The random amplified polymorphic DNA (RAPD) technique was used to amplify DNA fragment, aiming at finding markers linked to the sex determination in Ginkgo biloba L. One thousand and two hundred random decamers had be...The random amplified polymorphic DNA (RAPD) technique was used to amplify DNA fragment, aiming at finding markers linked to the sex determination in Ginkgo biloba L. One thousand and two hundred random decamers had been screened. Of the 8 372 RAPD bands, only a 682 bp RAPD marker generated by a primer (S1478) of random decamer sequence, named S1478-682, was found to be associated with the male plants. This marker was present in all male plants and absent in all female plants. Ginkgo trees collected in both Beijing and Shenyang, China were tested using primer S1478. Positive results were obtained, suggesting S1478-682 could be utilized as a reliable RAPD marker to detect the sexuality of Ginkgo.展开更多
In midday ginkgo ( Ginkgo biloba L.) leaves have to bear photon flux density over 1400 μmol·m -2 ·s -1 in combination with high temperatures around 35 ℃ at natural habitat. They show typical mi...In midday ginkgo ( Ginkgo biloba L.) leaves have to bear photon flux density over 1400 μmol·m -2 ·s -1 in combination with high temperatures around 35 ℃ at natural habitat. They show typical midday depression of stomatal conductance and of CO 2 assimilation rate. The zeaxanthin changes with light intensity during the day. The influence of the combination of strong light and temperature on photoinhibition was also examined in the laboratory. A low CO 2 internal conductance (31 mmol·m -2 ·s -1 ) was found in ginkgo leaves, which had been exposed to excessive light at temperature between 15 ℃ and 35 ℃ with reduced CO 2 (80 μL·L -1 ) or oxygen (2%) for 2 h, causing a low CO 2 concentration at the carboxylation site and a high proportion of photorespiration. The ratio of electron transport to CO 2 fixation was rather high in ginkgo (16 e -/CO 2 at 25 ℃) as compared with other plants. It increased with temperature also in 2% O 2 which could not be explained solely as due to change of photorespiration. The reduction of oxygen in 340 or 80 μL·L -1 CO 2 had no effect on the extent of photoinhibition at all temperatures, which indicated that electron flow caused by photorespiration in excess light was negligible in protective effect in ginkgo leaves. However, a decreased CO 2 concentration increased photoinhibition, especially at high temperature. It is concluded that the dissipation of excessive excitation energy in the PSⅡ antennae through the xanthophyll cycle may be the major protective mechanism to preventing from the deteriorated effects of strong light in ginkgo leaves.展开更多
Objective To study the central role of ginkgolide B （BN52021） in regulating cardiovascular function of nerve center by examining the effects of ginkgolide B on the electrical activity of rat paraventricular nucleus ...Objective To study the central role of ginkgolide B （BN52021） in regulating cardiovascular function of nerve center by examining the effects of ginkgolide B on the electrical activity of rat paraventricular nucleus （PVN） neurons in hypothalamic slice preparation and to elucidate the mechanism involved. Methods Extracellular single-unit discharge recording technique. Results （1） In response to the application of ginkgolide t3 （0.1, 1, 10 μmol/L; n = 27） into the perfusate for 2 rain, the spontaneous discharge rates （SDR） of 26 （26/27, 96.30%） neurons were significantly decreased in a dose-dependent manner. （2） Pretreatment with L-glutamate （L-Glu, 0.2 mmol/L） led to a marked increase in the SDR of all 8 （100%） neurons in an epileptiform pattern. The increased discharges were suppressed significantly after ginkgolide B （1 μmol/L） was applied into the perfusate for 2 min. （3） In 8 neurons, perfusion of the selective L-type calcium channel agonist, Bay K 8644 （0.1 μmol/L）, induced a significant increase in the discharge rates of 8 （8/8, 100%） neurons, while ginkgolide B （1μmol/L） applied into the perfusate, could inhibit the discharges of 8 （100%） neurons. （4） In 8 neurons, the broad potassium channels blocker, tetraethylammonium （TEA, 1 mmol/L） completely blocked the inhibitory effect of ginkgolide B （1 μmol/L）. Conclusion These results suggest that ginkgolide B can inhibit the electrical activity of paraventricular neurons. The inhibitory effect may be related to the blockade of L-type voltage-activated calcium channel and potentially concerned with delayed rectifier potassium channel （KDR）.展开更多
