Grain size is one of the most important factors affecting rice grain quality and yield,and attracts great attention from molecular biologists and breeders.In this study,we engineered a CRISPR/Cas9 system targeting the...Grain size is one of the most important factors affecting rice grain quality and yield,and attracts great attention from molecular biologists and breeders.In this study,we engineered a CRISPR/Cas9 system targeting the miR396 recognition site of the rice GS2 gene,which encodes growth-regulating factor 4(OsGRF4)and regulates multiple agronomic traits including grain size,grain quality,nitrogen use efficiency,abiotic stress response,and seed shattering.In contrast to most previous genome editing efforts in which indel mutations were chosen to obtain null mutants,a mutant named GS2^(E) carrying an in-frame 6-bp deletion and 1-bp substitution within the miR396-targeted sequence was identified.GS2^(E) plants showed increased expression of GS2 in consistent with impaired repression by miR396.As expected,the gain-of-function GS2^(E) mutant exhibited multiple beneficial traits including increased grain size and yield and bigger grain length/width ratio.Thousand grain weight and grain yield per plant of GS2^(E) plants were increased by 23.5%and 10.4%,respectively.These improved traits were passed to hybrids in a semidominant way,suggesting that the new GS2^(E) allele has great potential in rice improvement.Taken together,we report new GS2 germplasm and describe a novel gene-editing strategy that can be widely employed to improve grain size and yield in rice.This trait-improvement strategy could be applied to other genes containing miRNA target sites,in particular the conserved miR396-GRF/GIF module that governs plant growth,development and environmental response.展开更多
二十世纪五六十年代,半矮秆水稻和小麦新品种的育成和大面积推广,有效地解决了"高产与倒伏"之间的制约矛盾,这一历程即为众所周知的"绿色革命"。水稻半矮秆基因sd1(semi-dwarf1)既增加了水稻收获指数,又有效地解决...二十世纪五六十年代,半矮秆水稻和小麦新品种的育成和大面积推广,有效地解决了"高产与倒伏"之间的制约矛盾,这一历程即为众所周知的"绿色革命"。水稻半矮秆基因sd1(semi-dwarf1)既增加了水稻收获指数,又有效地解决了因密植和大量施肥导致的植株倒伏和减产问题,实现了水稻单产的大幅度提升。然而,半矮秆水稻和小麦品种也表现出其植物生长对氮肥响应的减弱,根系对氮肥的吸收速率下降,导致氮肥利用效率(nitrogen use efficiency,NUE)下降。因此,协同改良半矮化作物的高产与氮肥高效利用性状,对保障国家粮食安全和农业可持续发展具有重要意义。傅向东课题组的研究显示,水稻生长调节因子GRF4和生长抑制因子DELLA相互之间的反向平衡调节赋予了植物生长与碳-氮代谢之间的稳态共调节。DELLA蛋白高水平积累导致了作物半矮化,实现了植株耐高肥和抗倒伏的高产目标,但其伴随着氮肥利用效率的降低。相反,GRF4蛋白高水平积累能协同提高作物光合作用和氮肥利用效率,但并不改变"绿色革命"品种的半矮化优良性状,从而实现了在现有高产品种中协同提升产量和氮肥利用效率。DELLA-GRF4分子调控模块的解析为"少投入、多产出、保护环境"的绿色高产高效农作物新品种培育提供了理论基础和具有育种利用价值的新基因资源。展开更多
The LGS1(Large grain size 1)gene,also known as GS2/GL2/Os GRF4,is involved in regulating grain size and quality in rice,but the mechanism governing grain size has not been elucidated.We performed transcriptomic,proteo...The LGS1(Large grain size 1)gene,also known as GS2/GL2/Os GRF4,is involved in regulating grain size and quality in rice,but the mechanism governing grain size has not been elucidated.We performed transcriptomic,proteomic,and phosphoproteomic analyses of young rice panicles in Samba(a wild-type cultivar with extra-small grain)and NIL-LGS1(a nearly isogenic line of LGS1 with large grain in the Samba genetic background)at three developmental stages(4–6)to identify internal dynamic functional networks determining grain size that are mediated by LGS1.Differentially expressed proteins formed seven highly functionally correlated clusters.The concordant regulation of multiple functional clusters may be key features of the development of grain length in rice.In stage 5,16 and 24 phosphorylated proteins were significantly up-regulated and down-regulated,and