Anoectochilus roxburghii from different planting sources was used as the research object,and the biological activity of its extracts was compared.The results showed that the whole plant extracts of A.roxburghii plante...Anoectochilus roxburghii from different planting sources was used as the research object,and the biological activity of its extracts was compared.The results showed that the whole plant extracts of A.roxburghii planted in bag culture (BAG),greenhouse cultivation (GRE) and tissue culture seedlings (TIS) were rich in active ingredients such as A.roxburghii polysaccharides and A.roxburghii glycosides.In vitro experiments showed that Anoectochilus roxburghii extracts from different planting sources had good antioxidant capacity,tyrosinase inhibition ability and hyaluronidase inhibition ability.Among them,the GRE Anoectochilus roxburghii whole plant extract had the strongest scavenging ability on DPPH free radical,ABTS free radical and hydroxyl free radical,and the inhibitory effect on tyrosinase activity and hyaluronidase activity was significantly higher than that of BAG and TIS.At the same time,it also verified that the extract of Anoectochilus roxburghii has antioxidant,whitening,anti-inflammatory and other skin care effects,which provides a theoretical basis for the development and application of Anoectochilus roxburghii extract in the field of cosmetics.展开更多
Cosmetics made from natural ingredients are becoming more and more common because they contain bioactive compounds that provide many health benefits,are more environmentally friendly and sustainable.Anoectochilus roxb...Cosmetics made from natural ingredients are becoming more and more common because they contain bioactive compounds that provide many health benefits,are more environmentally friendly and sustainable.Anoectochilus roxburghii(AR)is a precious Chinese herb in China,possessing bioactive components including flavonoids,polysaccharides,volatile oils,kinsenoside,etc.Due to its unique bioactivity and high safety,the prospect of development and utilization is very broad.In this paper,the extraction,separation and purification techniques of AR’s flavonoid,polysaccharides,volatile oils and kinsenoside are briefly summarized.Meanwhile,we discusses their conformational relationships in cosmetics with antioxidant,whitening,antibacterial and soothing effect,and looks forward to the study of Anoectochilus roxburghii in the field of cosmetics,with a view to providing references to the development and utilization of this resource in the field of cosmetics.展开更多
This study employed single factor tests and an orthogonal deign to optimize the process for extracting triterpenoids from Anoectochilus roxburghii with ultrasound-assisted technology.The results showed that the optima...This study employed single factor tests and an orthogonal deign to optimize the process for extracting triterpenoids from Anoectochilus roxburghii with ultrasound-assisted technology.The results showed that the optimal extraction conditions were ultrasound treatment time of 30 min,a solid-to-liquid ratio of 1∶40,an ethanol volume fraction of 70%,and extraction for 40 min.Under these conditions,the yield of triterpenoids extracted from A.roxburghii was 1.621%,with a relative deviation of 0.0335%.The results provided a basis for the extraction and utilization of medicinal components in A.roxburghii.展开更多
Anoectochilus chapaensis Gagnep. （Orchidaceae）, an indigenous and valuable Chinese folk medicine, has been widely used in China to treat diabetes. However, few reports are available about its constituents and activi...Anoectochilus chapaensis Gagnep. （Orchidaceae）, an indigenous and valuable Chinese folk medicine, has been widely used in China to treat diabetes. However, few reports are available about its constituents and activity. The present experiment was conducted to investigate the active fractions from A. chapaensis in diabetic rat model induced by high-fat diet plus streptozotocin. The total EtOH extract from the whole herbs ofA. chapaensis, half of which was partitioned in sequence with petrol ether （PE）, ethyl acetate （EtOAc）, n-BuOH and H2O, thus yielding four fractions, all of them were orally administered with an identical dose amount to 4 g/kg dried crude herbs once a day for consecutive two weeks to further investigate