[Objective]The aim was to construct the fusion gene expression vector which consisted of GFP and TP gene of GGPPS from the Ginkgo biloba L.[Method]The transit-peptide（TP） sequence of GGPPS from cDNA of Ginkgo biloba...[Objective]The aim was to construct the fusion gene expression vector which consisted of GFP and TP gene of GGPPS from the Ginkgo biloba L.[Method]The transit-peptide（TP） sequence of GGPPS from cDNA of Ginkgo biloba L.was successfully cloned by using DNA recombination technology,which was then linked to the efficient plant expression vector p1304 ＋ to construct the fusion gene expression vector p1304 ＋-TP.Then engineering strain EHA105-p1304 ＋-TP was constructed by transformed p1304 ＋-TP to Agrobacterium rhizogenes EHA105 using freeze-thaw method.[Result]The fusion gene expression vector which consisted of GFP and TP gene of GGPPS from the Ginkgo biloba L.and engineering strain EHA105-p1304 ＋-TP were successfully constructed.[Conclusion]It lays a foundation for further study of subcellular localization of TP transit peptide,which can help to clarify the molecular mechanism of a key step in biosynthesis of ginkgolides precursors,and also provides an important basis for the research on metabolic engineering of ginkgolide.展开更多
The ginkgo terpenoids including bilobalide and ginkgolides are the main pharmaceutical components in the leaves or extracts of Ginkgo biloba L. In this paper, the analysis of bilobalide and ginkgolides in leaves of Gi...The ginkgo terpenoids including bilobalide and ginkgolides are the main pharmaceutical components in the leaves or extracts of Ginkgo biloba L. In this paper, the analysis of bilobalide and ginkgolides in leaves of Ginkgo biloba L. by high performance liquid chromatography (HPLC)-electrospray ionization (ESI)-mass spectrometry (MS) was carried out. The separation was performed on Inertsil ODS3 column with methanol-water (36:64) as mobile phase, with 1 mL·min -1 of flow rate at 35℃. Then the mass spectrum analysis was conducted by ZMD micromass electrospray ionization (ESI)-mass spectrometer (MS). The HPLC total ion chromatogram and selected ion chromatogram (with 325, 407, 423, 439 of m/z) of the sample and ESI-/MS mass spectra of the peaks in the chromatograms were obtained. So bilobalide, ginkgolide A, B, C and J in Ginkgo biloba L. leaves were identified. The method is easy and rapid, with a good accuracy.展开更多
文摘The random amplified polymorphic DNA (RAPD) technique was used to amplify DNA fragment, aiming at finding markers linked to the sex determination in Ginkgo biloba L. One thousand and two hundred random decamers had been screened. Of the 8 372 RAPD bands, only a 682 bp RAPD marker generated by a primer (S1478) of random decamer sequence, named S1478-682, was found to be associated with the male plants. This marker was present in all male plants and absent in all female plants. Ginkgo trees collected in both Beijing and Shenyang, China were tested using primer S1478. Positive results were obtained, suggesting S1478-682 could be utilized as a reliable RAPD marker to detect the sexuality of Ginkgo.
文摘In midday ginkgo ( Ginkgo biloba L.) leaves have to bear photon flux density over 1400 μmol·m -2 ·s -1 in combination with high temperatures around 35 ℃ at natural habitat. They show typical midday depression of stomatal conductance and of CO 2 assimilation rate. The zeaxanthin changes with light intensity during the day. The influence of the combination of strong light and temperature on photoinhibition was also examined in the laboratory. A low CO 2 internal conductance (31 mmol·m -2 ·s -1 ) was found in ginkgo leaves, which had been exposed to excessive light at temperature between 15 ℃ and 35 ℃ with reduced CO 2 (80 μL·L -1 ) or oxygen (2%) for 2 h, causing a low CO 2 concentration at the carboxylation site and a high proportion of photorespiration. The ratio of electron transport to CO 2 fixation was rather high in ginkgo (16 e -/CO 2 at 25 ℃) as compared with other plants. It increased with temperature also in 2% O 2 which could not be explained solely as due to change of photorespiration. The reduction of oxygen in 340 or 80 μL·L -1 CO 2 had no effect on the extent of photoinhibition at all temperatures, which indicated that electron flow caused by photorespiration in excess light was negligible in protective effect in ginkgo leaves. However, a decreased CO 2 concentration increased photoinhibition, especially at high temperature. It is concluded that the dissipation of excessive excitation energy in the PSⅡ antennae through the xanthophyll cycle may be the major protective mechanism to preventing from the deteriorated effects of strong light in ginkgo leaves.
