During the automated placement process of dry fibers,the positioning and fixation of dry fiber gauze belts are achieved by spraying setting agents.The amount of the setting agent is difficult to control when it is spr...During the automated placement process of dry fibers,the positioning and fixation of dry fiber gauze belts are achieved by spraying setting agents.The amount of the setting agent is difficult to control when it is sprayed manually.Furthermore,it can also affect the permeability of the preform,resin injection and the quality of the vacuum assisted resin infusion(VARI)molding,resulting in a decrease in the mechanical properties of composite materials.This study utilizes dry fiber automated placement equipment and an automated spraying system to manufacture preform structures,followed by VARI process to prepare composite samples with varying setting agent contents.Subsequently,mechanical characterization including interlaminar shear,bending and tensile testing is conducted to investigate the influence of setting agent content on both the manufacturing process and the mechanical properties of composite products.The results show that the interlaminar shear strength,bending strength and tensile strength of the sample gradually decrease with the increase of the content of the setting agent.The optimal setting agent content for automated laying of dry fiber is determined to be 4%-6%,balancing the preformed body’s layup quality and its impact on mechanical properties.Compared with agent-free samples,this range results in reductions of 3% in interlaminar shear strength,9% in bending strength,11% in bending modulus,and 13%-16% in tensile strength.展开更多
A computational fluid dynamics (CFD) study was carried out on a four stroke air cooled DI diesel engine, F912Q manufactured by Beinei Company, by using the KIVA 3V code. A three dimensional mesh was set up to mode...A computational fluid dynamics (CFD) study was carried out on a four stroke air cooled DI diesel engine, F912Q manufactured by Beinei Company, by using the KIVA 3V code. A three dimensional mesh was set up to model the cylinder, intake passage and exhaust passage. The calculated in cylinder pressure history and emissions are compared with the engine test data. The results show reasonable agreement. The effects of swirl ratio and spray angle on fuel spray, evaporation and mixing are investigated. It is found that there are optimum swirl ratio and spray angle for better evaporation and combustion.展开更多
在英语中,有些名词或名词短论既可看作单数.又可看作复数.在真体使用中.应该区别对待,确定适当的谓语形式。一、某些集体名词,如 audience, army,class,family,crowd,committe。,government等后着接单数动同.表示把该词的含义看作一个整...在英语中,有些名词或名词短论既可看作单数.又可看作复数.在真体使用中.应该区别对待,确定适当的谓语形式。一、某些集体名词,如 audience, army,class,family,crowd,committe。,government等后着接单数动同.表示把该词的含义看作一个整体,着接复数动词,则指该词表示的集体中的成员.试比较: His family is an old one.他们家庭是一个旧式家庭. My family are all well.我全家(人)都好. The class is v。ry biz.这个班很大.展开更多
一.Walls Have Ears隔墙有耳“隔墙有耳,”来源于下面的一个传说:在法国喀德琳·德·美第奇皇后(Catherine de Medici 1519—1589)摄政时代开始建造的卢浮宫,某些房间能够传声——在一个房间里讲话,在另一个房间可以听得一清二...一.Walls Have Ears隔墙有耳“隔墙有耳,”来源于下面的一个传说:在法国喀德琳·德·美第奇皇后(Catherine de Medici 1519—1589)摄政时代开始建造的卢浮宫,某些房间能够传声——在一个房间里讲话,在另一个房间可以听得一清二楚。生性多疑的美第奇皇后就是用这种手段获悉许多国家机密、发现许多人的阴谋的。在卢浮宫中能窃听他人谈话,这在当时是个秘密,后来秘密泄露,事情传开,于是就有“Walls have ears”的说法。展开更多
Based on the helix rotating screen and the digital micro-mirror device （DMD）, the former proto of volumetric-swept display system is improved. The 3-D display system adopting a helix rotating screen to construct an ...Based on the helix rotating screen and the digital micro-mirror device （DMD）, the former proto of volumetric-swept display system is improved. The 3-D display system adopting a helix rotating screen to construct an imaging space meliorate the defects, such as the smaller image space, the fewer voxels and the severer voxel overlap dead zone caused by planar rotating screen. DMD with spatial light modular （SLM） technology increases the transmission bandwidth of 3-D data in the voxel activation subsystem and activate multi-voxel once time. The volumetric-swept system based on helix rotating screen and DMD is developed. The experimental results show that the image space, the vision dead zone, the voxels on slice, and the voxel activation capacity of the designed proto are superior to the plane rotating screen system.展开更多
