谷子(Setaria italica P. Beauv.)作为C4模式植物,具有较强的抗旱性,但干旱仍是制约其生长和产量的重要因素。为探究谷子耐旱性突变体(Drought-resistant mutant)的抗旱生理机制,以野生型‘豫谷28’和其耐旱性突变体(drm1-1)为材料,采...谷子(Setaria italica P. Beauv.)作为C4模式植物,具有较强的抗旱性,但干旱仍是制约其生长和产量的重要因素。为探究谷子耐旱性突变体(Drought-resistant mutant)的抗旱生理机制,以野生型‘豫谷28’和其耐旱性突变体(drm1-1)为材料,采用盆栽法分析了干旱胁迫下野生型与drm1-1在孕穗期和花后10 d的光合特性和抗旱性。结果显示,在正常条件下,drm1-1诱导了谷子光合作用相关的磷酸烯醇式丙酮酸羧化酶基因pepc、丙酮酸磷酸双激酶基因ppdk以及苹果酸酶基因nadp-me的表达,显著提高了相应酶的活性,也使得drm1-1的光合速率较野生型显著提高。在干旱胁迫条件下,drm1-1的光合速率和叶绿素荧光参数与正常条件相比,下降幅度显著小于野生型,具有显著的光合优势,同时drm1-1叶片的相对含水量与正常条件相比,下降的幅度显著小于野生型,而渗透调节物含量及抗氧化物酶活性均较野生型显著增加;且drm1-1的持水能力及抗旱性能更强。产量性状分析表明,干旱胁迫条件下drm1-1具有突出的产量优势。展开更多
With increased cultivation of transgenic Bacillus thuringiensis(Bt) cotton in the saline alkaline soil of China, assessments of transgenic crop biosafety have focused on the effects of soil salinity on rhizosphere mic...With increased cultivation of transgenic Bacillus thuringiensis(Bt) cotton in the saline alkaline soil of China, assessments of transgenic crop biosafety have focused on the effects of soil salinity on rhizosphere microbes and Bt protein residues. In 2013 and 2014, investigations were conducted on the rhizosphere microbial biomass, soil enzyme activities and Bt protein contents of the soil under transgenic Bt cotton(variety GK19) and its parental non-transgenic cotton(Simian 3) cultivated at various salinity levels(1.15, 6.00 and 11.46 dS m^(-1)). Under soil salinity stress, trace amounts of Bt proteins were observed in the Bt cotton GK19 rhizosphere soil, although the protein content increased with cotton growth and increased soil salinity levels. The populations of slight halophilic bacteria, phosphate solubilizing bacteria, ammonifying bacteria, nitrifying bacteria and denitrifying bacteria decreased with increased soil salinity in the Bt and non-Bt cotton rhizosphere soil, and the microbial biomass carbon, microbial respiration and soil catalase, urease and alkaline phosphatase activity also decreased. Correlation analyses showed that the increased Bt protein content in the Bt cotton rhizosphere soil may have been caused by the slower decomposition of soil microorganisms, which suggests that salinity was the main factor influencing the relevant activities of the soil microorganisms and indicates that Bt proteins had no clear adverse effects on the soil microorganisms. The results of this study may provide a theoretical basis for risk assessments of genetically modified cotton in saline alkaline soil.展开更多
Aphis gossypii is a globally distributed species and therefore has a highly variable life cycle.Populations of A.gossypii in northern China exhibit greater genotypic diversity and a broader host range,yet the details ...Aphis gossypii is a globally distributed species and therefore has a highly variable life cycle.Populations of A.gossypii in northern China exhibit greater genotypic diversity and a broader host range,yet the details of life cycles of different biotypes is still unclear.In this study,the Cytb and 16S gene regions of A.gossypii collected from 5 common summer hosts and 4 primary hosts were analyzed.A total of 57 haplotypes were obtained from 1 046 individual A.gossypii sequences.The sequence included 44 variable sites,27 of which were parsimony informative sites and 17 of which were singleton variable sites.The most frequent 3 haplotypes were found in 896 individuals,representing a total of 85.7% of all individuals and 36 haplotypes were found in 1 individual.A neighbor-joining tree was constructed using 21 haplotypes that were found in more than 2 individuals.Considering the individual host plant,5 biotypes were identified.Type 1 corresponded exactly to the cucurbit host-race and the other 4 biotypes were found as cotton host-races.Type 3 was the most abundant biotype in cotton fields in northern China.展开更多
