners,and its activity was affected by pH.The treatment of powdered Pinellia Tuber suspension with oxalic acid significantly reduced its acrid irritation in gustatory test in human.Conclusions:We found that oxalic acid is the main active ingredient in ginger to reduce the acrid irritation of Pinellia Tuber.
基金Supported by Ankang Science and Technology Bureau,Shaanxi Province(2018ak03-ll)National Traditional Chinese Medicine Characteristic Technology Inheritor(t20194828003).
文摘Based on literature reviews and analysis of research reports on Pinellia ternata found locally and abroad in recent years,this article summarizes and arranges them.The research on Pinellia ternata mainly focuses on its cultivation,tissue culture,and so on.There are only a few research on its active components and its regulation mechanism.The wild resources of Pinellia ternata are gradually decreasing,hence it is urgent to take effective measures to protect these wild resources as well as to establish germplasm resources bank and nursery.In order to meet the needs of the domestic market,it is necessary to investigate the distribution of wild Pinellia ternata resources,explore the best growing environment and conditions,artificially cultivate Pinellia ternata,as well as implement resource industrialization,sustainable development,and utilization.
基金Supported by National Natural Science Foundation of China (30973963)Anhui Provincial Natural Science Foundation (090413252)Key Program of Natural Science Research of Anhui Provincial Universities (KJ2009A160)
文摘[ Objective] This study aimed to investigate the variation rule of total RNA concentration during formation of microtubers of PineUia ternata induced by sucrose. [ Method ] Plantlets of P. ternata were used as experimental materials. Total RNA concentration was determined with spectrophotometer, and the variation of total RNA was analyzed before and after formation of P. ternata microtubers treated with different concentrations of sucrose using DDRT-PCR technology. [ Result] Total RNA concentration of P. ternata treated with different concentrations of sucrose varied greatly, which showed an upward trend in the first 15 d, de- clined from 15~ d to 25th d, and slightly raised from 25＇h d to 30＇8 d. Microtubers formed under various sucrose treatments showed an increase in RNA concentration after induction, and it was assumed that the treatment of high concentrations of sucrose might have activated the expression of development-related＇ genes in P. ter- nata microtubers. [ Conclusion] This study provides the basis for revealing the related gene expression information and molecular mechanism during formation of P. ternata microtubers induced by sucrose.
文摘In this paper, Pinellia ternata lectin (PTL) purified from 95% saturated ammonium sulfate precipitation by applying mannose-Sepharose 4B affinity column chromatography was first described. The mannose-Sepharose 4B affinity adsorption gel has already been designed and prepared before the experiments according the combinative characterization between lectin and mannose. Mass spectrometry analysis result shows that PTL is a glycoprotein with the molecular weight of 12.165 kD. It was conclude that PTL had strong agglutination effects on mouse red blood cells, and the minimum reaction concentration was 25 μg/ml according the Hemagglutination. Results of automatic amino acid analysis indicated that PTL mainly contained 15 varieties of amino acids, of which the minimum content was cysteine and aspartate was the maximum. Cell experiment results suggested that PTL of low concentrations (0.004 mg/ml, 0.02 mg/ml and 0.1 mg/ml) promoted HeLa cell proliferation, but the effect weakened with the concentrations and treated-time increasing. However, the HeLa cells pro- liferation was intensively inhibited by higher PTL concentrations (0.5 mg/ml and 1 mg/ml), and the effect increased in a dose and time dependent manner.
基金supported by the Key Project of Natural Science Research for Colleges and Universities in Anhui Province(2024AH051661,2023AH050345)Excellent Scientific Research and Innovation Team of University in Anhui Province(2022AH010029).
