This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different deve...This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different developmental stages of 7th, 14th, 21st and 28th day after flowering （DAF） as wel as the root, stem, leaf at winter seedling stages of both the transgenic line W-4 and non-transgenic control Westar by real-time fluorescence quantitative PCR. [Results] The results showed the relative expression of fad2 gene was gradual y increasing with the days after flowering in the seeds of the control Westar, while it was found decreasing significantly since the 21st DAF in the seeds of the line W-4. The decline was up to 60% in comparison with the control Westar. However, no significant difference in the relative expression of fad2 gene in other organs like root, stem and leaf was observed between transgenic line W-4 and non-transgenic control Westar. Fatty acid composition analysis showed the oleic acid desaturation parameter（ODP） in seeds of the line W-4 was 0.07 in average, decreased by nearly 75% than control Westar which was 0.24 in average, while no significant difference in the seedling root, stem and leaf was measured between transgenic rapeseed and control. [Conclusion] The results above validated that RNA interference in transgenic rapeseed W-4 is at a seed-specific manner, not interfering with fad2 gene expression in organs such as the root, stem and leaf. The study also found that the period of fad2 gene expres-sion decline was wel coincided with the expression of napin gene, both appeared at the 21st DAF, indicating that the expression of dsRNA of fad2 gene is precisely control ed by the napin promoter.展开更多
The operation principle of a novel portable oil ingredient instrument is introduced. Using the theory of fluorescence measurement, this instrument can carry out fast and consecutive in-situ detection of oil ingredient...The operation principle of a novel portable oil ingredient instrument is introduced. Using the theory of fluorescence measurement, this instrument can carry out fast and consecutive in-situ detection of oil ingredient in sea water, confirming fast and correctly the situation of oil contamination such as the concentration and the area, etc. The lowest detectable concentration for the instrument is 1×10 -8 g/ml with the precision of less than 3 %. The results of experiment and practice show that this system has well water-sealed characteristic and repeatability.展开更多
Protoplasts of the pathogenic plant fungus,Sclerotinia sclerotiorum,were transformed using the pPGF plasmid,which contains green fluorescent protein gene,under the control of Aspergillus nidulans regulatory sequences....Protoplasts of the pathogenic plant fungus,Sclerotinia sclerotiorum,were transformed using the pPGF plasmid,which contains green fluorescent protein gene,under the control of Aspergillus nidulans regulatory sequences. The pPGF plasmid was introduced by PEG/CaCl2 treatment. Positive transformants were harvested with hygromycin B (HYG) resistance as selective marker,and then were observed with green fluorescence phenomena in response to blue light,which suggested that GFP gene was cloned into genome DNA of S. sclerotiorum. The transformants were verified mitotically stable by Southern blotting analysis and passage culturing. This study is developed as an initial step for further research into infection mechanisms of S. sclerotiorum to plants and interactions with bio-control fungus.展开更多
This paper presents the possibilities offered by fluorescence spectroscopy for the identification of vegetable oils such as soybean, sunflower, flax, walnut, corn, almond, sesame, olive and pumpkin oils. The probes un...This paper presents the possibilities offered by fluorescence spectroscopy for the identification of vegetable oils such as soybean, sunflower, flax, walnut, corn, almond, sesame, olive and pumpkin oils. The probes under study have been excited with two types of sources： a laser diode （LD） and light-emitting diodes （LEDs） emitting in the UV and in the visible range. Total luminescence spectra were recorded by measuring the emission spectra in the range 350-720 nm at excitation wavelengths from 375 to 450 nm. The excitation-emission matrices have been obtained and two basic fluorescence regions in the visible have been outlined. On this basis the fluorescence spectra of the oils have been subdivided into three categories depending on the prevalence of the fluorescence maxima. The samples show differences in their fluorescence spectra. The latter fact shows that fluorescence spectroscopy can be used for the quick identification of edible oils. The fatty acid, the tocopherol, the beta-carotene and chlorophyll contents in the analyzed oils have been studied. It is shown that some of the types of oils differ significantly from each other by the first derivatives of their fluorescence spectra. There also exist color differences between the groups of vegetable oils under study.展开更多
Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the...Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal Rhodophyta. Here, the effects of salinity and light intensity on the quantum yield of photosystem II in Py. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under different stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The cDNA was synthesized and the expression levels of seven potential internal control genes were evaluated using qRT-PCR method. Then, we used geNorm, a common statistical algorithm, to analyze the qRT-PCR data of seven reference genes. Potential genes that may constantly be expressed under different conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while tubulin, glyceraldehyde- 3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the geNorm screening. Based on the results of the software analysis and absolute quantification, we proposed that elongation factor 3 and 18S ribosomal RNA could be used as internal control genes when the Py. haitanensis blades were subjected to salinity stress, and that a-tubulin and 18S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis.展开更多
By using fluorescence lifetime image microscope （FLIM） and time-correlated single photon counting （TCSPC） technique, we measured fluorescence lifetime of crude oils with density of 0.9521-0.7606 g/cm3 and multiple...By using fluorescence lifetime image microscope （FLIM） and time-correlated single photon counting （TCSPC） technique, we measured fluorescence lifetime of crude oils with density of 0.9521-0.7606 g/cm3 and multiple petroleum inclusions from Tazhong uplift of Tarim Basin. As indicated by the test results, crude oil density is closely correlated with average fluorescence lifetime following the regression equation Y=-0.0319X＋0.9411, which can thus be used to calculate density of oil inclusions in relation to fluorescence lifetime and density of corresponding surface crude. For type A oil inclusions showing brown-yellow fluorescence from Tazhong 1 well in Tarim Basin, their average fluorescence lifetime was found to be 2.144-2.765 ns, so the density of surface crude corresponding to crude trapping these oil inclusions is 0.852-0.873 g/cm3, indicating that they are matured oil inclusions trapped at earlier stage ofoil formation. For type B oil inclusions with light yellow-white fluorescence, their average fluorescence lifetime was found to be 4.0294.919 ns, so the density of surface crude corresponding to crude trapping these oil inclusions is 0.784-0.812 g/cm3, indicating that they are higher matured oil inclusions trapped at the second stage of oil formation. For type C oil inclusions showing light blue-green fluorescence, their average fluorescence lifetime was found to be 5.0634.168 ns, so the density of surface crude corresponding to crude trapping these oil inclusions is 0.743-0.779 g/cm3, indicating that they are highly-matured light oil inclusions trapped at the third stage of oil formation.展开更多
When exposed to biotic or abiotic stress conditions, plants produce ethylene from its immediate precursor 1-aminocyclopropane-1- carboxylate （ACC）, leading to retarded root growth and senescence. Many plant growth-p...When exposed to biotic or abiotic stress conditions, plants produce ethylene from its immediate precursor 1-aminocyclopropane-1- carboxylate （ACC）, leading to retarded root growth and senescence. Many plant growth-promoting rhizobacteria contain the enzyme ACC deaminase and this enzyme can cleave ACC to form a-ketobutyrate and ammonium, thereby lowering levels of ethylene. The aim of this study was to isolate and characterize ACC deaminase-producing bacteria from the rhizosphere of salt-stressed canola （Brassica napus L.）. Out of 105 random bacterial isolates, 15 were able to utilize ACC as the sole source of nitrogen. These 15 isolates were also positive for indole acetic acid （IAA） production. Phylogenetic analysis based on partial 16S rDNA sequences showed that all isolates belonged to fluorescent Pseudomonas spp. In the canola rhizosphere investigated in this study, Pseudomonas fluorescens was the dominant ACC deaminase-producing species. Cluster analysis based on BOX-AIR-based repetitive extragenic palindromic-polymerase chain reaction （BOX-PCR） patterns suggested a high degree of genetic variability in ACC deaminase-producing P. fluorescens strains. The presence of indigenous ACC-degrading bacteria in the rhizosphere of canola grown in saline soils indicates that these bacteria may contribute to salinity tolerance.展开更多
