[Objective]The aim of this study was to identify transient expression of movement protein (MP) gene in Nicotinana benthaminana rapidly and further investigate the function of this exogenous gene. [Method]The movemen...[Objective]The aim of this study was to identify transient expression of movement protein (MP) gene in Nicotinana benthaminana rapidly and further investigate the function of this exogenous gene. [Method]The movement protein gene of barley yellow dwarf virus (BYDV) was cloned into potato virus X (PVX) viral vector of pGR107,and PVX-recombinant vector was obtained. After electroporation of Agrobacterium tumefaciens,PVX was inoculated into the lower leaves of tobacco by Agrobacterium infiltration assay to observe the infection of virus on tobacco. [Result]After infection for 7 days,upper non-inoculated leaves of tobacco infected by the PVX-recombinant vector showed the virus infection symptoms,while the control group had no viral infection phenomenon. Daily follow-up observations for two groups revealed that tobacco infected by PVX-recombinant vector had severe symptoms of virus infection and curling leaves,or even led to necrosis both in infiltrated and systemic leaves in late period. However,tobacco infected by PVX vector had only slight symptoms of virus infection and could recover from infection. RT-PCR of the infected tobacco indicated that exogenous gene BYDV-MP had a normal transcription and expression in tobacco. [Conclusion]As a determinant factor for viral disease,BYDV-MP promotes the systemic infection rate of PVX and its symptom. In addition,it is feasible to express exogenous MP gene in Nicotiana benthaminan via PVX expression vector.展开更多
The effect of sowing date on grain protein, hordein fraction content and malting quality of two-rowed spring barley was investigated by using ten commercial cultivars with different grain protein content and the relat...The effect of sowing date on grain protein, hordein fraction content and malting quality of two-rowed spring barley was investigated by using ten commercial cultivars with different grain protein content and the relationships among these traits were examined. The results showed that grain protein content and B hordein content increased as the sowing date postponed and were significantly affected by sowing date, while C and D hordein contents were less influenced by sowing date. There were significant differences in grain protein and hordein fraction content among the ten cultivars. The coefficient of variation of D hordein content was much larger than that of B and C hordein contents, suggesting its greater variation caused by different sowing dates. Beta-amylase activity and diastatic power were also significantly affected by sowing date, with malt extract being less affected. Significant differences in measured malt quality were found among the ten cultivars. Grain protein was significantly correlated with B hordein and malt extract positively and negatively, respectively. There was no significant correlation between beta-amylase activity or diastatic power and grain protein content. B hordein was negatively and significantly correlated with malt extract, but no significant correlations between C hordein, D hordein and malting quality traits.展开更多
AIM: To produce a recombinant protein rMBP-NAP, which was fusionally expressed by Helicobacter pylori(H pylori)neutrophil-activating protein (NAP) and E. coli maltosebinding protein (MBP) and to evaluate its immunorea...AIM: To produce a recombinant protein rMBP-NAP, which was fusionally expressed by Helicobacter pylori(H pylori)neutrophil-activating protein (NAP) and E. coli maltosebinding protein (MBP) and to evaluate its immunoreactivity and immunogenicity.METHODS: Neutrophil-activating protein gene of H pylori (HP-napA) was subcloned from the recombinant plasmid pNEB-napA, and fused to MalE gene of expressing vector pMAL-c2x. The recombinant plasmid pMAL-c2x-napA was confirmed by restriction enzyme digestion, and then transformed into E. coli TB1. Fusion protein rMBP-NAP was induced by IPTG and identified by SDS-PAGE analysis.Soluble rMBP-NAP was purified by amylose affinity chromatography. Immunoreactivity and immunogenicity of the fusion protein were evaluated by animal experiment,Western blotting with human H pylori anti-sera.RESULTS: E.coli TB1 carrying recombinant plasmid pMAL-c2x-napA was constructed and led to a high efficiency cytosol expression of fusion protein rBMP -NAP when induced by IPTG.The molecular weight of rBMP-NAP was about 57 kD,accounting for 37.55% of the total protein in the sonicated supematant of E. coli TB1 (pMAL-c2x-napA). The purity of the fusion protein after one-step affinity chromatography was 94% and the yield was 100 mg per liter of bacterial culture.The purified fusion protein could be specifically recognized by both human anti-sera from clinical patients with H pylori infection and rabbit sera immunized by rMBP-NAP itself.CONCLUSION: Recombinant protein rMBP-NAP might be a novel antigen for vaccine development against H pylori.展开更多
