Soybean is the primary source of plant protein for humans.Owing to the indigestibility of the raffinose family of oligosaccharides(RFO),raffinose and stachyose are considered anti-nutritive factors in soybean seeds.Lo...Soybean is the primary source of plant protein for humans.Owing to the indigestibility of the raffinose family of oligosaccharides(RFO),raffinose and stachyose are considered anti-nutritive factors in soybean seeds.Low-RFO soybean cultivars are generated by mutagenesis of RFO biosynthesis genes,but the carbohydrate profiles invite further modification to lower RFOs.This study employed a pooled multiplex genome editing approach to target four seed-specifically expressed genes mediating RFO biosynthesis,encoding three raffinose synthases(RS2,RS3,and RS4)and one stachyose synthase.In T1progeny,rs2/rs3 and rs4/sts homozygous double mutants and a rs2/rs3/rs4/sts quadruple mutant(rfo-4m)were characterized.The rs2/rs3 mutant showed reduced raffinose and stachyose contents,but the rs4/sts mutant showed only reduced stachyose in seeds.The RFO contents in the rfo-4m mutant were almost eliminated.Metabolomic analysis showed that the mutation of four RFO biosynthesis genes led to a shift of metabolic profile in the seeds,including the accumulation of several oligosaccharides-related metabolites.These mutants could contribute to precision breeding of soybean cultivars for soy food production.展开更多
Beany flavor induced by three lipoxygenases(LOXs, including LOX1, LOX2, and LOX3)restricts human consumption of soybean. It is desirable to generate lipoxygenase-free new mutant lines to improve the eating quality of ...Beany flavor induced by three lipoxygenases(LOXs, including LOX1, LOX2, and LOX3)restricts human consumption of soybean. It is desirable to generate lipoxygenase-free new mutant lines to improve the eating quality of soybean oil and protein products. In this study, a pooled clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPRassociated protein 9(Cas9) strategy targeting three GmLox genes(GmLox1, GmLox2, and GmLox3) was applied and 60 T_0 positive transgenic plants were generated, carrying combinations of sg RNAs and mutations. Among them, GmLox-28 and GmLox-60 were gmlox1 gmlox2 gmlox3 triple mutants and GmLox-40 was a gmlox1 gmlox2 double mutant.Sequencing of T_1 mutant plants derived from GmLox-28, GmLox-60, and GmLox-40 showed that mutation in the GmLox gene was inherited by the next generation. Colorimetric assay revealed that plants carrying different combinations of mutations lost the corresponding lipoxygenase activities. Transgene-free mutants were obtained by screening the T_2 generation of lipoxygenase-free mutant lines(GmLox-28 and GmLox-60). These transgeneand lipoxygenase-free mutants could be used for soybean beany flavor reduction without restriction by regulatory frameworks governing transgenic organisms.展开更多
Soybean(Glycine max[L.]Merr.)is an important source of human dietary protein and vegetable oil.A strong negative correlation between protein and oil contents has hindered efforts to improve soybean seed quality.The me...Soybean(Glycine max[L.]Merr.)is an important source of human dietary protein and vegetable oil.A strong negative correlation between protein and oil contents has hindered efforts to improve soybean seed quality.The metabolic and genetic bases of soybean seed composition remain elusive.We evaluated metabolic diversity in a soybean near-isogenic line(NIL)population derived from parents(JD12 and CMSD)with contrasting seed oil contents.Using GC-TOF/MS,we compared seed primary metabolites of high protein/low oil lines,low protein/high oil lines,and their parents.Principal-components analysis showed that metabolic profiles of all progeny lines could be discriminated based on protein and oil contents.Univariate analysis revealed wide variation and transgressive segregation of metabolites in the population.Twenty-eight annotated metabolites,in particular free asparagine,free 3-cyanoalanine,and L-malic acid,were correlated with seed protein content or seed oil content or seed protein and oil content.These results shed light on the metabolic and genetic basis of soybean seed composition.展开更多
Soybean is a photoperiod-sensitive short-day crop whose reproductive period and yield are markedly affected by day-length changes.Seed weight is one of the key traits determining the soybean yield;how-ever,the promine...Soybean is a photoperiod-sensitive short-day crop whose reproductive period and yield are markedly affected by day-length changes.Seed weight is one of the key traits determining the soybean yield;how-ever,the prominent genes that control the final seed weight of soybean and the mechanisms underlying the photoperiod's effect on this trait remain poorly understood.In this study,we identify SwW19 as a major locus controlling soybean seed weight by QTL mapping and determine Dt1,an orthologous gene of Arabidopsis TFL1 that is known to govern the soybean growth habit,as the causal gene of the SW19 locus.We showed that Dt1 is highly expressed in developing seeds and regulates photoperiod-dependent seed weight in soybean.Further analyses revealed that the Dt1 