Rice grain size and chalkiness are important traits that influence grain yield and quality,respectively.Mining of genes for grain yield and appearance quality and clarification of their action modes are of great impor...Rice grain size and chalkiness are important traits that influence grain yield and quality,respectively.Mining of genes for grain yield and appearance quality and clarification of their action modes are of great importance in rice breeding.In this study,a rice protein disulfide isomerase-like enzyme PDIL2-3 was characterized.Expression analysis revealed that PDIL2-3 was highly expressed in endosperm and spikelet hulls.The PDIL2-3-cri lines generated by CRISPR/Cas9 technology exhibited a chalky grain phenotype with altered storage substance accumulation and increased grain size and weight,whereas exactly opposite results were obtained for PDIL2-3 overexpression lines.Cytological experiments revealed that PDIL2-3-cri increased rice seed length mainly by increasing the cell number and rice seed width mainly by increasing the cell size in grains,implying that PDIL2-3 regulates the grain size by influencing both cell division and expansion of spikelet hulls.Further flow cytometric analysis validated that PDIL2-3 has a negative effect on cell proliferation,preventing DNA duplication and cell division in spikelet hulls.Moreover,q RT-PCR results showed that the expression levels of genes related to cell cycle and storage substance synthesis were significantly changed in PDIL2-3-cri transgenic lines.Thus,our results indicated that PDIL2-3 plays a pivotal role in influencing grain size and quality of rice by affecting cell division/expansion and storage substance accumulation,providing new insights into the function of PDIL family members in rice and enriching the genetic resources for rice breeding.展开更多
The genus Aquilegia(Ranunculaceae)has been cultivated as ornamental and medicinal plants for centuries.With petal spurs of strikingly diverse size and shape,Aquilegia has also been recognized as an excellent system fo...The genus Aquilegia(Ranunculaceae)has been cultivated as ornamental and medicinal plants for centuries.With petal spurs of strikingly diverse size and shape,Aquilegia has also been recognized as an excellent system for evolutionary studies.Pollinator‐mediated selection for longer spurs is believed to have shaped the evolution of this genus,especially the North American taxa.Recently,however,an opposite evolutionary trend was reported in an Asian lineage,where multiple origins of mini-or even nonspurred morphs have occurred.Interesting as it is,the lack of genomic resources has limited our ability to decipher the molecular and evolutionary mechanisms underlying spur reduction in this special lineage.Using long-read sequencing(PacBio Sequel),in combination with optical maps(BioNano DLS)and Hi–C,we assembled a high-quality reference genome of A.oxysepala var.kansuensis,a sister species to the nonspurred taxon.The final assembly is approximately 293.2 Mb,94.6%(277.4 Mb)of which has been anchored to 7 pseudochromosomes.A total of 25,571 protein-coding genes were predicted,with 97.2%being functionally annotated.When comparing this genome with that of A.coerulea,we detected a large rearrangement between Chr1 and Chr4,which might have caused the Chr4 of A.oxysepala var.kansuensis to partly deviate from the“decaying”path that was taken before the split of Aquilegia and Semiaquilegia.This high-quality reference genome is fundamental to further investigations on the development and evolution of petal spurs and provides a strong foundation for the breeding of new horticultural Aquilegia cultivars.展开更多
MicroRNAs(miRNAs)are important regulatory elements involved in the regulation of various plant developmental and physiological processes by blocking the expression of target genes.MiR156 and miR529 are two combinatori...MicroRNAs(miRNAs)are important regulatory elements involved in the regulation of various plant developmental and physiological processes by blocking the expression of target genes.MiR156 and miR529 are two combinatorial regulators,which cooperatively target the SQUAMOSA PROMOTER BINDING-LIKE(SPL)family genes.However,there has been no report about the functional conservation and divergence of miR156 and miR529 during plant development to date.In this study,the biological function and relationship of miR156,miR529 and their target Os SPL14 in rice were explored.Overexpression of miR156e or miR529a(miR156e-OE and miR529a-OE)increased the grain size and tiller number but decreased the plant height and panicle length,while an opposite phenotype was observed for their target mimicry(miR156-MIMIC and miR529a-MIMIC)transgenic plants.Stem-loop RT-PCR results revealed ubiquitous expression of miR156 in roots,axillary buds and leaves,while miR529 was preferentially expressed in the panicle.Accordingly,Os SPL14 could be preferentially and precisely cleaved by miR529a in young panicle but by miR156 in vegetative tissues.Transgenic plants generated by the target immune strategy exhibited obvious growth defects upon the blocking of miR156 and/or miR529 function in rice,confirming that both miR156 and miR529 play important roles in controlling rice growth and development.Moreover,the miR156/miR529-Os SPL14 module negatively controlled grain size by regulating the genes associated with grain size and cell cycling,and controlled plant height through a more complicated mechanism.Taken together,our results demonstrate that miR156 and miR529 respectively function dominantly in the vegetative stage and reproductive stage to control rice growth and development by regulating the accumulation of Os SPL14.These findings facilitate a better understanding of the functional conservation and divergence of miR156 and miR529 family in the miRNA combinatorial regulatory network of plants.展开更多