Branched glycerol dialkyl glycerol tetraethers （bGDGTs） are ubiquitous and abundant in soils, but their sources remain elusive. Recent studies demonstrate that the distributions of bGDGTs are sensitive to various en...Branched glycerol dialkyl glycerol tetraethers （bGDGTs） are ubiquitous and abundant in soils, but their sources remain elusive. Recent studies demonstrate that the distributions of bGDGTs are sensitive to various environmental factors. In an effort to understand how and to what extent soil moisture （expressed as soil water content （SWC） or mean annual precipita- tion （MAP）, pH and temperature may impact the distribution of bGDGTs, and to shed more light on the biological sources of bGDGTs in cold and arid regions, we investigated the distribution of bGDGTs as well as bacterial 16S rRNA gene and func- tional genes involved in the N cycle （including amoA, nirS and nirK） in 41 surface soil samples from around Lake Qinghai and east of Qaidam Basin on the Northeastern Qinghai-Tibetan Plateau. We found that lower soil moisture reduced the fractional concentrations of cyclic bGDGTs and thus the cyclisation ratio of branched tetraethers （CBT） index correlated negatively with SWC and MAP, suggesting that soil moisture is an important factor controlling bGDGT distributions in soils in this add and semi-add region. Two subgroups of bGDGTs were assigned on the basis of cluster analysis, and bGDGT indices behaved dif- ferently in the two groups, hinting at different biosynthetic mechanisms for bGDGTs under different environmental conditions. Real time PCR results showed that nirS and nirK genes correlated significantly with the concentration of bGDGTs, suggesting that the nirS- or nirK-encoding bacteria involved in denitrification might potentially be an additional biological source for soil bGDGTs （besides Acidobacteria）. Moreover, our results also support the application of new indices based on 5-methyl bGDGTs and 6-methyl bGDGTs in reconstructing past temperature and pH variations in this region.展开更多
基金Supported by Fund for National Rapeseed Research System(CARS-13)~~
文摘This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different developmental stages of 7th, 14th, 21st and 28th day after flowering （DAF） as wel as the root, stem, leaf at winter seedling stages of both the transgenic line W-4 and non-transgenic control Westar by real-time fluorescence quantitative PCR. [Results] The results showed the relative expression of fad2 gene was gradual y increasing with the days after flowering in the seeds of the control Westar, while it was found decreasing significantly since the 21st DAF in the seeds of the line W-4. The decline was up to 60% in comparison with the control Westar. However, no significant difference in the relative expression of fad2 gene in other organs like root, stem and leaf was observed between transgenic line W-4 and non-transgenic control Westar. Fatty acid composition analysis showed the oleic acid desaturation parameter（ODP） in seeds of the line W-4 was 0.07 in average, decreased by nearly 75% than control Westar which was 0.24 in average, while no significant difference in the seedling root, stem and leaf was measured between transgenic rapeseed and control. [Conclusion] The results above validated that RNA interference in transgenic rapeseed W-4 is at a seed-specific manner, not interfering with fad2 gene expression in organs such as the root, stem and leaf. The study also found that the period of fad2 gene expres-sion decline was wel coincided with the expression of napin gene, both appeared at the 21st DAF, indicating that the expression of dsRNA of fad2 gene is precisely control ed by the napin promoter.
文摘The operation principle of a novel portable oil ingredient instrument is introduced. Using the theory of fluorescence measurement, this instrument can carry out fast and consecutive in-situ detection of oil ingredient in sea water, confirming fast and correctly the situation of oil contamination such as the concentration and the area, etc. The lowest detectable concentration for the instrument is 1×10 -8 g/ml with the precision of less than 3 %. The results of experiment and practice show that this system has well water-sealed characteristic and repeatability.