A system architecture of solid-based NC simulation for milling machining is given, the function of which is composed of cutting simulation of different cutters, such as flat-end cutters, ball-end cutters, dome-end cut...A system architecture of solid-based NC simulation for milling machining is given, the function of which is composed of cutting simulation of different cutters, such as flat-end cutters, ball-end cutters, dome-end cutters, angle cutters and drill cutters, rapid pre-checking of interference, detection of collision, and visualization of over cut in the stock. A new method based on the design model is raised to detect the collision and pre-check the interference during NC milling machining. A special problem about the construction of a cutter′s swept volume, self-intersection is discussed. All the work on the construction, the subtraction and the display of solids is accomplished with the help of ACIS 3-D modeling.展开更多
Aiming at the impaction among granules of non obstructive particle damping(NOPD), the vibration absorption model for vertical impact of granules is established by adopting Hertz contact theory. The numerical simulati...Aiming at the impaction among granules of non obstructive particle damping(NOPD), the vibration absorption model for vertical impact of granules is established by adopting Hertz contact theory. The numerical simulation of the granules movement process is proceeded, and the vibration response of a free free uniform beam is obtained for the case when all granules act on it. Through this method, the effect on vibration absorption of impaction is investigated. The simulational data show that multi gra nule vertical impaction is not sensitive to the movement clearance. The vibration absorption is also very well when the clearance changes within a large range. Therefore, the phenomenon that the vibration magnitude may increase if the clearance in a single impact body is improperly selected will not happen. The effect of vibration suppression in the range of middle and high frequencies(2 500~6 000 Hz) is better than that in the range of low frequency(<2 500 Hz). It indicates that the effect on vibration absorption of multi granule can well restrain the vibration of middle and high frequencies.展开更多
The development process as well as the core theory of distributed interactive simulation and high level architecture are discussed, and combined with graphics features, a system of real time distributed visual simula...The development process as well as the core theory of distributed interactive simulation and high level architecture are discussed, and combined with graphics features, a system of real time distributed visual simulation is established. Based on computer network, simulation platform is built by installing related software and modeling object, and the interactive functions are extended by programming. A set of solutions for building a distributed visual simulation system that include both hardware and software are put forward, and a practical instance is also provided. The whole building process can be summarized into two steps that are scheme consideration and system realization.展开更多
Objective： To construct a human ether-a-go-go-related gene （HERG） nonsense mutant L539fs/47-558W into the autonomously fluorescent, eukaryotic expression vector pEGFP-C2, and to verify expression of the reconstruct...Objective： To construct a human ether-a-go-go-related gene （HERG） nonsense mutant L539fs/47-558W into the autonomously fluorescent, eukaryotic expression vector pEGFP-C2, and to verify expression of the reconstruct in human embryonic kidney-293 （HEK293） cells. Methods： The mutational fragment was subcloned into pEGFP-C2-HERG by double digestion of Sbf I, Eco91 1 and rejoining of T4 ligase. After verification, the recombinant pEGFP-C2-L539fs/47-558W and pEGFP-C2-HERG were respectively transfected into HEK293 cells for 48 h by the Lipofect method to observe the expression location of the fusion protein by laser confocal imaging scanning in vivo. pcDNA3 -L539fs/47-＊558W and pcDNA3-HERG were transfected to observe the expression location of the HERG protein by immunofluoresceoce. The mutant protein size was determined by Western blotting. Results： The about 1 kb-sized mutation region cDNA fragment from pcDNA3-L539fs/47-＊558Wand the about 7.2 kb-sized target vector fragment from pcDNA3-HERG were ligated after purification and gel recovery pEGFP-C2-L539fs/47＊-558W, approximately 8.2 kb, was demonstrated successfully been constructed under agarose gel electrophoresis and further sequencing. Laser confocal imaging showed that pEGFP-C2-HERG was mainly expressed in the membrane, whereas truncated mutant-type HERG in the pEGFP-C2 vector was partially located in the cytoplasm, the others were transported to the cell membrane in living HEK293 cells. The same as the immunofluoresceoce results after transfection of pcDNA3-HERG and pcDNA3-L539fs/47-558W. Wild-type HERG-GFP fusion protein expressed 160 and 180 kDa bands. The mutant and mutant-GFP fusion proteins were 70 and 100 kDa, respectively. Conclusion： pEGFP-C2-L539fs/47-＊558W was successfully constructed by double digestion method GFP had no effect on its protein expression and trafficking in HEK293 cells, which laid a foundation for the further study on L539fs/47-＊558W展开更多