The identification of seed development-related regulators is critical for the genetic improvement of yield and grain quality in cereal crops.SQUAMOSA PROMOTER BINDING PROTEIN-LIKE14(OsSPL14)is a well-studied,plant-spe...The identification of seed development-related regulators is critical for the genetic improvement of yield and grain quality in cereal crops.SQUAMOSA PROMOTER BINDING PROTEIN-LIKE14(OsSPL14)is a well-studied,plant-specific transcription factor;however,its roles in controlling rice grain appearance quality and the underlying molecular mechanisms have not been fully elucidated.In this study,we demon-strate that OsSPL14 positively regulates appearance quality by controlling grain chalkiness in rice.Genetic analysis revealed that knockdown or knockout of OsSPL14 leads to a chalky grain phenotype,which is associated with significant defects in compound starch granules and notable changes in both starch and protein contents in the endosperm.Transcript analysis identified multiple genes regulated by OsSPL14,including the key granule-bound starch synthase gene Waxy(Wx)and the protein disulfide isomerase-like enzyme-encoding gene PDIL1-1.Both in vitro and in vivo assays demonstrated that OsSPL14 directly binds to the GTAC-box motif in the Wx and PDIL1-1 promoters to enhance their expression.Protein-protein inter-action experiments further revealed that OsSPL14 interacts with the nuclear transcription factor Y(NF-Y)heterodimer OsNF-YB9/YC8-12 to promote the transcription of Wx and PDIL1-1,thereby enhancing rice grain appearance quality.Our findings uncover a novel regulatory pathway controlled by OsSPL14 and pro-vide new insights into the molecular mechanisms underlying rice grain appearance quality,with promising implications for genetic improvement in rice.展开更多
To promote and develop the theoretical basis and application of the wind-vehicle-bridge coupling vibration system,the corresponding research status and prospects are reviewed and discussed from five aspects,i.e.,the a...To promote and develop the theoretical basis and application of the wind-vehicle-bridge coupling vibration system,the corresponding research status and prospects are reviewed and discussed from five aspects,i.e.,the analytical framework,the aerodynamic interference,the evaluation criteria,the design loads of long-span bridge and the double-deck railcum-road bridge.The refining process of analysis system is reviewed from the aspects of simulation wind load,vehicle load and bridge structure,and the corresponding coupling relationship.For aerodynamic interference,the development process is summarized from the simulative precision of the elements(wind,vehicle and bridge),the load cases and the object of interference.For evaluation criteria,the corresponding development course is summarized from the certain evaluation method to uncertain one.For long-span bridge design load,the wind and vehicle loads are reviewed and summarized from current multinational codes and theoretical research.For double-deck rail-cum-road bridge,the mechanism of multi-element coupling relationship and corresponding aerodynamic interference are both reviewed.By comprehensive review and summary,the analytical framework is in the process from simplification to refinement.The simulation and consideration of the objects of structural interference gradually become complex.The corresponding simulation theory,wind tunnel scale,test equipment and technology are the key factors to limit its development.For systematic evaluation of vehicle and bridge,the structural and systemic security are the basis of the evaluation,and the auxiliary components and functional evaluation need to be paid more attention.The evaluation criterion will be developed from certain method to reliability assessment.For design load of long-span bridge,the vehicle load is gradually transferred from the simple application of the design load of small-medium span bridge into a complex model considering the load characteristics.For double-deck rail-cum-road bridge,the basic theory and experimental study on coupling mechanism and aerodynamic interference need to be developed.展开更多
基金supported in part by grants from the National Natural Science Foundation of China(32472141,32101746,and 32102231)Natural Science Foundation of Hubei Province(JCZRLH202500540,2021CFB033,and 2022CFB393)。
