In this study, we reported the repaid construction of a molecular marker linkage map of rice (Oryza sativa L.). An F-2 population from the cross between Annong S-1 and Nanjing 11 was used to construct a genetic linkag...In this study, we reported the repaid construction of a molecular marker linkage map of rice (Oryza sativa L.). An F-2 population from the cross between Annong S-1 and Nanjing 11 was used to construct a genetic linkage map of rice. Total of 142 newly screened AFLP markers and 30 anchor markers (25 SSR markers and 5 RFLP markers) were mapped on the 12 chromosomes covering 1537.4 cM of rice genome. The average interval between these markers was 9.0 cM. The total work which usually was finished in more than one year was finished within only 3 months by one person. This is the first plant AFLP map developed in China. A new thermosensitive genic male sterile gene in rice, tms5, was Egged and mapped onto chromosome 2 during the development of the linkage map.展开更多
Previous study indicated that the thermo-sensitive genic malesterile(TGMS) gene in rice was regulated by temperature.TGMS rice plays an important role in hybrid rice production,because the application of the TGMS syst...Previous study indicated that the thermo-sensitive genic malesterile(TGMS) gene in rice was regulated by temperature.TGMS rice plays an important role in hybrid rice production,because the application of the TGMS system in two-line breeding is laborsaving,timesaving,simple,inexpensive,efficient,and eliminating the limitations of the cytoplasmic male sterility(CMS) system.'AnnongS' is the first discovered and deeply studied TGMS rice lines in China.'AnnongS-1' and 'Y58S',two derivatives of TGMS line AnnongS,were both controlled by a single recessive gene named tms5,which was genetically mapped on chromosome 2.In this study,three populations('AnnongS-1' × 'Nanjing11','Y58S' × 'Q611',and 'Y58S' × 'Guanghui122') were developed and used for the molecular fine mapping of the tms5 gene.By analyzing recombination events in the sterile individuals using a total of 125 probes covering the tms5 region,the tms5 gene was physically mapped to a 19-kb DNA fragment between two markers 4039-1 and 4039-2,which were located on the BAC clone AP004039.After the construction of the physical map between two markers 4039-1 and 4039-2,a member(ONAC023) of the NAC(NAM-ATAF-CUC-related) gene family was identified as the candidate gene of the tms5 gene.展开更多
The TMS5e sequence from the Tumen section, at the southern edge of Tengger Desert in the northwestern China, is synchronous with Marine Isotope Stage 5e （MIS5e）. It consists of 16 layers of aeolian dune sands, 11 la...The TMS5e sequence from the Tumen section, at the southern edge of Tengger Desert in the northwestern China, is synchronous with Marine Isotope Stage 5e （MIS5e）. It consists of 16 layers of aeolian dune sands, 11 layers of lacustrine loess-like facies, and 5 layers of lacustrine facies. The results of grain-size analysis shows that the pa- laeo-mobile dune sands, palaeo-fixed to semi-fixed dune sands and loess-like sandy loams are mainly composed of sands, ranging from 70% to 96%; their silt contents ranged from 4% to 20%, and their clay contents ranged from 1% to 5%; the climate under which the aeolian dune sands were deposited is similar to that under which modern mobile dune sands form, which is caused by the dominance of the cold, dry East Asian winter monsoon. In contrast, the lacustrine loess-like facies and lacustrine facies had a lower sand contents than those of the three aeolian dune sands, but have higher silt and clay contents, most of their sand content ranged from 30% to 60%, their silt contents ranged from 35% to 55%, and their clay contents ranged from 6% to 20%. The lacustrine loess-like facies and lacustrine facies formed under the influence of the warm, humid East Asian summer monsoon based on their similarity with modern sediments. The grain-size indicator Mz （mean grain diameter） and the SC/D value in the TMS5e sequence indicate climatic insta- bility at the southern edge of the Tengger Desert during MIS5e, with at least 14 fluctuations between a warm, humid climate and a cold, dry climate, divided into five stages： TMS5e5 （139 kyr to 129.3 kyr B.P.）, TMS5e4 （129.3 kyr to 124 kyr B.P.）, TMS5e3 （124 kyr to 119.5 kyr B.P.）, TMS5e2 （119.5 kyr to 116.5 kyr B.P.）, and TMS5el （116.5 kyr to 113.7 kyr B.P.）. These correspond roughly to MIS5e5, MIS5e4, MIS5e3, MIS5e2, and MIS5el, respectively, in the GRIP ice core data.展开更多
