EV71-type virus is one of the main pathogens causing the occurrence of hand,foot and mouth disease(HFMD),and VP1 protein,a factor that directly determines the antigenicity of the virus,has been isolated.The tomato was...EV71-type virus is one of the main pathogens causing the occurrence of hand,foot and mouth disease(HFMD),and VP1 protein,a factor that directly determines the antigenicity of the virus,has been isolated.The tomato was selected as a bioreactor for the production of an edible EV71 vaccine designed for the VP1 capsid protein.Using molecular biology techniques,the fusion gene EV71-VP1 was cut from vector PGEX-4T-2,a vector containing the p2300-EV71 gene with CaMV35S promoter and TL regulatory elements was constructed,and the hypocotyl and cotyledons of tomato were transformed using Agrobacterium(EHA105)-mediated method,screened,elongated and rooted,and finally 20 resistant tomato plants were obtained.Five transgenic positive seedlings were obtained by digestion and PCR assay,among which three plants were detected by RT-PCR to be capable of transcriptional translation at the RNA level.The experimental results aimed to explore new material support for the preparation of transgenic plant oral vaccines against EV71 infection and provide a theoretical basis for accelerating the development of transgenic plant vaccines in the future.展开更多
We are associating the solutions of stochastic and deterministic vector borne plant disease model in this manuscript.The dynamics of plant model depends upon threshold number P^(∗).If P^(∗)<1 then condition helpful...We are associating the solutions of stochastic and deterministic vector borne plant disease model in this manuscript.The dynamics of plant model depends upon threshold number P^(∗).If P^(∗)<1 then condition helpful to eradicate the disease in plants while P^(∗)>1 explains the persistence of disease.Inappropriately,standard numerical systems do not behave well in certain scenarios.We have been proposed a structure preserving stochastic non-standard finite difference system to analyze the behavior of model.This system is dynamical consistent,positive and bounded as defined by Mickens.展开更多
Given the sequence of Chs gene promoter from Arabidopsis thaliana reported in GenBank （AF248988）, a pair of specific PCR primers was designed with the Primer Premier 5.0 software. PCR products of about 0.5 kb were s...Given the sequence of Chs gene promoter from Arabidopsis thaliana reported in GenBank （AF248988）, a pair of specific PCR primers was designed with the Primer Premier 5.0 software. PCR products of about 0.5 kb were successfully amplified with the genome DNA of A. thaliana as a DNA template and Taq polymerase as DNA polymerase. The purified PCR products were ligated to the pMD18-T vector. The sequencing result showed that the Chs promoter from A. thaliana was 531 bp long. Sequence alignment analysis based on the DNAMAN software revealed that the sequence similarity between the cloned promoter and target promoter （AF248988） was up to 100%. Online PLACE analysis indicated that the Chs promoter contained cis-elements such as TATA-box, CAAT-box, pollen-box, G-box, ACGT-containing element, R response element, Myb recognition element and TACPyAT-box. At the same time, a plant expression vectorpAtChs：：GUS which fused the Chs promoter and the marker gene GUS was successfully constructed.展开更多
A full-length cDNA sequence of xyloglucan endotransglycosylase gene （XET）, abundantly expressed in the cambium of Anthocephalus chinensis was cloned by conserved PCR, rapid-amplification of cDNA ends and by chromoso...A full-length cDNA sequence of xyloglucan endotransglycosylase gene （XET）, abundantly expressed in the cambium of Anthocephalus chinensis was cloned by conserved PCR, rapid-amplification of cDNA ends and by chromosome walking. Analytical results of the DNA sequence show that a 912 bp complete open reading frame （ORF） encoded a 303-amino acid protein was in the 1205 bp full cDNA sequence. The deduced amino acid sequence of AcXET, which contained the conserved specific EIDFE catalytic site sequence to XETs was homologous to the other known XET proteins. In order to study the gene function of AcXET and obtain transgenic plants, a plant expression vector pBIAcXET was constructed by recombinating the AcXET fragment from the cloning vector pMD19AcXET and the binary vector pBI121 between the XbaI and SmaI sites. The fragment ofAcXET gene was inserted between the CaMV 35S promotor and the coding region of the GUS gene in pBI121. The identification results show that the plant expression binary vector pBIAcXET was constructed successfully. These results lay the foundation for studying the molecular mechanism ofAcXET gene during wood formation.展开更多
