The class 2 clustered regularly interspaced short palindromic repeat （CRISPR） systems have been widely used for simultaneous modification of multiple loci in plants. Traditionally, the type II CRISPR-Cas9 or type V ...The class 2 clustered regularly interspaced short palindromic repeat （CRISPR） systems have been widely used for simultaneous modification of multiple loci in plants. Traditionally, the type II CRISPR-Cas9 or type V CRISPR-Cpfl （also known as Cas12a） system is a two-component transcriptional unit （TCTU） in which the Cas9 or Cpf1 protein is expressed from an RNA polymerase （pol） II promoter, whereas the single guide RNA （sgRNA） is typically expressed from a Pol III promoter, such as U6 or U3 promoter.展开更多
Of the more than 370000 species of higher plants in nature,fewer than 0.1%can be geneticallymodified due to limitations of the current gene delivery systems.Even for those that can be genetically modified,the modifica...Of the more than 370000 species of higher plants in nature,fewer than 0.1%can be geneticallymodified due to limitations of the current gene delivery systems.Even for those that can be genetically modified,the modification involves a tedious and costly tissue culture process.Here,we describe an extremely simple cut-dip-budding(CDB)delivery system,which uses Agrobacterium rhizogene to inoculate explants,generating transformed roots that produce transformed buds due to root suckering.We have successfully used CDB to achieve the heritable transformation of plant species inmultiple plant families,including two herbaceous plants(Taraxacum kok-saghyz and Coronilla varia),a tuberous root plant(sweet potato),and three woody plant species(Ailanthus altissima,Aralia elata,and Clerodendrum chinense).These plants have previously been difficult or impossible to transform,but the CDB method enabled efficient transformation or gene editing in them using a very simple explant dipping protocol,under non-sterile conditions and without the need for tissue culture.Our work suggests that large numbers of plants could be amenable to genetic modifications using the CDB method.展开更多
Base editing technologies enable precise base alterations in a target gene without inducing double-stranded breaks,and thus are powerful for targeted gene evolution in vivo(Zhan et al.,2021).The newly evolved cytosine...Base editing technologies enable precise base alterations in a target gene without inducing double-stranded breaks,and thus are powerful for targeted gene evolution in vivo(Zhan et al.,2021).The newly evolved cytosine deaminases such as evoAPOBEC1,evoFERNY and evoCDA1,adenine deaminase TadA8e,and a near-protospacer adjacent motif(PAM)less SpRYCas9 variant,have greatly increased base editing efficiency and expanded the editing window in plants(Zhang et al.,2021;Xu et al.,2021c;Tan et al.,2022).Dual cytosine and adenine editors enable concurrent C-to-T and A-to-G conversions and thus enrich the editing outcome.Although plant dual editors have been developed(Li et al.,2020;Xu et al.,2021a),they are rarely applied in evolving plant genes due to low efficiencies.展开更多
Maintenance of cell wall integrity is of great importance not only for plant growth and development,but also for the adaptation of plants to adverse environments.However,how the cell wall integrity is modulated under ...Maintenance of cell wall integrity is of great importance not only for plant growth and development,but also for the adaptation of plants to adverse environments.However,how the cell wall integrity is modulated under salt stress is still poorly understood.Here,we report that a nuclear-localized Agenet domain-containing protein SWO1(SWOLLEN 1)is required for the maintenance of cell wall integrity in Arabidopsis under salt stress.Mutation in SWO1 gene results in swollen root tips,disordered root cell morphology,and root elongation inhibition under salt stress.The swo1 mutant accumulates less cellulose and pectin but more lignin under high salinity.RNA-seq and ChIP-seq assays reveal that SWO1 binds to the promoter of several cell wall-related genes and regulates their expression under saline conditions.Further study indicates that SWO1 interacts with importinɑIMPA1 and IMPA2,which are required for the import of nuclear-localized proteins.The impa1 impa2 double mutant also exhibits root growth inhibition under salt stress and mutations of these two genes aggravate the salt-hypersensitive phenotype of the swo1 mutant.Taken together,our data suggest that SWO1 functions together with importinɑto regulate the expression of cell wall-related genes,which enables plants to maintain cell wall integrity under high salinity.展开更多
Deaminase-based cytosine base editors(CBEs)and adenine base editors(ABEs)induce C-to-T and A-to-G transitions,respectively,enabling single-nucleotide variants(SNVs)in plants for research and crop enhancement(Li et al....Deaminase-based cytosine base editors(CBEs)and adenine base editors(ABEs)induce C-to-T and A-to-G transitions,respectively,enabling single-nucleotide variants(SNVs)in plants for research and crop enhancement(Li et al.,2023a).The C-to-G base editors(CGBEs)and A-to-Y base editors(AYBEs),developed by incorporating glycosylases with CBEs and ABEs,expand the repertoire of base editing products,allowing C-to-T/G and A-to-T/G transversions in plants(Li et al.,2023b,2023c;Sretenovic et a.,2021).展开更多
Dear Editor, The class 2/type Ⅱ clustered regularly interspaced short palindromic repeat （CRISPR）/Cas9 system has been used successfully for simultaneous modification of multiple loci in plants. Two general strateg...Dear Editor, The class 2/type Ⅱ clustered regularly interspaced short palindromic repeat （CRISPR）/Cas9 system has been used successfully for simultaneous modification of multiple loci in plants. Two general strategies have been applied to coexpress multiple single guide RNAs （sgRNAs） to achieve multiplex gene editing in plant cells.展开更多
Dear Editor, Rice （Oryza sativa） is the staple food for more than half of the world＇s population. Technologies enabling precise and efficient DNA knock-in or replacement, hereinafter referred to as KI, have the pot...Dear Editor, Rice （Oryza sativa） is the staple food for more than half of the world＇s population. Technologies enabling precise and efficient DNA knock-in or replacement, hereinafter referred to as KI, have the potential to revolutionize the generation of crops by precision molecular breeding.展开更多
文摘The class 2 clustered regularly interspaced short palindromic repeat （CRISPR） systems have been widely used for simultaneous modification of multiple loci in plants. Traditionally, the type II CRISPR-Cas9 or type V CRISPR-Cpfl （also known as Cas12a） system is a two-component transcriptional unit （TCTU） in which the Cas9 or Cpf1 protein is expressed from an RNA polymerase （pol） II promoter, whereas the single guide RNA （sgRNA） is typically expressed from a Pol III promoter, such as U6 or U3 promoter.