文摘Rice grain size and chalkiness are important traits that influence grain yield and quality,respectively.Mining of genes for grain yield and appearance quality and clarification of their action modes are of great importance in rice breeding.In this study,a rice protein disulfide isomerase-like enzyme PDIL2-3 was characterized.Expression analysis revealed that PDIL2-3 was highly expressed in endosperm and spikelet hulls.The PDIL2-3-cri lines generated by CRISPR/Cas9 technology exhibited a chalky grain phenotype with altered storage substance accumulation and increased grain size and weight,whereas exactly opposite results were obtained for PDIL2-3 overexpression lines.Cytological experiments revealed that PDIL2-3-cri increased rice seed length mainly by increasing the cell number and rice seed width mainly by increasing the cell size in grains,implying that PDIL2-3 regulates the grain size by influencing both cell division and expansion of spikelet hulls.Further flow cytometric analysis validated that PDIL2-3 has a negative effect on cell proliferation,preventing DNA duplication and cell division in spikelet hulls.Moreover,q RT-PCR results showed that the expression levels of genes related to cell cycle and storage substance synthesis were significantly changed in PDIL2-3-cri transgenic lines.Thus,our results indicated that PDIL2-3 plays a pivotal role in influencing grain size and quality of rice by affecting cell division/expansion and storage substance accumulation,providing new insights into the function of PDIL family members in rice and enriching the genetic resources for rice breeding.
基金supported by grants from the National Natural Science Foundation of China(Nos.31930008,31870207,and 31570227)the CAS/SAFEA International Partnership Program for Creative Research Teams,and the National Ten Thousand Talents Program of China.
文摘The genus Aquilegia(Ranunculaceae)has been cultivated as ornamental and medicinal plants for centuries.With petal spurs of strikingly diverse size and shape,Aquilegia has also been recognized as an excellent system for evolutionary studies.Pollinator‐mediated selection for longer spurs is believed to have shaped the evolution of this genus,especially the North American taxa.Recently,however,an opposite evolutionary trend was reported in an Asian lineage,where multiple origins of mini-or even nonspurred morphs have occurred.Interesting as it is,the lack of genomic resources has limited our ability to decipher the molecular and evolutionary mechanisms underlying spur reduction in this special lineage.Using long-read sequencing(PacBio Sequel),in combination with optical maps(BioNano DLS)and Hi–C,we assembled a high-quality reference genome of A.oxysepala var.kansuensis,a sister species to the nonspurred taxon.The final assembly is approximately 293.2 Mb,94.6%(277.4 Mb)of which has been anchored to 7 pseudochromosomes.A total of 25,571 protein-coding genes were predicted,with 97.2%being functionally annotated.When comparing this genome with that of A.coerulea,we detected a large rearrangement between Chr1 and Chr4,which might have caused the Chr4 of A.oxysepala var.kansuensis to partly deviate from the“decaying”path that was taken before the split of Aquilegia and Semiaquilegia.This high-quality reference genome is fundamental to further investigations on the development and evolution of petal spurs and provides a strong foundation for the breeding of new horticultural Aquilegia cultivars.
基金supported by the National Natural Science Foundation of China(32101746,31872811)Hubei Provincial Natural Science Foundation of China(2021CFB033,2022CFB393)。
文摘MicroRNAs(miRNAs)are important regulatory elements involved in the regulation of various plant developmental and physiological processes by blocking the expression of target genes.MiR156 and miR529 are two combinatorial regulators,which cooperatively target the SQUAMOSA PROMOTER BINDING-LIKE(SPL)family genes.However,there has been no report about the functional conservation and divergence of miR156 and miR529 during plant development to date.In this study,the biological function and relationship of miR156,miR529 and their target Os SPL14 in rice were explored.Overexpression of miR156e or miR529a(miR156e-OE and miR529a-OE)increased the grain size and tiller number but decreased the plant height and panicle length,while an opposite phenotype was observed for their target mimicry(miR156-MIMIC and miR529a-MIMIC)transgenic plants.Stem-loop RT-PCR results revealed ubiquitous expression of miR156 in roots,axillary buds and leaves,while miR529 was preferentially expressed in the panicle.Accordingly,Os SPL14 could be preferentially and precisely cleaved by miR529a in young panicle but by miR156 in vegetative tissues.Transgenic plants generated by the target immune strategy exhibited obvious growth defects upon the blocking of miR156 and/or miR529 function in rice,confirming that both miR156 and miR529 play important roles in controlling rice growth and development.Moreover,the miR156/miR529-Os SPL14 module negatively controlled grain size by regulating the genes associated with grain size and cell cycling,and controlled plant height through a more complicated mechanism.Taken together,our results demonstrate that miR156 and miR529 respectively function dominantly in the vegetative stage and reproductive stage to control rice growth and development by regulating the accumulation of Os SPL14.These findings facilitate a better understanding of the functional conservation and divergence of miR156 and miR529 family in the miRNA combinatorial regulatory network of plants.