the antihyperglycemic activity. The EtOAc fraction caused a significant fall in the non-fasting blood glucose level of diabetic rats from （402.66±82.26） to （226.26±62.10） mg/dl, which may be attributed to ameliorating insulin resistance, modulating the activity of enzymatic antioxidants, reducing the content of NO, etc. Much more intact β cells in the islets of Langerhans in EtOAc fraction-treated groups than the negative control were observed, which greatly supported the morphological and functional elucidation. The OGTT evidenced that EtOAc fraction could promote the endurance capacity of acute glucose increase in diabetic rats. The EtOAc fraction of A. chapaensis contains some hypoglycemic and antioxidant principles with the potential to be developed further for the treatment of diabetes specifically associated with an insulin resistance state.展开更多
Anoectochilus chapaensis Gagnep. （Orchidaceae） was named as the ＂king of medicine＂ because of its excellent efficacy for the treatment of diabetes. However, the bioactive constituents are unknown. An ethanol extra...Anoectochilus chapaensis Gagnep. （Orchidaceae） was named as the ＂king of medicine＂ because of its excellent efficacy for the treatment of diabetes. However, the bioactive constituents are unknown. An ethanol extract from A. chapaensis showed significant stimulating effect on glucose consumption in HepG2 cells. The chemical composition was investigated by UPLC-MS/MS in negative electrospray ionization （ESI） mode, and 63 compounds including flavonoids, triterpenoids, and aliphatic acids were tentatively identified by accurate mass and characteristic fragments. Moreover, the method of hypoglycemic screening with insulin resistant HepG2 cells and UPLC-MS/MS might be potentially useful in rapid and efficient characterization and primary prediction of natural products prior to traditional isolation.展开更多
Kinsenoside is a main active component isolated from plants of the genus Anoectochilus,and exhibits many biological activities and pharmacological effects,including hepatoprotective,anti-hyperglycemic,anti-hyperliposi...Kinsenoside is a main active component isolated from plants of the genus Anoectochilus,and exhibits many biological activities and pharmacological effects,including hepatoprotective,anti-hyperglycemic,anti-hyperliposis,anti-inflammatory,vascular protective and anti-osteoporosis effects and so on,which is contributing to its promising potency in disease treatments.This review aims to recapitulate the pharmacological functions of kinsenoside,as well as its source,extraction,identification,quantitative analysis,pharmacokinetics,synthesis and patent information.The data reported in this work can confirm the therapeutic potential of kinsenoside and provide useful information for further new drug development.展开更多
[Objectives]To screen the tissue culture rapid propagation formula suitable for each tissue culture stage of Anoectochilus roxburghii( Wall) Lind. [Methods] The stem segments of Fujian A. roxburghii were used as expla...[Objectives]To screen the tissue culture rapid propagation formula suitable for each tissue culture stage of Anoectochilus roxburghii( Wall) Lind. [Methods] The stem segments of Fujian A. roxburghii were used as explant to study the tissue culture rapid propagation and transplantation techniques. The comparative experiment was carried out to study the effects of different hormone concentrations on the induction of stem segments,proliferation of cluster buds,rooting and seedling hardening of A. roxburghii,and study the effects of transplantation matrix on the transplantation of A. roxburghii. [Results]MS + 0. 5 mg/L NAA + 2 mg/L BA + 20 g/L sucrose + 6 g/L agar was suitable for induction of stem segments of A. roxburghii; MS + 0. 5 mg/L NAA + 2 mg/L BA + 1 mg/L KT + 25 g/L sucrose + 6 g/L agar was most suitable for proliferation of cluster buds of A. roxburghii; MS + 1. 0 mg/L IBA + 1. 0 mg/L NAA + 1 g/L activated carbon + 50 g/L mashed banana + 25 g/L sucrose + 6 g/L agar was most suitable for rooting and seedling hardening of A. roxburghii; using peat soil: fine sand( 3∶ 1) as transplantation matrix,the survival rate was the highest. [Conclusions] The experiment results are expected to provide references for factory production of A. roxburghii.展开更多