dynamic phosphorylation events may play accessory roles in determining rice grain size by participating in protein–protein interaction networks.Transcriptomic analysis in stage 5 showed that differentially expressed alternative splicing events and dynamic gene regulatory networks based on 39 transcription factors and their highly correlated target genes might contribute to rice grain development.Integrative multilevel omics analysis suggested that the regulatory network at the transcriptional and posttranscriptional levels could be directly manifested at the translational level,and this analysis also suggested a regulatory mechanism,regulation of protein translation levels,in the biological process that extends from transcript to protein to the development of grain.Functional analysis suggested that biological processes including MAPK signaling,calcium signaling,cell proliferation,cell wall,energy metabolism,hormone pathway,and ubiquitin-proteasome pathway might be involved in LGS1-mediated regulation of grain length.Thus,LGS1-mediated regulation of grain size is affected by dynamic transcriptional,posttranscriptional,translational and posttranslational changes.展开更多
在农业生产中,大量施用氮肥是农作物增产的重要措施之一.在长达半个多世纪的农作物育种史上一直占据主导地位的'绿色革命'半矮化品种具有耐高肥、抗倒伏和高产的优良特性,但同时也存在氮肥利用效率(nitrogen use efficiency, N...在农业生产中,大量施用氮肥是农作物增产的重要措施之一.在长达半个多世纪的农作物育种史上一直占据主导地位的'绿色革命'半矮化品种具有耐高肥、抗倒伏和高产的优良特性,但同时也存在氮肥利用效率(nitrogen use efficiency, NUE)低的局限性,其高产量对于高水肥投入的依赖性很大.因此,为了提高农作物产量,不得不大量施用氮肥.但是,持续大量的氮肥投入不仅增加了种植成本,还导致了日益严重的环境污染问题.面临粮食安全和生态安全的双重挑战,协同提升农作物NUE和产量已成为可持续农业发展的唯一出路.结合我国粮食安全和农业可持续发展的迫切需求,本研究团队在植物生长发育与氮素代谢协同作用机制的研究以及氮肥高效利用的新种质培育方面取得了突破性进展.这项研究为'少投入、多产出、保护环境'的农作物设计育种提供了理论依据和技术支撑.本文简单介绍近年来NUE相关研究进展以及本研究团队在GRF4-DELLA分子模块协同调控农作物NUE和产量方面的新发现,并对该领域的未来研究方向提出几点展望.展开更多
Modern semi-dwarf rice varieties of the“Green Revolution”require a high supply of nitrogen(N)fertilizer to produce high yields.A better understanding of the interplay between N metabolism and plant developmental pro...Modern semi-dwarf rice varieties of the“Green Revolution”require a high supply of nitrogen(N)fertilizer to produce high yields.A better understanding of the interplay between N metabolism and plant developmental processes is required for improved N-use efficiency and agricultural sustainability.Here,we show that strigolactones(SLs)modulate root metabolic and developmental adaptations to low N availability for ensuring efficient uptake and translocation of available N.The key repressor DWARF 53(D53)of the SL signaling pathway interacts with the transcription factor GROWTH-REGULATING FACTOR 4(GRF4)and prevents GRF4 from binding to its target gene promoters.N limitation induces the accumulation of SLs,which in turn promotes SL-mediated degradation of D53,leading to the release of GRF4 and thus promoting the expression of genes associated with N metabolism.N limitation also induces degradation of the DELLA protein SLENDER RICE 1(SLR1)in an D14-and D53-dependent manner,effectively releasing GRF4 from competitive inhibition caused by SLR1.Collectively,our findings reveal a previously unrecognized mechanism underlying SL and gibberellin crosstalk in response to N availability,advancing our understanding of plant growth–metabolic coordination and facilitating the design of the strategies for improving N-use efficiency in high-yield crops.展开更多
基金supported by the National Key Research and Development Program of China(2016YFD0102000)“Breeding of Major New Varieties of Main Grain Crops”Program(2020ABA016)from Department of Science and Technology of Hubei Province.
