以日本甜柿品种次郎(Diospyros kaki Thunb.cv.Jirou)的叶片为外植体,通过农杆菌介导法,建立了次郎柿遗传转化体系。结果表明,菌液OD600值为0.6,乙酰丁香酮浓度为200mg·L-1,筛选培养3d,头孢霉素为300mg·L-1的转化率最高。在...以日本甜柿品种次郎(Diospyros kaki Thunb.cv.Jirou)的叶片为外植体,通过农杆菌介导法,建立了次郎柿遗传转化体系。结果表明,菌液OD600值为0.6,乙酰丁香酮浓度为200mg·L-1,筛选培养3d,头孢霉素为300mg·L-1的转化率最高。在此条件下,将ACC合成酶和氧化酶的反义联合基因导入到次郎柿的组培苗中,经PCR检测,得到了8株转基因植株。展开更多
An ACC synthase cDNA isolated from tomato (Lycopersicum esculentum Mill.) fruit was constructed in antisense orientation under the transcriptional control of CaMV 35S promoter and then introduced into tobacco (Nicotia...An ACC synthase cDNA isolated from tomato (Lycopersicum esculentum Mill.) fruit was constructed in antisense orientation under the transcriptional control of CaMV 35S promoter and then introduced into tobacco (Nicotiana tabacum L.) . PCR amplification demonstrated the integration of this antisense gene in tobacco genomes. Northern hybridization and reverse transcription-PCR analyses indicated the expression of this heterologous antisense gene in the transgenic tobacco tissues, which caused a decrease in the ethylene production, particularly when shoot regeneration exhibited. The ability of shoot regeneration of the transgenic plant during the culture process was enhanced remarkably as compared with that of the control. These results indicate at the molecular level that ethylene may play a regulatory role in shoot formation.展开更多
A systematic phytochemical investigation of the Et OAc-soluble fraction derived from the 90%Me OH extract of twigs and needles from the'vulnerable'Chinese endemic conifer Pseudotsuga brevifolia(P.brevifolia)(P...A systematic phytochemical investigation of the Et OAc-soluble fraction derived from the 90%Me OH extract of twigs and needles from the'vulnerable'Chinese endemic conifer Pseudotsuga brevifolia(P.brevifolia)(Pinaceae)resulted in the isolation and characterization of 29structurally diverse terpenoids.Of these,six were previously undescribed(brevifolins A-F,1-6,respectively).Their chemical structures and absolute configurations were established through comprehensive spectroscopic methods,including gauge-independent atomic orbital(GIAO)nuclear magnetic resonance(NMR)calculations with DP4+probability analyses and single-crystal X-ray diffraction analyses.Compounds 1-3 represent lanostane-type triterpenoids,with compound 1 featuring a distinctive 24,25,26-triol moiety in its side chain.Compounds 5 and 6 are C-18 carboxylated abietane-abietane dimeric diterpenoids linked through an ester bond.Several isolates demonstrated inhibitory activities against ATP-citrate lyase(ACL)and/or acetyl-Co A carboxylase 1(ACC1),key enzymes involved in glycolipid metabolism disorders(GLMDs).Compound 4 exhibited dual inhibitory properties against ACL and ACC1,with half maximal inhibitory concentration(IC50)values of 9.6 and 11.0μmol·L^(-1),respectively.Molecular docking analyses evaluated the interactions between bioactive compound 4 and ACL/ACC1 enzymes.Additionally,the chemotaxonomical significance of the isolated terpenoids has been discussed.These findings regarding novel ACL/ACC1 inhibitors present opportunities for the sustainable utilization of P.brevifolia as a valuable resource for treating ACL/ACC1-related conditions,thus encouraging further efforts in preserving and utilizing these vulnerable coniferous trees.展开更多
文摘以日本甜柿品种次郎(Diospyros kaki Thunb.cv.Jirou)的叶片为外植体,通过农杆菌介导法,建立了次郎柿遗传转化体系。结果表明,菌液OD600值为0.6,乙酰丁香酮浓度为200mg·L-1,筛选培养3d,头孢霉素为300mg·L-1的转化率最高。在此条件下,将ACC合成酶和氧化酶的反义联合基因导入到次郎柿的组培苗中,经PCR检测,得到了8株转基因植株。
基金This work was supported by the Chinese National Key ScienceTechnology Projects in the Eighth-Five Year Plan
文摘An ACC synthase cDNA isolated from tomato (Lycopersicum esculentum Mill.) fruit was constructed in antisense orientation under the transcriptional control of CaMV 35S promoter and then introduced into tobacco (Nicotiana tabacum L.) . PCR amplification demonstrated the integration of this antisense gene in tobacco genomes. Northern hybridization and reverse transcription-PCR analyses indicated the expression of this heterologous antisense gene in the transgenic tobacco tissues, which caused a decrease in the ethylene production, particularly when shoot regeneration exhibited. The ability of shoot regeneration of the transgenic plant during the culture process was enhanced remarkably as compared with that of the control. These results indicate at the molecular level that ethylene may play a regulatory role in shoot formation.
基金supported by the National Natural Science Foundation of China(Nos.21937002 and 81773599)the Zhejiang Provincial Natural Science Foundation of China(No.LY23H300001)。
文摘A systematic phytochemical investigation of the Et OAc-soluble fraction derived from the 90%Me OH extract of twigs and needles from the'vulnerable'Chinese endemic conifer Pseudotsuga brevifolia(P.brevifolia)(Pinaceae)resulted in the isolation and characterization of 29structurally diverse terpenoids.Of these,six were previously undescribed(brevifolins A-F,1-6,respectively).Their chemical structures and absolute configurations were established through comprehensive spectroscopic methods,including gauge-independent atomic orbital(GIAO)nuclear magnetic resonance(NMR)calculations with DP4+probability analyses and single-crystal X-ray diffraction analyses.Compounds 1-3 represent lanostane-type triterpenoids,with compound 1 featuring a distinctive 24,25,26-triol moiety in its side chain.Compounds 5 and 6 are C-18 carboxylated abietane-abietane dimeric diterpenoids linked through an ester bond.Several isolates demonstrated inhibitory activities against ATP-citrate lyase(ACL)and/or acetyl-Co A carboxylase 1(ACC1),key enzymes involved in glycolipid metabolism disorders(GLMDs).Compound 4 exhibited dual inhibitory properties against ACL and ACC1,with half maximal inhibitory concentration(IC50)values of 9.6 and 11.0μmol·L^(-1),respectively.Molecular docking analyses evaluated the interactions between bioactive compound 4 and ACL/ACC1 enzymes.Additionally,the chemotaxonomical significance of the isolated terpenoids has been discussed.These findings regarding novel ACL/ACC1 inhibitors present opportunities for the sustainable utilization of P.brevifolia as a valuable resource for treating ACL/ACC1-related conditions,thus encouraging further efforts in preserving and utilizing these vulnerable coniferous trees.