protein physically interacts with the sucrose transporter GmSWEET10a to negatively regulate the import of sucrose from seed coat to the embryo,thus modulating seed weight under long days.However,Dt1 does not function in seed development under short days due to its very low expression.Importantly,we discovered a novel natural allelic variant of Dt1(H4 haplotype)that decouples its pleiotropic effects on seed size and growth habit;i.e.,this variant remains functional in seed development but fails to regulate the stem growth habit of soybean.Collectively,our findings provide new insights into how soybean seed development responds to photoperiod at different latitudes,offering an ideal genetic component for improving soybean's yield by manipulating its seed weightandgrowth habit.展开更多
Dear Editor,Despite myriad successful applications of gene editing in plant functional genomics research and precision breeding,many challenges persist around the efficiency of gene-editing tools for many plant specie...Dear Editor,Despite myriad successful applications of gene editing in plant functional genomics research and precision breeding,many challenges persist around the efficiency of gene-editing tools for many plant species.For instance,soybean(Glycine max)is a major crop providing oil and protein to human diets and feedstock,but its gene-editing efficiency remains relatively low(Bai et al.,2019).展开更多
Phytic acid(PA)in grain seeds reduces the bioavailability of nutrient elements in monogastric animals,and an important objective for crop seed biofortification is to decrease the seed PA content.Here,we employed CRISP...Phytic acid(PA)in grain seeds reduces the bioavailability of nutrient elements in monogastric animals,and an important objective for crop seed biofortification is to decrease the seed PA content.Here,we employed CRISPR/Cas9 to generate a PA mutant population targeting PA biosynthesis and transport genes,including two multi-drug-resistant protein 5(MRP5)and three inositol pentose-phosphate kinases(IPK1).We characterized a variety of lines containing mutations on multiple IPK and MRP5 genes.The seed PA was more significantly decreased in higher-order mutant lines with multiplex mutations.However,such mutants also exhibited poor agronomic performance.In the population,we identified two lines carrying single mutations in ipk1b and ipk1c,respectively.These mutants exhibited moderately reduced PA content,and regular agronomic performance compared to the wild type.Our study indicates that moderately decreasing PA by targeting single GmIPK1 genes,rather than multiplex mutagenesis toward ultra-low PA,is an optimal strategy for low-PA soybean with a minimal trade-off in yield performance.展开更多
Pollen tubes elongate rapidly at their tips through highly polarized cell growth known as tip growth. Tip growth requires intensive exocytosis at the tip, which is supported by a dynamic cytoskeleton and vesicle traff...Pollen tubes elongate rapidly at their tips through highly polarized cell growth known as tip growth. Tip growth requires intensive exocytosis at the tip, which is supported by a dynamic cytoskeleton and vesicle trafficking. Several signaling pathways have been demonstrated to coordinate pollen tube growth by regulating cellular activities such as actin dynamics, exocytosis, and endocytosis. These signaling pathways crosstalk to form a signaling network that coordinates the cellular processes required for tip growth. The homeostasis of key signaling molecules is critical for the proper elongation of the pollen tube tip, and is commonly fine-tuned by positive and negative regulations. In addition to the major signaling pathways, emerging evidence implies the roles of other signals in the regulation of pollen tube growth. Here we review and discuss how these signaling networks modulate the rapid growth of pollen tubes.展开更多
Dear Editor,Soybean(Glycine max)is a major source of plant-based protein for people worldwide,providing a healthy,affordable,and environmentally friendly alternative to animal-based protein(Montgomery,2003).It has bee...Dear Editor,Soybean(Glycine max)is a major source of plant-based protein for people worldwide,providing a healthy,affordable,and environmentally friendly alternative to animal-based protein(Montgomery,2003).It has been estimated that there are currently over 12000 food products that contain soy protein(Montgomery,2003).Different food applications of soy protein require distinct properties that affect its function and sensory features.For instance,the sensory quality of soymilk is significantly affected by protein emulsibility,whereas tofu quality is determined by protein-gelling ability(Kinsella,1979).展开更多