文摘Protoplasts of the pathogenic plant fungus,Sclerotinia sclerotiorum,were transformed using the pPGF plasmid,which contains green fluorescent protein gene,under the control of Aspergillus nidulans regulatory sequences. The pPGF plasmid was introduced by PEG/CaCl2 treatment. Positive transformants were harvested with hygromycin B (HYG) resistance as selective marker,and then were observed with green fluorescence phenomena in response to blue light,which suggested that GFP gene was cloned into genome DNA of S. sclerotiorum. The transformants were verified mitotically stable by Southern blotting analysis and passage culturing. This study is developed as an initial step for further research into infection mechanisms of S. sclerotiorum to plants and interactions with bio-control fungus.
文摘This paper presents the possibilities offered by fluorescence spectroscopy for the identification of vegetable oils such as soybean, sunflower, flax, walnut, corn, almond, sesame, olive and pumpkin oils. The probes under study have been excited with two types of sources： a laser diode （LD） and light-emitting diodes （LEDs） emitting in the UV and in the visible range. Total luminescence spectra were recorded by measuring the emission spectra in the range 350-720 nm at excitation wavelengths from 375 to 450 nm. The excitation-emission matrices have been obtained and two basic fluorescence regions in the visible have been outlined. On this basis the fluorescence spectra of the oils have been subdivided into three categories depending on the prevalence of the fluorescence maxima. The samples show differences in their fluorescence spectra. The latter fact shows that fluorescence spectroscopy can be used for the quick identification of edible oils. The fatty acid, the tocopherol, the beta-carotene and chlorophyll contents in the analyzed oils have been studied. It is shown that some of the types of oils differ significantly from each other by the first derivatives of their fluorescence spectra. There also exist color differences between the groups of vegetable oils under study.
基金Supported by the National Natural Science Foundation of China(Nos.41476140,41306151,41676157,41506172)the Strategic Leading Science and Technology Projects of Chinese Academy of Sciences(No.XDA11020404)+1 种基金the China Postdoctoral Science Foundation(No.2015M582153)the Science and Technology Plan of Jiangsu Province(No.BE2016330)
文摘Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal Rhodophyta. Here, the effects of salinity and light intensity on the quantum yield of photosystem II in Py. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under different stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The cDNA was synthesized and the expression levels of seven potential internal control genes were evaluated using qRT-PCR method. Then, we used geNorm, a common statistical algorithm, to analyze the qRT-PCR data of seven reference genes. Potential genes that may constantly be expressed under different conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while tubulin, glyceraldehyde- 3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the geNorm screening. Based on the results of the software analysis and absolute quantification, we proposed that elongation factor 3 and 18S ribosomal RNA could be used as internal control genes when the Py. haitanensis blades were subjected to salinity stress, and that a-tubulin and 18S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis.
基金supported by the National Natural Science Foundation of China(Grant Nos.41321002,41402116)
文摘By using fluorescence lifetime image microscope （FLIM） and time-correlated single photon counting （TCSPC） technique, we measured fluorescence lifetime of crude oils with density of 0.9521-0.7606 g/cm3 and multiple petroleum inclusions from Tazhong uplift of Tarim Basin. As indicated by the test results, crude oil density is closely correlated with average fluorescence lifetime following the regression equation Y=-0.0319X＋0.9411, which can thus be used to calculate density of oil inclusions in relation to fluorescence lifetime and density of corresponding surface crude. For type A oil inclusions showing brown-yellow fluorescence from Tazhong 1 well in Tarim Basin, their average fluorescence lifetime was found to be 2.144-2.765 ns, so the density of surface crude corresponding to crude trapping these oil inclusions is 0.852-0.873 g/cm3, indicating that they are matured oil inclusions trapped at earlier stage ofoil formation. For type B oil inclusions with light yellow-white fluorescence, their average fluorescence lifetime was found to be 4.0294.919 ns, so the density of surface crude corresponding to crude trapping these oil inclusions is 0.784-0.812 g/cm3, indicating that they are higher matured oil inclusions trapped at the second stage of oil formation. For type C oil inclusions showing light blue-green fluorescence, their average fluorescence lifetime was found to be 5.0634.168 ns, so the density of surface crude corresponding to crude trapping these oil inclusions is 0.743-0.779 g/cm3, indicating that they are highly-matured light oil inclusions trapped at the third stage of oil formation.
