Sedoheptulose-l,7-bisphosphatase （SBPase） is a Calvin cycle enzyme and functions in photosynthetic carbon fixation. We found that SBPase was rapidly carbonylated in response to methyl viologen （MV） treatments in d...Sedoheptulose-l,7-bisphosphatase （SBPase） is a Calvin cycle enzyme and functions in photosynthetic carbon fixation. We found that SBPase was rapidly carbonylated in response to methyl viologen （MV） treatments in detached leaves of Arabidopsis plants. In vitro activity analysis of the purified recombinant SBPase showed that SBPase was car- bonylated by hydroxyl radicals, which led to enzyme inactivation in an H202 dose-dependent manner. To determine the conformity with carbonylation-caused loss in enzymatic activity in response to stresses, we isolated a loss-of-function mutant sbp, which is deficient in SBPase-dependent carbon assimilation and starch biosynthesis, sbp mutant exhibited a severe growth retardation phenotype, especially for the developmental defects in leaves and flowers where SBPASE is highly expressed. The mutation of SBPASE caused growth retardation mainly through inhibition of cell division and ex- pansion, which can be partially rescued by exogenous application of sucrose. Our findings demonstrate that ROS-induced oxidative damage to SBPase affects growth, development, and chloroplast biogenesis in Arabidopsis through inhibiting carbon assimilation efficiency. The data presented here provide a case study that such inactivation of SBPase caused by carbonyl modification may be a kind of adaptation for plants to restrict the operation of the reductive pentose phosphate pathway under stress conditions.展开更多
Elevated activities of cytosolic fructose-1,6-bisphosphatase(cyFBPase) and sedoheptulose-1,7-bisphosphatase(SBPase)are associated with higher yields in plants. In this study, the expression levels of the cyFBPase and ...Elevated activities of cytosolic fructose-1,6-bisphosphatase(cyFBPase) and sedoheptulose-1,7-bisphosphatase(SBPase)are associated with higher yields in plants. In this study, the expression levels of the cyFBPase and SBPase genes were increased by overexpressing rape(Brassica napus) cDNA in tobacco(Nicotiana tabacum) plants. The transgenic plants coexpressing cy FBPase and SBPase(TpFS), or expressing single cy FBPase(TpF) or SBPase(TpS) had 1.77-, 1.55-, 1.23-fold cyFBPase and 1.45-, 1.12-, 1.36-fold SBPase activities as compared to the wild-type(WT), respectively. Photosynthesis rates of TpF, TpS and TpFS increased 4, 20 and 25% compared with WT plants. The SBPase and cyFBPase positively regulated each other and functioned synergistically in transgenic tobacco plants. In addition, the sucrose contents of the three transgenic plants were higher than that of WT plants. The starch accumulation of the TpFS and TpS plants was improved by 53 and 37%, but slightly decreased in TpF plants. Moreover, the transgenic tobacco plants harbouring SBPase and/or cyFBPase genes showed improvements in their growth, biomass, dry weight, plant height, stem diameter, leaf size,flower number, and pod weight. In conclusion, co-expression of SBPase and cyFBPase may pave a new way for improving crop yield in agricultural applications.展开更多
The Calvin Benson cycle(CBC)consists of three critical processes,including fixation of CO_(2) by Rubisco,reduction of 3-phosphoglycerate(3PGA)to triose phosphate(triose-P)with NADPH and ATP generated by the light reac...The Calvin Benson cycle(CBC)consists of three critical processes,including fixation of CO_(2) by Rubisco,reduction of 3-phosphoglycerate(3PGA)to triose phosphate(triose-P)with NADPH and ATP generated by the light reactions,and regeneration of ribulose 1,5-bisphosphate(RuBP)from triose-P.The activ-ities of photosynthesis-related proteins,mainly from the CBC,were found more significantly affected and regulated in plants challenged with high temperature stress,incuding Rubisco,Rubisco activase(RCA) and the enzymes involved in RuBP regeneration,such as sedoheptulose-1,7-bisphosphatase(SBPase).Over the past years,the regulatory mechanism of CBC,especially for redox-regulation,has attracted major interest,because balancing flux at the various enzymatic reactions and maintaining metabolite levels in a range are of critical importance for the optimal operation of CBC under high temperature stress,providing insights into the genetic manipulation of photosynthesis.Here,we summarize recent progress regarding the identification of various layers of regulation point to the key enzymes of CBC for acclimation to environmental temperature changes along with open questions are also discussed.展开更多