The unsteady performance of drag and double reverse propeller podded propulsors in open water was numerically simulated using a computational fluid dynamics （CFD） method. A moving mesh method was used to more realis...The unsteady performance of drag and double reverse propeller podded propulsors in open water was numerically simulated using a computational fluid dynamics （CFD） method. A moving mesh method was used to more realistically simulate propulsor working conditions, and the thrust, torque, and lateral force coefficients of both propulsors were compared and analyzed. Forces acting on different parts of the propulsors along with the flow field distribution of steady and unsteady results at different advance coefficients were compared. Moreover, the change of the lateral force and the difference between the abovementioned two methods were mainly analyzed. It was shown that the thrust and torque results of both methods were similar, with the lateral force results having the highest deviation展开更多
Objective： To facilitate manipulation of gene expression in different host cells, we used pEGFP-N1 as backbone to construct a versatile vector that can drive foreign gene expression in prokaryotic and eukaryotic cell...Objective： To facilitate manipulation of gene expression in different host cells, we used pEGFP-N1 as backbone to construct a versatile vector that can drive foreign gene expression in prokaryotic and eukaryotic cells. Methods： A cloning and expression vector, pEGFP-NI-lac, was constructed by inserting the prokaryotic lac promoter of pUC 19 into the eukaryotic expression vector, pEGFP-N1, between the eukaryotic PCMV promoter and enhanced green fluorescent protein （EGFP） open reading frames. To assess the function of pEGFP-NI-lac, the nucleotide sequence encoding the hepatitis C virus （HCV） core protein was cloned into the multiple cloning sites. Western blotting analysis was used to detect the expression of the HCV core protein in Escherichia coli DH5a and HepG2 cells. Results： Restriction enzyme digestion and sequence analysis indicated that pEGFP-NI-lac was successfully constructed and the HCV core gene was cloned into this vector. The Western blotting results showed that pEGFP-NI-lac promoted expression of HCV core gene in prokaryotic E. coli DH5a and eukaryotic HepG2 cells. Conclusion： The pEGFP-NI-lac vector has been successfully constructed and functions in both prokaryotic and eukaryotic cells. The EGFP reporter can be used as an insert-inactivation marker for clone selection or as an expression tag. This vector can be used for cloning and expression of genes in both prokaryotic and eukaryotic cells, making gene cloning, expression and functional studies convenient as well as time- and labor-efficient展开更多
The molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of inter...The molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of interest for improving crop management and developing new strategies for plant diseases control. The aim of this work is to investigate the components involved in this interaction and, for this purpose, a proteomic approach was used. 2-D maps of the protein extracts from the single components in various interactions between plants (potato, bean, tobacco or tomato), pathogens (Botrytis cinerea, Rhizoctonia solani or Pythium ultimum) and biocontrol fungi (Trichoderma atroviride strain P1 or Trichoderma harzianum strain T22) were obtained. The proteome of each partner was collected separately and extracted by acetone precipitation in presence of trichloroacetic acid and a reducing agent (DTT). The extracted proteins were separated by isoelectrofocusing (IEF), using IPG (Immobilized pH gradient) strips, followed by SDS-PAGE. In order to improve resolution the separations were performed both on wide than narrow pH range and on different gel lengths. Differential spots were noted in the proteome of the three-way interaction when compared to each single component. These were further characterized by mass spectrometry and in silico analysis with the aim of identifying and cloning the