文摘When exposed to biotic or abiotic stress conditions, plants produce ethylene from its immediate precursor 1-aminocyclopropane-1- carboxylate （ACC）, leading to retarded root growth and senescence. Many plant growth-promoting rhizobacteria contain the enzyme ACC deaminase and this enzyme can cleave ACC to form a-ketobutyrate and ammonium, thereby lowering levels of ethylene. The aim of this study was to isolate and characterize ACC deaminase-producing bacteria from the rhizosphere of salt-stressed canola （Brassica napus L.）. Out of 105 random bacterial isolates, 15 were able to utilize ACC as the sole source of nitrogen. These 15 isolates were also positive for indole acetic acid （IAA） production. Phylogenetic analysis based on partial 16S rDNA sequences showed that all isolates belonged to fluorescent Pseudomonas spp. In the canola rhizosphere investigated in this study, Pseudomonas fluorescens was the dominant ACC deaminase-producing species. Cluster analysis based on BOX-AIR-based repetitive extragenic palindromic-polymerase chain reaction （BOX-PCR） patterns suggested a high degree of genetic variability in ACC deaminase-producing P. fluorescens strains. The presence of indigenous ACC-degrading bacteria in the rhizosphere of canola grown in saline soils indicates that these bacteria may contribute to salinity tolerance.
基金supported by the National Natural Science Foundation of China (Grant Nos. 41373072, 40873011 &41030211)the Shanghai Bureau of Science and Technology (Grant No. 13JC1405200)the Thousand Talents Program through the State Key Laboratory of Marine Geology at Tongji University (C. L. Zhang)
文摘Branched glycerol dialkyl glycerol tetraethers （bGDGTs） are ubiquitous and abundant in soils, but their sources remain elusive. Recent studies demonstrate that the distributions of bGDGTs are sensitive to various environmental factors. In an effort to understand how and to what extent soil moisture （expressed as soil water content （SWC） or mean annual precipita- tion （MAP）, pH and temperature may impact the distribution of bGDGTs, and to shed more light on the biological sources of bGDGTs in cold and arid regions, we investigated the distribution of bGDGTs as well as bacterial 16S rRNA gene and func- tional genes involved in the N cycle （including amoA, nirS and nirK） in 41 surface soil samples from around Lake Qinghai and east of Qaidam Basin on the Northeastern Qinghai-Tibetan Plateau. We found that lower soil moisture reduced the fractional concentrations of cyclic bGDGTs and thus the cyclisation ratio of branched tetraethers （CBT） index correlated negatively with SWC and MAP, suggesting that soil moisture is an important factor controlling bGDGT distributions in soils in this add and semi-add region. Two subgroups of bGDGTs were assigned on the basis of cluster analysis, and bGDGT indices behaved dif- ferently in the two groups, hinting at different biosynthetic mechanisms for bGDGTs under different environmental conditions. Real time PCR results showed that nirS and nirK genes correlated significantly with the concentration of bGDGTs, suggesting that the nirS- or nirK-encoding bacteria involved in denitrification might potentially be an additional biological source for soil bGDGTs （besides Acidobacteria）. Moreover, our results also support the application of new indices based on 5-methyl bGDGTs and 6-methyl bGDGTs in reconstructing past temperature and pH variations in this region.
