Chlorophyll (Chl) content,especially Chl b content,and stomatal conductance (G_s) are the key factors affecting the net photosynthetic rate (P_n).Setaria italica,a diploid C_4 panicoid species with a simple genome and...Chlorophyll (Chl) content,especially Chl b content,and stomatal conductance (G_s) are the key factors affecting the net photosynthetic rate (P_n).Setaria italica,a diploid C_4 panicoid species with a simple genome and high transformation efficiency,has been widely accepted as a model in photosynthesis and drought-tolerance research.The current study characterized Chl content,G_s,and P_n of 48 Setaria mutants induced by ethyl methanesulfonate.A total of 24,34,and 35 mutants had significant variations in Chl content,G_s,and P_n,respectively.Correlation analysis showed a positive correlation between increased G_s and increased P_n,and a weak correlation between decreased Chl b content and decreased P_n was also found.Remarkably,two mutants behaved with significantly decreased Chl b content but increased P_n compared to Yugu 1.Seven mutants behaved with significantly decreased G_s but did not decrease P_(n )compared to Yugu 1.The current study thus identified various genetic lines,further exploration of which would be beneficial to elucidate the relationship between Chl content,G_s,and P_n and the mechanism underlying why C_4 species are efficient at photosynthesis and water saving.展开更多
Background:The goal of the current study was to identify the potential therapeutic targets by analyzing the production of oxidants and their effects on the antioxidant system of Setaria digitata,a filarial worm that w...Background:The goal of the current study was to identify the potential therapeutic targets by analyzing the production of oxidants and their effects on the antioxidant system of Setaria digitata,a filarial worm that was collected from the peritoneal cavity of recently butchered calves.Methods:S.digitata was collected and the mitochondria-like particles(MLP),somatic isolates and the post-nuclear fraction were prepared for investigation.The radical intensity and the antioxidant enzyme activity in these fractions were evaluated.The effect of diethylcarbamazine(DEC),Q0,omeprazole,and L-methyl arginine on NOS production in the worm was also evaluated.Results:Experiments showed that coelomic fluid and whole worm showed significant levels of Nitric oxide compared to other fractions.The cytosol of the worm showed higher NOS activity than MLP.Superoxide dismutase and nitric oxide(NO)levels were found to be higher in MLP fraction of the worm.It was also found that DEC did not have any effect on the prevention of NO,while Q0,omeprazole,and L-methyl arginine treatment showed a decrease in NO production significantly.Conclusion:The results showed that by using the existing drugs we could reduce the propagation of S.digitata by inhibiting the production of NO.展开更多
Breeding of male-sterile lines has become the mainstream for the heterosis utilization in foxtail millet,but the genetic basis of most male-sterile lines used for the hybrid is still an area to be elucidated.In this s...Breeding of male-sterile lines has become the mainstream for the heterosis utilization in foxtail millet,but the genetic basis of most male-sterile lines used for the hybrid is still an area to be elucidated.In this study,a highly male-sterile line Gao146A was investigated.Genetic analysis indicated that the highly male-sterile phenotype was controlled by a single recessive gene a single recessive gene.Using F 2 population derived from cross Gao146A/K103,one gene controlling the highly male- sterility,tentatively named as ms1,which linked to SSR marker b234 with genetic distance of 16.7 cM,was mapped on the chromosome VI.These results not only laid the foundation for fine mapping of this highly male-sterile gene,but also helped to accelerate the improvement of highly male-sterile lines by using molecular marker assisted breeding method.展开更多
Foxtail millet（Setaria italica（L.）P.Beauv）is a naturally stress tolerant crop.Compared to other gramineous crops,it has relatively stronger drought and lower nutrition stress tolerance traits.To date,the scope of ...Foxtail millet（Setaria italica（L.）P.Beauv）is a naturally stress tolerant crop.Compared to other gramineous crops,it has relatively stronger drought and lower nutrition stress tolerance traits.To date,the scope of functional genomics research in foxtail millet（S.italic L.）has been quite limited.NAC（NAM,ATAF1/2 and CUC2）-like transcription factors are known to be involved in various biological processes,including abiotic stress responses.In our previous foxtail millet（S.italic L.）RNA seq analysis,we found that