With the cDNA suppression subtraction hybridization method, a spikelet-specific cDNA library was constructed that expressed at meiosis stage in rice. A total of 121 cDNA fragments were selected from the library and us...With the cDNA suppression subtraction hybridization method, a spikelet-specific cDNA library was constructed that expressed at meiosis stage in rice. A total of 121 cDNA fragments were selected from the library and used as EST (expressed se-quence tags) markers to detect the polymorphism between Annong N, a normal fertile Indica rice line and Annong S-1, its spontaneous mutant with thermo-sensitive genic male sterility, using the RFLP (restriction fragment length polymorphism) technique. HN57, one of the EST probes, could detect poly-morphism between them. The results of segregation analysis with the F2 population developed from An-nong S-1 and Annong N indicate that HN57 co-seg- regates with the thermo-sensitive genic male-sterility controlled by tms5, the recessive gene in Annong S-1. This marker is located on the 31.2-cM region of the chromosome 2 of RGP (rice genome research pro-gram) genetic map. To further determine the location of tms5, 80 SSR (simple sequence repeat) markers around this region were developed, and 12 of them were polymorphic. And finally, the tms5 was mapped within region of 181 kb by using these new markers.展开更多
Background QacA, a main exporter mediating the multidrug-resistance of Staphylococcus aureus to a vanety of antiseptics and disinfectants, possesses a topology of 14 a-helical transmembrane segments （TMS）. Our study...Background QacA, a main exporter mediating the multidrug-resistance of Staphylococcus aureus to a vanety of antiseptics and disinfectants, possesses a topology of 14 a-helical transmembrane segments （TMS）. Our study aimed to determine the importance and topology of amino acid residues in and flanking the cytoplasmic end of TMS5. Methods Site-directed mutagenesis was used to mutate 5 residues, including L146, A147, V148, W149 and S150, into cysteine. A minimum inhibitory concentration （MIC） and transport assay with or without N-ethylmaleimide （NEM） were performed to analyse the function of these mutants. Results All of the mutants showed comparable protein expression levels. MIC analysis suggested that mutant W149C showed low resistance levels to the drugs, but the mutations at L146, A147, V148, and S150C had little or no effect on the resistance level. And the results of the fluorimetdc transport assay were in agreement with those of MIC analysis, that is to say, W149C did not allow transport to the substrates to be tested, while the other mutants retained significant transport ability. The reaction of the different mutant proteins with Fluorescein-NEM revealed that the mutant L146C was highly reactive with NEM; the W149C and S150C mutants were moderately reactive; A147C was barely reactive and V148C showed no reactivity. Conclusions The study identified that residues W149 and S150 situated at the interface of the aqueous： lipid junction as functionally important residues, probably involved in the substrate binding and translocation of QacA.展开更多
文摘In this study, we reported the repaid construction of a molecular marker linkage map of rice (Oryza sativa L.). An F-2 population from the cross between Annong S-1 and Nanjing 11 was used to construct a genetic linkage map of rice. Total of 142 newly screened AFLP markers and 30 anchor markers (25 SSR markers and 5 RFLP markers) were mapped on the 12 chromosomes covering 1537.4 cM of rice genome. The average interval between these markers was 9.0 cM. The total work which usually was finished in more than one year was finished within only 3 months by one person. This is the first plant AFLP map developed in China. A new thermosensitive genic male sterile gene in rice, tms5, was Egged and mapped onto chromosome 2 during the development of the linkage map.
文摘Previous study indicated that the thermo-sensitive genic malesterile(TGMS) gene in rice was regulated by temperature.TGMS rice plays an important role in hybrid rice production,because the application of the TGMS system in two-line breeding is laborsaving,timesaving,simple,inexpensive,efficient,and eliminating the limitations of the cytoplasmic male sterility(CMS) system.'AnnongS' is the first discovered and deeply studied TGMS rice lines in China.'AnnongS-1' and 'Y58S',two derivatives of TGMS line AnnongS,were both controlled by a single recessive gene named tms5,which was genetically mapped on chromosome 2.In this study,three populations('AnnongS-1' × 'Nanjing11','Y58S' × 'Q611',and 'Y58S' × 'Guanghui122') were developed and used for the molecular fine mapping of the tms5 gene.By analyzing recombination events in the sterile individuals using a total of 125 probes covering the tms5 region,the tms5 gene was physically mapped to a 19-kb DNA fragment between two markers 4039-1 and 4039-2,which were located on the BAC clone AP004039.After the construction of the physical map between two markers 4039-1 and 4039-2,a member(ONAC023) of the NAC(NAM-ATAF-CUC-related) gene family was identified as the candidate gene of the tms5 gene.