As a traditional precious Chinese herbal medicine,Anoectochilus roxburghii has the cooling and detoxifying effects,and can nourish yin to reduce pathogenic fire,and has anti-inflammatory and analgesic effects. In this...As a traditional precious Chinese herbal medicine,Anoectochilus roxburghii has the cooling and detoxifying effects,and can nourish yin to reduce pathogenic fire,and has anti-inflammatory and analgesic effects. In this paper,we summarized the research progress in detail about the biological characteristics,resource distribution,composition,medicinal value and rapid seedling breeding technology concerning the rare Chinese herbal medicine A. roxburghii,in order to provide a reference for the in-depth study and rational development and utilization of A.roxburghii.展开更多
[Objectives]To compare the differences in the total flavonoids between leaves and whole plants of Anoectochilus roxburghii.[Methods]The total flavonoids in the tissue cultured leaves and whole plants of A.roxburghii w...[Objectives]To compare the differences in the total flavonoids between leaves and whole plants of Anoectochilus roxburghii.[Methods]The total flavonoids in the tissue cultured leaves and whole plants of A.roxburghii were extracted by microwave-assisted extraction.The content of total flavonoids in the extracts was determined by UV spectrophotometry.[Results]The results of the content determination showed that there were significant differences in the flavonoid content between the leaves and the whole plants of A.roxburghii.[Conclusions]The content of total flavonoids in tissue cultured leaves was higher than that of whole plants.Thus,in the development and utilization of A.roxburghii,tissue cultured leaves can be used to take the place of the whole plants.展开更多
[Objective] This study aimed to investigate the diversity of community composition and diversity of endophytic bacteria in Anoectochilus roxburghii.[Method] By using Illumina MiSeq high-throughput sequencing technolo...[Objective] This study aimed to investigate the diversity of community composition and diversity of endophytic bacteria in Anoectochilus roxburghii.[Method] By using Illumina MiSeq high-throughput sequencing technology, the 16S rDNA-V4 region of endophytic bacteria in ^4. roxburghii was sequenced. Qiime and Mothur software packages were used to sort and count the number of sequences and operational taxonomic units （OTUs） and analyze the species abundance and distribution, Alpha diversity and differences of species abundance among samples. [Result] The number of effective sequences/OTUs for each sample was 110485/247 （stem） and 98524/259 （leaf） , respectively. The rarefaction curve showed that adequate sampling was performed. The number of OTUs had almost reached a saturation point. Endophytic bacteria in A. roxburghii mainly belonged to the genera Acidovorax, Pseudomonas, Sphingomonas and Halomonas. [Conclusions] Illumi-na MiSeq high-throughput sequencing technology provides more accurate and scientific data resources for the study of endophytic bacteria in A. roxburghii.展开更多
Mycorrhizal fungi promote the growth and development of plants, including medicinal plants. The mechanisms by which this growth promotion occurs are of theoretical interest and practical importance to agriculture. Her...Mycorrhizal fungi promote the growth and development of plants, including medicinal plants. The mechanisms by which this growth promotion occurs are of theoretical interest and practical importance to agriculture. Here, an endophytic fungus （AR-18） was isolated from roots of the orchid Anoectochilus roxburghii growing in the wild, and identified as Epulorhiza sp. Tissue-cultured seedlings of A. roxburghii were inoculated with AR-l 8 and co-cultured for 60 d. Endotrophic mycorrhiza formed and the growth of A. roxburghii was markedly promoted by the fungus. To identify genes in A. roxburghii that were differentially expressed during the symbiosis with AR-18, we used the differential display reverse transcription polymerase chain reac- tion （DDRT-PCR） method to compare the transcriptomes between seedlings inoculated with the fungus and control seedlings. We amplified 52 DDRT-PCR bands using 15 primer combinations of three anchor primers and five arbitrary primers, and nine bands were re-amplified by double primers. Reverse Northern blot analyses were used to further screen the bands. Five clones were up-regulated in the symbiotic interaction, including genes encoding a uracil phosphoribosyltransferase （UPRTs; EC 2.4.2.9） and a hypothetical protein. One gene encoding an amino acid transmembrane transporter was down-regulated, and one gene encoding a tRNA-Lys （trnK） and a maturase K （matK） pseudogene were expressed only in the inoculated seedlings. The possible roles of the above genes, especially the UPRTs and marK genes, are discussed in relation to the fungal interaction. This study is the first of its type in A. roxburghii.展开更多