The plant expression vector of choleratoxin B subunit(CTB)-human insulin(BA) fusion protein pBI121/(CTB-BA) was constructed first and then the Gynostemma Pentapyhllum Makino callus cell line that could express CTB-hum...The plant expression vector of choleratoxin B subunit(CTB)-human insulin(BA) fusion protein pBI121/(CTB-BA) was constructed first and then the Gynostemma Pentapyhllum Makino callus cell line that could express CTB-human insulin fusion protein was constructed and its hypoglycemic effect was evaluated in mice. The plant expression vector pBI121/(CTB-BA) was digested with both BamI and SacI. Agrobacterium tumerfaciens strain LBA4404 was transformed with previously constructed recombinant plasmid pBI121/(CTB-BA) via the freeze tha-wing method, then CTB-BA gene was integrated to G Pentapyhllum Makino callus cells by co-culturing the cells with the transformed LBA4404 strain. The transformed G Pentapyhllum Makino callus cells were identified by DNA se-quence assey and RT-PCR. The expressed product was identified by western-blot and its amount was tested by ELISA kit and its blood sugar decreasing effect was tested in mice.The sequences of synthetic CTB and human insulin genes(BA) were completely identical to those designed. Restriction map proved that the length of gene fragment in-serted into expression vector pBI121 was consistent with that expected. The sequence of genomic DNA of expressed product was completely identical to that designed. The result of RT-PCR was consistent with that expected. The ex-pressed product showed a specific band with a relative molecular mass of 17000 by Western-blot. The human insulin expression amount was 6.03 μIU/mL according to the ELISA result.The animal test showed that only the G Penta-pyhllum Makino callus cell line itself showed activity in decreasing the blood sugar of mice, however, the activity of the transformed G Pentapyhllum Makino callus cells was much higher. The plant expression vector pBI121/(CTB-BA) was constructed and expressed in the G Pentapyhllum Makino callus cells successfully for the first time. The trans-formed G Pentapyhllum Makino callus cells showed high activity in decreasing the blood sugar of mice. This study developed a new way for the development of oral administration insulin.展开更多
The cellular immunity of current influenza split vaccine is relatively low. It is necessary to develop a novel vaccine to improve the cellular immunity. Thes of this study prepared liposome-encapsulated influenza spli...The cellular immunity of current influenza split vaccine is relatively low. It is necessary to develop a novel vaccine to improve the cellular immunity. Thes of this study prepared liposome-encapsulated influenza split vaccine and tested it in BALB/c mice. The mice were immunized once with 4 μg of haemagglutinin of monovalent A/New Caledonia/20/99(H1N1) encapsulated with liposomes or the split virus vaccine only through intrastomach injection. In a comparative study, it was observed that the liposome-encapsulated vaccine elicited a higher neutralizing antibody response, more effectively stimulated spleen cell proliferation, increased cell subsets like CD4+ and CD4+/CD8+, and triggered IL-4 and IFN-γ production.展开更多