the expression of a NAC-like transcription factor,SiNAC110,could be induced by drought stress;additionally,other references have reported that SiNAC110 expression could be induced by abiotic stress.So,we here selected SiNAC110 for further characterization and functional analysis.First,the predicted SiNAC110 protein encoded indicated SiNAC110 has a conserved NAM（no apical meristem）domain between the 11–139 amino acid positions.Phylogenetic analysis then indicated that SiNAC110 belongs to subfamily III of the NAC gene family.Subcellular localization analysis revealed that the SiNAC110-GFP fusion protein was localized to the nucleus in Arabidopsis protoplasts.Gene expression profiling analysis indicated that expression of SiNAC110 was induced by dehydration,high salinity and other abiotic stresses.Gene functional analysis using SiNAC110 overexpressed Arabidopsis plants indicated that,under drought and high salt stress conditions,the seed germination rate,root length,root surface area,fresh weight,and dry weight of the SiNAC110 overexpressed lines were significantly higher than the wild type（WT）,suggesting that the SiNAC110 overexpressed lines had enhanced tolerance to drought and high salt stresses.However,overexpression of SiN AC110 did not affect the sensitivity of SiNAC110 overexpressed lines to abscisic acid（ABA）treatment.Expression analysis of genes involved in proline synthesis,Na＋/K＋transport,drought responses,and aqueous transport proteins were higher in the SiNAC110overexpressed lines than in the WT,whereas expression of ABA-dependent pathway genes did not change.These results indicated that overexpression of SiNAC110 conferred tolerance to drought and high salt stresses,likely through influencing the regulation of proline biosynthesis,ion homeostasis and osmotic balance.Therefore,SiNAC110 appears to function in the ABA-independent abiotic stress response pathway in plants.展开更多
Objective:To clone,express and purify a putative parasitic nematode specific protein of Setaria digitata(S.digitata),filarial nematode that infects livestock and cause significant economic losses in Far East and Asia ...Objective:To clone,express and purify a putative parasitic nematode specific protein of Setaria digitata(S.digitata),filarial nematode that infects livestock and cause significant economic losses in Far East and Asia to he used for structural and functional analyses.Methods:To characterize uneharacterized gene of,S.digitata(SDUG),the herterologous expression of SDUG was carried out in the pET[cloned into pET45b(+)]expression system initially and co-expression of SDUC using chaperoiie plasmids pG-KJE8,pGro 7,pKJE7,pG-Tf2 and pTf16 containing chapcrone proteins of dnaK-dnaJ-grpE-groES-gro-E,groES-groEL,dnaK-dnaJ-grpE,groES-groEL-tig,and tig respectively,was carried out subsequently.Results:Expression of SDUG was seen when Escherichia coli strain BI.21(DE3)is used,while concentrating protein largely into the insoluble fraction.The co-expression of SDUG using chaperoiie plasmid mediated system indicated a significant increase of the protein in the soluble fraction.Of the chaperon plasniid sets,the highest amount of recombinant SDUP in the soluble fraction was seen when pGro7 was used in the presence of2 mg/mL L-arabinosc and 0.6M IPTG concentration in the culture medium and for 3 h of incubation at the temperature of 28℃.Recombinant SDUG was purified both from soluble and insoluble fractions using Ni affinity chromatography.SDS-PAGE and western blot analyses of these proteins revealed a single band having expected size of^24 kDa.Conclusions:SDUG seems to be more aggregate-prone and hydrophobic in nature and such protein can make soluble by correct selecting the inducer concentrations and induction temperature and its duration.展开更多
Cell wall composition in monocotyledonous grasses has been identified as a key area of research for developing better feedstocks for forage and biofuel production.Setaria viridis and its close domesticated relative Se...Cell wall composition in monocotyledonous grasses has been identified as a key area of research for developing better feedstocks for forage and biofuel production.Setaria viridis and its close domesticated relative Setaria italica have been chosen as suitable monocotyledonous models for plants possessing the C4 pathway of photosynthesis including sorghum,maize,sugarcane,switchgrass and Miscanthus×giganteus.Accurate partial least squares regression（PLSR）models to predict S.italica stem composition have been generated,based upon Fourier transform mid-infrared（FTIR）spectra and calibrated with wet chemistry determinations of ground S.italica stem material measured using a modified version of the US