[目的]水稻Hd1(Heading date 1)在不同遗传背景的品种中存在多种单体型,利用CRISPR/Cas9基因编辑技术创制hd1突变体探究其对抽穗期的调控,解析Hd1单体型的功能特性。[方法]在Hd1的CDS设计靶位点,构建敲除载体,利用农杆菌介导法转入受体...[目的]水稻Hd1(Heading date 1)在不同遗传背景的品种中存在多种单体型,利用CRISPR/Cas9基因编辑技术创制hd1突变体探究其对抽穗期的调控,解析Hd1单体型的功能特性。[方法]在Hd1的CDS设计靶位点,构建敲除载体,利用农杆菌介导法转入受体品种龙粳11(LJ11)和松粳2(SJ2)中。测序鉴定转基因植株突变类型,与对照同时种植调查抽穗期,qRT-PCR检测突变体中成花素基因Hd3a和RFT1的表达量。[结果]Hd1的编码区在LJ11背景下第30个碱基后面插入G,在SJ2背景下第31个碱基后面插入T,获得纯合突变体hd1(LJ11)和hd1(SJ2),相较于野生型,hd1(LJ11)延迟了水稻开花,Hd3a和RFT1的表达量下降,而hd1(SJ2)提早了水稻开花伴随着Hd3a和RFT1的表达量上升。[结论]hd1(LJ11)延迟抽穗6.1 d,hd1(SJ2)提早开花2.8 d,说明两种遗传背景下Hd1发挥功能存在差异性,调控抽穗期具有背景依赖性。展开更多
Heading date determines the seasonal and regional adaptation of rice(Oryza sativa L.)varieties and is mainly controlled by photoperiod sensitivity(PS).The core heading date genes Hd1,Ghd7,DTH8,and PRR37 act synergisti...Heading date determines the seasonal and regional adaptation of rice(Oryza sativa L.)varieties and is mainly controlled by photoperiod sensitivity(PS).The core heading date genes Hd1,Ghd7,DTH8,and PRR37 act synergistically in regulating the PS.In this study,we systematically analyze the heading date,PS,and agronomic traits of eight homozygous lines with various combinations of Hd1,Ghd7,and DTH8 alleles in the prr37 background under long-day(LD)and short-day(SD)conditions,respectively.We find that Hd1 alone promotes heading,regardless of the day length.However,under LDs,Hd1 suppresses flowering,in coordination with functional Ghd7 or with Ghd7 and DTH8.These loci cooperate to negatively regulate the Ehd1-Hd3 a/RFT1 pathway and delay heading.Under SDs,Hd1 competes with various heading suppressors to promote heading.Therefore,the dual function of Hd1 is vital for PS.The lines carrying Hd1 alone show reduced plant height with fewer primary and secondary branches in panicles.Lines carrying Ghd7 and DTH8(with hd1)show delayed heading and improve agronomic traits.Overall,our results reveal the regulation of rice PS flowering by the core heading date genes and their effects on agronomic traits,providing valuable information for the selection of rice varieties for adaptation to different light and temperature conditions.展开更多
We investigated genetic variation of a rice HEADING DATE 1(HD1) homolog in foxtail millet.First, we searched for a rice HD1 homolog in a foxtail millet genome sequence and designed primers to amplify the entire coding...We investigated genetic variation of a rice HEADING DATE 1(HD1) homolog in foxtail millet.First, we searched for a rice HD1 homolog in a foxtail millet genome sequence and designed primers to amplify the entire coding sequence of the gene. We compared full HD1 gene sequences of 11 accessions(including Yugu 1, a Chinese cultivar used for genome sequencing) from various regions in Europe and Asia, found a nucleotide substitution at a putative splice site of intron 1, and designated the accessions with the nucleotide substitution as carrying a splicing variant. We verified by RT-PCR that this single nucleotide substitution causes aberrant splicing of intron 1. We investigated the geographical distribution of the splicing variant in 480 accessions of foxtail millet from various regions of Europe and Asia and part of Africa by d CAPS and found that the splicing variant is broadly distributed in Europe and Asia. Differences of heading times between accessions with wild type allele of the HD1 gene and those with the splicing variant allele were unclear. We also investigated variation in 13 accessions of ssp. viridis, the wild ancestor, and the results suggested that the wild type is predominant in the wild ancestor.展开更多
