Grain size is one of the key factors determining grain weight and yield.However,it remains largely unknown how the auxin signal regulates grain size in rice.Here,a quantitative trait locus qGL1 for grain length was id...Grain size is one of the key factors determining grain weight and yield.However,it remains largely unknown how the auxin signal regulates grain size in rice.Here,a quantitative trait locus qGL1 for grain length was identified with recombinant inbred lines 9311 and Nipponbare(NIP),and fine-mapped to the OsABCB4 gene,which encodes a member of the ATP Binding Cassette B(ABCB)subfamily.Compared to NIP,loss of OsABCB4 function leads to longer and heavier grains,while over-expression of OsABCB4 causes shorter and lighter grains,demonstrating a negative control of grain length and weight in rice.Haplotype analyses using 3024 rice-sequenced genomes indicated that SNP25 and SNP55 in OsABCB4 are correlated with grain length in rice.OsbHLH166,a basic helix-loop-helix transcription factor,was screened from a yeast one-hybrid library and confirmed to directly bind the promoter of OsABCB4 by electrophoretic mobility shift assay and yeast one hybridization validation and to activate the expression of OsABCB4 by dual-luciferase reporter assay.And OsbHLH166 expressions matching OsABCB4 were all higher expressed in stems and glumes.OsABCB4 was subcellularly localized on the plasma membrane,which was verified as an auxin efflux transporter.Histological analysis,IAA content measurement and DR5:GUS activity analyses demonstrated that OsbHLH166 and OsABCB4 control grain length by regulating cell expansion and auxin contents in glume cells and altering the expression of GL7,GS2,TGW3 and GS3 genes during rice grain development.This newly identified OsbHLH166-OsABCB4 module sheds light on our understanding of molecular mechanisms underlying the regulation of grain size and weight via auxin and provides a new gene resource for molecular design breeding of grain type.展开更多
Among the five members of AUX1/LAX genes coding for auxin carriers in rice,only OsAUX1 and OsAUX3 have been reported.To understand the function of the other AUX1/LAX genes,two independent alleles of osaux4 mutants,osa...Among the five members of AUX1/LAX genes coding for auxin carriers in rice,only OsAUX1 and OsAUX3 have been reported.To understand the function of the other AUX1/LAX genes,two independent alleles of osaux4 mutants,osaux4-1 and osaux4-2,were constructed using the CRISPR/Cas9 editing system.Homozygous osaux4-1 or osaux4-2 exhibited shorter primary root(PR)and longer root hair(RH)compared to the wild-type Dongjin(WT/DJ),and lost response to indoleacetic acid(IAA)treatment.OsAUX4 is intensively expressed in roots and localized on the plasma membrane,suggesting that OsAUX4 might function in the regulation of root development.The decreased meristem cell division activity and the downregulated expression of cell cycle genes in root apices of osaux4 mutants supported the hypothesis that OsAUX4 positively regulates PR elongation.OsAUX4 is expressed in RH,and osaux4 mutants showing longer RH compared to WT/DJ implies that OsAUX4 negatively regulates RH development.Furthermore,osaux4 mutants are insensitive to Pi starvation(-Pi)and OsAUX4 effects on the-Pi response is associated with altered expression levels of Pi starvation-regulated genes,and auxin distribution/contents.This study revealed that OsAUX4 not only regulates PR and RH development but also plays a regulatory role in crosstalk between auxin and-Pi signaling.展开更多
基金supported by the National Natural Science Foundation of China(32188102,32372073,and 32301911)the Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-CSNCB-202301)+4 种基金the Natural Science Foundation of Nei Mongol(2022ZD11)the Science Technology Plan Project of Nei Mongol(2023YFDZ0007)the Swiss National Funds(310030_197563)the 2024 Science and Technology Support Project of the Nei Mongol Innovation Center of Biological Breeding Technology(2024NSZC06)the Opening/Autonomous Fund of Key Laboratory of Herbage and Endemic Crop Biology,Ministry of Education,IMU(HECB2024AF01).
文摘Grain size is one of the key factors determining grain weight and yield.However,it remains largely unknown how the auxin signal regulates grain size in rice.Here,a quantitative trait locus qGL1 for grain length was identified with recombinant inbred lines 9311 and Nipponbare(NIP),and fine-mapped to the OsABCB4 gene,which encodes a member of the ATP Binding Cassette B(ABCB)subfamily.Compared to NIP,loss of OsABCB4 function leads to longer and heavier grains,while over-expression of OsABCB4 causes shorter and lighter grains,demonstrating a negative control of grain length and weight in rice.Haplotype analyses using 3024 rice-sequenced genomes indicated that SNP25 and SNP55 in OsABCB4 are correlated with grain length in rice.OsbHLH166,a basic helix-loop-helix transcription factor,was screened from a yeast one-hybrid library and confirmed to directly bind the promoter of OsABCB4 by electrophoretic mobility shift assay and yeast one hybridization validation and to activate the expression of OsABCB4 by dual-luciferase reporter assay.And OsbHLH166 expressions matching OsABCB4 were all higher expressed in stems and glumes.OsABCB4 was subcellularly localized on the plasma membrane,which was verified as an auxin efflux transporter.Histological analysis,IAA content measurement and DR5:GUS activity analyses demonstrated that OsbHLH166 and OsABCB4 control grain length by regulating cell expansion and auxin contents in glume cells and altering the expression of GL7,GS2,TGW3 and GS3 genes during rice grain development.This newly identified OsbHLH166-OsABCB4 module sheds light on our understanding of molecular mechanisms underlying the regulation of grain size and weight via auxin and provides a new gene resource for molecular design breeding of grain type.
基金This project was funded by grants from the National Key Research and Development Program of China(2016YFD0100400)the Zhejiang Provincial Natural Science Foundation of China(LZ19C020001)the National Natural Science Foundation of China(32060451).
文摘Among the five members of AUX1/LAX genes coding for auxin carriers in rice,only OsAUX1 and OsAUX3 have been reported.To understand the function of the other AUX1/LAX genes,two independent alleles of osaux4 mutants,osaux4-1 and osaux4-2,were constructed using the CRISPR/Cas9 editing system.Homozygous osaux4-1 or osaux4-2 exhibited shorter primary root(PR)and longer root hair(RH)compared to the wild-type Dongjin(WT/DJ),and lost response to indoleacetic acid(IAA)treatment.OsAUX4 is intensively expressed in roots and localized on the plasma membrane,suggesting that OsAUX4 might function in the regulation of root development.The decreased meristem cell division activity and the downregulated expression of cell cycle genes in root apices of osaux4 mutants supported the hypothesis that OsAUX4 positively regulates PR elongation.OsAUX4 is expressed in RH,and osaux4 mutants showing longer RH compared to WT/DJ implies that OsAUX4 negatively regulates RH development.Furthermore,osaux4 mutants are insensitive to Pi starvation(-Pi)and OsAUX4 effects on the-Pi response is associated with altered expression levels of Pi starvation-regulated genes,and auxin distribution/contents.This study revealed that OsAUX4 not only regulates PR and RH development but also plays a regulatory role in crosstalk between auxin and-Pi signaling.