relative genes. During the in vitro interaction of T. harzianum strain T22 with tomato and the culture filtrate or cell walls of pathogens, the spot number was higher than in the presence of pathogen biomass. In terms of Trichoderma differential proteins displayed on 2D gels, the most important changes were obtained in the presence of P. ultimum . During the in vivo interaction with tomato, the antagonist proteome changed much more in presence of soilborne fungi R. solani and P. ultimum than with the foliar fungus B. cinerea, both in terms of total and increased or novel spots. In silico analysis of some of those spots revealed homology with intracellular enzymes (GTPases, hydrolases) and with stress-related proteins (heat shock proteins HSP70, bacteriocin cloacin). Specific proteins in the plant proteome, i.e. pathogenesis-related proteins, have been identified during the in vivo interaction of bean with R. solani and T. atroviride strain P1. This is in agreement with the demonstrated ability of these beneficial fungi to induce plant systemic disease resistance by activating expression of defence-related genes. Proteins extracted from T. atrovride strain P1 which were analysed by mass spectrometry, revealed some interesting homologies with a fungal hydrophobin of Pleurotus ostreatus and an ABC transporter of Ralstonia metallidurans. These could represent molecular factors involved in the antagonistic mechanisms of Trichoderma and play a role in the three-way interaction with the plant and other microbes.展开更多
Since the first terpenoid synthase cDNA was obtained by the reverse genetic approach from grand fir, great progress in the molecular genetics of terpenoid formation has been made with angiosperms and genes encoding a ...Since the first terpenoid synthase cDNA was obtained by the reverse genetic approach from grand fir, great progress in the molecular genetics of terpenoid formation has been made with angiosperms and genes encoding a monoterpene synthase, a sesquiterpene synthase, and a diterpene synthase. Tree killing bark beetles and their vectored fungal pathogens are the most destructive agents of conifer forests worldwide. Conifers defend against attack by the constitutive and inducible production of oleoresin that accumulates at the wound site to kill invaders and both flush and seal the injury. Although toxic to the bark beetle and fungal pathogen, oleoresin also plays a central role in the chemical ecology of these boring insects. Recent advances in the molecular genetics of terpenoid biosynthesis provide evidence for the evolutionary origins of oleoresin and permit consideration of genetic engineering strategies to improve conifer defenses as a component of modern forest biotechnology. This review described enzymes of resin biosynthesis, structural feathers of genes genomic intron and exon organization, pathway organization and evolution, resin production and accumulation, interactions between conifer and bark beetle, and engineering strategies to improve conifer defenses.展开更多
基金supported by Jiangsu Provincial Key Research and Development Program(No.BE2023014-4).
文摘During the automated placement process of dry fibers,the positioning and fixation of dry fiber gauze belts are achieved by spraying setting agents.The amount of the setting agent is difficult to control when it is sprayed manually.Furthermore,it can also affect the permeability of the preform,resin injection and the quality of the vacuum assisted resin infusion(VARI)molding,resulting in a decrease in the mechanical properties of composite materials.This study utilizes dry fiber automated placement equipment and an automated spraying system to manufacture preform structures,followed by VARI process to prepare composite samples with varying setting agent contents.Subsequently,mechanical characterization including interlaminar shear,bending and tensile testing is conducted to investigate the influence of setting agent content on both the manufacturing process and the mechanical properties of composite products.The results show that the interlaminar shear strength,bending strength and tensile strength of the sample gradually decrease with the increase of the content of the setting agent.The optimal setting agent content for automated laying of dry fiber is determined to be 4%-6%,balancing the preformed body’s layup quality and its impact on mechanical properties.Compared with agent-free samples,this range results in reductions of 3% in interlaminar shear strength,9% in bending strength,11% in bending modulus,and 13%-16% in tensile strength.