The microstructural evolution and Vickers hardness measurement in the welding heat-affected zone （HAZ） of HD15 Nil MnMoNbCu steel for nuclear power station were investigated by Gleeble-3180 thermal mechanical simula...The microstructural evolution and Vickers hardness measurement in the welding heat-affected zone （HAZ） of HD15 Nil MnMoNbCu steel for nuclear power station were investigated by Gleeble-3180 thermal mechanical simulator, and the simulated HAZ continuous cooling transformation curves （SH-CCT） were measured simultaneously. With ts/5 inereasing from 3.75 s to 15 000 s, the product was obtained martensite, bainite, ferrite and pearlite, successively. The result of microstructure and Vickers hardness in the heat-affected zone was in good agreement with those measured by SH-CCT diagram with the heat input 16. 2 kJ/cm as an example to weld the HD15Ni1MnMoNbCu steel pipe using TIG/SMAW/SAW welding methods.展开更多
Photoperiodic flowering is one of the most important pathways to govern flowering in rice （Oryza sativa）, in which Heading date 1 （Hdl）, an ortholog of the Arabidopsis CONSTANS gene, encodes a pivotal regulator. H...Photoperiodic flowering is one of the most important pathways to govern flowering in rice （Oryza sativa）, in which Heading date 1 （Hdl）, an ortholog of the Arabidopsis CONSTANS gene, encodes a pivotal regulator. Hdl promotes flowering under short-day conditions （SD） but represses flowering under long-day conditions （LD） by regulating the expression of Heading date 3a （Hd3a）, the FLOWERING LOCUS T （FT） ortholog in rice. However, the molecular mechanism of how Hdl changes its regulatory activity in response to day length remains largely unknown. In this study, we demonstrated that the repression of flowering in LD by Hdl is dependent on the transcription factor DAYS TO HEADING 8 （DTHS）. Loss of DTH8 function results in the activation of Hd3a by Hdl, leading to early flowering. We found that Hdl directly interacts with DTH8 and that the formation of the DTH8-Hd1 complex is necessary for the transcriptional repression of Hd3a by Hdl in LD, implicating that the switch of Hdl function is mediated by DTH8 in LD rather than in SD. Furthermore, we revealed that DTH8 associates with the Hd3a promoter to modulate the level of H3K27 trimethylation （H3K27me3） at the Hd3a locus. In the presence of the DTH8-Hdl complex, the H3K27me3 level was increased at Hd3a, whereas loss of DTH8 function resulted in decreased H3K27me3 level at Hd3a. Taken together, our findings indicate that, in response to day length, DTH8 plays a critical role in mediating the transcriptional regulation of Hd3a by Hdl through the DTH8-Hd1 module to shape epigenetic modifications in photoperiodic flowering.展开更多
Flowering time(heading date)is a critical agronomic trait that determines the yield and regional adaptability of crops.Heading date 1(Hd1)is a central regulator of photoperiodic flowering in rice(Oryza sativa).However...Flowering time(heading date)is a critical agronomic trait that determines the yield and regional adaptability of crops.Heading date 1(Hd1)is a central regulator of photoperiodic flowering in rice(Oryza sativa).However,how the homeostasis of Hd1 protein is achieved is poorly understood.Here,we report that the nuclear autophagy pathway mediates Hd1 degradation in the dark to regulate flowering.Loss of autophagy function results in an accumulation of Hd1 and delays flowering under both short-day and long-day conditions.In the dark,nucleus-localized Hd1 is recognized as a substrate for autophagy and is subjected to vacuolar degradation via the autophagy protein OsATG8.The Hd1-0sATG8 interaction is required for autophagic degradation of Hd1 in the dark.Our study reveals a new mechanism by which Hd1 protein homeostasis is regulated by autophagy to control rice flowering.Our study also indicates that the regulation of flowering by autophagic degradation of Hd1 orthologs may have arisen over the course of mesangiosperm evolution,which would have increased their flexibility and adaptability to the environment by modulating flowering time.展开更多
