Background:Serratia ureilytica DW2 is a highly efficient phosphate-solubilizing bacteria isolated from Codonopsis pilosula rhizosphere soil that can promote the growth of C.pilosula;nonetheless,until now,no validated ...Background:Serratia ureilytica DW2 is a highly efficient phosphate-solubilizing bacteria isolated from Codonopsis pilosula rhizosphere soil that can promote the growth of C.pilosula;nonetheless,until now,no validated reference genes from the genus Serratia have been reported that can be used for the normalization of quantitative real-time polymerase chain reaction(RT–qPCR)data.Methods:To screen stable reference genes of S.ureilytica DW2,the expression of its eight candidate reference genes(16S rRNA,ftsZ,ftsA,mreB,recA,slyD,thiC,and zipA)under different treatment conditions(pH,temperature,culture time,and salt content)was assayed by RT–qPCR.The expression stability of these genes was analyzed using different algorithms(geNorm,NormFinder,and BestKeeper).To verify the reliability of the data,the expression of the glucose dehydrogenase(gdh)gene under different soluble phosphate levels was quantified using the most stably expressed reference gene.Results:The results showed that the zipA and 16S rRNA genes were the most stable reference genes,and the least stable genes were thiC and recA.The expression of gdh was consistent with the phosphate solubilization ability on plates containing the National Botanical Research Institute phosphate growth medium.Conclusion:Therefore,this study provides a stable and reliable reference gene of Serratia for the accurate quantification of functional gene expression in future studies.展开更多
Grain size is determined by the size and number of cells in the grain.The regulation of grain size is crucial for improving crop yield;however,the genes and molecular mechanisms that control grain size remain elusive....Grain size is determined by the size and number of cells in the grain.The regulation of grain size is crucial for improving crop yield;however,the genes and molecular mechanisms that control grain size remain elusive.Here,we report that a member of the detoxification efflux carrier/Multidrug and Toxic Compound Extrusion(DTX/MATE)family transporters,BIG RICE GRAIN 1(BIRG1),negatively influences grain size in rice(Oryza sativa L.).BIRG1 is highly expressed in reproductive organs and roots.In birg1 grain,the outer parenchyma layer cells of spikelet hulls are larger than in wild-type(WT)grains,but the cell number is unaltered.When expressed in Xenopus laevis oocytes,BIRG1 exhibits chloride efflux activity.Consistent with this role of BIRG1,the birg1 mutant shows reduced tolerance to salt stress at a toxic chloride level.Moreover,grains from birg1 plants contain a higher level of chloride than those of WT plants when grown under normal paddy field conditions,and the roots of birg1 accumulate more chloride than those of WT under saline conditions.Collectively,the data suggest that BIRG1 in rice functions as a chloride efflux transporter that is involved in mediating grain size and salt tolerance by controlling chloride homeostasis.展开更多
基金supported by the General Program of Natural Science Foundation of China(32071770)the Fundamental Research Program of Shanxi Province(Award No.202103021223380)the Fund for Shanxi“1331 Project”Key Subjects Construction(1331KSC).
文摘Background:Serratia ureilytica DW2 is a highly efficient phosphate-solubilizing bacteria isolated from Codonopsis pilosula rhizosphere soil that can promote the growth of C.pilosula;nonetheless,until now,no validated reference genes from the genus Serratia have been reported that can be used for the normalization of quantitative real-time polymerase chain reaction(RT–qPCR)data.Methods:To screen stable reference genes of S.ureilytica DW2,the expression of its eight candidate reference genes(16S rRNA,ftsZ,ftsA,mreB,recA,slyD,thiC,and zipA)under different treatment conditions(pH,temperature,culture time,and salt content)was assayed by RT–qPCR.The expression stability of these genes was analyzed using different algorithms(geNorm,NormFinder,and BestKeeper).To verify the reliability of the data,the expression of the glucose dehydrogenase(gdh)gene under different soluble phosphate levels was quantified using the most stably expressed reference gene.Results:The results showed that the zipA and 16S rRNA genes were the most stable reference genes,and the least stable genes were thiC and recA.The expression of gdh was consistent with the phosphate solubilization ability on plates containing the National Botanical Research Institute phosphate growth medium.Conclusion:Therefore,this study provides a stable and reliable reference gene of Serratia for the accurate quantification of functional gene expression in future studies.
基金supported by grants from the National Key Research and Development Program of China(YFD0300102-3 to L.G.L.)the General Program of National Natural Science Foundation of China(No.31872170 to L.L.and No.31900234 to C.H.)+1 种基金the Key Program of the National Natural Science Foundation of China(31930010 to L.L.)the Capacity Building for Sci-Tech Innovation-Fundamental Scientific Research Funds(19530050165 to L.L.)。
文摘Grain size is determined by the size and number of cells in the grain.The regulation of grain size is crucial for improving crop yield;however,the genes and molecular mechanisms that control grain size remain elusive.Here,we report that a member of the detoxification efflux carrier/Multidrug and Toxic Compound Extrusion(DTX/MATE)family transporters,BIG RICE GRAIN 1(BIRG1),negatively influences grain size in rice(Oryza sativa L.).BIRG1 is highly expressed in reproductive organs and roots.In birg1 grain,the outer parenchyma layer cells of spikelet hulls are larger than in wild-type(WT)grains,but the cell number is unaltered.When expressed in Xenopus laevis oocytes,BIRG1 exhibits chloride efflux activity.Consistent with this role of BIRG1,the birg1 mutant shows reduced tolerance to salt stress at a toxic chloride level.Moreover,grains from birg1 plants contain a higher level of chloride than those of WT plants when grown under normal paddy field conditions,and the roots of birg1 accumulate more chloride than those of WT under saline conditions.Collectively,the data suggest that BIRG1 in rice functions as a chloride efflux transporter that is involved in mediating grain size and salt tolerance by controlling chloride homeostasis.