文摘A computational fluid dynamics (CFD) study was carried out on a four stroke air cooled DI diesel engine, F912Q manufactured by Beinei Company, by using the KIVA 3V code. A three dimensional mesh was set up to model the cylinder, intake passage and exhaust passage. The calculated in cylinder pressure history and emissions are compared with the engine test data. The results show reasonable agreement. The effects of swirl ratio and spray angle on fuel spray, evaporation and mixing are investigated. It is found that there are optimum swirl ratio and spray angle for better evaporation and combustion.
文摘在英语中,有些名词或名词短论既可看作单数.又可看作复数.在真体使用中.应该区别对待,确定适当的谓语形式。一、某些集体名词,如 audience, army,class,family,crowd,committe。,government等后着接单数动同.表示把该词的含义看作一个整体,着接复数动词,则指该词表示的集体中的成员.试比较: His family is an old one.他们家庭是一个旧式家庭. My family are all well.我全家(人)都好. The class is v。ry biz.这个班很大.
文摘一.Walls Have Ears隔墙有耳“隔墙有耳,”来源于下面的一个传说:在法国喀德琳·德·美第奇皇后(Catherine de Medici 1519—1589)摄政时代开始建造的卢浮宫,某些房间能够传声——在一个房间里讲话,在另一个房间可以听得一清二楚。生性多疑的美第奇皇后就是用这种手段获悉许多国家机密、发现许多人的阴谋的。在卢浮宫中能窃听他人谈话,这在当时是个秘密,后来秘密泄露,事情传开,于是就有“Walls have ears”的说法。
基金Supported by the National High Technology Research and Development Program of China(″863″Program)(2007AA01Z338)the National Science Foundation for Post-doctoral Scientists of China(20080441051)the Jiangsu Province Science Foundation for Post-doctoral Scientists(0802014c)~~
文摘Based on the helix rotating screen and the digital micro-mirror device （DMD）, the former proto of volumetric-swept display system is improved. The 3-D display system adopting a helix rotating screen to construct an imaging space meliorate the defects, such as the smaller image space, the fewer voxels and the severer voxel overlap dead zone caused by planar rotating screen. DMD with spatial light modular （SLM） technology increases the transmission bandwidth of 3-D data in the voxel activation subsystem and activate multi-voxel once time. The volumetric-swept system based on helix rotating screen and DMD is developed. The experimental results show that the image space, the vision dead zone, the voxels on slice, and the voxel activation capacity of the designed proto are superior to the plane rotating screen system.
文摘A system architecture of solid-based NC simulation for milling machining is given, the function of which is composed of cutting simulation of different cutters, such as flat-end cutters, ball-end cutters, dome-end cutters, angle cutters and drill cutters, rapid pre-checking of interference, detection of collision, and visualization of over cut in the stock. A new method based on the design model is raised to detect the collision and pre-check the interference during NC milling machining. A special problem about the construction of a cutter′s swept volume, self-intersection is discussed. All the work on the construction, the subtraction and the display of solids is accomplished with the help of ACIS 3-D modeling.
文摘Aiming at the impaction among granules of non obstructive particle damping(NOPD), the vibration absorption model for vertical impact of granules is established by adopting Hertz contact theory. The numerical simulation of the granules movement process is proceeded, and the vibration response of a free free uniform beam is obtained for the case when all granules act on it. Through this method, the effect on vibration absorption of impaction is investigated. The simulational data show that multi gra nule vertical impaction is not sensitive to the movement clearance. The vibration absorption is also very well when the clearance changes within a large range. Therefore, the phenomenon that the vibration magnitude may increase if the clearance in a single impact body is improperly selected will not happen. The effect of vibration suppression in the range of middle and high frequencies(2 500~6 000 Hz) is better than that in the range of low frequency(<2 500 Hz). It indicates that the effect on vibration absorption of multi granule can well restrain the vibration of middle and high frequencies.