Eight compounds were isolated from the ethyl acetate- and n-butanol-soluble fractions of the ethanollc extract of the whole plant of Anoectochllus roxburghll（Wali.） Lindi. （Orchidaceee）. On the basis of spectrosco...Eight compounds were isolated from the ethyl acetate- and n-butanol-soluble fractions of the ethanollc extract of the whole plant of Anoectochllus roxburghll（Wali.） Lindi. （Orchidaceee）. On the basis of spectroscopic methods, the structures of these compounds were elucidated as quercetin-7-O-β-D-[6＂-O-（trans-feruloyi）]- giucopyranosids （compound 1）, 8-C-p-HydroxybenzyiquerceUn （compound 2）, isorhamnetin-7-O-β-D- giucopyranoside （compound 3）, isorhamnetin-3-O-β-D-giucopyranoside （compound 4）, kaempferoi-3-O-β-D- giucopyranoside （compound 5）, kaempferoi-7-O-β-D-giucopyranoside （compound 6）, 5-hydroxy-3＇,4＇,7- trimethoxyfiavonoi-3-O-β-D-rutinoside （compound 7）, and isorhamnetin-3-O-β-D-rutinoside （compound 8）. Of the compounds isoisted, compound 1 was a new flavonoid giucoside and exhibited strong scavenging activity against the 1,1-diphenyi-2-picryihydrazyi free radical, whereas the ethanolic extract showed weak activity. Compounds 2-8 were obtained from this family for the first time.展开更多
Objective:Diterpenoids with a wide variety of biological activities from Anoectochilus roxburghii,a precious medicinal plant,are important active components.However,due to the lack of genetic information on the metabo...Objective:Diterpenoids with a wide variety of biological activities from Anoectochilus roxburghii,a precious medicinal plant,are important active components.However,due to the lack of genetic information on the metabolic process of diterpenoids in A.roxburghii,the genes involved in the molecular regulation mechanism of diterpenoid metabolism are still unclear.This study revealed the complex metabolic genes for diterpenoids biosynthesis in different organs of A.roxburghii by combining analysis of transcriptomics and metabolomics.Methods:The differences in diterpenoid accumulation in roots,stems and leaves of A.roxburghii were analyzed by metabonomic analysis,and its metabolic gene information was obtained by transcriptome sequencing.Then,the molecular mechanism of differential diterpenoid accumulation in different organs of A.roxburghii was analyzed from the perspective of gene expression patterns.Results:A total of 296 terpenoid metabolites were identified in the five terpenoid metabolic pathways in A.roxburghii.There were 38,34,and 18 diterpenoids with different contents between roots and leaves,between leaves and stems,and between roots and stems,respectively.Twenty-nine metabolic enzyme genes with 883 unigenes in the diterpenoid synthesis process were identified,and the DXS and FDPS in the terpenoid backbone biosynthesis stage and CPA,GA20ox,GA3ox,GA2ox,and MAS in the diterpenoid biosynthesis stage were predicted to be the key metabolic enzymes for the accumulation of diterpenoids.In addition,14 key transcription factor coding genes were predicted to be involved in the regulation of the diterpenoid biosynthesis.The expression of genes such as GA2ox,MAS,CPA,GA20ox and GA3ox might be activated by some of the 14 transcription factors.The transcription factor NTF-Y and PRE6 were predicted to be the most important transcription factors.Conclusion:This study determined 29 metabolic enzyme genes and predicted 14 transcription factors involved in the molecular regulation mechanism of diterpenoid metabolism in A.roxburghii,which provided a reference for the further study of the molecular regulation mechanism of the accumulation of diterpenoids in different organs of A.roxburghii.展开更多