Based on meteorological data including daily sunshine duration, temperature and precipitation from 97 meteorological stations in Hunan province during the period of 1981—2010, in combination with the field experiment...Based on meteorological data including daily sunshine duration, temperature and precipitation from 97 meteorological stations in Hunan province during the period of 1981—2010, in combination with the field experiment in different places at different sowing dates, the precise climatic risk zoning of double cropping super rice cultivation has been studied by using the spatial interpolation method and other Geographical Information System(GIS)technologies. Three key climatic factors were selected including chilling in May, high temperature heat damage during July to early August and low temperature damage in autumn in this study. Furthermore, based on the analysis of climatic conditions suitable for double cropping super rice cultivation and climatic disasters, 8-22 ℃ active accumulated temperature, sunshine duration from late March to October, climatic risk index of the low temperature in autumn, and climatic risk index of chilling in May were selected as key climatic factors to study the precise agro-meteorological regionalization of double cropping super rice in Hunan province. The results showed that: the high-yielding zones of double cropping super rice in Hunan were mainly located in Zhuzhou, Hengyang, Yongzhou and Chenzhou City, the moderate-yielding zones were primarily located in the east and north reaches of Dongting Lake,together with most of Changsha, Zhuzhou and Xiangtan City, and other regions in Hunan were not suitable for double cropping super rice. These findings can provide valuable information for the large-scale cultivation of double cropping super rice in Hunan province.展开更多
Plants reshape their transcriptomes, proteomes and metabolomes in response to insect damage. In this study, we used suppression subtractive hybridization to investigate the transcriptomes of two cotton varieties(CCRI4...Plants reshape their transcriptomes, proteomes and metabolomes in response to insect damage. In this study, we used suppression subtractive hybridization to investigate the transcriptomes of two cotton varieties(CCRI41 and CCRI23) under Apolygus lucorum damage. From the CCRI23 libraries we obtained 92 transcripts and from the CCRI41 libraries we obtained 96 transcripts. 26 and 63 of the transcripts from CCRI23 and CCRI41, respectively, had known functions. Using reverse transcription PCR, we detected expression profile of genes with known functions. Ultimately, we identified eight significantly regulated genes, including one downregulated and four upregulated genes from the CCRI41 libraries, and one downregulated and two upregulated genes from the CCRI23 libraries. Only the gene encoding the polyphenol oxidase(PPO) is involved in plant defense against insect herbivores, and the others are related to improving tolerance to insect damage. Quantitative real-time PCR was used to study changes in expression levels during A. lucorum damage in CCRI23 and CCRI41. Significantly regulated genes from CCRI23 showed a response in CCRI23 but not response in CCRI41. Similarly, significantly regulated genes from CCRI41 showed a response in CCRI41 but not response in CCRI23. The results showed that, among transcriptomes of cotton varieties, there are different responses to A. lucorum damage.展开更多
The human interferon α 2b(hIFN-α2b) gene was cloned into binary vector pBI121 to obtain plant expression vector pBIFN. The recombinant plasmid pBIFN was transferred into Agrobacterium tumefaciens strain LBA4404. The...The human interferon α 2b(hIFN-α2b) gene was cloned into binary vector pBI121 to obtain plant expression vector pBIFN. The recombinant plasmid pBIFN was transferred into Agrobacterium tumefaciens strain LBA4404. Then the hIFN-α2b gene was introduced into ginseng callus cells via Agrobacterium-mediated transformation and the ginseng cell line carrying hIFN-α2b gene was selected on G418-containing medium. The presence of target gene in transformed cells was confirmed by PCR and RT-PCR. The results indicate that hIFN-α2b gene has been integrated into the ginseng cells’ genome, with transcription products. hIFN-α2b expressed by the transgenic ginseng cells was detected by Western blot. It was shown that a specific protein band at 19000 could be observed. Cytopathic effect(CPE) inhibition assay using the WISH-VSV system shows that the mean antiviral activity of expressed hIFN-α2b was 6.0×104 IU/mL.展开更多