基金supported by grants from the National Natural Science Foundation of China(32472141 and 32101746)the Natural Science Foundation of Hubei Province(JCZRLH202500540,2021CFB033).
文摘The identification of seed development-related regulators is critical for the genetic improvement of yield and grain quality in cereal crops.SQUAMOSA PROMOTER BINDING PROTEIN-LIKE14(OsSPL14)is a well-studied,plant-specific transcription factor;however,its roles in controlling rice grain appearance quality and the underlying molecular mechanisms have not been fully elucidated.In this study,we demon-strate that OsSPL14 positively regulates appearance quality by controlling grain chalkiness in rice.Genetic analysis revealed that knockdown or knockout of OsSPL14 leads to a chalky grain phenotype,which is associated with significant defects in compound starch granules and notable changes in both starch and protein contents in the endosperm.Transcript analysis identified multiple genes regulated by OsSPL14,including the key granule-bound starch synthase gene Waxy(Wx)and the protein disulfide isomerase-like enzyme-encoding gene PDIL1-1.Both in vitro and in vivo assays demonstrated that OsSPL14 directly binds to the GTAC-box motif in the Wx and PDIL1-1 promoters to enhance their expression.Protein-protein inter-action experiments further revealed that OsSPL14 interacts with the nuclear transcription factor Y(NF-Y)heterodimer OsNF-YB9/YC8-12 to promote the transcription of Wx and PDIL1-1,thereby enhancing rice grain appearance quality.Our findings uncover a novel regulatory pathway controlled by OsSPL14 and pro-vide new insights into the molecular mechanisms underlying rice grain appearance quality,with promising implications for genetic improvement in rice.
基金supported by the National Key Research and Development Program of China(2019YFB1600702)General Program of National Natural Science Foundation of China(51878058)+5 种基金National Natural Science Foundation of China(52008027)the General Project Supported by Natural Science Basic Research Plan in Shaanxi Province of China for Young Scientists(2021JQ-269)Fundamental Research Funds for the Central Universities,CHD(300102211304)National Natural Science Foundation of China(51908178)Top Young Talent Program of Higher Learning Institutions of Hebei(BJ2020012)Basic Research Program of Natural Science in Shaanxi Province of China(2019JZ-02)。
文摘To promote and develop the theoretical basis and application of the wind-vehicle-bridge coupling vibration system,the corresponding research status and prospects are reviewed and discussed from five aspects,i.e.,the analytical framework,the aerodynamic interference,the evaluation criteria,the design loads of long-span bridge and the double-deck railcum-road bridge.The refining process of analysis system is reviewed from the aspects of simulation wind load,vehicle load and bridge structure,and the corresponding coupling relationship.For aerodynamic interference,the development process is summarized from the simulative precision of the elements(wind,vehicle and bridge),the load cases and the object of interference.For evaluation criteria,the corresponding development course is summarized from the certain evaluation method to uncertain one.For long-span bridge design load,the wind and vehicle loads are reviewed and summarized from current multinational codes and theoretical research.For double-deck rail-cum-road bridge,the mechanism of multi-element coupling relationship and corresponding aerodynamic interference are both reviewed.By comprehensive review and summary,the analytical framework is in the process from simplification to refinement.The simulation and consideration of the objects of structural interference gradually become complex.The corresponding simulation theory,wind tunnel scale,test equipment and technology are the key factors to limit its development.For systematic evaluation of vehicle and bridge,the structural and systemic security are the basis of the evaluation,and the auxiliary components and functional evaluation need to be paid more attention.The evaluation criterion will be developed from certain method to reliability assessment.For design load of long-span bridge,the vehicle load is gradually transferred from the simple application of the design load of small-medium span bridge into a complex model considering the load characteristics.For double-deck rail-cum-road bridge,the basic theory and experimental study on coupling mechanism and aerodynamic interference need to be developed.