文摘Anoectochilus roxburghii from different planting sources was used as the research object,and the biological activity of its extracts was compared.The results showed that the whole plant extracts of A.roxburghii planted in bag culture (BAG),greenhouse cultivation (GRE) and tissue culture seedlings (TIS) were rich in active ingredients such as A.roxburghii polysaccharides and A.roxburghii glycosides.In vitro experiments showed that Anoectochilus roxburghii extracts from different planting sources had good antioxidant capacity,tyrosinase inhibition ability and hyaluronidase inhibition ability.Among them,the GRE Anoectochilus roxburghii whole plant extract had the strongest scavenging ability on DPPH free radical,ABTS free radical and hydroxyl free radical,and the inhibitory effect on tyrosinase activity and hyaluronidase activity was significantly higher than that of BAG and TIS.At the same time,it also verified that the extract of Anoectochilus roxburghii has antioxidant,whitening,anti-inflammatory and other skin care effects,which provides a theoretical basis for the development and application of Anoectochilus roxburghii extract in the field of cosmetics.
文摘Cosmetics made from natural ingredients are becoming more and more common because they contain bioactive compounds that provide many health benefits,are more environmentally friendly and sustainable.Anoectochilus roxburghii(AR)is a precious Chinese herb in China,possessing bioactive components including flavonoids,polysaccharides,volatile oils,kinsenoside,etc.Due to its unique bioactivity and high safety,the prospect of development and utilization is very broad.In this paper,the extraction,separation and purification techniques of AR’s flavonoid,polysaccharides,volatile oils and kinsenoside are briefly summarized.Meanwhile,we discusses their conformational relationships in cosmetics with antioxidant,whitening,antibacterial and soothing effect,and looks forward to the study of Anoectochilus roxburghii in the field of cosmetics,with a view to providing references to the development and utilization of this resource in the field of cosmetics.
文摘This study employed single factor tests and an orthogonal deign to optimize the process for extracting triterpenoids from Anoectochilus roxburghii with ultrasound-assisted technology.The results showed that the optimal extraction conditions were ultrasound treatment time of 30 min,a solid-to-liquid ratio of 1∶40,an ethanol volume fraction of 70%,and extraction for 40 min.Under these conditions,the yield of triterpenoids extracted from A.roxburghii was 1.621%,with a relative deviation of 0.0335%.The results provided a basis for the extraction and utilization of medicinal components in A.roxburghii.
基金National Natural Science Foundation of China(Grant No.81001628)Guangdong Natural Science Foundation(Grant No.S2013010014771)
文摘Anoectochilus chapaensis Gagnep. （Orchidaceae）, an indigenous and valuable Chinese folk medicine, has been widely used in China to treat diabetes. However, few reports are available about its constituents and activity. The present experiment was conducted to investigate the active fractions from A. chapaensis in diabetic rat model induced by high-fat diet plus streptozotocin. The total EtOH extract from the whole herbs ofA. chapaensis, half of which was partitioned in sequence with petrol ether （PE）, ethyl acetate （EtOAc）, n-BuOH and H2O, thus yielding four fractions, all of them were orally administered with an identical dose amount to 4 g/kg dried crude herbs once a day for consecutive two weeks to further investigate the antihyperglycemic activity. The EtOAc fraction caused a significant fall in the non-fasting blood glucose level of diabetic rats from （402.66±82.26） to （226.26±62.10） mg/dl, which may be attributed to ameliorating insulin resistance, modulating the activity of enzymatic antioxidants, reducing the content of NO, etc. Much more intact β cells in the islets of Langerhans in EtOAc fraction-treated groups than the negative control were observed, which greatly supported the morphological and functional elucidation. The OGTT evidenced that EtOAc fraction could promote the endurance capacity of acute glucose increase in diabetic rats. The EtOAc fraction of A. chapaensis contains some hypoglycemic and antioxidant principles with the potential to be developed further for the treatment of diabetes specifically associated with an insulin resistance state.
