Grain size is one of the most important factors that control rice yield,as it is associated with grain weight(GW).To date,dozens of rice genes that regulate grain size have been isolated;however,the regulatory mechani...Grain size is one of the most important factors that control rice yield,as it is associated with grain weight(GW).To date,dozens of rice genes that regulate grain size have been isolated;however,the regulatory mechanism underlying GW control is not fully understood.Here,the quantitative trait locus qGL5 for grain length(GL)and GW was identified in recombinant inbred lines of 9311 and Nipponbare(NPB)and fine mapped to a candidate gene,OsAUX3.Sequence variations between 9311 and NPB in the OsAUX3 promoter and loss of function of OsAUX3 led to higher GL and GW.RNA sequencing,gene expression quantification,dual-luciferase reporter assays,chromatin immunoprecipitation-quantitative PCR,and yeast one-hybrid assays demonstrated that OsARF6 is an upstream transcription factor regulating the expression of OsAUX3.OsARF6 binds directly to the auxin response elements of the OsAUX3 promoter,covering a single-nucleotide polymorphism site between 9311 and NPB/Dongjin/Hwayoung,and thereby controls GL by altering longitudinal expansion and auxin distribution/content in glume cells.Furthermore,we showed that miR167a positively regulate GL and GW by directing OsARF6 mRNA silencing.Taken together,our study reveals that a novel miR167a-OsARF6-OsAUX3 module regulates GL and GW in rice,providing a potential target for the improvement of rice yield.展开更多
Grain weight and quality are always determined by grain filling.Plant microRNAs have drawn attention as key targets for regulation of grain size and yield.However,the mechanisms that underlie grain size regulation rem...Grain weight and quality are always determined by grain filling.Plant microRNAs have drawn attention as key targets for regulation of grain size and yield.However,the mechanisms that underlie grain size regulation remain largely unclear because of the complex networks that control this trait.Our earlier studies demonstrated that suppressed expression of miR167(STTM/MIM167)substantially increased grain weight.In a field test,the yield increased up to 12.90%-21.94% because of a significantly enhanced grain filling rate.Here,biochemical and genetic analyses revealed the regulatory effects of miR159 on miR167 expression.Further analysis indicated that OsARF12 is the major mediator by which miR167 regulates rice grain filling.Overexpression of OsARF12 produced grain weight and grain filling phenotypes resembling those of STTM/MIM167 plants.Upon in-depth analysis,we found that OsARF12 activates OsCDKF;2 expression by directly binding to the TGTCGG motif in its promoter region.Flow cytometry analysis of young panicles from OsARF12-overexpressing plants and examination of cell number in cdkf;2 mutants verified that OsARF12 positively regulates grain filling and grain size by targeting OsCDKF;2.Moreover,RNA sequencing results suggested that the miR167-OsARF12 module is involved in the cell development process and hormone pathways.OsARF12-overexpressing plants and cdkf;2 mutants exhibited enhanced and reduced sensitivity to exogenous auxin and brassinosteroid(BR)treatment,confirming that targeting of OsCDKF;2 by OsARF12 mediates auxin and BR signaling.Our results reveal that the miR167-OsARF12 module works downstream of miR159 to regulate rice grain filling and grain size via OsCDKF;2 by controlling cell division and mediating auxin and BR signals.展开更多
Bacterial blight(BB), which is caused by Xanthomonas oryzae pv. oryzae(Xoo), is one of the most destructive bacterial diseases of rice(Oryza sativa L.). During plant defense responses, micro RNAs(mi RNAs) play importa...Bacterial blight(BB), which is caused by Xanthomonas oryzae pv. oryzae(Xoo), is one of the most destructive bacterial diseases of rice(Oryza sativa L.). During plant defense responses, micro RNAs(mi RNAs) play important roles in regulating disease resistance. However, the functions of mi RNAs in the interaction between rice and Xoo remain relatively uncharacterized. In this study, we compared the mi RNA profiles of the BB resistant rice introgression line F329 and its susceptible recurrent parent Huang-Hua-Zhan(HHZ) at multiple time points after inoculation with Xoo. A total of 538 known and 312 novel mi RNAs were identified, among which only 17 and 26 were responsive to Xoo infection in F329 and HHZ, respectively. Compared with the expression levels in HHZ, 37 up-regulated and 53 down-regulated mi RNAs were detected in F329. The predicted target genes for the mi RNAs differentially expressed between F329 and HHZ were revealed to be associated with flavonoid synthesis, the reactive oxygen species regulatory pathway, plant hormone signal transduction, defense responses, and growth and development.Additionally, the patterns of interactions between osa-mi R390-3 p, novel_mi R_104, novel_mi R_238,osa-mi R166 k-5 p, osa-mi R529 b, and osa-mi R167 h-3 p and their target genes were further validated by quantitative real-time PCR. Furthermore, we overexpressed osa-mi R167 h-3 p in transgenic plants and proved that this mi RNA positively regulates the resistance of rice to BB. These results provide novel information regarding the mi RNA-based molecular mechanisms underlying the disease resistance of rice. The data presented herein may be useful for engineering rice BB resistance via mi RNAs.展开更多
基金This project was funded by grants from the National Key Research and Development Program of China(2016YFD0100400)the National Natural Science Foundation of China(32060451)the Zhejiang Provincial Nat-ural Science Foundation of China(grant no.L Z19C020001).
