Flower and fruit abscission is a highly programmed physiological process,which is closely related to the yield of horticultural plants.The coordination of many regulatory factors associated with metabolic and signalin...Flower and fruit abscission is a highly programmed physiological process,which is closely related to the yield of horticultural plants.The coordination of many regulatory factors associated with metabolic and signaling pathways plays a key role in the flower and fruit shedding.Hormones,peptides,carbohydrates,polyamines or cell wall modifying proteins regulate flower and fruit shedding.This article reviewed the recent studies of flower and fruit abscission,including the molecular regulation mechanism of abscission zone formation,typical structure and location of abscission zones,and other factors affecting flower and fruit abscission,such as stresses,hormones,peptides,carbohydrates,polyamines and cell wall modifying proteins.Overall,the review summarizes the developmental mechanism and the diversity of abscission zones,and the key factors affecting flower and fruit abscission of horticultural plants,aiming to provide guidance for studying the molecular regulatory mechanism of flower and fruit abscission.展开更多
GRAS proteins are plant-specific transcription factors that play crucial roles in plant development and stress responses.However,their involvement in the ripening of economically important fruits and their transcripti...GRAS proteins are plant-specific transcription factors that play crucial roles in plant development and stress responses.However,their involvement in the ripening of economically important fruits and their transcriptional regulatory mechanisms remain largely unclear.Here,we demonstrated that SlGRAS4,encoding a transcription factor of the GRAS family,was induced by the tomato ripening process and regulated by ethylene.Overexpression of SlGRAS4 accelerated fruit ripening,increased the total carotenoid content and increased PSY1 expression in SlGRAS4-OE fruit compared to wild-type fruit.The expression levels of key ethylene biosynthesis genes(SlACS2,SlACS4,SlACO1,and SlACO3)and crucial ripening regulators(RIN and NOR)were increased in SlGRAS4-OE fruit.The negative regulator of tomato fruit ripening,SlMADS1,was repressed in OE fruit.Exogenous ethylene and 1-MCP treatment revealed that more endogenous ethylene was derived in SlGRAS4-OE fruit.More obvious phenotypes were observed in OE seedlings after ACC treatment.Yeast one-hybrid and dual-luciferase assays confirmed that SlGRAS4 can directly bind SlACO1 and SlACO3 promoters to activate their transcription,and SlGRAS4 can also directly repress SlMADS1 expression.Our study identified that SlGRAS4 acts as a new regulator of fruit ripening by regulating ethylene biosynthesis genes in a direct manner.This provides new knowledge of GRAS transcription factors involved in regulating fruit ripening.展开更多
Chlorophylls and carotenoids are essential and bene fi cial substances for both plant and human health.Identifying the regulatory network of these pigments is necessary for improving fruit quality.In a previous study,...Chlorophylls and carotenoids are essential and bene fi cial substances for both plant and human health.Identifying the regulatory network of these pigments is necessary for improving fruit quality.In a previous study,we identi fi ed an R2R3-MYB transcription factor,SlMYB72,that plays an important role in chlorophyll and carotenoid metabolism in tomato fruit.Here,we demonstrated that the SlMYB72-interacting protein SlZHD17,which belongs to the zinc-fi nger homeodomain transcription factor family,also functions in chlorophyll and carotenoid metabolism.Silencing SIZHD 17 in tomato improved multiple bene fi cial agronomic traits,including dwar fi sm,accelerated fl owering,and earlier fruit harvest.More importantly,downregulating SIZHD17 in fruits resulted in larger chloroplasts and a higher chlorophyll content.Dual-luciferase,yeast one-hybrid and electrophoretic mobility shift assays clari fi ed that SlZHD17 regulates the chlorophyll biosynthesis gene SIPOR-B and chloroplast developmental regulator SITKN2 in a direct manner.Chlorophyll degradation and plastid transformation were also retarded after suppression of SIZHD17 in fruits,which was caused by the inhibition of SISGR1,a crucial factor in chlorophyll degradation.On the other hand,the expression of the carotenoid biosynthesis genes SIPSY1 and SIZISO was also suppressed and directly regulated by SlZHD17,which induced uneven pigmentation and decreased the lycopene content in fruits with SIZHD17 suppression at the ripe stage.Furthermore,the protein-protein interactions between SlZHD17 and other pigment regulators,including SlARF4,SlBEL11,and SlTAGL1,were also presented.This study provides new insight into the complex pigment regulatory network and provides new options for breeding strategies aiming to improve fruit quality.展开更多
