Activities of several key enzymes of C-4 photosynthesis pathway and stable carbon isotope discrimination were investigated in flag leaves of a super high-yield hybrid rice (Oryza sativa L.) cv. Peiai 64S/E32 and a tra...Activities of several key enzymes of C-4 photosynthesis pathway and stable carbon isotope discrimination were investigated in flag leaves of a super high-yield hybrid rice (Oryza sativa L.) cv. Peiai 64S/E32 and a traditional hybrid rice cv. Shanyou 63 at different developing stages. Results show that the activity of PEP carboxylase (PEPCase) increased with age of flag leave; the activity of NADP-malate dehydrogenase (NADP-MDH) increased and reached to a peak value at grain filling stage (68-75 d after transplanting), then fell down; the activity of NADP-MDH in cv. Peiai 64S/E32 was much higher than that in cv. Shanyou 63. Before ripening stage (95 d after transplanting), NADP-malic enzyme activity rose gradually. The level of stable carbon isotope discrimination (Delta(13)C) in flag leaves and grains at different developing stages were similar and exhibited a comparative high value at ripening stage. The average Delta(13)C in leaf of cv. Peiai 64S/E32 during different developing stages was 0.43parts per thousand more than that in cv. Shanyou 63.展开更多
The green-tide-forming macroalga Ulva linza was profiled by transcriptome sequencing to ascertain whether the alga carries both C3 and C4 photosynthesis genes.The key enzymes involved in C4 metabolism including pyruva...The green-tide-forming macroalga Ulva linza was profiled by transcriptome sequencing to ascertain whether the alga carries both C3 and C4 photosynthesis genes.The key enzymes involved in C4 metabolism including pyruvate orthophosphate dikinase(PPDK),phosphoenolpyruvate carboxylase(PEPC),and phosphoenolpyruvate carboxykinase(PCK) were found.When measured under normal and different stress conditions,expression of rbcL was higher under normal conditions and lower under the adverse conditions,whereas that of PPDK was higher under some adverse conditions,namely desiccation,high salinity,and low salinity.Both ribulose-1,5-biphosphate carboxylase(RuBPCase) and PPDK were found to play a role in carbon fixation,with significantly higher PPDK activity across the stress conditions.These results suggest that elevated PPDK activity alters carbon metabolism in U.linza leading to partial operation of the C4 carbon metabolism,a pathway that,under stress conditions,probably contributes to the hardy character of U.linza and thus to its wide distribution.展开更多
We applied a resistance split-fusion strategy to increase the in vivo direct cloning efficiency mediated by Red recombination. The cat cassette was divided into two parts： cma （which has a homologous sequence with ...We applied a resistance split-fusion strategy to increase the in vivo direct cloning efficiency mediated by Red recombination. The cat cassette was divided into two parts： cma （which has a homologous sequence with cmb） and cmb, each of which has no resistance separately unless the two parts are fused together. The crab sequence was integrated into one flank of a target clon- ing region in the chromosome, and a linear vector containing the cma sequence was electroporated into the cells to directly capture the target region. Based on this strategy, we successfully cloned an approximately 48 kb DNA fragment from the E. coli DH1-Z chromosome with a positive frequency of approximately 80%. Combined with double-strand breakage-stimulated homologous recombination, we applied this strategy to successfully replace the corresponding region of the E. coli DH36 chromosome and knock out four non-essential genomic regions in one step. This strategy could provide a powerful tool for the heterologous expression of microbial natural product biosynthetic pathways for genome assembly and for the functional study of DNA sequences dozens of kilobases in length.展开更多
文摘Activities of several key enzymes of C-4 photosynthesis pathway and stable carbon isotope discrimination were investigated in flag leaves of a super high-yield hybrid rice (Oryza sativa L.) cv. Peiai 64S/E32 and a traditional hybrid rice cv. Shanyou 63 at different developing stages. Results show that the activity of PEP carboxylase (PEPCase) increased with age of flag leave; the activity of NADP-malate dehydrogenase (NADP-MDH) increased and reached to a peak value at grain filling stage (68-75 d after transplanting), then fell down; the activity of NADP-MDH in cv. Peiai 64S/E32 was much higher than that in cv. Shanyou 63. Before ripening stage (95 d after transplanting), NADP-malic enzyme activity rose gradually. The level of stable carbon isotope discrimination (Delta(13)C) in flag leaves and grains at different developing stages were similar and exhibited a comparative high value at ripening stage. The average Delta(13)C in leaf of cv. Peiai 64S/E32 during different developing stages was 0.43parts per thousand more than that in cv. Shanyou 63.
基金supported by Shandong Science and Technology Plan Project (2011GHY11528)the Specialized Fund for the Basic Research Operating Expenses Program (20603022012004)+4 种基金National Natural Science Foundation of China (41176153)Natural Science Foundation of Shandong Province (2009ZRA02075)Qingdao Municipal Science and Technology Plan Project (11-3-1-5-hy)Qingdao Municipal Science and Technology Plan Project (10-3-4-11-1-jch)National Marine Public Welfare Research Project (200805069)
文摘The green-tide-forming macroalga Ulva linza was profiled by transcriptome sequencing to ascertain whether the alga carries both C3 and C4 photosynthesis genes.The key enzymes involved in C4 metabolism including pyruvate orthophosphate dikinase(PPDK),phosphoenolpyruvate carboxylase(PEPC),and phosphoenolpyruvate carboxykinase(PCK) were found.When measured under normal and different stress conditions,expression of rbcL was higher under normal conditions and lower under the adverse conditions,whereas that of PPDK was higher under some adverse conditions,namely desiccation,high salinity,and low salinity.Both ribulose-1,5-biphosphate carboxylase(RuBPCase) and PPDK were found to play a role in carbon fixation,with significantly higher PPDK activity across the stress conditions.These results suggest that elevated PPDK activity alters carbon metabolism in U.linza leading to partial operation of the C4 carbon metabolism,a pathway that,under stress conditions,probably contributes to the hardy character of U.linza and thus to its wide distribution.
基金supported by the National Natural Science Foundation of China(81373286)National Basic Research Program of China(2011CBA00800)
文摘We applied a resistance split-fusion strategy to increase the in vivo direct cloning efficiency mediated by Red recombination. The cat cassette was divided into two parts： cma （which has a homologous sequence with cmb） and cmb, each of which has no resistance separately unless the two parts are fused together. The crab sequence was integrated into one flank of a target clon- ing region in the chromosome, and a linear vector containing the cma sequence was electroporated into the cells to directly capture the target region. Based on this strategy, we successfully cloned an approximately 48 kb DNA fragment from the E. coli DH1-Z chromosome with a positive frequency of approximately 80%. Combined with double-strand breakage-stimulated homologous recombination, we applied this strategy to successfully replace the corresponding region of the E. coli DH36 chromosome and knock out four non-essential genomic regions in one step. This strategy could provide a powerful tool for the heterologous expression of microbial natural product biosynthetic pathways for genome assembly and for the functional study of DNA sequences dozens of kilobases in length.