文摘Grain size is one of the most important factors that control rice yield,as it is associated with grain weight(GW).To date,dozens of rice genes that regulate grain size have been isolated;however,the regulatory mechanism underlying GW control is not fully understood.Here,the quantitative trait locus qGL5 for grain length(GL)and GW was identified in recombinant inbred lines of 9311 and Nipponbare(NPB)and fine mapped to a candidate gene,OsAUX3.Sequence variations between 9311 and NPB in the OsAUX3 promoter and loss of function of OsAUX3 led to higher GL and GW.RNA sequencing,gene expression quantification,dual-luciferase reporter assays,chromatin immunoprecipitation-quantitative PCR,and yeast one-hybrid assays demonstrated that OsARF6 is an upstream transcription factor regulating the expression of OsAUX3.OsARF6 binds directly to the auxin response elements of the OsAUX3 promoter,covering a single-nucleotide polymorphism site between 9311 and NPB/Dongjin/Hwayoung,and thereby controls GL by altering longitudinal expansion and auxin distribution/content in glume cells.Furthermore,we showed that miR167a positively regulate GL and GW by directing OsARF6 mRNA silencing.Taken together,our study reveals that a novel miR167a-OsARF6-OsAUX3 module regulates GL and GW in rice,providing a potential target for the improvement of rice yield.
基金funded by the National Natural Science Foundation of China(NSFC,32272014,32001440,31971846,and 31871554)the Natural Science Foundation of Henan Province-Excellent Youth Fund(222300420049)+2 种基金the Central Plains Talents Program of Henan Province(Talent Training Series)-Top Young Talents in Central Plains(ZYY-CYU202012170)the Support Plan for Scientific and Technological Innovation Talents in Colleges and Universities of Henan Province(21HAS-TIT037)the China Postdoctoral Science Foundation(2020M682294).
文摘Grain weight and quality are always determined by grain filling.Plant microRNAs have drawn attention as key targets for regulation of grain size and yield.However,the mechanisms that underlie grain size regulation remain largely unclear because of the complex networks that control this trait.Our earlier studies demonstrated that suppressed expression of miR167(STTM/MIM167)substantially increased grain weight.In a field test,the yield increased up to 12.90%-21.94% because of a significantly enhanced grain filling rate.Here,biochemical and genetic analyses revealed the regulatory effects of miR159 on miR167 expression.Further analysis indicated that OsARF12 is the major mediator by which miR167 regulates rice grain filling.Overexpression of OsARF12 produced grain weight and grain filling phenotypes resembling those of STTM/MIM167 plants.Upon in-depth analysis,we found that OsARF12 activates OsCDKF;2 expression by directly binding to the TGTCGG motif in its promoter region.Flow cytometry analysis of young panicles from OsARF12-overexpressing plants and examination of cell number in cdkf;2 mutants verified that OsARF12 positively regulates grain filling and grain size by targeting OsCDKF;2.Moreover,RNA sequencing results suggested that the miR167-OsARF12 module is involved in the cell development process and hormone pathways.OsARF12-overexpressing plants and cdkf;2 mutants exhibited enhanced and reduced sensitivity to exogenous auxin and brassinosteroid(BR)treatment,confirming that targeting of OsCDKF;2 by OsARF12 mediates auxin and BR signaling.Our results reveal that the miR167-OsARF12 module works downstream of miR159 to regulate rice grain filling and grain size via OsCDKF;2 by controlling cell division and mediating auxin and BR signals.
基金supported by grants from the National Natural Science Foundation of China(31571632 and 31661143009)the CAAS Innovative Team Award,and the Bill&Melinda Gates Foundation(OPP51587)。
文摘Bacterial blight(BB), which is caused by Xanthomonas oryzae pv. oryzae(Xoo), is one of the most destructive bacterial diseases of rice(Oryza sativa L.). During plant defense responses, micro RNAs(mi RNAs) play important roles in regulating disease resistance. However, the functions of mi RNAs in the interaction between rice and Xoo remain relatively uncharacterized. In this study, we compared the mi RNA profiles of the BB resistant rice introgression line F329 and its susceptible recurrent parent Huang-Hua-Zhan(HHZ) at multiple time points after inoculation with Xoo. A total of 538 known and 312 novel mi RNAs were identified, among which only 17 and 26 were responsive to Xoo infection in F329 and HHZ, respectively. Compared with the expression levels in HHZ, 37 up-regulated and 53 down-regulated mi RNAs were detected in F329. The predicted target genes for the mi RNAs differentially expressed between F329 and HHZ were revealed to be associated with flavonoid synthesis, the reactive oxygen species regulatory pathway, plant hormone signal transduction, defense responses, and growth and development.Additionally, the patterns of interactions between osa-mi R390-3 p, novel_mi R_104, novel_mi R_238,osa-mi R166 k-5 p, osa-mi R529 b, and osa-mi R167 h-3 p and their target genes were further validated by quantitative real-time PCR. Furthermore, we overexpressed osa-mi R167 h-3 p in transgenic plants and proved that this mi RNA positively regulates the resistance of rice to BB. These results provide novel information regarding the mi RNA-based molecular mechanisms underlying the disease resistance of rice. The data presented herein may be useful for engineering rice BB resistance via mi RNAs.