DNA vaccines encoding a viral protein have been shown to induce antiviral immune responses and provide protection against subsequent viral challenge.The present article deals with the efficacy of a DNA vaccine greatly...DNA vaccines encoding a viral protein have been shown to induce antiviral immune responses and provide protection against subsequent viral challenge.The present article deals with the efficacy of a DNA vaccine greatly improved by the simultaneous expression of HBsAg and interferon-γ gene.We constructed a dual expression vector pHIN encoding the HBsAg of Hepatitis B virus and murine IFN-γ which are connected with Internal Ribosome Entry Site(IRES).Mice immunized with this dual expression DNA vaccine exhibited the enhancement of cellular immune response and increased the production of anti-HBV surface antibody,compared with the mice of single gene expression control.Taken together,these results demonstrate that the application of a cytokine gene in a DNA vaccine formulation as an adjuvant can improve its immunigenicity.展开更多
文摘谷子(Setaria italica P. Beauv.)作为C4模式植物,具有较强的抗旱性,但干旱仍是制约其生长和产量的重要因素。为探究谷子耐旱性突变体(Drought-resistant mutant)的抗旱生理机制,以野生型‘豫谷28’和其耐旱性突变体(drm1-1)为材料,采用盆栽法分析了干旱胁迫下野生型与drm1-1在孕穗期和花后10 d的光合特性和抗旱性。结果显示,在正常条件下,drm1-1诱导了谷子光合作用相关的磷酸烯醇式丙酮酸羧化酶基因pepc、丙酮酸磷酸双激酶基因ppdk以及苹果酸酶基因nadp-me的表达,显著提高了相应酶的活性,也使得drm1-1的光合速率较野生型显著提高。在干旱胁迫条件下,drm1-1的光合速率和叶绿素荧光参数与正常条件相比,下降幅度显著小于野生型,具有显著的光合优势,同时drm1-1叶片的相对含水量与正常条件相比,下降的幅度显著小于野生型,而渗透调节物含量及抗氧化物酶活性均较野生型显著增加;且drm1-1的持水能力及抗旱性能更强。产量性状分析表明,干旱胁迫条件下drm1-1具有突出的产量优势。
基金supported by the National Natural Science Foundation of China (31501253)the Project for the Development of Genetically Modified Crops, Ministry of Agriculture, China (2016ZX08011-002)
文摘With increased cultivation of transgenic Bacillus thuringiensis(Bt) cotton in the saline alkaline soil of China, assessments of transgenic crop biosafety have focused on the effects of soil salinity on rhizosphere microbes and Bt protein residues. In 2013 and 2014, investigations were conducted on the rhizosphere microbial biomass, soil enzyme activities and Bt protein contents of the soil under transgenic Bt cotton(variety GK19) and its parental non-transgenic cotton(Simian 3) cultivated at various salinity levels(1.15, 6.00 and 11.46 dS m^(-1)). Under soil salinity stress, trace amounts of Bt proteins were observed in the Bt cotton GK19 rhizosphere soil, although the protein content increased with cotton growth and increased soil salinity levels. The populations of slight halophilic bacteria, phosphate solubilizing bacteria, ammonifying bacteria, nitrifying bacteria and denitrifying bacteria decreased with increased soil salinity in the Bt and non-Bt cotton rhizosphere soil, and the microbial biomass carbon, microbial respiration and soil catalase, urease and alkaline phosphatase activity also decreased. Correlation analyses showed that the increased Bt protein content in the Bt cotton rhizosphere soil may have been caused by the slower decomposition of soil microorganisms, which suggests that salinity was the main factor influencing the relevant activities of the soil microorganisms and indicates that Bt proteins had no clear adverse effects on the soil microorganisms. The results of this study may provide a theoretical basis for risk assessments of genetically modified cotton in saline alkaline soil.