基金Pearl River Nova Program of Guangzhou 2015(Grant No.201506010061)Foundation for Distinguished Young Teachers in Higher Education of Guangdong(Grant No.YQ2015097)National Natural Science Foundation of China(Grant No.81001628)
文摘Anoectochilus chapaensis Gagnep. （Orchidaceae） was named as the ＂king of medicine＂ because of its excellent efficacy for the treatment of diabetes. However, the bioactive constituents are unknown. An ethanol extract from A. chapaensis showed significant stimulating effect on glucose consumption in HepG2 cells. The chemical composition was investigated by UPLC-MS/MS in negative electrospray ionization （ESI） mode, and 63 compounds including flavonoids, triterpenoids, and aliphatic acids were tentatively identified by accurate mass and characteristic fragments. Moreover, the method of hypoglycemic screening with insulin resistant HepG2 cells and UPLC-MS/MS might be potentially useful in rapid and efficient characterization and primary prediction of natural products prior to traditional isolation.
文摘Kinsenoside is a main active component isolated from plants of the genus Anoectochilus,and exhibits many biological activities and pharmacological effects,including hepatoprotective,anti-hyperglycemic,anti-hyperliposis,anti-inflammatory,vascular protective and anti-osteoporosis effects and so on,which is contributing to its promising potency in disease treatments.This review aims to recapitulate the pharmacological functions of kinsenoside,as well as its source,extraction,identification,quantitative analysis,pharmacokinetics,synthesis and patent information.The data reported in this work can confirm the therapeutic potential of kinsenoside and provide useful information for further new drug development.
基金Supported by Science and Technology Project of Huizhou City(2014C040007001)
文摘[Objectives]To screen the tissue culture rapid propagation formula suitable for each tissue culture stage of Anoectochilus roxburghii( Wall) Lind. [Methods] The stem segments of Fujian A. roxburghii were used as explant to study the tissue culture rapid propagation and transplantation techniques. The comparative experiment was carried out to study the effects of different hormone concentrations on the induction of stem segments,proliferation of cluster buds,rooting and seedling hardening of A. roxburghii,and study the effects of transplantation matrix on the transplantation of A. roxburghii. [Results]MS + 0. 5 mg/L NAA + 2 mg/L BA + 20 g/L sucrose + 6 g/L agar was suitable for induction of stem segments of A. roxburghii; MS + 0. 5 mg/L NAA + 2 mg/L BA + 1 mg/L KT + 25 g/L sucrose + 6 g/L agar was most suitable for proliferation of cluster buds of A. roxburghii; MS + 1. 0 mg/L IBA + 1. 0 mg/L NAA + 1 g/L activated carbon + 50 g/L mashed banana + 25 g/L sucrose + 6 g/L agar was most suitable for rooting and seedling hardening of A. roxburghii; using peat soil: fine sand( 3∶ 1) as transplantation matrix,the survival rate was the highest. [Conclusions] The experiment results are expected to provide references for factory production of A. roxburghii.
基金Supported by Special Chinese Herbal Medicine Breeding Project of Provincial Department of Science and Technology(2016C02058)Key Provincial Research and Development Project(2017C02019)
文摘As a traditional precious Chinese herbal medicine,Anoectochilus roxburghii has the cooling and detoxifying effects,and can nourish yin to reduce pathogenic fire,and has anti-inflammatory and analgesic effects. In this paper,we summarized the research progress in detail about the biological characteristics,resource distribution,composition,medicinal value and rapid seedling breeding technology concerning the rare Chinese herbal medicine A. roxburghii,in order to provide a reference for the in-depth study and rational development and utilization of A.roxburghii.
