Insertion mutagenesis has become one of the most popular methods for gene functions analysis.Here we report a two-element Ac/Ds transposon system containing enhancer trap and gene trap for gene tagging in rice.The exc...Insertion mutagenesis has become one of the most popular methods for gene functions analysis.Here we report a two-element Ac/Ds transposon system containing enhancer trap and gene trap for gene tagging in rice.The excision of Ds element was examined by PCR amplification.The excision frequency of Ds element varied from 0% to 40% among 20 F2 populations derived from 11 different Ds parents.Southern blot analysis revealed that more than 70% of excised Ds elements reinserted into rice genome and above 70% of the reinserted Ds elements were located at different positions of the chromosome in rice.The result of histochemical GUS analysis indicated that 28% of enhancer trap and 22% of gene trap tagging plants displayed GUS activity in leaves, roots,flowers or seeds.The GUS positive lines will be useful for identifying gene function in rice.展开更多
Since the hallmark discovery of Aequorea victoria＇s Green Fluorescent Protein （GFP） and its adaptation for efficient use in plants, fluorescent protein tags marking expression profiles or genuine proteins of intere...Since the hallmark discovery of Aequorea victoria＇s Green Fluorescent Protein （GFP） and its adaptation for efficient use in plants, fluorescent protein tags marking expression profiles or genuine proteins of interest have been used to recognize plant tissues and cell types, to monitor dynamic cell fate selection processes, and to obtain cell type-specific transcriptomes. Fluorescent tagging enabled visualization in living tissues and the precise recordings of dy- namic expression pattern changes. The resulting accurate recording of cell fate acquisition kinetics in space and time has strongly stimulated mathematical modeling of self-organizing feedback mechanisms. In developmental studies, the use of fluorescent proteins has become critical, where morphological markers of tissues, cell types, or differentiation stages are either not known or not easily recognizable. In this review, we focus on the use of fluorescent markers to identify and illuminate otherwise invisible cell states in plant development.展开更多
The fully sequenced genomes of Arabidopsis, rice, tomato, potato, ma ize, wheat, and soybean offer large amounts of information about cellular and de velopmental biology. It is a central challenge of genomics to use t...The fully sequenced genomes of Arabidopsis, rice, tomato, potato, ma ize, wheat, and soybean offer large amounts of information about cellular and de velopmental biology. It is a central challenge of genomics to use this informati on in discovering the function of proteins and identifying developmentally impor tant genes. Although classical genetic approaches to gene identification which r ely on disruption of a gene leading to a recognizable phenotype continues to be an extremely successful one, T-DNA mediated gene trap tagging which has been dev eloped that utilize random integration of reporter gene constructs has also prov en to be an extremely powerful tool in plant cellular developmental biology. In this review, how gene trap tagging, promoter trap tagging, and enhancer trap tag ging detection systems have been applied to plant biology is described and these gene identification techniques could be useful to the plant molecular biology a nd plant biotechnology community.展开更多
Plants employ several strategies to maintain cellular ion homeostasis under salinity stress, including mediat-ing ion fluxes by transmembrane transport proteins and adjusting osmotic pressure by accumulating osmolytes...Plants employ several strategies to maintain cellular ion homeostasis under salinity stress, including mediat-ing ion fluxes by transmembrane transport proteins and adjusting osmotic pressure by accumulating osmolytes. The HKT （high-affinity potassium transporter） gene family comprises Na^＋ and Na^＋/K^＋ transporters in diverse plant species, with HKT1,1 as the only member in Arabidopsis thaliana. Cell-type-specific overexpression of AtHKT1;1 has been shown to prevent shoot Na^＋ overaccumulation under salinity stress. Here, we analyzed a broad range of metabolites and elements in shoots and roots of different AtHKT1;1 genotypes and their parental strains before and after salinity stress, revealing a reciprocal relationship of metabolite differences between an AtHKT1;1 knockout line （hktl;1） and the AtHKT1;1 overex- pressing lines （E2586 UASGAL4：HKT1;1 and J2731＊UASGAL4：HKT1;1）. Although levels of root sugars were increased after salt stress in both AtHKTI,1 overexpressing lines, E2586 UASGAL4：HKT1;1 showed higher accumulation of the osmopro-tectants trehalose, gentiobiose, and melibiose, whereas J2731＊UASGAL4：HKT1;1 showed higher levels of sucrose and raffinose, compared with their parental lines, respectively. In contrast, the knockout line hktl,1 showed strong increases in the levels of the tricarboxylic acid （TCA） cycle intermediates in the shoots after salt treatment. This coincided with a significant depletion of sugars, suggesting that there is an increased rate of carbon influx into the TCA cycle at a constant rate of C-efflux from the cycle, which might be needed to support plant survival during salt stress. Using correlation analysis, we identified associations between the Na^＋ content and several sugars, suggesting that regulation of sugar metabolism is important in plant responses to salinity stress.展开更多
基金supported by The National Natural Science Foundation of China (30721064, 30730056, 30620120101)National BasicResearch Program of China (2005CB522504, 2006CB943801,2007CB914502)~~
文摘Insertion mutagenesis has become one of the most popular methods for gene functions analysis.Here we report a two-element Ac/Ds transposon system containing enhancer trap and gene trap for gene tagging in rice.The excision of Ds element was examined by PCR amplification.The excision frequency of Ds element varied from 0% to 40% among 20 F2 populations derived from 11 different Ds parents.Southern blot analysis revealed that more than 70% of excised Ds elements reinserted into rice genome and above 70% of the reinserted Ds elements were located at different positions of the chromosome in rice.The result of histochemical GUS analysis indicated that 28% of enhancer trap and 22% of gene trap tagging plants displayed GUS activity in leaves, roots,flowers or seeds.The GUS positive lines will be useful for identifying gene function in rice.