文摘[目的]水稻Hd1(Heading date 1)在不同遗传背景的品种中存在多种单体型,利用CRISPR/Cas9基因编辑技术创制hd1突变体探究其对抽穗期的调控,解析Hd1单体型的功能特性。[方法]在Hd1的CDS设计靶位点,构建敲除载体,利用农杆菌介导法转入受体品种龙粳11(LJ11)和松粳2(SJ2)中。测序鉴定转基因植株突变类型,与对照同时种植调查抽穗期,qRT-PCR检测突变体中成花素基因Hd3a和RFT1的表达量。[结果]Hd1的编码区在LJ11背景下第30个碱基后面插入G,在SJ2背景下第31个碱基后面插入T,获得纯合突变体hd1(LJ11)和hd1(SJ2),相较于野生型,hd1(LJ11)延迟了水稻开花,Hd3a和RFT1的表达量下降,而hd1(SJ2)提早了水稻开花伴随着Hd3a和RFT1的表达量上升。[结论]hd1(LJ11)延迟抽穗6.1 d,hd1(SJ2)提早开花2.8 d,说明两种遗传背景下Hd1发挥功能存在差异性,调控抽穗期具有背景依赖性。
基金supported by grants from the National Natural Science Foundation of China (31921004 and 31871596)the Major Program of Guangdong Basic and Applied Research (2019B030302006)the Laboratory of Lingnan Modern Agriculture Project (NT2021002)
文摘Heading date determines the seasonal and regional adaptation of rice(Oryza sativa L.)varieties and is mainly controlled by photoperiod sensitivity(PS).The core heading date genes Hd1,Ghd7,DTH8,and PRR37 act synergistically in regulating the PS.In this study,we systematically analyze the heading date,PS,and agronomic traits of eight homozygous lines with various combinations of Hd1,Ghd7,and DTH8 alleles in the prr37 background under long-day(LD)and short-day(SD)conditions,respectively.We find that Hd1 alone promotes heading,regardless of the day length.However,under LDs,Hd1 suppresses flowering,in coordination with functional Ghd7 or with Ghd7 and DTH8.These loci cooperate to negatively regulate the Ehd1-Hd3 a/RFT1 pathway and delay heading.Under SDs,Hd1 competes with various heading suppressors to promote heading.Therefore,the dual function of Hd1 is vital for PS.The lines carrying Hd1 alone show reduced plant height with fewer primary and secondary branches in panicles.Lines carrying Ghd7 and DTH8(with hd1)show delayed heading and improve agronomic traits.Overall,our results reveal the regulation of rice PS flowering by the core heading date genes and their effects on agronomic traits,providing valuable information for the selection of rice varieties for adaptation to different light and temperature conditions.
基金supported by the NIAS Genebank Project,NIAS,Japan
文摘We investigated genetic variation of a rice HEADING DATE 1(HD1) homolog in foxtail millet.First, we searched for a rice HD1 homolog in a foxtail millet genome sequence and designed primers to amplify the entire coding sequence of the gene. We compared full HD1 gene sequences of 11 accessions(including Yugu 1, a Chinese cultivar used for genome sequencing) from various regions in Europe and Asia, found a nucleotide substitution at a putative splice site of intron 1, and designated the accessions with the nucleotide substitution as carrying a splicing variant. We verified by RT-PCR that this single nucleotide substitution causes aberrant splicing of intron 1. We investigated the geographical distribution of the splicing variant in 480 accessions of foxtail millet from various regions of Europe and Asia and part of Africa by d CAPS and found that the splicing variant is broadly distributed in Europe and Asia. Differences of heading times between accessions with wild type allele of the HD1 gene and those with the splicing variant allele were unclear. We also investigated variation in 13 accessions of ssp. viridis, the wild ancestor, and the results suggested that the wild type is predominant in the wild ancestor.