Chinese cabbage was cultivated in upland soil with the addition of biochar in order to investigate the potential for reduction of greenhouse gas emissions. Barley straw biochar(BSB) was introduced in a Wagner pot(1/50...Chinese cabbage was cultivated in upland soil with the addition of biochar in order to investigate the potential for reduction of greenhouse gas emissions. Barley straw biochar(BSB) was introduced in a Wagner pot(1/5000a) in amounts of 0(BSB0, control), 100(BSB100), 300(BSB300), and 500(BSB500) kg 10a^(-1). After the addition of BSB into the upland soil, carbon dioxide(CO_2) emission increased while methane(CH_4) and nitrous oxide(N_2O) emissions decreased. The highest CO2 flux was measured for the BSB500 sample,(84.6 g m^(-2)) followed by BSB300, BSB100, and BSB0 in decreasing order. Relative to those of control, the total CH_4 flux and N_2O flux for the BSB500 treatment were lower by 31.6% and 26.1%, respectively. The global warming potential(GWP) of the treatment without biochar was 281.4 g CO_2 m-2 and those for treatments with biochar were in the range from 194.1 to 224.9 g CO_2 m^(-2). Therefore, introducing BSB into upland soil to cultivate Chinese cabbages can reduce the global warming potential.展开更多
Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing plasmid pRP22-GFP was constru...Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing plasmid pRP22-GFP was constructed for monitoring the colonization of two biocontrol agents, Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116, on the spikes of barley and their effect on suppression of FHB. Survival and colonization of the Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116 strains on spikes of barley were observed by tracking the bacterial transformants with GFP expression. Our field study revealed that plasmid pRP22-GFP was stably maintained in the bacterial strains without selective pressure. The retrieved GFP-tagged strains showed that the bacterial population fluctuation accorded with that of the rain events. Furthermore, both biocontrol strains gave significant protection against FHB on spikes of barley in fields. The greater suppression of barley FHB disease was resulted from the treat-ment of barley spikes with biocontrol agents before inoculation with F. graminearum.展开更多
The authors described a novel submerged batch culture system that produced high levels of amylase by Aspergillus kawachii using whole barley (WB), the surface of which is covered by its husk. In this study, detailed...The authors described a novel submerged batch culture system that produced high levels of amylase by Aspergillus kawachii using whole barley (WB), the surface of which is covered by its husk. In this study, detailed analyses determining the amylase activities, residual sugars, fungal morphology and expression levels of genes were performed in a submerged culture using WB to address the mechanism underlying high amylase productivity in A. kawachii. High levels of glucoamylase and acid-stable u-amylase were produced in this culture, and expression levels of amylases, as well as glucose-repressive genes including high-affinity glucose transporter and peroxidase/catalase were also high. On the other hand, the morphology of mycelia was altered, with swollen, bulbous, multi-septum hyphae and conidiophores that normally form in a solid culture being partially generated. Furthermore, cell cycle and post-translational modification-related gene expression levels were altered, and were similar to those in the solid culture. These findings suggest that high amylase productivity in the submerged culture using WB is accompanied by both the up-regulation of amylase genes and activation of post-translational modifications due to fungal morphological changes being brought closer to those in the solid culture.展开更多
The difference in accumulation of high molecular weight glutenin subunits (HMW-GS) in superior and inferior grains results in the non-uniformity of grain quality in winter wheat (Triticum aestivum L.). The HMW-GS ...The difference in accumulation of high molecular weight glutenin subunits (HMW-GS) in superior and inferior grains results in the non-uniformity of grain quality in winter wheat (Triticum aestivum L.). The HMW-GS accumulation and glutenin macropolymer (GMP) content were studied in superior and inferior grains during the grain filling period, using the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Compared to inferior grains, HMW-GS was formed earlier and total accumulation amount was higher in superior grains. The total HMW-GS content was higher in superior grain than that in inferior grain except at maturity. For individual HMW-GS types, the accumulation and content of subunit 7 were the highest, followed by subunit 12, while that of subunit 8 were the lowest, followed by subunit 2 in superior grain. In contrary, the accumulation and content of subunit 7 at maturity were significantly higher than subunit 8, but they were similar between subunit 2 and subunit 12 in inferior grain. Moreover, the accumulation of subunit 7 and 12 in superior grain were significantly higher than that in inferior grain. However, compared to the inferior grain, the GMP accumulation was higher, while content was lower in superior grain at maturity.展开更多
