With two-year-old Amorpha fruticosa L. cv. Jinye and A. fruticosa as the research objects, the relative conductivity, MDA content, soluble sugar content, solu- ble protein content and proline content in leaf were dete...With two-year-old Amorpha fruticosa L. cv. Jinye and A. fruticosa as the research objects, the relative conductivity, MDA content, soluble sugar content, solu- ble protein content and proline content in leaf were determined during natural drop in temperature, and the SOD, POD and COD activity in leaf were determined under artificial low temperature to explore the adaptability of A. fruticosa L. cv. Jinye to low temperature. The results showed that with the decrease in natural temperature, the content of MDA in A. fruticosa L. cv. Jinye was lower than that in the control, and the contents of osmotic adjustment substances （soluble sugar, soluble protein and proline） increased, indicating that A. fruticosa L. cv. Jinye had a stronger toler- ance to low temperature compared with A. fruticosa. The activity of SOD, POD and CAT in A. fruticosa L. cv. Jinye increased continuously, higher than that in the con- trol, indicating that the resisting ability of A. fruticosa L. cv. Jinye against low tem- perature was superior to that of the control. There was no significant difference in relative conductivity between A. fruticosa L. cv. Jinye and A. fruticosa.展开更多
In this paper,the ultrasonic-assisted extraction process of flavonoid compounds from leaves of Amorpha fruticosa is optimized.In single factor experiments,solid/liquid ratios,ultrasonic power,ethanol concentrations an...In this paper,the ultrasonic-assisted extraction process of flavonoid compounds from leaves of Amorpha fruticosa is optimized.In single factor experiments,solid/liquid ratios,ultrasonic power,ethanol concentrations and extraction cycles were experimental factors.Box–Behnken central composite design and RSM analyzed the effects of the four factors on the yield of total flavonoids.The optimal extraction parameters were solid/liquid ratio 1:50 g/mL,ultrasonic power 316 W,ethanol concentration 50%,4 extraction cycles.In the optimized condition,the estimated value of the regression model was 66.6456 mg/g while the measured value was 66.4329 mg/g.展开更多
We investigated the combined effects of soil moisture and light intensity on the growth, development and ecophysiological characteristics of one-year old Amorpha fruticosa seedlings. Soil moisture and light intensity ...We investigated the combined effects of soil moisture and light intensity on the growth, development and ecophysiological characteristics of one-year old Amorpha fruticosa seedlings. Soil moisture and light intensity influenced the ecophysiological characteristics of Amorpha fruticosa seedlings. Soil moisture resulted in the decreases of growth rate, individual size, net photosynthetic rate, transpiration rate, leaf water loss rate （WLR）, and biomass accumulation of plant parts, and led to increased leaf water saturation deficit （WSD）. Under water stress, more photosynthetic products were allocated to root growth. With decreasing light intensity, net photosynthetic rate, transpiration rate, chla/b, water saturation deficit, water use efficiency, water loss rate and biomass accumulation declined, while Chla, Chlb, Chla＋b and carotenoids （Car） increased and more photosynthetic products were allocated to stem and leaf growth. Maximum growth vigor, net photosynthetic rate and total biomass accumulation in Amorpha fruticosa seedlings was recorded at 75 80% soil water-holding capacity and 100% light density in greenhouse environments.展开更多
An Amorpha fruticosa cDNA encoding 4 coumarate:CoA ligase (4CL), a key enzyme of phenylpropanoid metabolism related to lignin forming, was cloned by degenerating oligo primed polymerase chain reaction (PCR) and ...An Amorpha fruticosa cDNA encoding 4 coumarate:CoA ligase (4CL), a key enzyme of phenylpropanoid metabolism related to lignin forming, was cloned by degenerating oligo primed polymerase chain reaction (PCR) and rapid amplification of cDNA end (RACE) PCR. We designed 5′RACE primers based on 4CLA1 fragment which obtained from degenerate PCR. Inverse PCR and nested PCR enabled cloning of the remainder fragments of the gene included 5′ and 3′ end sequence. The ORF encodes a polypeptide of 540 amino acids. The predicted amino acid sequence exhibits significant homology with those of other cloned 4CL genes, contain domains typical of predicted 4CL proteins, in particular a postulated AMP binding site, catalytic domain, and conserved Cys residues.展开更多
