The Brassica polima cytoplasmic male sterility(pol CMS)line causes complete and stable sterility and is most extensively used in breeding.The regulatory pathway,however,is not clear.In this work,we studied molecular i...The Brassica polima cytoplasmic male sterility(pol CMS)line causes complete and stable sterility and is most extensively used in breeding.The regulatory pathway,however,is not clear.In this work,we studied molecular interaction among several key genes involved in pol CMS.Firstly,we found that the multicellular organelle RNA-editing factor protein(Bna.MORF1)interacted with the pol CMS-restorer protein RFP using the yeast two-hybrid system.Secondly,knock down of Bna.MORF1 using CRISPR/Cas9 editing resulted in sterile transgenic lines.The function of the pol CMS sterility gene orf224 was further confirmed by ectopic expression of the gene in both Arabidopsis and Brassica.Furthermore,using CRISPR/Cas9 we determined that an anther-specific proline-rich protein(APG)was also involved in sterility.We propose a working model for pol CMS in Brassica napus that may expedite the utilization of this popular CMS line in Brassica breeding.展开更多
Chloroplasts are essential for plant growth and development,as they play a key role in photosynthesis.The chloroplast biogenesis process is complex and its regulatory mechanism remains elusive.We characterized a spont...Chloroplasts are essential for plant growth and development,as they play a key role in photosynthesis.The chloroplast biogenesis process is complex and its regulatory mechanism remains elusive.We characterized a spontaneous Brassica napus(rapeseed)mutant,ytg,that showed a delayed greening phenotype in all green organs and retarded growth.We identified Bna A02.YTG1 encoding a chloroplastlocalized tetratricopeptide repeat protein widely expressed in rapeseed tissues.We speculated that the ytg phenotype was caused by the deletion of Bna A02.YTG1 based on sequence comparison of 4608(with normal green leaves,isolated from the elite Chinese rapeseed cultivar ZS11)and ytg combined with transcriptome data and CRISPR/Cas9 gene editing results.The homologous gene(Bna C02.YTG1)restored the phenotype of the mutant.Bna A02.YTG1 interacted with MORF2,MORF8,and OZ1.RNA editing of the ndh D-2,ndh F-290,pet L-5,and ndh G-50 plastid transcripts was affected in ytg.These findings suggested that Bna A02.YTG1 participates in RNA editing events.We predicted 29 RNA editing sites in the chloroplast of Brassica napus by comparison with the Arabidopsis chloroplast genome.We conclude that Bna A02.YTG1 affects the posttranscriptional regulation of plastid gene expression and suggest that a tetratricopeptide repeat protein is involved in the chloroplast RNA editing in rapeseed.展开更多
In the chloroplasts/plastids and mitochondria of flowering plants,RNA editing alters hundreds of cytidines to uridines at specific sites mediated by the editosome.Over the past decade,Multiple Organellar RNA Editing F...In the chloroplasts/plastids and mitochondria of flowering plants,RNA editing alters hundreds of cytidines to uridines at specific sites mediated by the editosome.Over the past decade,Multiple Organellar RNA Editing Factor(MORF)proteins have emerged as essential regulators that affect the editing efficiency of most editing sites in plastids and mitochondria.In Arabidopsis,the MORF family consists of nine members,each possessing a single conserved MORF-box that is distributed among flowering plants.Accumulating studies have demonstrated that MORF proteins interact with many other factors,including the PPR proteins and enzymes in different biosynthetic pathways,indicating that the MORF proteins play a more extensive role in regulating organellar development than RNA editing.Recent studies reveal that MORF2 and MORF9 possess holdase activity and may act as chaperones and that MORF8 undergoes heatdependent phase separation to inhibit RNA editing in chloroplasts.In this review,we provide an overview of our current knowledge of the MORF family proteins and discuss the biological and molecular functions of this family in plants.展开更多
Fast and accurate prediction of urban flood is of considerable practical importance to mitigate the effects of frequent flood disasters in advance.To improve urban flood prediction efficiency and accuracy,we proposed ...Fast and accurate prediction of urban flood is of considerable practical importance to mitigate the effects of frequent flood disasters in advance.To improve urban flood prediction efficiency and accuracy,we proposed a framework for fast mapping of urban flood:a coupled model based on physical mechanisms was first constructed,a rainfall-inundation database was generated,and a hybrid flood mapping model was finally proposed using the multi-objective random forest(MORF)method.The results show that the coupled model had good reliability in modelling urban flood,and 48 rainfall-inundation scenarios were then specified.The proposed hybrid MORF model in the framework also demonstrated good performance in predicting inundated depth under the observed and scenario rainfall events.The spatial inundated depths predicted by the MORF model were close to those of the coupled model,with differences typically less than 0.1 m and an average correlation coefficient reaching 0.951.The MORF model,however,achieved a computational speed of 200 times faster than the coupled model.The overall prediction performance of the MORF model was also better than that of the k-nearest neighbor model.Our research provides a novel approach to rapid urban flood mapping and flood early warning.展开更多