基金Supported by National Natural Science Foundation of China(30870109)~~
文摘[Objective]The aim of this study was to identify transient expression of movement protein (MP) gene in Nicotinana benthaminana rapidly and further investigate the function of this exogenous gene. [Method]The movement protein gene of barley yellow dwarf virus (BYDV) was cloned into potato virus X (PVX) viral vector of pGR107,and PVX-recombinant vector was obtained. After electroporation of Agrobacterium tumefaciens,PVX was inoculated into the lower leaves of tobacco by Agrobacterium infiltration assay to observe the infection of virus on tobacco. [Result]After infection for 7 days,upper non-inoculated leaves of tobacco infected by the PVX-recombinant vector showed the virus infection symptoms,while the control group had no viral infection phenomenon. Daily follow-up observations for two groups revealed that tobacco infected by PVX-recombinant vector had severe symptoms of virus infection and curling leaves,or even led to necrosis both in infiltrated and systemic leaves in late period. However,tobacco infected by PVX vector had only slight symptoms of virus infection and could recover from infection. RT-PCR of the infected tobacco indicated that exogenous gene BYDV-MP had a normal transcription and expression in tobacco. [Conclusion]As a determinant factor for viral disease,BYDV-MP promotes the systemic infection rate of PVX and its symptom. In addition,it is feasible to express exogenous MP gene in Nicotiana benthaminan via PVX expression vector.
基金Project supported by the National Natural Science Foundation of China (Nos. 30270779 and 30471022) and Specialized Research Fund for the Doctoral Program of Higher Education (SRFDP) (No. 20020335028), China
文摘The effect of sowing date on grain protein, hordein fraction content and malting quality of two-rowed spring barley was investigated by using ten commercial cultivars with different grain protein content and the relationships among these traits were examined. The results showed that grain protein content and B hordein content increased as the sowing date postponed and were significantly affected by sowing date, while C and D hordein contents were less influenced by sowing date. There were significant differences in grain protein and hordein fraction content among the ten cultivars. The coefficient of variation of D hordein content was much larger than that of B and C hordein contents, suggesting its greater variation caused by different sowing dates. Beta-amylase activity and diastatic power were also significantly affected by sowing date, with malt extract being less affected. Significant differences in measured malt quality were found among the ten cultivars. Grain protein was significantly correlated with B hordein and malt extract positively and negatively, respectively. There was no significant correlation between beta-amylase activity or diastatic power and grain protein content. B hordein was negatively and significantly correlated with malt extract, but no significant correlations between C hordein, D hordein and malting quality traits.
基金Supported by the Medical Science Foundation for Distinguished Scholars of Henan Province, No. 200084
文摘AIM: To produce a recombinant protein rMBP-NAP, which was fusionally expressed by Helicobacter pylori(H pylori)neutrophil-activating protein (NAP) and E. coli maltosebinding protein (MBP) and to evaluate its immunoreactivity and immunogenicity.METHODS: Neutrophil-activating protein gene of H pylori (HP-napA) was subcloned from the recombinant plasmid pNEB-napA, and fused to MalE gene of expressing vector pMAL-c2x. The recombinant plasmid pMAL-c2x-napA was confirmed by restriction enzyme digestion, and then transformed into E. coli TB1. Fusion protein rMBP-NAP was induced by IPTG and identified by SDS-PAGE analysis.Soluble rMBP-NAP was purified by amylose affinity chromatography. Immunoreactivity and immunogenicity of the fusion protein were evaluated by animal experiment,Western blotting with human H pylori anti-sera.RESULTS: E.coli TB1 carrying recombinant plasmid pMAL-c2x-napA was constructed and led to a high efficiency cytosol expression of fusion protein rBMP -NAP when induced by IPTG.The molecular weight of rBMP-NAP was about 57 kD,accounting for 37.55% of the total protein in the sonicated supematant of E. coli TB1 (pMAL-c2x-napA). The purity of the fusion protein after one-step affinity chromatography was 94% and the yield was 100 mg per liter of bacterial culture.The purified fusion protein could be specifically recognized by both human anti-sera from clinical patients with H pylori infection and rabbit sera immunized by rMBP-NAP itself.CONCLUSION: Recombinant protein rMBP-NAP might be a novel antigen for vaccine development against H pylori.