基金supported by the National Natural Science Foundation of China(31572015)the National Special Transgenic Project of China(2016ZX08012-004)
文摘Aphis gossypii is a globally distributed species and therefore has a highly variable life cycle.Populations of A.gossypii in northern China exhibit greater genotypic diversity and a broader host range,yet the details of life cycles of different biotypes is still unclear.In this study,the Cytb and 16S gene regions of A.gossypii collected from 5 common summer hosts and 4 primary hosts were analyzed.A total of 57 haplotypes were obtained from 1 046 individual A.gossypii sequences.The sequence included 44 variable sites,27 of which were parsimony informative sites and 17 of which were singleton variable sites.The most frequent 3 haplotypes were found in 896 individuals,representing a total of 85.7% of all individuals and 36 haplotypes were found in 1 individual.A neighbor-joining tree was constructed using 21 haplotypes that were found in more than 2 individuals.Considering the individual host plant,5 biotypes were identified.Type 1 corresponded exactly to the cucurbit host-race and the other 4 biotypes were found as cotton host-races.Type 3 was the most abundant biotype in cotton fields in northern China.
基金Supported by the Science and Technology Development Planning Foundation of Jilin Province of China(No.20030405)the Party and Government Administration "First-Leader-Hand" Science and Technology Project of Yunnan Province of Chi-na(No.2008QA028)
文摘The plant expression vector of choleratoxin B subunit(CTB)-human insulin(BA) fusion protein pBI121/(CTB-BA) was constructed first and then the Gynostemma Pentapyhllum Makino callus cell line that could express CTB-human insulin fusion protein was constructed and its hypoglycemic effect was evaluated in mice. The plant expression vector pBI121/(CTB-BA) was digested with both BamI and SacI. Agrobacterium tumerfaciens strain LBA4404 was transformed with previously constructed recombinant plasmid pBI121/(CTB-BA) via the freeze tha-wing method, then CTB-BA gene was integrated to G Pentapyhllum Makino callus cells by co-culturing the cells with the transformed LBA4404 strain. The transformed G Pentapyhllum Makino callus cells were identified by DNA se-quence assey and RT-PCR. The expressed product was identified by western-blot and its amount was tested by ELISA kit and its blood sugar decreasing effect was tested in mice.The sequences of synthetic CTB and human insulin genes(BA) were completely identical to those designed. Restriction map proved that the length of gene fragment in-serted into expression vector pBI121 was consistent with that expected. The sequence of genomic DNA of expressed product was completely identical to that designed. The result of RT-PCR was consistent with that expected. The ex-pressed product showed a specific band with a relative molecular mass of 17000 by Western-blot. The human insulin expression amount was 6.03 μIU/mL according to the ELISA result.The animal test showed that only the G Penta-pyhllum Makino callus cell line itself showed activity in decreasing the blood sugar of mice, however, the activity of the transformed G Pentapyhllum Makino callus cells was much higher. The plant expression vector pBI121/(CTB-BA) was constructed and expressed in the G Pentapyhllum Makino callus cells successfully for the first time. The trans-formed G Pentapyhllum Makino callus cells showed high activity in decreasing the blood sugar of mice. This study developed a new way for the development of oral administration insulin.
基金Supported by Innovation Fund for Technology Based Firms of China(No06C26212200804)
文摘The cellular immunity of current influenza split vaccine is relatively low. It is necessary to develop a novel vaccine to improve the cellular immunity. Thes of this study prepared liposome-encapsulated influenza split vaccine and tested it in BALB/c mice. The mice were immunized once with 4 μg of haemagglutinin of monovalent A/New Caledonia/20/99(H1N1) encapsulated with liposomes or the split virus vaccine only through intrastomach injection. In a comparative study, it was observed that the liposome-encapsulated vaccine elicited a higher neutralizing antibody response, more effectively stimulated spleen cell proliferation, increased cell subsets like CD4+ and CD4+/CD8+, and triggered IL-4 and IFN-γ production.