文摘Grain size is one of the most important factors affecting rice grain quality and yield,and attracts great attention from molecular biologists and breeders.In this study,we engineered a CRISPR/Cas9 system targeting the miR396 recognition site of the rice GS2 gene,which encodes growth-regulating factor 4(OsGRF4)and regulates multiple agronomic traits including grain size,grain quality,nitrogen use efficiency,abiotic stress response,and seed shattering.In contrast to most previous genome editing efforts in which indel mutations were chosen to obtain null mutants,a mutant named GS2^(E) carrying an in-frame 6-bp deletion and 1-bp substitution within the miR396-targeted sequence was identified.GS2^(E) plants showed increased expression of GS2 in consistent with impaired repression by miR396.As expected,the gain-of-function GS2^(E) mutant exhibited multiple beneficial traits including increased grain size and yield and bigger grain length/width ratio.Thousand grain weight and grain yield per plant of GS2^(E) plants were increased by 23.5%and 10.4%,respectively.These improved traits were passed to hybrids in a semidominant way,suggesting that the new GS2^(E) allele has great potential in rice improvement.Taken together,we report new GS2 germplasm and describe a novel gene-editing strategy that can be widely employed to improve grain size and yield in rice.This trait-improvement strategy could be applied to other genes containing miRNA target sites,in particular the conserved miR396-GRF/GIF module that governs plant growth,development and environmental response.
文摘二十世纪五六十年代,半矮秆水稻和小麦新品种的育成和大面积推广,有效地解决了"高产与倒伏"之间的制约矛盾,这一历程即为众所周知的"绿色革命"。水稻半矮秆基因sd1(semi-dwarf1)既增加了水稻收获指数,又有效地解决了因密植和大量施肥导致的植株倒伏和减产问题,实现了水稻单产的大幅度提升。然而,半矮秆水稻和小麦品种也表现出其植物生长对氮肥响应的减弱,根系对氮肥的吸收速率下降,导致氮肥利用效率(nitrogen use efficiency,NUE)下降。因此,协同改良半矮化作物的高产与氮肥高效利用性状,对保障国家粮食安全和农业可持续发展具有重要意义。傅向东课题组的研究显示,水稻生长调节因子GRF4和生长抑制因子DELLA相互之间的反向平衡调节赋予了植物生长与碳-氮代谢之间的稳态共调节。DELLA蛋白高水平积累导致了作物半矮化,实现了植株耐高肥和抗倒伏的高产目标,但其伴随着氮肥利用效率的降低。相反,GRF4蛋白高水平积累能协同提高作物光合作用和氮肥利用效率,但并不改变"绿色革命"品种的半矮化优良性状,从而实现了在现有高产品种中协同提升产量和氮肥利用效率。DELLA-GRF4分子调控模块的解析为"少投入、多产出、保护环境"的绿色高产高效农作物新品种培育提供了理论基础和具有育种利用价值的新基因资源。
基金the National Key Research and Development Program of China(2017YFD0100103)the Seed Industry Innovation and Industrialization Project of Fujian Province(fjzycxny2017004,zycxny2021004)+1 种基金the Program on Technology of Fujian Province(2020NZ08016,2020N0049)the Open Program of State Key Laboratory of Rice Biology of China(170101)。
文摘The LGS1(Large grain size 1)gene,also known as GS2/GL2/Os GRF4,is involved in regulating grain size and quality in rice,but the mechanism governing grain size has not been elucidated.We performed transcriptomic,proteomic,and phosphoproteomic analyses of young rice panicles in Samba(a wild-type cultivar with extra-small grain)and NIL-LGS1(a nearly isogenic line of LGS1 with large grain in the Samba genetic background)at three developmental stages(4–6)to identify internal dynamic functional networks determining grain size that are mediated by LGS1.Differentially expressed proteins formed seven highly functionally correlated clusters.The concordant regulation of multiple functional clusters may be key features of the development of grain length in rice.In stage 5,16 and 24 