文摘Since the hallmark discovery of Aequorea victoria＇s Green Fluorescent Protein （GFP） and its adaptation for efficient use in plants, fluorescent protein tags marking expression profiles or genuine proteins of interest have been used to recognize plant tissues and cell types, to monitor dynamic cell fate selection processes, and to obtain cell type-specific transcriptomes. Fluorescent tagging enabled visualization in living tissues and the precise recordings of dy- namic expression pattern changes. The resulting accurate recording of cell fate acquisition kinetics in space and time has strongly stimulated mathematical modeling of self-organizing feedback mechanisms. In developmental studies, the use of fluorescent proteins has become critical, where morphological markers of tissues, cell types, or differentiation stages are either not known or not easily recognizable. In this review, we focus on the use of fluorescent markers to identify and illuminate otherwise invisible cell states in plant development.
文摘The fully sequenced genomes of Arabidopsis, rice, tomato, potato, ma ize, wheat, and soybean offer large amounts of information about cellular and de velopmental biology. It is a central challenge of genomics to use this informati on in discovering the function of proteins and identifying developmentally impor tant genes. Although classical genetic approaches to gene identification which r ely on disruption of a gene leading to a recognizable phenotype continues to be an extremely successful one, T-DNA mediated gene trap tagging which has been dev eloped that utilize random integration of reporter gene constructs has also prov en to be an extremely powerful tool in plant cellular developmental biology. In this review, how gene trap tagging, promoter trap tagging, and enhancer trap tag ging detection systems have been applied to plant biology is described and these gene identification techniques could be useful to the plant molecular biology a nd plant biotechnology community.
文摘Plants employ several strategies to maintain cellular ion homeostasis under salinity stress, including mediat-ing ion fluxes by transmembrane transport proteins and adjusting osmotic pressure by accumulating osmolytes. The HKT （high-affinity potassium transporter） gene family comprises Na^＋ and Na^＋/K^＋ transporters in diverse plant species, with HKT1,1 as the only member in Arabidopsis thaliana. Cell-type-specific overexpression of AtHKT1;1 has been shown to prevent shoot Na^＋ overaccumulation under salinity stress. Here, we analyzed a broad range of metabolites and elements in shoots and roots of different AtHKT1;1 genotypes and their parental strains before and after salinity stress, revealing a reciprocal relationship of metabolite differences between an AtHKT1;1 knockout line （hktl;1） and the AtHKT1;1 overex- pressing lines （E2586 UASGAL4：HKT1;1 and J2731＊UASGAL4：HKT1;1）. Although levels of root sugars were increased after salt stress in both AtHKTI,1 overexpressing lines, E2586 UASGAL4：HKT1;1 showed higher accumulation of the osmopro-tectants trehalose, gentiobiose, and melibiose, whereas J2731＊UASGAL4：HKT1;1 showed higher levels of sucrose and raffinose, compared with their parental lines, respectively. In contrast, the knockout line hktl,1 showed strong increases in the levels of the tricarboxylic acid （TCA） cycle intermediates in the shoots after salt treatment. This coincided with a significant depletion of sugars, suggesting that there is an increased rate of carbon influx into the TCA cycle at a constant rate of C-efflux from the cycle, which might be needed to support plant survival during salt stress. Using correlation analysis, we identified associations between the Na^＋ content and several sugars, suggesting that regulation of sugar metabolism is important in plant responses to salinity stress.