基金China Special Fund for Meteorological Research in the Public Interest(201206045)
文摘Based on meteorological data including daily sunshine duration, temperature and precipitation from 97 meteorological stations in Hunan province during the period of 1981—2010, in combination with the field experiment in different places at different sowing dates, the precise climatic risk zoning of double cropping super rice cultivation has been studied by using the spatial interpolation method and other Geographical Information System(GIS)technologies. Three key climatic factors were selected including chilling in May, high temperature heat damage during July to early August and low temperature damage in autumn in this study. Furthermore, based on the analysis of climatic conditions suitable for double cropping super rice cultivation and climatic disasters, 8-22 ℃ active accumulated temperature, sunshine duration from late March to October, climatic risk index of the low temperature in autumn, and climatic risk index of chilling in May were selected as key climatic factors to study the precise agro-meteorological regionalization of double cropping super rice in Hunan province. The results showed that: the high-yielding zones of double cropping super rice in Hunan were mainly located in Zhuzhou, Hengyang, Yongzhou and Chenzhou City, the moderate-yielding zones were primarily located in the east and north reaches of Dongting Lake,together with most of Changsha, Zhuzhou and Xiangtan City, and other regions in Hunan were not suitable for double cropping super rice. These findings can provide valuable information for the large-scale cultivation of double cropping super rice in Hunan province.
基金supported by the National Natural Science Foundation of China (31201518)
文摘Plants reshape their transcriptomes, proteomes and metabolomes in response to insect damage. In this study, we used suppression subtractive hybridization to investigate the transcriptomes of two cotton varieties(CCRI41 and CCRI23) under Apolygus lucorum damage. From the CCRI23 libraries we obtained 92 transcripts and from the CCRI41 libraries we obtained 96 transcripts. 26 and 63 of the transcripts from CCRI23 and CCRI41, respectively, had known functions. Using reverse transcription PCR, we detected expression profile of genes with known functions. Ultimately, we identified eight significantly regulated genes, including one downregulated and four upregulated genes from the CCRI41 libraries, and one downregulated and two upregulated genes from the CCRI23 libraries. Only the gene encoding the polyphenol oxidase(PPO) is involved in plant defense against insect herbivores, and the others are related to improving tolerance to insect damage. Quantitative real-time PCR was used to study changes in expression levels during A. lucorum damage in CCRI23 and CCRI41. Significantly regulated genes from CCRI23 showed a response in CCRI23 but not response in CCRI41. Similarly, significantly regulated genes from CCRI41 showed a response in CCRI41 but not response in CCRI23. The results showed that, among transcriptomes of cotton varieties, there are different responses to A. lucorum damage.
基金Supported by the Science and Technology Development Planning Foundation of Jilin Province of China(No.20030405)
文摘The human interferon α 2b(hIFN-α2b) gene was cloned into binary vector pBI121 to obtain plant expression vector pBIFN. The recombinant plasmid pBIFN was transferred into Agrobacterium tumefaciens strain LBA4404. Then the hIFN-α2b gene was introduced into ginseng callus cells via Agrobacterium-mediated transformation and the ginseng cell line carrying hIFN-α2b gene was selected on G418-containing medium. The presence of target gene in transformed cells was confirmed by PCR and RT-PCR. The results indicate that hIFN-α2b gene has been integrated into the ginseng cells’ genome, with transcription products. hIFN-α2b expressed by the transgenic ginseng cells was detected by Western blot. It was shown that a specific protein band at 19000 could be observed. Cytopathic effect(CPE) inhibition assay using the WISH-VSV system shows that the mean antiviral activity of expressed hIFN-α2b was 6.0×104 IU/mL.
文摘DNA vaccines encoding a viral protein have been shown to induce antiviral immune responses and provide protection against subsequent viral challenge.The present article deals with the efficacy of a DNA vaccine greatly improved by the simultaneous expression of HBsAg and interferon-γ gene.We constructed a dual expression vector pHIN encoding the HBsAg of Hepatitis B virus and murine IFN-γ which are connected with Internal Ribosome Entry Site(IRES).Mice immunized with this dual expression DNA vaccine exhibited the enhancement of cellular immune response and increased the production of anti-HBV surface antibody,compared with the mice of single gene expression control.Taken together,these results demonstrate that the application of a cytokine gene in a DNA vaccine formulation as an adjuvant can improve its immunigenicity.