This paper proposes a vector-borne plant disease model with discontinuous treatment strategies. Constructing Lyapunov function and applying non-smooth theory to analyze discontinuous differential equations, the basic ...This paper proposes a vector-borne plant disease model with discontinuous treatment strategies. Constructing Lyapunov function and applying non-smooth theory to analyze discontinuous differential equations, the basic reproductive number R0 is proved, which determines whether the plant disease will be extinct or not. If R0 R0 > 1 , there exists a unique endemic equilibrium which is globally stable. The numerical simulations are provided to verify our theoretical results, which indicate that after infective individuals reach some level, strengthening treatment measures is proved to be beneficial in controlling disease transmission.展开更多
[ Objective ] Heat shock factors （HSFs） are the major transcription factors of eukaryotic heat shock responses. This study aims to investigate the adversity stress tolerance functions of Arabidopsis heat shock facto...[ Objective ] Heat shock factors （HSFs） are the major transcription factors of eukaryotic heat shock responses. This study aims to investigate the adversity stress tolerance functions of Arabidopsis heat shock factor AtHsfAla, which has important significance for in-depth understanding of adversity stress tolerance mechanisms of plants and further utilization of heat shock factor genes. [Method] Genomic DNA of Arabidopsis was extracted with CTAB method and purified to obtain Arabidopsis DNA samples for in vitro site-specific recombination cloning （ Gateway cloning） to construct plant expression vector of heat shock factor AtHs- fAla. Firstly, donor vector pDONR 201/AtHsfAla was constructed based on attB and attP site-specific recombination method （BP reaction）, to identify E. coli transformants harboring correct sequence of AtHsfAla by sequencing; secondly, plant expression vector pBTWG2/AttlsfAla overexpressing Arabidopsis heat shock factor AtHsfAla was constructed based on attL and attR site-specific recombination method （LR reaction）, to screen E. coli transformants harboring target plasmid. [ Result] Plant expression vector of Arabidopsis heat shock factor gene AtHsfAla was constructed successfully. [ Conclusion] This study not only provided experimental materials for acquiring transgenic plants overexpressing heat shock transcription factor AtHsfAla, but also laid the foundation for further investigation of the diversity of adversity stress tolerance functions reanlated by HSFs.展开更多
[Objective] This study was conducted to clone and analyze ERECTA-LIKE1 gene in Zea mays by PCR and bioinformatics methods and to construct plant expression vector p Cambia3301-zm ERECTA-LIKE1. [Method] zm ERECTA-LIKE1...[Objective] This study was conducted to clone and analyze ERECTA-LIKE1 gene in Zea mays by PCR and bioinformatics methods and to construct plant expression vector p Cambia3301-zm ERECTA-LIKE1. [Method] zm ERECTA-LIKE1(zm ERL1)gene was obtained using RT-PCR, and physical-chemical properties were analyzed by bioinformatics methods, including domains,transmembrane regions, N-Glycosylation potential sites phosphorylation sites, and etc. [Result] Bioinformatics results showed that zm ERL1 gene was 2 169 bp, which encoded a protein consisting of 722 amino acids, 11 N-glycosylation potential sites and 42 kinase specific phosphorylation sites. According to CDD2.23 and TMHMM Server v. 2.0 software, there were leucine-rich repeats,a PKC domain and a transmembrane region in this protein. The theoretical p I and molecular weight of zm ERL1 encoded protein was 6.20 and 79 184.8 using Compute PI/Mw tool. Furthermore, we constructed the plant expression vector p Cambia3301-zm ERECTA-LIKE1 by subcloning zm ERL1 gene into p Cambia3301 instead of GUS. [Conclusion] The results provide a theoretical basis for the application of zm ERL1 gene in future study.展开更多