Crop breeding during the Green Revolution resulted in high yields largely due to the creation of plants with semi-dwarf architectures that could tolerate high-density planting.Although semi-dwarf varieties have been d...Crop breeding during the Green Revolution resulted in high yields largely due to the creation of plants with semi-dwarf architectures that could tolerate high-density planting.Although semi-dwarf varieties have been developed in rice,wheat and maize,none was reported in soybean(Glycine max),and few genes controlling plant architecture have been characterized in soybean.Here,we demonstrate that the auxin efflux transporter PINFORMED1(GmPIN1),which determines polar auxin transport,regulates the leaf petiole angle in soybean.CRISPR-Cas9-induced Gmpin1abc and Gmpin1bc multiple mutants displayed a compact architecture with a smaller petiole angle than wildtype plants.GmPIN1 transcripts and auxin were distributed asymmetrically in the petiole base,with high levels of GmPIN1a/c transcript and auxin in the lower cells,which resulted in asymmetric cell expansion.By contrast,the(iso)flavonoid content was greater in the upper petiole cells than in the lower cells.Our results suggest that(iso)flavonoids inhibit GmPIN1a/c expression to regulate the petiole angle.Overall,our study demonstrates that a signal cascade that integrates(iso)flavonoid biosynthesis,GmPIN1a/c expression,auxin accumulation,and cell expansion in an asymmetric manner creates a desirable petiole curvature in soybean.This study provides a genetic resource for improving soybean plant architecture.展开更多
Pod shattering can lead to devastating yield loss of soybean and has been a negatively selected trait in soybean domestication and breeding.Nevertheless,a significant portion of soybean cultivars are still pod shatter...Pod shattering can lead to devastating yield loss of soybean and has been a negatively selected trait in soybean domestication and breeding.Nevertheless,a significant portion of soybean cultivars are still pod shattering-susceptible,limiting their regional and climatic adaptabilities.Here we performed genetic diagnosis on the shattering-susceptible trait of a national registered cultivar,Huachun6(HC6),and found that HC6 carries the susceptible genotype of a candidate Pod dehiscence 1(PDH1)gene,which exists in a significant portion of soybean cultivars.We next performed genome editing on PDH1 gene by clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPR-associated protein 9(Cas9).In T2 progenies,several transgene-free lines with pdh1 mutations were characterized without affecting major agronomic traits.The pdh1 mutation significantly improved the pod shattering resistance which is associated with aberrant lignin distribution in inner sclerenchyma.Our work demonstrated that precision breeding by genome editing on PDH1 holds great potential for precisely improving pod shattering resistance and adaptability of soybean cultivars.展开更多
基金supported by the Chuying Scholar Project in Fujian Province and the Innovative Research Project at Fujian Agriculture and Forestry University。
文摘Soybean is the primary source of plant protein for humans.Owing to the indigestibility of the raffinose family of oligosaccharides(RFO),raffinose and stachyose are considered anti-nutritive factors in soybean seeds.Low-RFO soybean cultivars are generated by mutagenesis of RFO biosynthesis genes,but the carbohydrate profiles invite further modification to lower RFOs.This study employed a pooled multiplex genome editing approach to target four seed-specifically expressed genes mediating RFO biosynthesis,encoding three raffinose synthases(RS2,RS3,and RS4)and one stachyose synthase.In T1progeny,rs2/rs3 and rs4/sts homozygous double mutants and a rs2/rs3/rs4/sts quadruple mutant(rfo-4m)were characterized.The rs2/rs3 mutant showed reduced raffinose and stachyose contents,but the rs4/sts mutant showed only reduced stachyose in seeds.The RFO contents in the rfo-4m mutant were almost eliminated.Metabolomic analysis showed that the mutation of four RFO biosynthesis genes led to a shift of metabolic profile in the seeds,including the accumulation of several oligosaccharides-related metabolites.These mutants could contribute to precision breeding of soybean cultivars for soy food production.
基金supported by funds from the National Key Research and Development Program of China(2016YFD0100700)to Y.G。
文摘Beany flavor induced by three lipoxygenases(LOXs, including LOX1, LOX2, and LOX3)restricts human consumption of soybean. It is desirable to generate lipoxygenase-free new mutant lines to improve the eating quality of soybean oil and protein products. In this study, a pooled clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPRassociated protein 9(Cas9) strategy targeting three GmLox genes(GmLox1, GmLox2, and GmLox3) was applied and 60 T_0 positive transgenic plants were generated, carrying combinations of sg RNAs and mutations. Among them, GmLox-28 and GmLox-60 were gmlox1 gmlox2 gmlox3 triple mutants and GmLox-40 was a gmlox1 gmlox2 double mutant.Sequencing of T_1 mutant plants derived from GmLox-28, GmLox-60, and GmLox-40 showed that mutation in the GmLox gene was inherited by the next generation. Colorimetric assay revealed that plants carrying different combinations of mutations lost the corresponding lipoxygenase activities. Transgene-free mutants were obtained by screening the T_2 generation of lipoxygenase-free mutant lines(GmLox-28 and GmLox-60). These transgeneand lipoxygenase-free mutants could be used for soybean beany flavor reduction without restriction by regulatory frameworks governing transgenic organisms.
