Pyrus pyrifolia Nakai‘Whangkeumbae'is a sand pear fruit with excellent nutritional quality and taste.However,the industrial development of pear fruit is significantly limited by its short shelf life.Salicylic aci...Pyrus pyrifolia Nakai‘Whangkeumbae'is a sand pear fruit with excellent nutritional quality and taste.However,the industrial development of pear fruit is significantly limited by its short shelf life.Salicylic acid(SA),a well-known phytohormone,can delay fruit senescence and improve shelf life.However,the mechanism by which SA regulates CONSTANS-LIKE genes(COLs)during fruit senescence and the role of COL genes in mediating fruit senescence in sand pear are poorly understood.In this study,22 COL genes were identified in sand pear,including four COLs(Pp COL8,Pp COL9a,Pp COL9b,and Pp COL14)identified via transcriptome analysis and 18 COLs through genome-wide analysis.These COL genes were divided into three subgroups according to the structural domains of the COL protein.Pp COL8,with two B-box motifs and one CCT domain,belonged to the first subgroup.In contrast,the other three Pp COLs,Pp COL9a,Pp COL9b,and Pp COL14,with similar conserved protein domains and gene structures,were assigned to the third subgroup.The four COLs showed different expression patterns in pear tissues and were preferentially expressed at the early stage of fruit development.Moreover,the expression of Pp COL8 was inhibited by exogenous SA treatment,while SA up-regulated the expression of Pp COL9a and Pp COL9b.Interestingly,Pp COL8 interacts with Pp MADS,a MADS-box protein preferentially expressed in fruit,and SA up-regulated its expression.While the production of ethylene and the content of malondialdehyde(MDA)were increased in Pp COL8-overexpression sand pear fruit,the antioxidant enzyme(POD and SOD)activity and the expression of Pp POD1 and Pp SOD1 in the sand pear fruits were down-regulated,which showed that Pp COL8 promoted sand pear fruit senescence.In contrast,the corresponding changes were the opposite in Pp MADS-overexpression sand pear fruits,suggesting that Pp MADS delayed sand pear fruit senescence.The co-transformation of Pp COL8 and Pp MADS also delayed sand pear fruit senescence.The results of this study revealed that Pp COL8 can play a key role in pear fruit senescence by interacting with Pp MADS through the SA signaling pathway.展开更多
Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system has been widely used for precise gene editing in plants.However,simultaneous gene editing of multiple homoeoalleles r...Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system has been widely used for precise gene editing in plants.However,simultaneous gene editing of multiple homoeoalleles remains challenging,especially in self-incompatible polyploid plants.Here,we simultaneously introduced targeted mutations in all three homoeoalleles of two genes in the self-incompatible allohexaploid tall fescue,using both CRISPR/Cas9 and LbCas12a(LbCpf1)systems.Loss-of-function mutants of FaPDS exhibited albino leaves,while knockout of FaHSP17.9 resulted in impaired heat resistance in T0 generation of tall fescue.Moreover,these mutations were inheritable.Our findings demonstrate the feasibility of generating loss-of-function mutants in T0 generation polyploid perennial grasses using CRISPR/Cas systems.展开更多
Bioengineering of photorespiratory bypasses is an effective strategy for improving plant productivity by modulating photosynthesis.In previouswork,two photorespiratory bypasses,theGOC andGCGT bypasses,increased photos...Bioengineering of photorespiratory bypasses is an effective strategy for improving plant productivity by modulating photosynthesis.In previouswork,two photorespiratory bypasses,theGOC andGCGT bypasses,increased photosynthetic rates but decreased seed-setting rate in rice(Oryza sativa),probably owing to excess photosynthate accumulation in the stem.To solve this bottleneck,we successfully developed a newsynthetic photorespiratory bypass(called theGMAbypass)in rice chloroplasts by introducing Oryza sativa glycolate oxidase 1(OsGLO1),Cucurbita maxima malate synthase(CmMS),and Oryza sativa ascorbate peroxidase7(OsAPX7)into the rice genome using a high-efficiency transgene stacking system.Unlike the GOC and GCGT bypass genes driven by constitutive promoters,OsGLO1 in GMA plants was driven by a light-inducible Rubisco small subunit promoter(pRbcS);its expression dynamically changed in response to light,producing a more moderate increase in photosynthate.Photosynthetic rates were significantly increased inGMA plants,and grain yieldswere significantly improved under greenhouse and field conditions.Transgenic GMA rice showed no reduction in seed-setting rate under either test condition,unlike previous photorespiratory-bypass rice,probably reflecting proper modulation of the photorespiratory bypass.Together,these results imply that appropriate engineering of the GMA bypass can enhance rice growth and grain yield without affecting seed-setting rate.展开更多