文摘The microstructural evolution and Vickers hardness measurement in the welding heat-affected zone （HAZ） of HD15 Nil MnMoNbCu steel for nuclear power station were investigated by Gleeble-3180 thermal mechanical simulator, and the simulated HAZ continuous cooling transformation curves （SH-CCT） were measured simultaneously. With ts/5 inereasing from 3.75 s to 15 000 s, the product was obtained martensite, bainite, ferrite and pearlite, successively. The result of microstructure and Vickers hardness in the heat-affected zone was in good agreement with those measured by SH-CCT diagram with the heat input 16. 2 kJ/cm as an example to weld the HD15Ni1MnMoNbCu steel pipe using TIG/SMAW/SAW welding methods.
基金This work was supported by the National Natural Science Foundation （31501278, 31671378 to X.O.）, the National Key Research and Development Program of China （2017YFA0506100 to X.H.） and the Fundamental Research Funds for the Central Universities （20720170068 to X.O. and 20720150098 to X.H.）.
文摘Photoperiodic flowering is one of the most important pathways to govern flowering in rice （Oryza sativa）, in which Heading date 1 （Hdl）, an ortholog of the Arabidopsis CONSTANS gene, encodes a pivotal regulator. Hdl promotes flowering under short-day conditions （SD） but represses flowering under long-day conditions （LD） by regulating the expression of Heading date 3a （Hd3a）, the FLOWERING LOCUS T （FT） ortholog in rice. However, the molecular mechanism of how Hdl changes its regulatory activity in response to day length remains largely unknown. In this study, we demonstrated that the repression of flowering in LD by Hdl is dependent on the transcription factor DAYS TO HEADING 8 （DTHS）. Loss of DTH8 function results in the activation of Hd3a by Hdl, leading to early flowering. We found that Hdl directly interacts with DTH8 and that the formation of the DTH8-Hd1 complex is necessary for the transcriptional repression of Hd3a by Hdl in LD, implicating that the switch of Hdl function is mediated by DTH8 in LD rather than in SD. Furthermore, we revealed that DTH8 associates with the Hd3a promoter to modulate the level of H3K27 trimethylation （H3K27me3） at the Hd3a locus. In the presence of the DTH8-Hdl complex, the H3K27me3 level was increased at Hd3a, whereas loss of DTH8 function resulted in decreased H3K27me3 level at Hd3a. Taken together, our findings indicate that, in response to day length, DTH8 plays a critical role in mediating the transcriptional regulation of Hd3a by Hdl through the DTH8-Hd1 module to shape epigenetic modifications in photoperiodic flowering.
基金supported by the National Key Research and Development Program of China(2020YFA0907602)the National Natural Science Foundation of China(31730102).
文摘Flowering time(heading date)is a critical agronomic trait that determines the yield and regional adaptability of crops.Heading date 1(Hd1)is a central regulator of photoperiodic flowering in rice(Oryza sativa).However,how the homeostasis of Hd1 protein is achieved is poorly understood.Here,we report that the nuclear autophagy pathway mediates Hd1 degradation in the dark to regulate flowering.Loss of autophagy function results in an accumulation of Hd1 and delays flowering under both short-day and long-day conditions.In the dark,nucleus-localized Hd1 is recognized as a substrate for autophagy and is subjected to vacuolar degradation via the autophagy protein OsATG8.The Hd1-0sATG8 interaction is required for autophagic degradation of Hd1 in the dark.Our study reveals a new mechanism by which Hd1 protein homeostasis is regulated by autophagy to control rice flowering.Our study also indicates that the regulation of flowering by autophagic degradation of Hd1 orthologs may have arisen over the course of mesangiosperm evolution,which would have increased their flexibility and adaptability to the environment by modulating flowering time.