Artemisia annua L. produces small amounts of the sesquiterpenoid artemisinin, which is used for treatment of malaria. A worldwide shortage of the drug has led to intense research to increase the yield of artemisinin i...Artemisia annua L. produces small amounts of the sesquiterpenoid artemisinin, which is used for treatment of malaria. A worldwide shortage of the drug has led to intense research to increase the yield of artemisinin in the plant. In order to study the regulation of expression of a key enzyme of artemisinin biosynthesis, the promoter region of the key enzyme amorpha-4,11-diene synthase (ADS) was cloned and fused with the β-glucuronidase (GUS) reporter gene. Transgenic plants of A. annua expressing this fusion were generated and studied. Transgenic plants expressing the GUS gene were used to establish the activity of the cloned promoter by a GUS activity staining procedure. GUS under the control of the ADS promoter showed specific expression in glandular trichomes. The activity of the ADS promoter varies temporally and in old tissues essentially no GUS staining could be observed. The expression pattern of GUS and ADS in aerial parts of the transgenic plant was essentially the same indicating that the cis-elements controlling glandular trichome specific expression are included in the cloned promoter. However, some cis-element(s) that control expression in root and old leaf appears to be missing in the cloned promoter. Furthermore, qPCR was used to compare the activity of the wild-type ADS promoter with that of the cloned ADS promoter. The latter promoter showed a considerably lower activity than the wild-type promoter as judged from the levels of GUS and ADS transcripts, respectively, which may be due to the removal of an enhancing cis-element from the ADS promoter. The ADS gene is specifically expressed in stalk and secretory cells of glandular trichomes of A. annua.展开更多
A new isoflavone was obstained from the medicinal herb Amorpha fruticosa. It was elucidated asbenzopyran- 12-one, 1,4,10,11 - tetrahydro-6′-[8′-（hydroxymethy1）etheny1]-2,3-dimethoxy-8′-O-β-glucopyranosy1-O-α-D-...A new isoflavone was obstained from the medicinal herb Amorpha fruticosa. It was elucidated asbenzopyran- 12-one, 1,4,10,11 - tetrahydro-6′-[8′-（hydroxymethy1）etheny1]-2,3-dimethoxy-8′-O-β-glucopyranosy1-O-α-D-arabinoside by spectroscopic methods including UV, IR, 1D NMR and 2D NMR techniques. And the activity against acetaminophen-induced hepatotoxicity of this compound was also studied, and found this compound can protect liver obviously from hepatotoxicity induced by acetaminophen （AAP）. ？2009 Ting Guo Kang. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.展开更多
Arbuscular mycorrhizal fungi(AMF) can colonize and form associations with the roots of Amorpha fruticosa L.(desert false indigo). Various genes are induced during the symbiotic process. In this study, de novo transcri...Arbuscular mycorrhizal fungi(AMF) can colonize and form associations with the roots of Amorpha fruticosa L.(desert false indigo). Various genes are induced during the symbiotic process. In this study, de novo transcriptome sequencing using RNA-seq was conducted for the first time for a comprehensive analysis of AMF-A. fruticosa symbionts at the transcript level. We obtained 12 G of raw data from illumina sequencing and recovered 115,786 unigenes with an average length of547 bp, among them 41,848 of significance. A total of2460 diffexpression genes were identified, including 1579 down-regulated and 881 up-regulated genes. A threshold for false discovery rate of \ 0.001 and fold change of [ 1 determined significant differences in gene expression.Using these criteria, we screened 285 significant differentially expressed genes, of which 82 were up-regulated and203 down-regulated. The 82 up-regulated genes were classified according to their functions and assigned into seven categories: stress and defense, metabolism, signaling transduction, protein folding and degradation, energy,protein synthesis, and transcription. The 203 down-regulated genes were screened according to fold change [ 2,and 50 highly significant down-regulated genes were obtained related to stress and defense. The results of this study will provide a useful foundation for further investigation on the metabolic characteristics and molecular mechanisms of AMF associations with leguminous woody shrubs.展开更多