文摘As its tuberous alkaloids induce valuable pharmacological effects,Pinellia ternata has considerable clinical value.However,its production currently fails to meet its demand.In vitro microtuber culture,combined with salicylic acid(SA)elicitation,may provide an effective alternative to traditional field production.In this study,an in vitro P.ternata microtuber induction system was developed and used to evaluate SA-induced elicitation of alkaloid accumulation.The quality of in vitro microtubers was assessed by total alkaloid measurement,a cytotoxicity assay,and transcriptomic analysis.With or without SA treatment,P.ternata microtuber induction was achieved within 60 d using petiole-derived explants,with a microtuber proliferation rate of approximately 17 microtubers per explant.The total alkaloid content of in vitro microtubers elicited with 100μM SA was equivalent to that of field-grown tubers,while those not treated with SA contained lower alkaloid content.The cytotoxicity assay showed preliminary cytotoxic effects of SA-treated microtubers against the breast cancer cell line SUM159,comparable to field-grown tubers.Transcriptomic analysis revealed many differentially expressed genes(DEGs)in SA-treated in vitro microtubers.Six and twelve DEGs were annotated to the tropane,piperidine,and pyridine alkaloid pathway and the isoquinoline alkaloid pathway,respectively.RT-qPCR confirmed that the genes encoding spermidine synthase(c64642_g1),hyoscyamine(6S)-dioxygenase(c62620_g1),catechol oxidase(c61704_g3),monoamine oxidase(c65996_g3),and aspartate transaminase(c71069_g1)were significantly induced by SA.This study advances the production of P.ternata microtubers without field cultivation and provides considerable genetic information regarding SA-promoted alkaloid accumulation in P.ternata.
基金supported by the National Natural Science Foundation of China (No. 81460595)
文摘Objective: To investigate the toxicity difference between raw and processed Pinelliae Rhizoma(Banxia in Chinese, BX), the rhizoma of Pinellia ternata, from the view of chemical composition.Methods: Sixteen samples of raw and processed BX were prepared and analyzed by UPLC/Q-TOF-MS/MS.The discrimination(chemical marker) between the two group was investigated by principal component analysis(PCA) and T-test analysis. According to the accurate charge-to-mass ratio, MS/MS fragments, and comparison of corresponding data with the reference or database, the chemical markers were identified preliminarily.Results: Liquiritin, liquiritigenin, and lysophosphatidylcholine(LPC) were identified as the characteristic markers. The reducing of LPC in processed BX was one of the main reasons for detoxification because LPC could induce the inflammatory response;Liquiritin and liquiritigenin showed the anti-inflammatory effect and reduced liver injury, therefore the appearance of them in processed BX was an another reason for detoxification.Conclusion: An approach to explain the mechanisms of reducing the toxicity in medicinal plants by processing was proposed. Moreover, the chemical markers of toxicity could be used to differentiate the raw material from processed herbs for the quality control and safety application in clinical practice.
基金Project supported by the National Natural Science Foundation of China.
文摘By means of the two-compartment system. PTL-channels, the first lectin channels formed on planar lipid bilayers by Pinellia ternata lectin (PTL) have been studied. The results show (i) PTL-channels are voltage-independent and have apparent subunits; (ii) in 50 mmol/L KC1 and 25 mmol/L BaCl2 solutions, a single channel has unit conductance of 21 pS and 42 pS, respectively; (iii) the channel exhibits a slightly higher permeability to divalent than monovalent cation (PBa/PK=4.1), and (iv) the selectivity among divalent or monovalent cations is poor. The cation selectivity sequence for the channel will follow PBa(7. 0)-Psr (6. 4)~PMg(6. 4)>PK(1. 7)>PN.(1. 0)~PLi(l. 0).Moreover, these data also give explanation to the facilitatory action of PTL on the release of acetyl-choline from motor nerve terminals.
基金Project supported by the National Natural Science Foundation of China.
文摘Pinellia ternata lectin (PTL) extracted from the fresh juice of rhizome of pinellia ternata used as a traditional Chinese medicine facilitated the quantal release of acetylcholine (ACh) in the mouse motor nerve terminals and formed cation channels in artificial lipid bilayer. Here we report the action of PTL on presynaptic membrane currents of motor nerve terminals.The experiments were performed on the intercostal nerve triangularis sterni muscle preparations. By means of the perineurial recording, the effects of PTL on the sodium current in the preterminal part , three potassium currents and two calcium currents generated from the nerve terminals were investigated. The results show that PTL increases voltage-dependent fast Ca2+ current (ICa,f), Na+ current (INa) and Ca2+-acti-vated K+ current (IK,Ca) without action on either the voltage-dependent fast K+ current (IK,f) or the slow K+ current (IK,S). These effects are irreversible, but can be reversed by mannan, the specific binding sugar for PTL.The total action of PTL can be considered to raise [Ca2+], by promoting Ca2+ influx, and then facilitate the neurotransmitter release. To our knowledge, this is the first report about the influence of lectin on presynaptic membrane currents at mammalian motor nerve terminals.