Plastid biogenesis and the coordination of plastid and nuclear genome expression through anterograde and retrograde signaling are essential for plant development.GENOMES UNCOUPLED1(GUN1)plays a central role in retrogr...Plastid biogenesis and the coordination of plastid and nuclear genome expression through anterograde and retrograde signaling are essential for plant development.GENOMES UNCOUPLED1(GUN1)plays a central role in retrograde signaling during early plant development.The putative function of GUN1 has been extensively studied,but its molecular function remains controversial.Here,we evaluate published transcriptome data and generate our own data from gun1 mutants grown under signaling-relevant condi-tions to show that editing and splicing are not relevant for GUN1-dependent retrograde signaling.Our study of the plastid(post)transcriptome of gun1 seedlings with white and pale cotyledons demonstrates that GUN1 deficiency significantly alters the entire plastid transcriptome.By combining this result with a penta-tricopeptide repeat code-based prediction and experimental validation by RNA immunoprecipitation ex-periments,we identified several putative targets of GUN1,including tRNAs and RNAs derived from ycf1.2,rpoC1,and rpoC2 and the ndhH–ndhA–ndhI–ndhG–ndhE–psaC–ndhD gene cluster.The absence of plastid rRNAs and the significant reduction of almost all plastid transcripts in white gun1 mutants ac-count for the cotyledon phenotype.Our study provides evidence for RNA binding and maturation as the long-sought molecular function of GUN1 and resolves long-standing controversies.We anticipate that ourfindings will serve as a basis for subsequent studies on mechanisms of plastid gene expression and will help to elucidate the function of GUN1 in retrograde signaling.展开更多
基金supported by the Wuhan Science and Technology Major Project on Key Techniques of Biological Breeding and Breeding of New Varieties(2022021302024851)the National Key Research and Development Program of China(2021YFD1600500)the National Natural Science Foundation of China(32201883)。
文摘The Brassica polima cytoplasmic male sterility(pol CMS)line causes complete and stable sterility and is most extensively used in breeding.The regulatory pathway,however,is not clear.In this work,we studied molecular interaction among several key genes involved in pol CMS.Firstly,we found that the multicellular organelle RNA-editing factor protein(Bna.MORF1)interacted with the pol CMS-restorer protein RFP using the yeast two-hybrid system.Secondly,knock down of Bna.MORF1 using CRISPR/Cas9 editing resulted in sterile transgenic lines.The function of the pol CMS sterility gene orf224 was further confirmed by ectopic expression of the gene in both Arabidopsis and Brassica.Furthermore,using CRISPR/Cas9 we determined that an anther-specific proline-rich protein(APG)was also involved in sterility.We propose a working model for pol CMS in Brassica napus that may expedite the utilization of this popular CMS line in Brassica breeding.
基金the National Key Research and Development Program of China(2016YFD0100305)。
文摘Chloroplasts are essential for plant growth and development,as they play a key role in photosynthesis.The chloroplast biogenesis process is complex and its regulatory mechanism remains elusive.We characterized a spontaneous Brassica napus(rapeseed)mutant,ytg,that showed a delayed greening phenotype in all green organs and retarded growth.We identified Bna A02.YTG1 encoding a chloroplastlocalized tetratricopeptide repeat protein widely expressed in rapeseed tissues.We speculated that the ytg phenotype was caused by the deletion of Bna A02.YTG1 based on sequence comparison of 4608(with normal green leaves,isolated from the elite Chinese rapeseed cultivar ZS11)and ytg combined with transcriptome data and CRISPR/Cas9 gene editing results.The homologous gene(Bna C02.YTG1)restored the phenotype of the mutant.Bna A02.YTG1 interacted with MORF2,MORF8,and OZ1.RNA editing of the ndh D-2,ndh F-290,pet L-5,and ndh G-50 plastid transcripts was affected in ytg.These findings suggested that Bna A02.YTG1 participates in RNA editing events.We predicted 29 RNA editing sites in the chloroplast of Brassica napus by comparison with the Arabidopsis chloroplast genome.We conclude that Bna A02.YTG1 affects the posttranscriptional regulation of plastid gene expression and suggest that a tetratricopeptide repeat protein is involved in the chloroplast RNA editing in rapeseed.