A method based on degenerate Oligo primed polymerase chain reaction (PCR) and random amplification of cDNA end (RACE) PCR for cloning a full length cDNA is described. An Amorpha fruticosa cDNA clone encoding UDP gluco...A method based on degenerate Oligo primed polymerase chain reaction (PCR) and random amplification of cDNA end (RACE) PCR for cloning a full length cDNA is described. An Amorpha fruticosa cDNA clone encoding UDP glucose pyrophosphorylase (UGP), a key enzyme producing UDP glucose in the synthesis of sucrose and cellulose, is cloned by using this method. We design 5’ RACE primers based on UGPA1 fragment, which obtains from degenerate PCR. Inverse PCR and nested PCR enable cloning of the remainder 5’ and 3’ end fragments of the gene. The deduced amino acid sequence exhibits significant homology with the other UGP genes cloned. This method is more simple and inexpensive than screening cDNA library, and can be easily adapted to clone other genes.展开更多
increasing demand of artemisinin in the treatment of malaria has placed substantial stress on the total artemisinin supplies world-wide, so more attention has been paid to increasing the content of artemisinin in the ...increasing demand of artemisinin in the treatment of malaria has placed substantial stress on the total artemisinin supplies world-wide, so more attention has been paid to increasing the content of artemisinin in the Artemisia annua L. plant. In this study, amorpha-4, 11-diene synthase （ADS） cDNA （ads1） and genomics gene （gads1） were cloned from a high-yield A. annua strain 001. The activity of ADS1 was confirmed by heterogeneous overexpression of ads I and in vitro enzymatic incubation. Reverse transcript-polymerase chain reaction results demonstrated that ads1 expressed in leaves, flowers and young stems, but not in roots. This organ-specific expression pattern of ads1 is consistent with that of artemisinin accumulation in the plant. The gads1 has a complex organization including seven exons and six introns, and belongs to class III terpene synthase. DNA gel blotting revealed that the ADS gene has at least four copies in the genome of strain 001. The higher copy numbers might be one of the reasons for its high artemisinin content.展开更多
In order to explore the feasibility of cryopreservation storage on forest tree seeds, germination of Amorpha fruticosa and Platycladus orientalis seeds with moisture content (MC) changing was tested after storage in l...In order to explore the feasibility of cryopreservation storage on forest tree seeds, germination of Amorpha fruticosa and Platycladus orientalis seeds with moisture content (MC) changing was tested after storage in liquid nitrogen of -196 oC for 30 d. The results showed that both species of seeds can keep high germination percentage after cryopreservation. For A. fruticosa, the germination percentage of seeds was the highest when 4.1% MC was used for storage, and for P. orientalis it was the highest when 6.0% MC was used for storage. It is feasible for both species of seeds to be stored in liquid nitrogen.展开更多