基金supported by the National Natural Science Foundation of China (31201234, 31871652)the China Agriculture Research System (CARS-004-PS06)
文摘Soybean(Glycine max[L.]Merr.)is an important source of human dietary protein and vegetable oil.A strong negative correlation between protein and oil contents has hindered efforts to improve soybean seed quality.The metabolic and genetic bases of soybean seed composition remain elusive.We evaluated metabolic diversity in a soybean near-isogenic line(NIL)population derived from parents(JD12 and CMSD)with contrasting seed oil contents.Using GC-TOF/MS,we compared seed primary metabolites of high protein/low oil lines,low protein/high oil lines,and their parents.Principal-components analysis showed that metabolic profiles of all progeny lines could be discriminated based on protein and oil contents.Univariate analysis revealed wide variation and transgressive segregation of metabolites in the population.Twenty-eight annotated metabolites,in particular free asparagine,free 3-cyanoalanine,and L-malic acid,were correlated with seed protein content or seed oil content or seed protein and oil content.These results shed light on the metabolic and genetic basis of soybean seed composition.
基金supported by the National Natural Science Foundation of China(grant no.32090064 to F.K.)the National Key R&D Program of China(2021YFF1001203 to X.H.)+3 种基金the“Strategic Priority Research Program"of the Chinese Academy of Sciences(grant no.XDA24010105 to X.H.)the National Natural Science Foundation of China(grant nos.32230078 and 31871643 to X.H.)the Major Program of Guangdong Basic and Applied Research(grant no.2019B030302006 to F.K.)the Guangzhou Municipal Science and Technology Project(grant no.202201010641toY.H.).
文摘Soybean is a photoperiod-sensitive short-day crop whose reproductive period and yield are markedly affected by day-length changes.Seed weight is one of the key traits determining the soybean yield;how-ever,the prominent genes that control the final seed weight of soybean and the mechanisms underlying the photoperiod's effect on this trait remain poorly understood.In this study,we identify SwW19 as a major locus controlling soybean seed weight by QTL mapping and determine Dt1,an orthologous gene of Arabidopsis TFL1 that is known to govern the soybean growth habit,as the causal gene of the SW19 locus.We showed that Dt1 is highly expressed in developing seeds and regulates photoperiod-dependent seed weight in soybean.Further analyses revealed that the Dt1 protein physically interacts with the sucrose transporter GmSWEET10a to negatively regulate the import of sucrose from seed coat to the embryo,thus modulating seed weight under long days.However,Dt1 does not function in seed development under short days due to its very low expression.Importantly,we discovered a novel natural allelic variant of Dt1(H4 haplotype)that decouples its pleiotropic effects on seed size and growth habit;i.e.,this variant remains functional in seed development but fails to regulate the stem growth habit of soybean.Collectively,our findings provide new insights into how soybean seed development responds to photoperiod at different latitudes,offering an ideal genetic component for improving soybean's yield by manipulating its seed weightandgrowth habit.
文摘Dear Editor,Despite myriad successful applications of gene editing in plant functional genomics research and precision breeding,many challenges persist around the efficiency of gene-editing tools for many plant species.For instance,soybean(Glycine max)is a major crop providing oil and protein to human diets and feedstock,but its gene-editing efficiency remains relatively low(Bai et al.,2019).
基金supported by the Guangzhou Science Grant(2024A03J0010)to YG.
文摘Phytic acid(PA)in grain seeds reduces the bioavailability of nutrient elements in monogastric animals,and an important objective for crop seed biofortification is to decrease the seed PA content.Here,we employed CRISPR/Cas9 to generate a PA mutant population targeting PA biosynthesis and transport genes,including two multi-drug-resistant protein 5(MRP5)and three inositol pentose-phosphate kinases(IPK1).We characterized a variety of lines containing mutations on multiple IPK and MRP5 genes.The seed PA was more significantly decreased in higher-order mutant lines with multiplex mutations.However,such mutants also exhibited poor agronomic performance.In the population,we identified two lines carrying single mutations in ipk1b and ipk1c,respectively.These mutants exhibited moderately reduced PA content,and regular agronomic performance compared to the wild type.Our study indicates that moderately decreasing PA by targeting single GmIPK1 genes,rather than multiplex mutagenesis toward ultra-low PA,is an optimal strategy for low-PA soybean with a minimal trade-off in yield performance.