CRISPR/Cas9-induced genome editing is a powerful tool for studying gene function in a variety of organisms,including plants.Using multi-sgRNAs to target one or more genes is helpful to improve the efficacy of gene edi...CRISPR/Cas9-induced genome editing is a powerful tool for studying gene function in a variety of organisms,including plants.Using multi-sgRNAs to target one or more genes is helpful to improve the efficacy of gene editing and facilitate multi-gene editing.Here,we describe a CRISPR/Cas9 system which can be conveniently developed as a CRISPR kit.SgRNA expression cassettes can be rapidly generated by one-step PCR using our CRISPR kit.In our kit,there are two binary vectors pHNCas9 and pHNCas9HT.The binary vector pHNCas9 was constructed to allow to assemble up to eight sgRNA expression cassettes by one-step Golden Gate cloning.Another binary vector pHNCas9HT can be used to generate a large number of single target constructs by directly transforming ligation reactions products into A.tumefaciens without several procedures,such as PCR and plasmid extraction.The two binary vectors are designed according to the principles of standard BioBrick assembly to be used as an open-source tool.For example,we used BioBrick as a visual T-DNA tag.We also developed a primer design aid to complement the system.With this primer design aid,researchers can rapidly obtain primers and GC content,and sgRNA sequence of target site.Our CRISPR/Cas9 system can perform single-and multi-site editing and multiple gene editing to produce various types of mutations in tomato.This rapid and user-friendly CRISPR/Cas9 system for tomato can be potentially used for mutagenesis of important crop species for genetic improvement and is suitable for research into the function of genes.展开更多
基金funded by the National Natural Science Foundation of China(Grant Nos.31972470,32002100,31772370)Key research and development project of Sichuan Provincial Science and Technology Department(Grant No.2021YFQ0071)。
文摘Flower and fruit abscission is a highly programmed physiological process,which is closely related to the yield of horticultural plants.The coordination of many regulatory factors associated with metabolic and signaling pathways plays a key role in the flower and fruit shedding.Hormones,peptides,carbohydrates,polyamines or cell wall modifying proteins regulate flower and fruit shedding.This article reviewed the recent studies of flower and fruit abscission,including the molecular regulation mechanism of abscission zone formation,typical structure and location of abscission zones,and other factors affecting flower and fruit abscission,such as stresses,hormones,peptides,carbohydrates,polyamines and cell wall modifying proteins.Overall,the review summarizes the developmental mechanism and the diversity of abscission zones,and the key factors affecting flower and fruit abscission of horticultural plants,aiming to provide guidance for studying the molecular regulatory mechanism of flower and fruit abscission.
基金the Natural Science Foundation of Chongqing,China(cstc2019jcyj-bshX0008 to Y.L.)funding provided by the National Key Research and Development Program(2016YFD0400101)the National Natural Science Foundation of China(No.31772370,31972470)to Z.L.
文摘GRAS proteins are plant-specific transcription factors that play crucial roles in plant development and stress responses.However,their involvement in the ripening of economically important fruits and their transcriptional regulatory mechanisms remain largely unclear.Here,we demonstrated that SlGRAS4,encoding a transcription factor of the GRAS family,was induced by the tomato ripening process and regulated by ethylene.Overexpression of SlGRAS4 accelerated fruit ripening,increased the total carotenoid content and increased PSY1 expression in SlGRAS4-OE fruit compared to wild-type fruit.The expression levels of key ethylene biosynthesis genes(SlACS2,SlACS4,SlACO1,and SlACO3)and crucial ripening regulators(RIN and NOR)were increased in SlGRAS4-OE fruit.The negative regulator of tomato fruit ripening,SlMADS1,was repressed in OE fruit.Exogenous ethylene and 1-MCP treatment revealed that more endogenous ethylene was derived in SlGRAS4-OE fruit.More obvious phenotypes were observed in OE seedlings after ACC treatment.Yeast one-hybrid and dual-luciferase assays confirmed that SlGRAS4 can directly bind SlACO1 and SlACO3 promoters to activate their transcription,and SlGRAS4 can also directly repress SlMADS1 expression.Our study identified that SlGRAS4 acts as a new regulator of fruit ripening by regulating ethylene biosynthesis genes in a direct manner.This provides new knowledge of GRAS transcription factors involved in regulating fruit ripening.