The karyotype analysis and physical locations of 45S rDNA were carried out by means of fluorescence in situ hybridization in three species,and two forms of Sophora,two species of Robina,and one species of Amorpha.S.ja...The karyotype analysis and physical locations of 45S rDNA were carried out by means of fluorescence in situ hybridization in three species,and two forms of Sophora,two species of Robina,and one species of Amorpha.S.japonica L.,S.japonica L.f.oligophylla Franch.,S.japonica L.f.pendula Loud.,and S.xanthantha C.Y.Ma.are all tetraploids with 2n=28.There were four 45S rDNA sites in pericentromeric regions of two pairs of chromosomes in each of them.S.rubriflora Tsoong.is a triploid with 2n=21,and three sites were located in each satellite of group 5 chromosomes.In R.pseudoacacia L.(2n=2x=22),we examined four intensive signals in te-lomeric regions of two pairs of satellite chromosomes.In R.hispida L.(2n=2x=30),there were four other signals in centromeric regions besides those like in R.pseudoacacia.Amorpha fruticosa L.has most chromosomes(2n=40)among the eight materials,however,there were only six 45S rDNA loci and they laid in centromeric regions,and satellites of three pairs of chromosomes.45S rDNA is a valuable chromosomal landmark in karyotype analysis.The distribution and genomic organization of rDNA in the three genera were also discussed.展开更多
文摘With two-year-old Amorpha fruticosa L. cv. Jinye and A. fruticosa as the research objects, the relative conductivity, MDA content, soluble sugar content, solu- ble protein content and proline content in leaf were determined during natural drop in temperature, and the SOD, POD and COD activity in leaf were determined under artificial low temperature to explore the adaptability of A. fruticosa L. cv. Jinye to low temperature. The results showed that with the decrease in natural temperature, the content of MDA in A. fruticosa L. cv. Jinye was lower than that in the control, and the contents of osmotic adjustment substances （soluble sugar, soluble protein and proline） increased, indicating that A. fruticosa L. cv. Jinye had a stronger toler- ance to low temperature compared with A. fruticosa. The activity of SOD, POD and CAT in A. fruticosa L. cv. Jinye increased continuously, higher than that in the con- trol, indicating that the resisting ability of A. fruticosa L. cv. Jinye against low tem- perature was superior to that of the control. There was no significant difference in relative conductivity between A. fruticosa L. cv. Jinye and A. fruticosa.
基金supported by Central University Basic Research Funds(2572014CA27),(2572018DB01)Heilongjiang Province Natural Fund(C200913)
文摘In this paper,the ultrasonic-assisted extraction process of flavonoid compounds from leaves of Amorpha fruticosa is optimized.In single factor experiments,solid/liquid ratios,ultrasonic power,ethanol concentrations and extraction cycles were experimental factors.Box–Behnken central composite design and RSM analyzed the effects of the four factors on the yield of total flavonoids.The optimal extraction parameters were solid/liquid ratio 1:50 g/mL,ultrasonic power 316 W,ethanol concentration 50%,4 extraction cycles.In the optimized condition,the estimated value of the regression model was 66.6456 mg/g while the measured value was 66.4329 mg/g.
基金supported by National Science Foundation of China (No.31270374)Independent Innovation Foundation of Shandong University (No.2011DX008)+1 种基金Natural Science Foundation of Shandong Province,China (No.2009ZRB01875ZR2010CM062)
文摘We investigated the combined effects of soil moisture and light intensity on the growth, development and ecophysiological characteristics of one-year old Amorpha fruticosa seedlings. Soil moisture and light intensity influenced the ecophysiological characteristics of Amorpha fruticosa seedlings. Soil moisture resulted in the decreases of growth rate, individual size, net photosynthetic rate, transpiration rate, leaf water loss rate （WLR）, and biomass accumulation of plant parts, and led to increased leaf water saturation deficit （WSD）. Under water stress, more photosynthetic products were allocated to root growth. With decreasing light intensity, net photosynthetic rate, transpiration rate, chla/b, water saturation deficit, water use efficiency, water loss rate and biomass accumulation declined, while Chla, Chlb, Chla＋b and carotenoids （Car） increased and more photosynthetic products were allocated to stem and leaf growth. Maximum growth vigor, net photosynthetic rate and total biomass accumulation in Amorpha fruticosa seedlings was recorded at 75 80% soil water-holding capacity and 100% light density in greenhouse environments.