基金supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF)funded by the Ministry of Education,Science and Technology (2012R1A2A2A01015706)the support of "Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ011227042016)" Rural Development Administration, Republic of Korea
文摘Chinese cabbage was cultivated in upland soil with the addition of biochar in order to investigate the potential for reduction of greenhouse gas emissions. Barley straw biochar(BSB) was introduced in a Wagner pot(1/5000a) in amounts of 0(BSB0, control), 100(BSB100), 300(BSB300), and 500(BSB500) kg 10a^(-1). After the addition of BSB into the upland soil, carbon dioxide(CO_2) emission increased while methane(CH_4) and nitrous oxide(N_2O) emissions decreased. The highest CO2 flux was measured for the BSB500 sample,(84.6 g m^(-2)) followed by BSB300, BSB100, and BSB0 in decreasing order. Relative to those of control, the total CH_4 flux and N_2O flux for the BSB500 treatment were lower by 31.6% and 26.1%, respectively. The global warming potential(GWP) of the treatment without biochar was 281.4 g CO_2 m-2 and those for treatments with biochar were in the range from 194.1 to 224.9 g CO_2 m^(-2). Therefore, introducing BSB into upland soil to cultivate Chinese cabbages can reduce the global warming potential.
基金Project supported by the National Natural Science Foundation of China (No. 30230250)Science and Technology Committee of Zhejiang Province (No. 2003C22029), China
文摘Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing plasmid pRP22-GFP was constructed for monitoring the colonization of two biocontrol agents, Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116, on the spikes of barley and their effect on suppression of FHB. Survival and colonization of the Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116 strains on spikes of barley were observed by tracking the bacterial transformants with GFP expression. Our field study revealed that plasmid pRP22-GFP was stably maintained in the bacterial strains without selective pressure. The retrieved GFP-tagged strains showed that the bacterial population fluctuation accorded with that of the rain events. Furthermore, both biocontrol strains gave significant protection against FHB on spikes of barley in fields. The greater suppression of barley FHB disease was resulted from the treat-ment of barley spikes with biocontrol agents before inoculation with F. graminearum.
文摘The authors described a novel submerged batch culture system that produced high levels of amylase by Aspergillus kawachii using whole barley (WB), the surface of which is covered by its husk. In this study, detailed analyses determining the amylase activities, residual sugars, fungal morphology and expression levels of genes were performed in a submerged culture using WB to address the mechanism underlying high amylase productivity in A. kawachii. High levels of glucoamylase and acid-stable u-amylase were produced in this culture, and expression levels of amylases, as well as glucose-repressive genes including high-affinity glucose transporter and peroxidase/catalase were also high. On the other hand, the morphology of mycelia was altered, with swollen, bulbous, multi-septum hyphae and conidiophores that normally form in a solid culture being partially generated. Furthermore, cell cycle and post-translational modification-related gene expression levels were altered, and were similar to those in the solid culture. These findings suggest that high amylase productivity in the submerged culture using WB is accompanied by both the up-regulation of amylase genes and activation of post-translational modifications due to fungal morphological changes being brought closer to those in the solid culture.
基金This study is supported by the projects of Priority Academic Program Developmem (PAPD), the National Natural Science Foundation of China (31171484, 31471445), the National Natural Science Foundation for Distinguished Young Scientists (31325020), Jiangsu Agriculture Science and Technology Innovation Fund (CX (14) 2002), the Specialized Research Fund for the Doctoral Program of Higher Education (20090097110009), the National Non-profit Program by Ministry of Agriculture (200903003), and the China Agriculture Research System (CARS-03).
文摘The difference in accumulation of high molecular weight glutenin subunits (HMW-GS) in superior and inferior grains results in the non-uniformity of grain quality in winter wheat (Triticum aestivum L.). The HMW-GS accumulation and glutenin macropolymer (GMP) content were studied in superior and inferior grains during the grain filling period, using the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Compared to inferior grains, HMW-GS was formed earlier and total accumulation amount was higher in superior grains. The total HMW-GS content was higher in superior grain than that in inferior grain except at maturity. For individual HMW-GS types, the accumulation and content of subunit 7 were the highest, followed by subunit 12, while that of subunit 8 were the lowest, followed by subunit 2 in superior grain. In contrary, the accumulation and content of subunit 7 at maturity were significantly higher than subunit 8, but they were similar between subunit 2 and subunit 12 in inferior grain. Moreover, the accumulation of subunit 7 and 12 in superior grain were significantly higher than that in inferior grain. However, compared to the inferior grain, the GMP accumulation was higher, while content was lower in superior grain at maturity.