基金This work was sponsored by the National Natural Science Foundation of China(No.32002100,31772370,31972470).
文摘Chlorophylls and carotenoids are essential and bene fi cial substances for both plant and human health.Identifying the regulatory network of these pigments is necessary for improving fruit quality.In a previous study,we identi fi ed an R2R3-MYB transcription factor,SlMYB72,that plays an important role in chlorophyll and carotenoid metabolism in tomato fruit.Here,we demonstrated that the SlMYB72-interacting protein SlZHD17,which belongs to the zinc-fi nger homeodomain transcription factor family,also functions in chlorophyll and carotenoid metabolism.Silencing SIZHD 17 in tomato improved multiple bene fi cial agronomic traits,including dwar fi sm,accelerated fl owering,and earlier fruit harvest.More importantly,downregulating SIZHD17 in fruits resulted in larger chloroplasts and a higher chlorophyll content.Dual-luciferase,yeast one-hybrid and electrophoretic mobility shift assays clari fi ed that SlZHD17 regulates the chlorophyll biosynthesis gene SIPOR-B and chloroplast developmental regulator SITKN2 in a direct manner.Chlorophyll degradation and plastid transformation were also retarded after suppression of SIZHD17 in fruits,which was caused by the inhibition of SISGR1,a crucial factor in chlorophyll degradation.On the other hand,the expression of the carotenoid biosynthesis genes SIPSY1 and SIZISO was also suppressed and directly regulated by SlZHD17,which induced uneven pigmentation and decreased the lycopene content in fruits with SIZHD17 suppression at the ripe stage.Furthermore,the protein-protein interactions between SlZHD17 and other pigment regulators,including SlARF4,SlBEL11,and SlTAGL1,were also presented.This study provides new insight into the complex pigment regulatory network and provides new options for breeding strategies aiming to improve fruit quality.
基金supported by the National Key Research and Development Program of China(2016YFD0400101),the National Basic Research Program of China(2013CB127101)the National Natural Science Foundation of China(31572175,31401924)the Project of Chongqing Science and Technology Commission(cstckjcxljrc15).
文摘CRISPR/Cas9-induced genome editing is a powerful tool for studying gene function in a variety of organisms,including plants.Using multi-sgRNAs to target one or more genes is helpful to improve the efficacy of gene editing and facilitate multi-gene editing.Here,we describe a CRISPR/Cas9 system which can be conveniently developed as a CRISPR kit.SgRNA expression cassettes can be rapidly generated by one-step PCR using our CRISPR kit.In our kit,there are two binary vectors pHNCas9 and pHNCas9HT.The binary vector pHNCas9 was constructed to allow to assemble up to eight sgRNA expression cassettes by one-step Golden Gate cloning.Another binary vector pHNCas9HT can be used to generate a large number of single target constructs by directly transforming ligation reactions products into A.tumefaciens without several procedures,such as PCR and plasmid extraction.The two binary vectors are designed according to the principles of standard BioBrick assembly to be used as an open-source tool.For example,we used BioBrick as a visual T-DNA tag.We also developed a primer design aid to complement the system.With this primer design aid,researchers can rapidly obtain primers and GC content,and sgRNA sequence of target site.Our CRISPR/Cas9 system can perform single-and multi-site editing and multiple gene editing to produce various types of mutations in tomato.This rapid and user-friendly CRISPR/Cas9 system for tomato can be potentially used for mutagenesis of important crop species for genetic improvement and is suitable for research into the function of genes.