National Renewable Energy Laboratory（NREL）acid hydrolysis protocol.The models facilitated a high-throughput screening analysis for glucan,xylan,Klason lignin and acid soluble lignin（ASL）in a collection of 183 natural S.italica variants and clustered them into classes,some possessing unique chemotypes.The predictive models provide a highly efficient screening tool for large scale breeding programs aimed at identifying lines or mutants possessing unique cell wall chemotypes.Genes encoding key catalytic enzymes of the lignin biosynthesis pathway exhibit a high level of conservation with matching expression profiles,measured by RT-q PCR,among accessions of S.italica,which closely mirror profiles observed in the different developmental regions of an elongating internode of S.viridis by RNASeq.展开更多
In this study, the plant biomass production, biomass translocation rates across tissues and the lodging resistant-associated traits of millet （ Setaria italica L.） in North China were investigated. Among the four su...In this study, the plant biomass production, biomass translocation rates across tissues and the lodging resistant-associated traits of millet （ Setaria italica L.） in North China were investigated. Among the four summer millet cultivars, Baogu 19 exhibited improved plant biomass （PB） production at flowering and maturity stages, biomass translocation amount （BTA） from vegetative tissues to seeds during filling period, and lodging resistant-associated （LRA） traits compared with other cultivars, including enhanced stem lignin contents, increased anti-broken resistance （ABR）, anti-puncturing resistance （APR）, and stem diameter （SD） of plants. Compared with treatment regular cultivation （RC）, high fertility treatment （HF） increased the plant BP, BTA from vegetative tissue to seed at filling stage, and the plant LRA traits; whereas high density treatment （HD） decreased the plant BP at plant level, plant BTA from vegetative tissues to seeds at filling stage, and the plant LRA traits. Correlation analysis revealed that stem ABR is significantly correlated with the plant lodging resistant-associated traits including APR and SD in the summer millet cultivars examined under various cultivation treatments. Our investigation indicates that cultivar Baogu 19 together with suitable fertilization and density can promote the plant biomass production, enhance vegetative tissue biomass translocation to seeds, and improve the lodging resistance of summer millet plants in North China.展开更多
Salinity,a major abiotic stress,reduces plant growth and severely limits agricultural productivity.Plants regulate salt uptake via calcineurin B-like proteins(CBLs).Although extensive studies of the functions of CBLs ...Salinity,a major abiotic stress,reduces plant growth and severely limits agricultural productivity.Plants regulate salt uptake via calcineurin B-like proteins(CBLs).Although extensive studies of the functions of CBLs in response to salt stress have been conducted in Arabidopsis,their functions in Setaria italica are still poorly understood.The foxtail millet genome encodes seven CBLs,of which only SiCBL4 was shown to be involved in salt response.Overexpression of SiCBL5 in Arabidopsis thaliana sos3-1 mutant rescued its salt hypersensitivity phenotype,but that of other SiCBLs(SiCBL1,SiCBL2,SiCBL3,SiCBL6,and SiCBL7)did not rescue the salt hypersensitivity of the Atsos3-1 mutant.SiCBL5 harbors an N-myristoylation motif and is located in the plasma membrane.Overexpression of SiCBL5 in foxtail millet increased its salt tolerance,but its knockdown increased salt hypersensitivity.Yeast two-hybrid and firefly luciferase complementation imaging assays showed that SiCBL5 physically interacted with SiCIPK24 in vitro and in vivo.Cooverexpression of SiCBL5,SiCIPK24,and SiSOS1 in yeast conferred a high-salt-tolerance phenotype.Compared to wild-type plants under salt stress conditions,SiCBL5 overexpressors showed lower accumulations of Na^(+) and stronger Na^(+) efflux,whereas RNAi-SiCBL5 plants showed higher accumulations of Na^(+) and weaker Na^(+) efflux.These results indicate that SiCBL5 confers salt tolerance in foxtail millet by modulating Na^(+) homeostasis.展开更多