基金supported by grants from the Key Research and Development Program of Zhejiang Province(2024SSYS0100)the National Natural Science Foundation of China(32030009)。
文摘In the chloroplasts/plastids and mitochondria of flowering plants,RNA editing alters hundreds of cytidines to uridines at specific sites mediated by the editosome.Over the past decade,Multiple Organellar RNA Editing Factor(MORF)proteins have emerged as essential regulators that affect the editing efficiency of most editing sites in plastids and mitochondria.In Arabidopsis,the MORF family consists of nine members,each possessing a single conserved MORF-box that is distributed among flowering plants.Accumulating studies have demonstrated that MORF proteins interact with many other factors,including the PPR proteins and enzymes in different biosynthetic pathways,indicating that the MORF proteins play a more extensive role in regulating organellar development than RNA editing.Recent studies reveal that MORF2 and MORF9 possess holdase activity and may act as chaperones and that MORF8 undergoes heatdependent phase separation to inhibit RNA editing in chloroplasts.In this review,we provide an overview of our current knowledge of the MORF family proteins and discuss the biological and molecular functions of this family in plants.
基金financial or data support of the National Key R&D Program of China(2021YFC3001000)the National Natural Science Foundation of China(U1911204,51879107)+1 种基金the Natural Science Foundation of Guangdong Province(2023B1515020087,2022A1515010019)the Fund of Science and Technology Program of Guangzhou(202102020216)。
文摘Fast and accurate prediction of urban flood is of considerable practical importance to mitigate the effects of frequent flood disasters in advance.To improve urban flood prediction efficiency and accuracy,we proposed a framework for fast mapping of urban flood:a coupled model based on physical mechanisms was first constructed,a rainfall-inundation database was generated,and a hybrid flood mapping model was finally proposed using the multi-objective random forest(MORF)method.The results show that the coupled model had good reliability in modelling urban flood,and 48 rainfall-inundation scenarios were then specified.The proposed hybrid MORF model in the framework also demonstrated good performance in predicting inundated depth under the observed and scenario rainfall events.The spatial inundated depths predicted by the MORF model were close to those of the coupled model,with differences typically less than 0.1 m and an average correlation coefficient reaching 0.951.The MORF model,however,achieved a computational speed of 200 times faster than the coupled model.The overall prediction performance of the MORF model was also better than that of the k-nearest neighbor model.Our research provides a novel approach to rapid urban flood mapping and flood early warning.
基金Deutsche Forschungsgemeinschaft to C.S.-L.,D.L.,and T.K.(TRR175,projects A02,C01,and C05)Research in the Hua laboratory was supported by a US NSF CAREER award(MCB-1750361).
文摘Plastid biogenesis and the coordination of plastid and nuclear genome expression through anterograde and retrograde signaling are essential for plant development.GENOMES UNCOUPLED1(GUN1)plays a central role in retrograde signaling during early plant development.The putative function of GUN1 has been extensively studied,but its molecular function remains controversial.Here,we evaluate published transcriptome data and generate our own data from gun1 mutants grown under signaling-relevant condi-tions to show that editing and splicing are not relevant for GUN1-dependent retrograde signaling.Our study of the plastid(post)transcriptome of gun1 seedlings with white and pale cotyledons demonstrates that GUN1 deficiency significantly alters the entire plastid transcriptome.By combining this result with a penta-tricopeptide repeat code-based prediction and experimental validation by RNA immunoprecipitation ex-periments,we identified several putative targets of GUN1,including tRNAs and RNAs derived from ycf1.2,rpoC1,and rpoC2 and the ndhH–ndhA–ndhI–ndhG–ndhE–psaC–ndhD gene cluster.The absence of plastid rRNAs and the significant reduction of almost all plastid transcripts in white gun1 mutants ac-count for the cotyledon phenotype.Our study provides evidence for RNA binding and maturation as the long-sought molecular function of GUN1 and resolves long-standing controversies.We anticipate that ourfindings will serve as a basis for subsequent studies on mechanisms of plastid gene expression and will help to elucidate the function of GUN1 in retrograde signaling.