文摘Pollen tubes elongate rapidly at their tips through highly polarized cell growth known as tip growth. Tip growth requires intensive exocytosis at the tip, which is supported by a dynamic cytoskeleton and vesicle trafficking. Several signaling pathways have been demonstrated to coordinate pollen tube growth by regulating cellular activities such as actin dynamics, exocytosis, and endocytosis. These signaling pathways crosstalk to form a signaling network that coordinates the cellular processes required for tip growth. The homeostasis of key signaling molecules is critical for the proper elongation of the pollen tube tip, and is commonly fine-tuned by positive and negative regulations. In addition to the major signaling pathways, emerging evidence implies the roles of other signals in the regulation of pollen tube growth. Here we review and discuss how these signaling networks modulate the rapid growth of pollen tubes.
基金supported by the ChuYing scholar program of Fujian province to Y.G.
文摘Dear Editor,Soybean(Glycine max)is a major source of plant-based protein for people worldwide,providing a healthy,affordable,and environmentally friendly alternative to animal-based protein(Montgomery,2003).It has been estimated that there are currently over 12000 food products that contain soy protein(Montgomery,2003).Different food applications of soy protein require distinct properties that affect its function and sensory features.For instance,the sensory quality of soymilk is significantly affected by protein emulsibility,whereas tofu quality is determined by protein-gelling ability(Kinsella,1979).
基金supported by the National Key Research and Development Program of China(2017YFA0506100)Major Program of Natural Science Foundation in Fujian Province(2021J02011)+4 种基金Training Program for Excellent Young Scholars of Fujian Agriculture and Forestry University(KSYLX011)to Xu ChenNational Natural Science Foundation of China(31900213)Natural Science Foundation of Fujian(2019J01420)Distinguished Young Scholar Program of Fujian Agriculture and Forestry University(xjq201921)and to Zhen GaoNational Natural Science Foundation of China(32001571)to Le Gao。
文摘Crop breeding during the Green Revolution resulted in high yields largely due to the creation of plants with semi-dwarf architectures that could tolerate high-density planting.Although semi-dwarf varieties have been developed in rice,wheat and maize,none was reported in soybean(Glycine max),and few genes controlling plant architecture have been characterized in soybean.Here,we demonstrate that the auxin efflux transporter PINFORMED1(GmPIN1),which determines polar auxin transport,regulates the leaf petiole angle in soybean.CRISPR-Cas9-induced Gmpin1abc and Gmpin1bc multiple mutants displayed a compact architecture with a smaller petiole angle than wildtype plants.GmPIN1 transcripts and auxin were distributed asymmetrically in the petiole base,with high levels of GmPIN1a/c transcript and auxin in the lower cells,which resulted in asymmetric cell expansion.By contrast,the(iso)flavonoid content was greater in the upper petiole cells than in the lower cells.Our results suggest that(iso)flavonoids inhibit GmPIN1a/c expression to regulate the petiole angle.Overall,our study demonstrates that a signal cascade that integrates(iso)flavonoid biosynthesis,GmPIN1a/c expression,auxin accumulation,and cell expansion in an asymmetric manner creates a desirable petiole curvature in soybean.This study provides a genetic resource for improving soybean plant architecture.
基金supported by Innovative Research Groups of the Natural Science Foundation of Hebei province(C2020301020).
文摘Pod shattering can lead to devastating yield loss of soybean and has been a negatively selected trait in soybean domestication and breeding.Nevertheless,a significant portion of soybean cultivars are still pod shattering-susceptible,limiting their regional and climatic adaptabilities.Here we performed genetic diagnosis on the shattering-susceptible trait of a national registered cultivar,Huachun6(HC6),and found that HC6 carries the susceptible genotype of a candidate Pod dehiscence 1(PDH1)gene,which exists in a significant portion of soybean cultivars.We next performed genome editing on PDH1 gene by clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPR-associated protein 9(Cas9).In T2 progenies,several transgene-free lines with pdh1 mutations were characterized without affecting major agronomic traits.The pdh1 mutation significantly improved the pod shattering resistance which is associated with aberrant lignin distribution in inner sclerenchyma.Our work demonstrated that precision breeding by genome editing on PDH1 holds great potential for precisely improving pod shattering resistance and adaptability of soybean cultivars.