文摘The development process as well as the core theory of distributed interactive simulation and high level architecture are discussed, and combined with graphics features, a system of real time distributed visual simulation is established. Based on computer network, simulation platform is built by installing related software and modeling object, and the interactive functions are extended by programming. A set of solutions for building a distributed visual simulation system that include both hardware and software are put forward, and a practical instance is also provided. The whole building process can be summarized into two steps that are scheme consideration and system realization.
基金Supported by the National Natural Science Foundation of China(No.30800473)
文摘Objective： To construct a human ether-a-go-go-related gene （HERG） nonsense mutant L539fs/47-558W into the autonomously fluorescent, eukaryotic expression vector pEGFP-C2, and to verify expression of the reconstruct in human embryonic kidney-293 （HEK293） cells. Methods： The mutational fragment was subcloned into pEGFP-C2-HERG by double digestion of Sbf I, Eco91 1 and rejoining of T4 ligase. After verification, the recombinant pEGFP-C2-L539fs/47-558W and pEGFP-C2-HERG were respectively transfected into HEK293 cells for 48 h by the Lipofect method to observe the expression location of the fusion protein by laser confocal imaging scanning in vivo. pcDNA3 -L539fs/47-＊558W and pcDNA3-HERG were transfected to observe the expression location of the HERG protein by immunofluoresceoce. The mutant protein size was determined by Western blotting. Results： The about 1 kb-sized mutation region cDNA fragment from pcDNA3-L539fs/47-＊558Wand the about 7.2 kb-sized target vector fragment from pcDNA3-HERG were ligated after purification and gel recovery pEGFP-C2-L539fs/47＊-558W, approximately 8.2 kb, was demonstrated successfully been constructed under agarose gel electrophoresis and further sequencing. Laser confocal imaging showed that pEGFP-C2-HERG was mainly expressed in the membrane, whereas truncated mutant-type HERG in the pEGFP-C2 vector was partially located in the cytoplasm, the others were transported to the cell membrane in living HEK293 cells. The same as the immunofluoresceoce results after transfection of pcDNA3-HERG and pcDNA3-L539fs/47-558W. Wild-type HERG-GFP fusion protein expressed 160 and 180 kDa bands. The mutant and mutant-GFP fusion proteins were 70 and 100 kDa, respectively. Conclusion： pEGFP-C2-L539fs/47-＊558W was successfully constructed by double digestion method GFP had no effect on its protein expression and trafficking in HEK293 cells, which laid a foundation for the further study on L539fs/47-＊558W
基金Supported by National Natural Science Foundation of China （41176074, 51209048,51379043,51409063） High tech ship research project of Ministry of industry and technology （G014613002） The support plan for youth backbone teachers of Harbin Engineering University （HEUCFQ1408）
文摘The unsteady performance of drag and double reverse propeller podded propulsors in open water was numerically simulated using a computational fluid dynamics （CFD） method. A moving mesh method was used to more realistically simulate propulsor working conditions, and the thrust, torque, and lateral force coefficients of both propulsors were compared and analyzed. Forces acting on different parts of the propulsors along with the flow field distribution of steady and unsteady results at different advance coefficients were compared. Moreover, the change of the lateral force and the difference between the abovementioned two methods were mainly analyzed. It was shown that the thrust and torque results of both methods were similar, with the lateral force results having the highest deviation
基金Supported by the National High Technology Research and Development Program of China (863 Program, 2009AA02Z111)the National Natural Science Foundation of China (30872223)the Funds of the State Key Laboratory of Pathogen and Biosecurity
文摘Objective： To facilitate manipulation of gene expression in different host cells, we used pEGFP-N1 as backbone to construct a versatile vector that can drive foreign gene expression in prokaryotic and eukaryotic cells. Methods： A cloning and expression vector, pEGFP-NI-lac, was constructed by inserting the prokaryotic lac promoter of pUC 19 into the eukaryotic expression vector, pEGFP-N1, between the eukaryotic PCMV promoter and enhanced green fluorescent protein （EGFP） open reading frames. To assess the function of pEGFP-NI-lac, the nucleotide sequence encoding the hepatitis C virus （HCV） core protein was cloned into the multiple cloning sites. Western blotting analysis was used to detect the expression of the HCV core protein in Escherichia coli DH5a and HepG2 cells. Results： Restriction