文摘An Amorpha fruticosa cDNA encoding 4 coumarate:CoA ligase (4CL), a key enzyme of phenylpropanoid metabolism related to lignin forming, was cloned by degenerating oligo primed polymerase chain reaction (PCR) and rapid amplification of cDNA end (RACE) PCR. We designed 5′RACE primers based on 4CLA1 fragment which obtained from degenerate PCR. Inverse PCR and nested PCR enabled cloning of the remainder fragments of the gene included 5′ and 3′ end sequence. The ORF encodes a polypeptide of 540 amino acids. The predicted amino acid sequence exhibits significant homology with those of other cloned 4CL genes, contain domains typical of predicted 4CL proteins, in particular a postulated AMP binding site, catalytic domain, and conserved Cys residues.
文摘Artemisia annua L. produces small amounts of the sesquiterpenoid artemisinin, which is used for treatment of malaria. A worldwide shortage of the drug has led to intense research to increase the yield of artemisinin in the plant. In order to study the regulation of expression of a key enzyme of artemisinin biosynthesis, the promoter region of the key enzyme amorpha-4,11-diene synthase (ADS) was cloned and fused with the β-glucuronidase (GUS) reporter gene. Transgenic plants of A. annua expressing this fusion were generated and studied. Transgenic plants expressing the GUS gene were used to establish the activity of the cloned promoter by a GUS activity staining procedure. GUS under the control of the ADS promoter showed specific expression in glandular trichomes. The activity of the ADS promoter varies temporally and in old tissues essentially no GUS staining could be observed. The expression pattern of GUS and ADS in aerial parts of the transgenic plant was essentially the same indicating that the cis-elements controlling glandular trichome specific expression are included in the cloned promoter. However, some cis-element(s) that control expression in root and old leaf appears to be missing in the cloned promoter. Furthermore, qPCR was used to compare the activity of the wild-type ADS promoter with that of the cloned ADS promoter. The latter promoter showed a considerably lower activity than the wild-type promoter as judged from the levels of GUS and ADS transcripts, respectively, which may be due to the removal of an enhancing cis-element from the ADS promoter. The ADS gene is specifically expressed in stalk and secretory cells of glandular trichomes of A. annua.
基金supported by the Science and Technology of Dalian(No.2008E11SF168).
文摘A new isoflavone was obstained from the medicinal herb Amorpha fruticosa. It was elucidated asbenzopyran- 12-one, 1,4,10,11 - tetrahydro-6′-[8′-（hydroxymethy1）etheny1]-2,3-dimethoxy-8′-O-β-glucopyranosy1-O-α-D-arabinoside by spectroscopic methods including UV, IR, 1D NMR and 2D NMR techniques. And the activity against acetaminophen-induced hepatotoxicity of this compound was also studied, and found this compound can protect liver obviously from hepatotoxicity induced by acetaminophen （AAP）. ？2009 Ting Guo Kang. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.
基金supported by the National Natural Science Foundation of China(31070576 and 31270535)the Natural Science Foundation of Heilongjiang Province(No.ZD201206)+1 种基金the Excellent Youth Foundation of Heilongjiang Province(No.JC201306)High-level Talents Support Program of Heilongjiang University(Ecological Restoration Team)
文摘Arbuscular mycorrhizal fungi(AMF) can colonize and form associations with the roots of Amorpha fruticosa L.(desert false indigo). Various genes are induced during the symbiotic process. In this study, de novo transcriptome sequencing using RNA-seq was conducted for the first time for a comprehensive analysis of AMF-A. fruticosa symbionts at the transcript level. We obtained 12 G of raw data from illumina sequencing and recovered 115,786 unigenes with an average length of547 bp, among them 41,848 of significance. A total of2460 diffexpression genes were identified, including 1579 down-regulated and 881 up-regulated genes. A threshold for false discovery rate of \ 0.001 and fold change of [ 1 determined significant differences in gene expression.Using these criteria, we screened 285 significant differentially expressed genes, of which 82 were up-regulated and203 down-regulated. The 82 up-regulated genes were classified according to their functions and assigned into seven categories: stress and defense, metabolism, signaling transduction, protein folding and degradation, energy,protein synthesis, and transcription. The 203 down-regulated genes were screened according to fold change [ 2,and 50 highly significant down-regulated genes were obtained related to stress and defense. The results of this study will provide a useful foundation for further investigation on the metabolic characteristics and molecular mechanisms of AMF associations with leguminous woody shrubs.