文摘Objective To study the central role of ginkgolide B （BN52021） in regulating cardiovascular function of nerve center by examining the effects of ginkgolide B on the electrical activity of rat paraventricular nucleus （PVN） neurons in hypothalamic slice preparation and to elucidate the mechanism involved. Methods Extracellular single-unit discharge recording technique. Results （1） In response to the application of ginkgolide t3 （0.1, 1, 10 μmol/L; n = 27） into the perfusate for 2 rain, the spontaneous discharge rates （SDR） of 26 （26/27, 96.30%） neurons were significantly decreased in a dose-dependent manner. （2） Pretreatment with L-glutamate （L-Glu, 0.2 mmol/L） led to a marked increase in the SDR of all 8 （100%） neurons in an epileptiform pattern. The increased discharges were suppressed significantly after ginkgolide B （1 μmol/L） was applied into the perfusate for 2 min. （3） In 8 neurons, perfusion of the selective L-type calcium channel agonist, Bay K 8644 （0.1 μmol/L）, induced a significant increase in the discharge rates of 8 （8/8, 100%） neurons, while ginkgolide B （1μmol/L） applied into the perfusate, could inhibit the discharges of 8 （100%） neurons. （4） In 8 neurons, the broad potassium channels blocker, tetraethylammonium （TEA, 1 mmol/L） completely blocked the inhibitory effect of ginkgolide B （1 μmol/L）. Conclusion These results suggest that ginkgolide B can inhibit the electrical activity of paraventricular neurons. The inhibitory effect may be related to the blockade of L-type voltage-activated calcium channel and potentially concerned with delayed rectifier potassium channel （KDR）.
基金Supported by The Cloning and Analysis of Key Enzyme Genes in the Biosynthesis Pathway of Lactone Precursor of Ginkgo biloba(30500303)~~
文摘[Objective]The aim was to construct the fusion gene expression vector which consisted of GFP and TP gene of GGPPS from the Ginkgo biloba L.[Method]The transit-peptide（TP） sequence of GGPPS from cDNA of Ginkgo biloba L.was successfully cloned by using DNA recombination technology,which was then linked to the efficient plant expression vector p1304 ＋ to construct the fusion gene expression vector p1304 ＋-TP.Then engineering strain EHA105-p1304 ＋-TP was constructed by transformed p1304 ＋-TP to Agrobacterium rhizogenes EHA105 using freeze-thaw method.[Result]The fusion gene expression vector which consisted of GFP and TP gene of GGPPS from the Ginkgo biloba L.and engineering strain EHA105-p1304 ＋-TP were successfully constructed.[Conclusion]It lays a foundation for further study of subcellular localization of TP transit peptide,which can help to clarify the molecular mechanism of a key step in biosynthesis of ginkgolides precursors,and also provides an important basis for the research on metabolic engineering of ginkgolide.
文摘The ginkgo terpenoids including bilobalide and ginkgolides are the main pharmaceutical components in the leaves or extracts of Ginkgo biloba L. In this paper, the analysis of bilobalide and ginkgolides in leaves of Ginkgo biloba L. by high performance liquid chromatography (HPLC)-electrospray ionization (ESI)-mass spectrometry (MS) was carried out. The separation was performed on Inertsil ODS3 column with methanol-water (36:64) as mobile phase, with 1 mL·min -1 of flow rate at 35℃. Then the mass spectrum analysis was conducted by ZMD micromass electrospray ionization (ESI)-mass spectrometer (MS). The HPLC total ion chromatogram and selected ion chromatogram (with 325, 407, 423, 439 of m/z) of the sample and ESI-/MS mass spectra of the peaks in the chromatograms were obtained. So bilobalide, ginkgolide A, B, C and J in Ginkgo biloba L. leaves were identified. The method is easy and rapid, with a good accuracy.
