Rice(Oryza sativa L.),a thermophilic crop,is highly sensitive to cold stress,particularly during the seedling stage.Developing cold-tolerant rice varieties is a possible strategy to mitigate yield losses caused by low...Rice(Oryza sativa L.),a thermophilic crop,is highly sensitive to cold stress,particularly during the seedling stage.Developing cold-tolerant rice varieties is a possible strategy to mitigate yield losses caused by low temperatures.However,few genes for cold tolerance have been identified.In this study,we identified OsALA4(Aminophospholipid ATPase 4),encoding a plasma membrane-localized P4-ATPase,from a chromosomal segment substitution line(CSSL-K2832-2)harboring cold-tolerance QTL qLTS5(Low Temperature Sensitive 5).Genetic and subcellular localization analyses revealed that OsALA4 regulates cold tolerance by maintaining plasma membrane fluidity and cellular homeostasis.Physiological assessments showed that OsALA4 reduces malondialdehyde(MDA),electrolyte leakage,reactive oxygen species(ROS),and cell death under cold stress.Promoter activity assays indicated that stronger OsALA4 expression in Nipponbare(OsALA4Nip)correlated with enhanced cold tolerance.Further experiments demonstrated that SNP sites within the promoter regions(-1500 bp to-700 bp)of OsALA4Nipand OsALA49311influenced their activity.This study highlights Os ALA4 as a valuable genetic target for breeding cold tolerant rice.展开更多
Temperature is an important environmental factor affecting heading date of rice.Despite its importance,genes responsible for temperature-sensitive heading in rice have remained elusive.Our previous study identified a ...Temperature is an important environmental factor affecting heading date of rice.Despite its importance,genes responsible for temperature-sensitive heading in rice have remained elusive.Our previous study identified a quantitative trait locus qHd1 which advances heading date under high temperatures.A 9.5-kb insertion was found in the first intron of OsMADS51 in indica variety Zhenshan 97(ZS97).However,the function of this natural variant in controlling temperature sensitivity has not been verified.In this study,we used CRISPR/Cas9 to knock out the 9.5-kb insertion in ZS97.Experiments conducted under cotrolled conditions in phytotrons confirmed that deletion increased temperature sensitivity and advanced heading by downregulating the expression level of OsMADS51.One-hybrid assays in yeast,ChIP-quantitative polymerase chain reaction,electrophoretic mobility shift,and luciferase-based transient transactivation assays collectively confirmed that OsMADS51 affects heading date by regulation of heading date gene Ehd1.We further determined that the long non-coding RNA HEATINR is generated from the first intron of OsMADS51,offering an explanation for how the 9.5-kb insertion affects temperature sensitivity.We also found that OsMADS51 was strongly selected in early/late-season rice varieties in South China,possibly accounting for their strong temperature sensitivity.These insights not only advance our understanding of the molecular mechanisms underlying the temperature-responsive regulation of heading date in rice but also provide a valuable genetic target for molecular breeding.展开更多
Flower organ identity in rice is mainly determined by the A-,B-,C-and E-class genes,with the majority encoding MADS-box transcription factors.However,few studies have investigated how the expression of these floral or...Flower organ identity in rice is mainly determined by the A-,B-,C-and E-class genes,with the majority encoding MADS-box transcription factors.However,few studies have investigated how the expression of these floral organ identity genes is regulated during flower development.In this study,we identified a gene named SUPER WOMAN 2(SPW2),which is necessary for spikelet/floret development in rice by participating in the regulation of the expression of pistil identity genes such as OsMADS3,OsMADS13,OsMADS58 and DL.In the spw2 mutant,ectopic stigma/ovary-like tissues were observed in the non-pistil organs,including sterile lemma,lemma,palea,lodicule,and stamen,suggesting that the identities of these organs were severely affected by mutations in SPW2.SPW2 was shown to encode a plant-specific EMF1-like protein that is involved in H3K27me3 modification as an important component of the PRC2 complex.Expression analysis showed that the SPW2 mutation led to the ectopic expression of OsMADS3,OsMADS13,OsMADS58,and DL in non-pistil organs of the spikelet.The ChIP-qPCR results showed significant reductions in the levels of H3K27me3 modification on the chromatin of these genes.Thus,we demonstrated that SPW2 can mediate the process of H3K27me3 modification of pistil-related genes to regulate their expression in non-pistil organs of spikelets in rice.The results of this study expand our understanding of the molecular mechanism by which SPW2 regulates floral organ identity genes through epigenetic regulation.展开更多
基金supported by grants from the National Natural Science Foundation of China(32121003,32325038)Key Research and Development Projects of Sichuan(2021YFYZ0016)Open Project Program(SKL-ZD202207)of State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China。
文摘Rice(Oryza sativa L.),a thermophilic crop,is highly sensitive to cold stress,particularly during the seedling stage.Developing cold-tolerant rice varieties is a possible strategy to mitigate yield losses caused by low temperatures.However,few genes for cold tolerance have been identified.In this study,we identified OsALA4(Aminophospholipid ATPase 4),encoding a plasma membrane-localized P4-ATPase,from a chromosomal segment substitution line(CSSL-K2832-2)harboring cold-tolerance QTL qLTS5(Low Temperature Sensitive 5).Genetic and subcellular localization analyses revealed that OsALA4 regulates cold tolerance by maintaining plasma membrane fluidity and cellular homeostasis.Physiological assessments showed that OsALA4 reduces malondialdehyde(MDA),electrolyte leakage,reactive oxygen species(ROS),and cell death under cold stress.Promoter activity assays indicated that stronger OsALA4 expression in Nipponbare(OsALA4Nip)correlated with enhanced cold tolerance.Further experiments demonstrated that SNP sites within the promoter regions(-1500 bp to-700 bp)of OsALA4Nipand OsALA49311influenced their activity.This study highlights Os ALA4 as a valuable genetic target for breeding cold tolerant rice.