文摘A method based on degenerate Oligo primed polymerase chain reaction (PCR) and random amplification of cDNA end (RACE) PCR for cloning a full length cDNA is described. An Amorpha fruticosa cDNA clone encoding UDP glucose pyrophosphorylase (UGP), a key enzyme producing UDP glucose in the synthesis of sucrose and cellulose, is cloned by using this method. We design 5’ RACE primers based on UGPA1 fragment, which obtains from degenerate PCR. Inverse PCR and nested PCR enable cloning of the remainder 5’ and 3’ end fragments of the gene. The deduced amino acid sequence exhibits significant homology with the other UGP genes cloned. This method is more simple and inexpensive than screening cDNA library, and can be easily adapted to clone other genes.
基金Supported by the National Natural Science Foundation of China （30171740 and 30672623）.
文摘increasing demand of artemisinin in the treatment of malaria has placed substantial stress on the total artemisinin supplies world-wide, so more attention has been paid to increasing the content of artemisinin in the Artemisia annua L. plant. In this study, amorpha-4, 11-diene synthase （ADS） cDNA （ads1） and genomics gene （gads1） were cloned from a high-yield A. annua strain 001. The activity of ADS1 was confirmed by heterogeneous overexpression of ads I and in vitro enzymatic incubation. Reverse transcript-polymerase chain reaction results demonstrated that ads1 expressed in leaves, flowers and young stems, but not in roots. This organ-specific expression pattern of ads1 is consistent with that of artemisinin accumulation in the plant. The gads1 has a complex organization including seven exons and six introns, and belongs to class III terpene synthase. DNA gel blotting revealed that the ADS gene has at least four copies in the genome of strain 001. The higher copy numbers might be one of the reasons for its high artemisinin content.
基金subsidized by the project of "Study on Coming back of Vegetation and Technology of Afforestation in Northwest Farming and Pasturing Mixed Area with Serious Corrosion(2006BAD03A0308)"which is a part of "Study on Key Technology of Afforestation in Barren Site (2006BAD03A03)".
文摘In order to explore the feasibility of cryopreservation storage on forest tree seeds, germination of Amorpha fruticosa and Platycladus orientalis seeds with moisture content (MC) changing was tested after storage in liquid nitrogen of -196 oC for 30 d. The results showed that both species of seeds can keep high germination percentage after cryopreservation. For A. fruticosa, the germination percentage of seeds was the highest when 4.1% MC was used for storage, and for P. orientalis it was the highest when 6.0% MC was used for storage. It is feasible for both species of seeds to be stored in liquid nitrogen.
文摘The karyotype analysis and physical locations of 45S rDNA were carried out by means of fluorescence in situ hybridization in three species,and two forms of Sophora,two species of Robina,and one species of Amorpha.S.japonica L.,S.japonica L.f.oligophylla Franch.,S.japonica L.f.pendula Loud.,and S.xanthantha C.Y.Ma.are all tetraploids with 2n=28.There were four 45S rDNA sites in pericentromeric regions of two pairs of chromosomes in each of them.S.rubriflora Tsoong.is a triploid with 2n=21,and three sites were located in each satellite of group 5 chromosomes.In R.pseudoacacia L.(2n=2x=22),we examined four intensive signals in te-lomeric regions of two pairs of satellite chromosomes.In R.hispida L.(2n=2x=30),there were four other signals in centromeric regions besides those like in R.pseudoacacia.Amorpha fruticosa L.has most chromosomes(2n=40)among the eight materials,however,there were only six 45S rDNA loci and they laid in centromeric regions,and satellites of three pairs of chromosomes.45S rDNA is a valuable chromosomal landmark in karyotype analysis.The distribution and genomic organization of rDNA in the three genera were also discussed.