基金This research was supported in part by the Ministry of Science and Technology of China，the National Natural Science Foundation of China，the CAS/SAFEA International Partnership Program for Creative Research Teams,and the Bairen Project of Chinese Academy of Sciences
文摘Sedoheptulose-l,7-bisphosphatase （SBPase） is a Calvin cycle enzyme and functions in photosynthetic carbon fixation. We found that SBPase was rapidly carbonylated in response to methyl viologen （MV） treatments in detached leaves of Arabidopsis plants. In vitro activity analysis of the purified recombinant SBPase showed that SBPase was car- bonylated by hydroxyl radicals, which led to enzyme inactivation in an H202 dose-dependent manner. To determine the conformity with carbonylation-caused loss in enzymatic activity in response to stresses, we isolated a loss-of-function mutant sbp, which is deficient in SBPase-dependent carbon assimilation and starch biosynthesis, sbp mutant exhibited a severe growth retardation phenotype, especially for the developmental defects in leaves and flowers where SBPASE is highly expressed. The mutation of SBPASE caused growth retardation mainly through inhibition of cell division and ex- pansion, which can be partially rescued by exogenous application of sucrose. Our findings demonstrate that ROS-induced oxidative damage to SBPase affects growth, development, and chloroplast biogenesis in Arabidopsis through inhibiting carbon assimilation efficiency. The data presented here provide a case study that such inactivation of SBPase caused by carbonyl modification may be a kind of adaptation for plants to restrict the operation of the reductive pentose phosphate pathway under stress conditions.
基金supported by the National Major Program on Transgenic Organisms from Ministry of Agriculture,China(2016ZX08005-004)。
文摘Elevated activities of cytosolic fructose-1,6-bisphosphatase(cyFBPase) and sedoheptulose-1,7-bisphosphatase(SBPase)are associated with higher yields in plants. In this study, the expression levels of the cyFBPase and SBPase genes were increased by overexpressing rape(Brassica napus) cDNA in tobacco(Nicotiana tabacum) plants. The transgenic plants coexpressing cy FBPase and SBPase(TpFS), or expressing single cy FBPase(TpF) or SBPase(TpS) had 1.77-, 1.55-, 1.23-fold cyFBPase and 1.45-, 1.12-, 1.36-fold SBPase activities as compared to the wild-type(WT), respectively. Photosynthesis rates of TpF, TpS and TpFS increased 4, 20 and 25% compared with WT plants. The SBPase and cyFBPase positively regulated each other and functioned synergistically in transgenic tobacco plants. In addition, the sucrose contents of the three transgenic plants were higher than that of WT plants. The starch accumulation of the TpFS and TpS plants was improved by 53 and 37%, but slightly decreased in TpF plants. Moreover, the transgenic tobacco plants harbouring SBPase and/or cyFBPase genes showed improvements in their growth, biomass, dry weight, plant height, stem diameter, leaf size,flower number, and pod weight. In conclusion, co-expression of SBPase and cyFBPase may pave a new way for improving crop yield in agricultural applications.
基金This work was supported by grants from the Minitry of Science and Technology of China(National Key R&D Program of China,2020YFA0907604)the National Natural Science Founda-tion of China(U1812401,31770314,32000211 and 31600225)+1 种基金Science and Technology Commission of Shanghai Municipality(19ZR1466100)the Chinese Academy of Sciences(The Stra-tegic Priority Research Program,XDB27040105).
文摘The Calvin Benson cycle(CBC)consists of three critical processes,including fixation of CO_(2) by Rubisco,reduction of 3-phosphoglycerate(3PGA)to triose phosphate(triose-P)with NADPH and ATP generated by the light reactions,and regeneration of ribulose 1,5-bisphosphate(RuBP)from triose-P.The activ-ities of photosynthesis-related proteins,mainly from the CBC,were found more significantly affected and regulated in plants challenged with high temperature stress,incuding Rubisco,Rubisco activase(RCA) and the enzymes involved in RuBP regeneration,such as sedoheptulose-1,7-bisphosphatase(SBPase).Over the past years,the regulatory mechanism of CBC,especially for redox-regulation,has attracted major interest,because balancing flux at the various enzymatic reactions and maintaining metabolite levels in a range are of critical importance for the optimal operation of CBC under high temperature stress,providing insights into the genetic manipulation of photosynthesis.Here,we summarize recent progress regarding the identification of various layers of regulation point to the key enzymes of CBC for acclimation to environmental temperature changes along with open questions are also discussed.