基金supported by the Laboratory of Lingnan Modern Agriculture Project(NT2021001)Guangdong Province Basic and Applied Basic Research Fund-Provincial and Municipal Joint Fund Project(2023A1515110882)+3 种基金Guangzhou Science and Technology Plan Project(2023A04J0811)Guangdong Province Rural Revitalization Strategy Special Fund Seed Industry Revitalization Project(2022-NPY-00-013)Natural Science Foundation of Zhejiang Pro-vince(LY22C130006)Key Laboratory of New Rice Breeding Technologies in Guangdong Province(2023B1212060042).
文摘Temperature is an important environmental factor affecting heading date of rice.Despite its importance,genes responsible for temperature-sensitive heading in rice have remained elusive.Our previous study identified a quantitative trait locus qHd1 which advances heading date under high temperatures.A 9.5-kb insertion was found in the first intron of OsMADS51 in indica variety Zhenshan 97(ZS97).However,the function of this natural variant in controlling temperature sensitivity has not been verified.In this study,we used CRISPR/Cas9 to knock out the 9.5-kb insertion in ZS97.Experiments conducted under cotrolled conditions in phytotrons confirmed that deletion increased temperature sensitivity and advanced heading by downregulating the expression level of OsMADS51.One-hybrid assays in yeast,ChIP-quantitative polymerase chain reaction,electrophoretic mobility shift,and luciferase-based transient transactivation assays collectively confirmed that OsMADS51 affects heading date by regulation of heading date gene Ehd1.We further determined that the long non-coding RNA HEATINR is generated from the first intron of OsMADS51,offering an explanation for how the 9.5-kb insertion affects temperature sensitivity.We also found that OsMADS51 was strongly selected in early/late-season rice varieties in South China,possibly accounting for their strong temperature sensitivity.These insights not only advance our understanding of the molecular mechanisms underlying the temperature-responsive regulation of heading date in rice but also provide a valuable genetic target for molecular breeding.
基金supported by the Chongqing Modern Agricultural Industry Technology System,China(CQMAITS202301)the National Natural Science Foundation of China(32100287 and 31971919)+2 种基金the Natural Science Foundation of Chongqing,China(cstc2020jcyj-jq X0020 and cstc2021ycjh-bgzxm0066)the China Postdoctoral Science Foundation Funded Project(2020M683219)the Fundamental Research Funds for the Central Universities,China(SWU-XDJH202315)。
文摘Flower organ identity in rice is mainly determined by the A-,B-,C-and E-class genes,with the majority encoding MADS-box transcription factors.However,few studies have investigated how the expression of these floral organ identity genes is regulated during flower development.In this study,we identified a gene named SUPER WOMAN 2(SPW2),which is necessary for spikelet/floret development in rice by participating in the regulation of the expression of pistil identity genes such as OsMADS3,OsMADS13,OsMADS58 and DL.In the spw2 mutant,ectopic stigma/ovary-like tissues were observed in the non-pistil organs,including sterile lemma,lemma,palea,lodicule,and stamen,suggesting that the identities of these organs were severely affected by mutations in SPW2.SPW2 was shown to encode a plant-specific EMF1-like protein that is involved in H3K27me3 modification as an important component of the PRC2 complex.Expression analysis showed that the SPW2 mutation led to the ectopic expression of OsMADS3,OsMADS13,OsMADS58,and DL in non-pistil organs of the spikelet.The ChIP-qPCR results showed significant reductions in the levels of H3K27me3 modification on the chromatin of these genes.Thus,we demonstrated that SPW2 can mediate the process of H3K27me3 modification of pistil-related genes to regulate their expression in non-pistil organs of spikelets in rice.The results of this study expand our understanding of the molecular mechanism by which SPW2 regulates floral organ identity genes through epigenetic regulation.