enzyme digestion and sequence analysis indicated that pEGFP-NI-lac was successfully constructed and the HCV core gene was cloned into this vector. The Western blotting results showed that pEGFP-NI-lac promoted expression of HCV core gene in prokaryotic E. coli DH5a and eukaryotic HepG2 cells. Conclusion： The pEGFP-NI-lac vector has been successfully constructed and functions in both prokaryotic and eukaryotic cells. The EGFP reporter can be used as an insert-inactivation marker for clone selection or as an expression tag. This vector can be used for cloning and expression of genes in both prokaryotic and eukaryotic cells, making gene cloning, expression and functional studies convenient as well as time- and labor-efficient
文摘The molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of interest for improving crop management and developing new strategies for plant diseases control. The aim of this work is to investigate the components involved in this interaction and, for this purpose, a proteomic approach was used. 2-D maps of the protein extracts from the single components in various interactions between plants (potato, bean, tobacco or tomato), pathogens (Botrytis cinerea, Rhizoctonia solani or Pythium ultimum) and biocontrol fungi (Trichoderma atroviride strain P1 or Trichoderma harzianum strain T22) were obtained. The proteome of each partner was collected separately and extracted by acetone precipitation in presence of trichloroacetic acid and a reducing agent (DTT). The extracted proteins were separated by isoelectrofocusing (IEF), using IPG (Immobilized pH gradient) strips, followed by SDS-PAGE. In order to improve resolution the separations were performed both on wide than narrow pH range and on different gel lengths. Differential spots were noted in the proteome of the three-way interaction when compared to each single component. These were further characterized by mass spectrometry and in silico analysis with the aim of identifying and cloning the relative genes. During the in vitro interaction of T. harzianum strain T22 with tomato and the culture filtrate or cell walls of pathogens, the spot number was higher than in the presence of pathogen biomass. In terms of Trichoderma differential proteins displayed on 2D gels, the most important changes were obtained in the presence of P. ultimum . During the in vivo interaction with tomato, the antagonist proteome changed much more in presence of soilborne fungi R. solani and P. ultimum than with the foliar fungus B. cinerea, both in terms of total and increased or novel spots. In silico analysis of some of those spots revealed homology with intracellular enzymes (GTPases, hydrolases) and with stress-related proteins (heat shock proteins HSP70, bacteriocin cloacin). Specific proteins in the plant proteome, i.e. pathogenesis-related proteins, have been identified during the in vivo interaction of bean with R. solani and T. atroviride strain P1. This is in agreement with the demonstrated ability of these beneficial fungi to induce plant systemic disease resistance by activating expression of defence-related genes. Proteins extracted from T. atrovride strain P1 which were analysed by mass spectrometry, revealed some interesting homologies with a fungal hydrophobin of Pleurotus ostreatus and an ABC transporter of Ralstonia metallidurans. These could represent molecular factors involved in the antagonistic mechanisms of Trichoderma and play a role in the three-way interaction with the plant and other microbes.
文摘Since the first terpenoid synthase cDNA was obtained by the reverse genetic approach from grand fir, great progress in the molecular genetics of terpenoid formation has been made with angiosperms and genes encoding a monoterpene synthase, a sesquiterpene synthase, and a diterpene synthase. Tree killing bark beetles and their vectored fungal pathogens are the most destructive agents of conifer forests worldwide. Conifers defend against attack by the constitutive and inducible production of oleoresin that accumulates at the wound site to kill invaders and both flush and seal the injury. Although toxic to the bark beetle and fungal pathogen, oleoresin also plays a central role in the chemical ecology of these boring insects. Recent advances in the molecular genetics of terpenoid biosynthesis provide evidence for the evolutionary origins of oleoresin and permit consideration of genetic engineering strategies to improve conifer defenses as a component of modern forest biotechnology. This review described enzymes of resin biosynthesis, structural feathers of genes genomic intron and exon organization, pathway organization and evolution, resin production and accumulation, interactions between conifer and bark beetle, and engineering strategies to improve conifer defenses.