Objective:To evaluate the larvicidal and pupicidal potential of the methanolic extracts from Moringa oleifera(M.oleifera) plant seeds against malarial vector Anopheles stephensi(A.stephensi) mosquitoes at different co...Objective:To evaluate the larvicidal and pupicidal potential of the methanolic extracts from Moringa oleifera(M.oleifera) plant seeds against malarial vector Anopheles stephensi(A.stephensi) mosquitoes at different concentrations(20,40,60,80 and 100 ppm).Methods:M.oleifera was collected from the area of around Bharathiar University,Coimbatore.The dried plant materials were powdered by an electrical blender.From each sample,100 g of the plant material were extracted with 300 mL of methanol for 8 h in a Soxhlet apparatus.The extracts were evaporated to dryness in rotary vacuum evaporator to yield 122 mg and 110 mg of dark greenish material(residue) from Arcang amara and Ocimum basilicum,respectively.One gram of the each plant residue was dissolved separately in 100 mL of acetone(stock solution) from which different concentrations, i.e.,20,40,60,80 and 100 ppm were prepared.Results:Larvicidal activity of M.oleifera exhibited in the first to fourth instar larvae of the A.stephensi,and the LC_(50) and LC_(90) values were 57.79 ppm and 125.93 ppm for the first instar,63.90 ppm and 133.07 ppm for the second inslar,72.45 ppm and 139.82 ppm for the third instar,78.93 ppm and 143.20 ppm for the fourth instar,respectively. During the pupal stage the methanolic extract of M.oleifera showed that the LC_(50),and LC_(90) values were 67.77 ppm and 141.00 ppm,respectively.Conclusions:The present study indicates that the phytochemicals derived from M.oleifera seeds extracts are effective mosquito vector control agents and the plant extracts may be used for further integrated pest management programs.展开更多
Rxol cloned from maize is a non-host gene resistant to bacterial leaf streak of rice. pCAMBIA1305-1 with Rxo1 was digested with Sca I and NgoM IV and the double right-border binary vector pMNDRBBin6 was digested with ...Rxol cloned from maize is a non-host gene resistant to bacterial leaf streak of rice. pCAMBIA1305-1 with Rxo1 was digested with Sca I and NgoM IV and the double right-border binary vector pMNDRBBin6 was digested with Hpa I and Xma I. pMNDRBBin6 carrying the gene Rxo1 was acquired by ligation of blunt-end and cohesive end. The results of PCR, restriction enzyme analysis and sequencing indicated that the Rxo1 gene had been cloned into pMNDRBBin6. This double right-border binary vector, named as pMNDRBBin6-Rxol, will play a role in breeding marker-free plants resistant to bacterial leaf streak of rice by genetic transformation.展开更多
基金Supported by the Natural Science Foundation of Heilongjiang Province(LH2021C032)。
文摘EV71-type virus is one of the main pathogens causing the occurrence of hand,foot and mouth disease(HFMD),and VP1 protein,a factor that directly determines the antigenicity of the virus,has been isolated.The tomato was selected as a bioreactor for the production of an edible EV71 vaccine designed for the VP1 capsid protein.Using molecular biology techniques,the fusion gene EV71-VP1 was cut from vector PGEX-4T-2,a vector containing the p2300-EV71 gene with CaMV35S promoter and TL regulatory elements was constructed,and the hypocotyl and cotyledons of tomato were transformed using Agrobacterium(EHA105)-mediated method,screened,elongated and rooted,and finally 20 resistant tomato plants were obtained.Five transgenic positive seedlings were obtained by digestion and PCR assay,among which three plants were detected by RT-PCR to be capable of transcriptional translation at the RNA level.The experimental results aimed to explore new material support for the preparation of transgenic plant oral vaccines against EV71 infection and provide a theoretical basis for accelerating the development of transgenic plant vaccines in the future.
基金The first author thanks Prince Sultan University for supporting this paper through the research group Nonlinear Analysis Methods in Applied Mathematics(NAMAM),group number RG-DES-2017-01-17.
文摘We are associating the solutions of stochastic and deterministic vector borne plant disease model in this manuscript.The dynamics of plant model depends upon threshold number P^(∗).If P^(∗)<1 then condition helpful to eradicate the disease in plants while P^(∗)>1 explains the persistence of disease.Inappropriately,standard numerical systems do not behave well in certain scenarios.We have been proposed a structure preserving stochastic non-standard finite difference system to analyze the behavior of model.This system is dynamical consistent,positive and bounded as defined by Mickens.