基金Supported by the Forestry Science and Technology Extension Project of Central Finance"Tissue Culture Rapid Propagation and Cultivation Techniques for Anoectochilus roxburghii"(JXTG[2016]-05)
文摘[Objectives]To compare the differences in the total flavonoids between leaves and whole plants of Anoectochilus roxburghii.[Methods]The total flavonoids in the tissue cultured leaves and whole plants of A.roxburghii were extracted by microwave-assisted extraction.The content of total flavonoids in the extracts was determined by UV spectrophotometry.[Results]The results of the content determination showed that there were significant differences in the flavonoid content between the leaves and the whole plants of A.roxburghii.[Conclusions]The content of total flavonoids in tissue cultured leaves was higher than that of whole plants.Thus,in the development and utilization of A.roxburghii,tissue cultured leaves can be used to take the place of the whole plants.
基金Supported by National Natural Science Foundation of China(31460491)Youth Project of Applied Basic Research Program of Yunnan Provincial Science and Technology Department(2013FD040)+2 种基金Scientific Research Project of Education Department of Yunnan Province(2014Y390)Talent Introduction Project of Kunming University(YJL14005)Project of Key Characteristic Discipline(Ecology)Construction for Colleges and Universities in Yunnan Province(05000511311)
文摘[Objective] This study aimed to investigate the diversity of community composition and diversity of endophytic bacteria in Anoectochilus roxburghii.[Method] By using Illumina MiSeq high-throughput sequencing technology, the 16S rDNA-V4 region of endophytic bacteria in ^4. roxburghii was sequenced. Qiime and Mothur software packages were used to sort and count the number of sequences and operational taxonomic units （OTUs） and analyze the species abundance and distribution, Alpha diversity and differences of species abundance among samples. [Result] The number of effective sequences/OTUs for each sample was 110485/247 （stem） and 98524/259 （leaf） , respectively. The rarefaction curve showed that adequate sampling was performed. The number of OTUs had almost reached a saturation point. Endophytic bacteria in A. roxburghii mainly belonged to the genera Acidovorax, Pseudomonas, Sphingomonas and Halomonas. [Conclusions] Illumi-na MiSeq high-throughput sequencing technology provides more accurate and scientific data resources for the study of endophytic bacteria in A. roxburghii.
基金supported by the National Natural Science Foundation of China (Grant Nos. 31070300, 31170314 and 31100265)the Chinese Postdoctoral Science Fund (Grant No. 20080440328)+1 种基金the Natural Science Foundation of Chongqing (Grant No. CSTC2008BB5410)the Educational Committee Science & Technology Foundation of Chongqing (Grant No. KJ090504)
文摘Mycorrhizal fungi promote the growth and development of plants, including medicinal plants. The mechanisms by which this growth promotion occurs are of theoretical interest and practical importance to agriculture. Here, an endophytic fungus （AR-18） was isolated from roots of the orchid Anoectochilus roxburghii growing in the wild, and identified as Epulorhiza sp. Tissue-cultured seedlings of A. roxburghii were inoculated with AR-l 8 and co-cultured for 60 d. Endotrophic mycorrhiza formed and the growth of A. roxburghii was markedly promoted by the fungus. To identify genes in A. roxburghii that were differentially expressed during the symbiosis with AR-18, we used the differential display reverse transcription polymerase chain reac- tion （DDRT-PCR） method to compare the transcriptomes between seedlings inoculated with the fungus and control seedlings. We amplified 52 DDRT-PCR bands using 15 primer combinations of three anchor primers and five arbitrary primers, and nine bands were re-amplified by double primers. Reverse Northern blot analyses were used to further screen the bands. Five clones were up-regulated in the symbiotic interaction, including genes encoding a uracil phosphoribosyltransferase （UPRTs; EC 2.4.2.9） and a hypothetical protein. One gene encoding an amino acid transmembrane transporter was down-regulated, and one gene encoding a tRNA-Lys （trnK） and a maturase K （matK） pseudogene were expressed only in the inoculated seedlings. The possible roles of the above genes, especially the UPRTs and marK genes, are discussed in relation to the fungal interaction. This study is the first of its type in A. roxburghii.