phosphorylated proteins were significantly up-regulated and down-regulated,and dynamic phosphorylation events may play accessory roles in determining rice grain size by participating in protein–protein interaction networks.Transcriptomic analysis in stage 5 showed that differentially expressed alternative splicing events and dynamic gene regulatory networks based on 39 transcription factors and their highly correlated target genes might contribute to rice grain development.Integrative multilevel omics analysis suggested that the regulatory network at the transcriptional and posttranscriptional levels could be directly manifested at the translational level,and this analysis also suggested a regulatory mechanism,regulation of protein translation levels,in the biological process that extends from transcript to protein to the development of grain.Functional analysis suggested that biological processes including MAPK signaling,calcium signaling,cell proliferation,cell wall,energy metabolism,hormone pathway,and ubiquitin-proteasome pathway might be involved in LGS1-mediated regulation of grain length.Thus,LGS1-mediated regulation of grain size is affected by dynamic transcriptional,posttranscriptional,translational and posttranslational changes.
文摘在农业生产中,大量施用氮肥是农作物增产的重要措施之一.在长达半个多世纪的农作物育种史上一直占据主导地位的'绿色革命'半矮化品种具有耐高肥、抗倒伏和高产的优良特性,但同时也存在氮肥利用效率(nitrogen use efficiency, NUE)低的局限性,其高产量对于高水肥投入的依赖性很大.因此,为了提高农作物产量,不得不大量施用氮肥.但是,持续大量的氮肥投入不仅增加了种植成本,还导致了日益严重的环境污染问题.面临粮食安全和生态安全的双重挑战,协同提升农作物NUE和产量已成为可持续农业发展的唯一出路.结合我国粮食安全和农业可持续发展的迫切需求,本研究团队在植物生长发育与氮素代谢协同作用机制的研究以及氮肥高效利用的新种质培育方面取得了突破性进展.这项研究为'少投入、多产出、保护环境'的农作物设计育种提供了理论依据和技术支撑.本文简单介绍近年来NUE相关研究进展以及本研究团队在GRF4-DELLA分子模块协同调控农作物NUE和产量方面的新发现,并对该领域的未来研究方向提出几点展望.
基金supported by the National Natural Science Foundation of China(grant nos.31830082,31972501,31672225,and 31601821)the National Key R&D Programme of China(2022YFD1200010-02 and 2022YFD1900702).
文摘Modern semi-dwarf rice varieties of the“Green Revolution”require a high supply of nitrogen(N)fertilizer to produce high yields.A better understanding of the interplay between N metabolism and plant developmental processes is required for improved N-use efficiency and agricultural sustainability.Here,we show that strigolactones(SLs)modulate root metabolic and developmental adaptations to low N availability for ensuring efficient uptake and translocation of available N.The key repressor DWARF 53(D53)of the SL signaling pathway interacts with the transcription factor GROWTH-REGULATING FACTOR 4(GRF4)and prevents GRF4 from binding to its target gene promoters.N limitation induces the accumulation of SLs,which in turn promotes SL-mediated degradation of D53,leading to the release of GRF4 and thus promoting the expression of genes associated with N metabolism.N limitation also induces degradation of the DELLA protein SLENDER RICE 1(SLR1)in an D14-and D53-dependent manner,effectively releasing GRF4 from competitive inhibition caused by SLR1.Collectively,our findings reveal a previously unrecognized mechanism underlying SL and gibberellin crosstalk in response to N availability,advancing our understanding of plant growth–metabolic coordination and facilitating the design of the strategies for improving N-use efficiency in high-yield crops.