基金supported by the National Natural Science Foundation of China (Grant No.30740013)the Key Laboratory for Genetics and Breeding in Forestry Trees and Ornamental Plants,Ministry of Education (03-05)
文摘Given the sequence of Chs gene promoter from Arabidopsis thaliana reported in GenBank （AF248988）, a pair of specific PCR primers was designed with the Primer Premier 5.0 software. PCR products of about 0.5 kb were successfully amplified with the genome DNA of A. thaliana as a DNA template and Taq polymerase as DNA polymerase. The purified PCR products were ligated to the pMD18-T vector. The sequencing result showed that the Chs promoter from A. thaliana was 531 bp long. Sequence alignment analysis based on the DNAMAN software revealed that the sequence similarity between the cloned promoter and target promoter （AF248988） was up to 100%. Online PLACE analysis indicated that the Chs promoter contained cis-elements such as TATA-box, CAAT-box, pollen-box, G-box, ACGT-containing element, R response element, Myb recognition element and TACPyAT-box. At the same time, a plant expression vectorpAtChs：：GUS which fused the Chs promoter and the marker gene GUS was successfully constructed.
基金supported by the National Natural Science Foundation of China (Grant No. 30901158)the Key Project of Chinese Ministry of Education (Grant No. 104243)
文摘A full-length cDNA sequence of xyloglucan endotransglycosylase gene （XET）, abundantly expressed in the cambium of Anthocephalus chinensis was cloned by conserved PCR, rapid-amplification of cDNA ends and by chromosome walking. Analytical results of the DNA sequence show that a 912 bp complete open reading frame （ORF） encoded a 303-amino acid protein was in the 1205 bp full cDNA sequence. The deduced amino acid sequence of AcXET, which contained the conserved specific EIDFE catalytic site sequence to XETs was homologous to the other known XET proteins. In order to study the gene function of AcXET and obtain transgenic plants, a plant expression vector pBIAcXET was constructed by recombinating the AcXET fragment from the cloning vector pMD19AcXET and the binary vector pBI121 between the XbaI and SmaI sites. The fragment ofAcXET gene was inserted between the CaMV 35S promotor and the coding region of the GUS gene in pBI121. The identification results show that the plant expression binary vector pBIAcXET was constructed successfully. These results lay the foundation for studying the molecular mechanism ofAcXET gene during wood formation.
文摘This paper proposes a vector-borne plant disease model with discontinuous treatment strategies. Constructing Lyapunov function and applying non-smooth theory to analyze discontinuous differential equations, the basic reproductive number R0 is proved, which determines whether the plant disease will be extinct or not. If R0 R0 > 1 , there exists a unique endemic equilibrium which is globally stable. The numerical simulations are provided to verify our theoretical results, which indicate that after infective individuals reach some level, strengthening treatment measures is proved to be beneficial in controlling disease transmission.
基金Supported by National Natural Science Foundation of China(31060039,31260061)Natural Science Foundation of Yunnan Province(2010ZC163)+1 种基金Project of Kunming University(YJL11025)Fund for Key Discipline Construction of Kunming University
文摘[ Objective ] Heat shock factors （HSFs） are the major transcription factors of eukaryotic heat shock responses. This study aims to investigate the adversity stress tolerance functions of Arabidopsis heat shock factor AtHsfAla, which has important significance for in-depth understanding of adversity stress tolerance mechanisms of plants and further utilization of heat shock factor genes. [Method] Genomic DNA of Arabidopsis was extracted with CTAB method and purified to obtain Arabidopsis DNA samples for in vitro site-specific recombination cloning （ Gateway cloning） to construct plant expression vector of heat shock factor AtHs- fAla. Firstly, donor vector pDONR 201/AtHsfAla was constructed based on attB and attP site-specific recombination method （BP reaction）, to identify E. coli transformants harboring correct sequence of AtHsfAla by sequencing; secondly, plant expression vector pBTWG2/AttlsfAla overexpressing Arabidopsis heat shock factor AtHsfAla was constructed based on attL and attR site-specific recombination method （LR reaction）, to screen E. coli transformants harboring target plasmid. [ Result] Plant expression vector of Arabidopsis heat shock factor gene AtHsfAla was constructed successfully. [ Conclusion] This study not only provided experimental materials for acquiring transgenic plants overexpressing heat shock transcription factor AtHsfAla, but also laid the foundation for further investigation of the diversity of adversity stress tolerance functions reanlated by HSFs.