基金supported by Shandong Shunfeng BiotechnologyCo.Ltd.,Jinan,China.
文摘Of the more than 370000 species of higher plants in nature,fewer than 0.1%can be geneticallymodified due to limitations of the current gene delivery systems.Even for those that can be genetically modified,the modification involves a tedious and costly tissue culture process.Here,we describe an extremely simple cut-dip-budding(CDB)delivery system,which uses Agrobacterium rhizogene to inoculate explants,generating transformed roots that produce transformed buds due to root suckering.We have successfully used CDB to achieve the heritable transformation of plant species inmultiple plant families,including two herbaceous plants(Taraxacum kok-saghyz and Coronilla varia),a tuberous root plant(sweet potato),and three woody plant species(Ailanthus altissima,Aralia elata,and Clerodendrum chinense).These plants have previously been difficult or impossible to transform,but the CDB method enabled efficient transformation or gene editing in them using a very simple explant dipping protocol,under non-sterile conditions and without the need for tissue culture.Our work suggests that large numbers of plants could be amenable to genetic modifications using the CDB method.
基金funded by the National Natural Science Foundation of China(31901046)Youth Innovation Promotion Association,Chinese Academy of Science(2020272)。
文摘Base editing technologies enable precise base alterations in a target gene without inducing double-stranded breaks,and thus are powerful for targeted gene evolution in vivo(Zhan et al.,2021).The newly evolved cytosine deaminases such as evoAPOBEC1,evoFERNY and evoCDA1,adenine deaminase TadA8e,and a near-protospacer adjacent motif(PAM)less SpRYCas9 variant,have greatly increased base editing efficiency and expanded the editing window in plants(Zhang et al.,2021;Xu et al.,2021c;Tan et al.,2022).Dual cytosine and adenine editors enable concurrent C-to-T and A-to-G conversions and thus enrich the editing outcome.Although plant dual editors have been developed(Li et al.,2020;Xu et al.,2021a),they are rarely applied in evolving plant genes due to low efficiencies.
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences,Grant XDB27040101(to J-K.Z.)Shanghai Pujiang Program,Grant 20PJ1414800(to C.Z.),and National Natural Science Foundation of China,Grant No.32070295(to C.Z.).
文摘Maintenance of cell wall integrity is of great importance not only for plant growth and development,but also for the adaptation of plants to adverse environments.However,how the cell wall integrity is modulated under salt stress is still poorly understood.Here,we report that a nuclear-localized Agenet domain-containing protein SWO1(SWOLLEN 1)is required for the maintenance of cell wall integrity in Arabidopsis under salt stress.Mutation in SWO1 gene results in swollen root tips,disordered root cell morphology,and root elongation inhibition under salt stress.The swo1 mutant accumulates less cellulose and pectin but more lignin under high salinity.RNA-seq and ChIP-seq assays reveal that SWO1 binds to the promoter of several cell wall-related genes and regulates their expression under saline conditions.Further study indicates that SWO1 interacts with importinɑIMPA1 and IMPA2,which are required for the import of nuclear-localized proteins.The impa1 impa2 double mutant also exhibits root growth inhibition under salt stress and mutations of these two genes aggravate the salt-hypersensitive phenotype of the swo1 mutant.Taken together,our data suggest that SWO1 functions together with importinɑto regulate the expression of cell wall-related genes,which enables plants to maintain cell wall integrity under high salinity.
基金supported by the National Key R&D Program of China(2021YFA1300404 to J.-K.Z.)the National Natural Science Foundation of China(32188102 to J.-K.Z.)+2 种基金the China Postdoctoral Science Foundation(BX20220098 and 2022M720973 to Y.T.)the Hainan Seed Industry Laboratory(B22C1000P to Y.T.)Nanfan special project,CAAS(ZDXM2314 to M.W.)。
文摘Deaminase-based cytosine base editors(CBEs)and adenine base editors(ABEs)induce C-to-T and A-to-G transitions,respectively,enabling single-nucleotide variants(SNVs)in plants for research and crop enhancement(Li et al.,2023a).The C-to-G base editors(CGBEs)and A-to-Y base editors(AYBEs),developed by incorporating glycosylases with CBEs and ABEs,expand the repertoire of base editing products,allowing C-to-T/G and A-to-T/G transversions in plants(Li et al.,2023b,2023c;Sretenovic et a.,2021).
文摘Dear Editor, The class 2/type Ⅱ clustered regularly interspaced short palindromic repeat （CRISPR）/Cas9 system has been used successfully for simultaneous modification of multiple loci in plants. Two general strategies have been applied to coexpress multiple single guide RNAs （sgRNAs） to achieve multiplex gene editing in plant cells.
文摘Dear Editor, Rice （Oryza sativa） is the staple food for more than half of the world＇s population. Technologies enabling precise and efficient DNA knock-in or replacement, hereinafter referred to as KI, have the potential to revolutionize the generation of crops by precision molecular breeding.