基金Supported by the National Natural Science Foundation of China （30270148） and the Chinese National Science Fund for 0utstanding Youths （30325047）.
文摘Eight compounds were isolated from the ethyl acetate- and n-butanol-soluble fractions of the ethanollc extract of the whole plant of Anoectochllus roxburghll（Wali.） Lindi. （Orchidaceee）. On the basis of spectroscopic methods, the structures of these compounds were elucidated as quercetin-7-O-β-D-[6＂-O-（trans-feruloyi）]- giucopyranosids （compound 1）, 8-C-p-HydroxybenzyiquerceUn （compound 2）, isorhamnetin-7-O-β-D- giucopyranoside （compound 3）, isorhamnetin-3-O-β-D-giucopyranoside （compound 4）, kaempferoi-3-O-β-D- giucopyranoside （compound 5）, kaempferoi-7-O-β-D-giucopyranoside （compound 6）, 5-hydroxy-3＇,4＇,7- trimethoxyfiavonoi-3-O-β-D-rutinoside （compound 7）, and isorhamnetin-3-O-β-D-rutinoside （compound 8）. Of the compounds isoisted, compound 1 was a new flavonoid giucoside and exhibited strong scavenging activity against the 1,1-diphenyi-2-picryihydrazyi free radical, whereas the ethanolic extract showed weak activity. Compounds 2-8 were obtained from this family for the first time.
基金This work was financially supported by grants from the Department of Technology and Science of the Fujian Provincial Government(No.2021J01132,No.2020J01732)the Fujian Agriculture and Forestry University Science and Technology Innovation Special Fund(No.KFA18071A).
文摘Objective:Diterpenoids with a wide variety of biological activities from Anoectochilus roxburghii,a precious medicinal plant,are important active components.However,due to the lack of genetic information on the metabolic process of diterpenoids in A.roxburghii,the genes involved in the molecular regulation mechanism of diterpenoid metabolism are still unclear.This study revealed the complex metabolic genes for diterpenoids biosynthesis in different organs of A.roxburghii by combining analysis of transcriptomics and metabolomics.Methods:The differences in diterpenoid accumulation in roots,stems and leaves of A.roxburghii were analyzed by metabonomic analysis,and its metabolic gene information was obtained by transcriptome sequencing.Then,the molecular mechanism of differential diterpenoid accumulation in different organs of A.roxburghii was analyzed from the perspective of gene expression patterns.Results:A total of 296 terpenoid metabolites were identified in the five terpenoid metabolic pathways in A.roxburghii.There were 38,34,and 18 diterpenoids with different contents between roots and leaves,between leaves and stems,and between roots and stems,respectively.Twenty-nine metabolic enzyme genes with 883 unigenes in the diterpenoid synthesis process were identified,and the DXS and FDPS in the terpenoid backbone biosynthesis stage and CPA,GA20ox,GA3ox,GA2ox,and MAS in the diterpenoid biosynthesis stage were predicted to be the key metabolic enzymes for the accumulation of diterpenoids.In addition,14 key transcription factor coding genes were predicted to be involved in the regulation of the diterpenoid biosynthesis.The expression of genes such as GA2ox,MAS,CPA,GA20ox and GA3ox might be activated by some of the 14 transcription factors.The transcription factor NTF-Y and PRE6 were predicted to be the most important transcription factors.Conclusion:This study determined 29 metabolic enzyme genes and predicted 14 transcription factors involved in the molecular regulation mechanism of diterpenoid metabolism in A.roxburghii,which provided a reference for the further study of the molecular regulation mechanism of the accumulation of diterpenoids in different organs of A.roxburghii.