基金Foundation item:Under the auspices of State Key Laboratory of Loess and Quaternary Geology,Institute of Earth Environment,Chinese Academy of Sciences (No.SKLLQG0901,SKLLQG1013)Research Fund for the Doctoral Program of Higher Education of China (No.20094407120004)
文摘The TMS5e sequence from the Tumen section, at the southern edge of Tengger Desert in the northwestern China, is synchronous with Marine Isotope Stage 5e （MIS5e）. It consists of 16 layers of aeolian dune sands, 11 layers of lacustrine loess-like facies, and 5 layers of lacustrine facies. The results of grain-size analysis shows that the pa- laeo-mobile dune sands, palaeo-fixed to semi-fixed dune sands and loess-like sandy loams are mainly composed of sands, ranging from 70% to 96%; their silt contents ranged from 4% to 20%, and their clay contents ranged from 1% to 5%; the climate under which the aeolian dune sands were deposited is similar to that under which modern mobile dune sands form, which is caused by the dominance of the cold, dry East Asian winter monsoon. In contrast, the lacustrine loess-like facies and lacustrine facies had a lower sand contents than those of the three aeolian dune sands, but have higher silt and clay contents, most of their sand content ranged from 30% to 60%, their silt contents ranged from 35% to 55%, and their clay contents ranged from 6% to 20%. The lacustrine loess-like facies and lacustrine facies formed under the influence of the warm, humid East Asian summer monsoon based on their similarity with modern sediments. The grain-size indicator Mz （mean grain diameter） and the SC/D value in the TMS5e sequence indicate climatic insta- bility at the southern edge of the Tengger Desert during MIS5e, with at least 14 fluctuations between a warm, humid climate and a cold, dry climate, divided into five stages： TMS5e5 （139 kyr to 129.3 kyr B.P.）, TMS5e4 （129.3 kyr to 124 kyr B.P.）, TMS5e3 （124 kyr to 119.5 kyr B.P.）, TMS5e2 （119.5 kyr to 116.5 kyr B.P.）, and TMS5el （116.5 kyr to 113.7 kyr B.P.）. These correspond roughly to MIS5e5, MIS5e4, MIS5e3, MIS5e2, and MIS5el, respectively, in the GRIP ice core data.
文摘With the cDNA suppression subtraction hybridization method, a spikelet-specific cDNA library was constructed that expressed at meiosis stage in rice. A total of 121 cDNA fragments were selected from the library and used as EST (expressed se-quence tags) markers to detect the polymorphism between Annong N, a normal fertile Indica rice line and Annong S-1, its spontaneous mutant with thermo-sensitive genic male sterility, using the RFLP (restriction fragment length polymorphism) technique. HN57, one of the EST probes, could detect poly-morphism between them. The results of segregation analysis with the F2 population developed from An-nong S-1 and Annong N indicate that HN57 co-seg- regates with the thermo-sensitive genic male-sterility controlled by tms5, the recessive gene in Annong S-1. This marker is located on the 31.2-cM region of the chromosome 2 of RGP (rice genome research pro-gram) genetic map. To further determine the location of tms5, 80 SSR (simple sequence repeat) markers around this region were developed, and 12 of them were polymorphic. And finally, the tms5 was mapped within region of 181 kb by using these new markers.
基金This project was supported by a grant from National Natural Science Foundation of China （No. 30400373）.
文摘Background QacA, a main exporter mediating the multidrug-resistance of Staphylococcus aureus to a vanety of antiseptics and disinfectants, possesses a topology of 14 a-helical transmembrane segments （TMS）. Our study aimed to determine the importance and topology of amino acid residues in and flanking the cytoplasmic end of TMS5. Methods Site-directed mutagenesis was used to mutate 5 residues, including L146, A147, V148, W149 and S150, into cysteine. A minimum inhibitory concentration （MIC） and transport assay with or without N-ethylmaleimide （NEM） were performed to analyse the function of these mutants. Results All of the mutants showed comparable protein expression levels. MIC analysis suggested that mutant W149C showed low resistance levels to the drugs, but the mutations at L146, A147, V148, and S150C had little or no effect on the resistance level. And the results of the fluorimetdc transport assay were in agreement with those of MIC analysis, that is to say, W149C did not allow transport to the substrates to be tested, while the other mutants retained significant transport ability. The reaction of the different mutant proteins with Fluorescein-NEM revealed that the mutant L146C was highly reactive with NEM; the W149C and S150C mutants were moderately reactive; A147C was barely reactive and V148C showed no reactivity. Conclusions The study identified that residues W149 and S150 situated at the interface of the aqueous： lipid junction as functionally important residues, probably involved in the substrate binding and translocation of QacA.