基金supported by the National Natural Science Foundation of China(32272654)the Natural Science Foundation of Hebei Province China(C2023204016)+2 种基金the Hebei Province Introduced Overseas-Scholar Fund China(C20220361)the S&T Program of Hebei China(20326330D)the Hebei Province Outstanding Youth Fund China(2016,2019)。
文摘Pyrus pyrifolia Nakai‘Whangkeumbae'is a sand pear fruit with excellent nutritional quality and taste.However,the industrial development of pear fruit is significantly limited by its short shelf life.Salicylic acid(SA),a well-known phytohormone,can delay fruit senescence and improve shelf life.However,the mechanism by which SA regulates CONSTANS-LIKE genes(COLs)during fruit senescence and the role of COL genes in mediating fruit senescence in sand pear are poorly understood.In this study,22 COL genes were identified in sand pear,including four COLs(Pp COL8,Pp COL9a,Pp COL9b,and Pp COL14)identified via transcriptome analysis and 18 COLs through genome-wide analysis.These COL genes were divided into three subgroups according to the structural domains of the COL protein.Pp COL8,with two B-box motifs and one CCT domain,belonged to the first subgroup.In contrast,the other three Pp COLs,Pp COL9a,Pp COL9b,and Pp COL14,with similar conserved protein domains and gene structures,were assigned to the third subgroup.The four COLs showed different expression patterns in pear tissues and were preferentially expressed at the early stage of fruit development.Moreover,the expression of Pp COL8 was inhibited by exogenous SA treatment,while SA up-regulated the expression of Pp COL9a and Pp COL9b.Interestingly,Pp COL8 interacts with Pp MADS,a MADS-box protein preferentially expressed in fruit,and SA up-regulated its expression.While the production of ethylene and the content of malondialdehyde(MDA)were increased in Pp COL8-overexpression sand pear fruit,the antioxidant enzyme(POD and SOD)activity and the expression of Pp POD1 and Pp SOD1 in the sand pear fruits were down-regulated,which showed that Pp COL8 promoted sand pear fruit senescence.In contrast,the corresponding changes were the opposite in Pp MADS-overexpression sand pear fruits,suggesting that Pp MADS delayed sand pear fruit senescence.The co-transformation of Pp COL8 and Pp MADS also delayed sand pear fruit senescence.The results of this study revealed that Pp COL8 can play a key role in pear fruit senescence by interacting with Pp MADS through the SA signaling pathway.
基金This work was supported by the National Natural Science Foundation of China(31772349,31672482,and 31401915)the Major Science and Technology Innovation Project of Shandong Province(2019JZZY010726).
文摘Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system has been widely used for precise gene editing in plants.However,simultaneous gene editing of multiple homoeoalleles remains challenging,especially in self-incompatible polyploid plants.Here,we simultaneously introduced targeted mutations in all three homoeoalleles of two genes in the self-incompatible allohexaploid tall fescue,using both CRISPR/Cas9 and LbCas12a(LbCpf1)systems.Loss-of-function mutants of FaPDS exhibited albino leaves,while knockout of FaHSP17.9 resulted in impaired heat resistance in T0 generation of tall fescue.Moreover,these mutations were inheritable.Our findings demonstrate the feasibility of generating loss-of-function mutants in T0 generation polyploid perennial grasses using CRISPR/Cas systems.
基金supported by the National Natural Science Foundation of China(3110019,32271757)the Natural Science Foundation of Henan Province(21010338)the Gansu Provincial Science and Technology Major Projects(22ZD6NA007).
文摘Bioengineering of photorespiratory bypasses is an effective strategy for improving plant productivity by modulating photosynthesis.In previouswork,two photorespiratory bypasses,theGOC andGCGT bypasses,increased photosynthetic rates but decreased seed-setting rate in rice(Oryza sativa),probably owing to excess photosynthate accumulation in the stem.To solve this bottleneck,we successfully developed a newsynthetic photorespiratory bypass(called theGMAbypass)in rice chloroplasts by introducing Oryza sativa glycolate oxidase 1(OsGLO1),Cucurbita maxima malate synthase(CmMS),and Oryza sativa ascorbate peroxidase7(OsAPX7)into the rice genome using a high-efficiency transgene stacking system.Unlike the GOC and GCGT bypass genes driven by constitutive promoters,OsGLO1 in GMA plants was driven by a light-inducible Rubisco small subunit promoter(pRbcS);its expression dynamically changed in response to light,producing a more moderate increase in photosynthate.Photosynthetic rates were significantly increased inGMA plants,and grain yieldswere significantly improved under greenhouse and field conditions.Transgenic GMA rice showed no reduction in seed-setting rate under either test condition,unlike previous photorespiratory-bypass rice,probably reflecting proper modulation of the photorespiratory bypass.Together,these results imply that appropriate engineering of the GMA bypass can enhance rice growth and grain yield without affecting seed-setting rate.