基金Supported by the Distinguished Young Scientists Project of Beijing(CIT&TCD201304096)Academic Degrees and Graduate Education Reform and Development Program of Beijing University of Agriculture(5056516002\016)
文摘[Objective] This study was conducted to clone and analyze ERECTA-LIKE1 gene in Zea mays by PCR and bioinformatics methods and to construct plant expression vector p Cambia3301-zm ERECTA-LIKE1. [Method] zm ERECTA-LIKE1(zm ERL1)gene was obtained using RT-PCR, and physical-chemical properties were analyzed by bioinformatics methods, including domains,transmembrane regions, N-Glycosylation potential sites phosphorylation sites, and etc. [Result] Bioinformatics results showed that zm ERL1 gene was 2 169 bp, which encoded a protein consisting of 722 amino acids, 11 N-glycosylation potential sites and 42 kinase specific phosphorylation sites. According to CDD2.23 and TMHMM Server v. 2.0 software, there were leucine-rich repeats,a PKC domain and a transmembrane region in this protein. The theoretical p I and molecular weight of zm ERL1 encoded protein was 6.20 and 79 184.8 using Compute PI/Mw tool. Furthermore, we constructed the plant expression vector p Cambia3301-zm ERECTA-LIKE1 by subcloning zm ERL1 gene into p Cambia3301 instead of GUS. [Conclusion] The results provide a theoretical basis for the application of zm ERL1 gene in future study.
文摘Objective:To evaluate the larvicidal and pupicidal potential of the methanolic extracts from Moringa oleifera(M.oleifera) plant seeds against malarial vector Anopheles stephensi(A.stephensi) mosquitoes at different concentrations(20,40,60,80 and 100 ppm).Methods:M.oleifera was collected from the area of around Bharathiar University,Coimbatore.The dried plant materials were powdered by an electrical blender.From each sample,100 g of the plant material were extracted with 300 mL of methanol for 8 h in a Soxhlet apparatus.The extracts were evaporated to dryness in rotary vacuum evaporator to yield 122 mg and 110 mg of dark greenish material(residue) from Arcang amara and Ocimum basilicum,respectively.One gram of the each plant residue was dissolved separately in 100 mL of acetone(stock solution) from which different concentrations, i.e.,20,40,60,80 and 100 ppm were prepared.Results:Larvicidal activity of M.oleifera exhibited in the first to fourth instar larvae of the A.stephensi,and the LC_(50) and LC_(90) values were 57.79 ppm and 125.93 ppm for the first instar,63.90 ppm and 133.07 ppm for the second inslar,72.45 ppm and 139.82 ppm for the third instar,78.93 ppm and 143.20 ppm for the fourth instar,respectively. During the pupal stage the methanolic extract of M.oleifera showed that the LC_(50),and LC_(90) values were 67.77 ppm and 141.00 ppm,respectively.Conclusions:The present study indicates that the phytochemicals derived from M.oleifera seeds extracts are effective mosquito vector control agents and the plant extracts may be used for further integrated pest management programs.
文摘Rxol cloned from maize is a non-host gene resistant to bacterial leaf streak of rice. pCAMBIA1305-1 with Rxo1 was digested with Sca I and NgoM IV and the double right-border binary vector pMNDRBBin6 was digested with Hpa I and Xma I. pMNDRBBin6 carrying the gene Rxo1 was acquired by ligation of blunt-end and cohesive end. The results of PCR, restriction enzyme analysis and sequencing indicated that the Rxo1 gene had been cloned into pMNDRBBin6. This double right-border binary vector, named as pMNDRBBin6-Rxol, will play a role in breeding marker-free plants resistant to bacterial leaf streak of rice by genetic transformation.