Male sterility is a common biological phenomenon in plant kingdom and has been used to generate male-sterile lines, which are important genetic resources for commercial hybrid seed production. Although increasing numb...Male sterility is a common biological phenomenon in plant kingdom and has been used to generate male-sterile lines, which are important genetic resources for commercial hybrid seed production. Although increasing numbers of male-sterility genes have been identified in rice(Oryza sativa) and Arabidopsis(Arabidopsis thaliana), few male-sterility-related genes have been characterized in foxtail millet(Setaria italica). In this study, we isolated a male-sterile ethyl methanesulfonate-generated mutant in foxtail millet, no pollen 1(sinp1), which displayed abnormal Ubisch bodies, defective pollen exine and complete male sterility. Using bulk segregation analysis, we cloned SiNP1 and confirmed its function with CRISPR/Cas9 genome editing. SiNP1 encoded a putative glucose-methanol-choline oxidoreductase.Subcellular localization showed that the SiNP1 protein was preferentially localized to the endoplasmic reticulum and was predominantly expressed in panicle. Transcriptome analysis revealed that many genes were differentially expressed in the sinp1 mutant, some of which encoded proteins putatively involved in carbohydrate metabolism, fatty acid biosynthesis, and lipid transport and metabolism, which were closely associated with pollen wall development. Metabolome analysis revealed the disturbance of flavonoids metabolism and fatty acid biosynthesis in the mutant. In conclusion, identification of SiNP1 provides a candidate male-sterility gene for heterosis utilization in foxtail millet and gives further insight into the mechanism of pollen reproduction in plants.展开更多
Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important...Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important cereal crops in China. Evaluation of germplasm and genetic diversity of foxtail millet is still in its infancy, but prolamin could play an important role as a protein marker. To investigate the genetic diversity and population structure of foxtail millet from different ecological zones of China, 90 accessions of foxtail millet were collected from three major ecological areas: North, Northwest, and Northeast China. The prolamin contents were examined by acid polyacrylamide gel electrophoresis (acid-PAGE). Five to twenty-two prolamin bands appeared in tested varieties, of which were polymorphic, so prolamin patterns of foxtail millet varieties can be used in variety identification and evaluation. Structure analysis identified six groups, which matches their pedigree information but not their geographic origins. This indicated a high degree (87.78%) of consistency with a phylogenetic classification based on SSR. The results showed prolamin banding patterns were an effective method for analyzing foxtail millet genetic variability.展开更多
Objective: To analyse the efficacy of diethylcarbamazine(DEC), tetramisole and chlorpromazine on the longevity and activity of glucose-6-phosphatase and succinate dehydrogenase in the microi lariae recovered from the ...Objective: To analyse the efficacy of diethylcarbamazine(DEC), tetramisole and chlorpromazine on the longevity and activity of glucose-6-phosphatase and succinate dehydrogenase in the microi lariae recovered from the peripheral circulation of the rats before and after the treatment.Methods: Setaria cer vi worms were implanted in white rats via laparotomy and microfilaraemic rats were divided into 4 groups. Groups 1, 2 and 3 were treated with DEC, tetramisole and chlorpromazine respectively, while Group 4 served as infected control. Longevity of microi lariae and dif erential leucocyte counts were recorded till the disappearance of microi lariae from peripheral blood. Glucose-6-phosphatase and succinate dehydrogenase enzymes were localized in the microi lariae recovered from normal and treated rats.Results: The microi lariae survived for 48 days in untreated rats while survival was reduced to 15, 21 and 27 days after treatment with DEC, tetramisole and chlorpromazine, respectively. Eosinophils and neutrophils increased during 2nd and 3rd weeks, whereas the lymphocytes increased during 4-7 weeks. DEC treatment resulted in slight decrease in the localization of succinate dehydrogenase but not in glucose-6-phosphatase. Tetramisole and chlorpromazine treatment did not show any appreciable change in the localization of both the above enzymes. Conclusions: DEC proved the most ef ective drug which cleared the microi laraemia within 15 days and reduced the activity of succinate dehydrogenase to some extent followed by tetramisole and chlorpromazine which took more time for the clearance of microi lariae and had no ef ect on the localization of both glucose-6-phosphatase and succinate dehydrogenase.展开更多
Foxtail millet(Setaria italica),a vital drought-resistant crop,plays a significant role in ensuring food and nutritional security.However,its drought resistance mechanism is not fully understood.N6-methyladenosine(m^(...Foxtail millet(Setaria italica),a vital drought-resistant crop,plays a significant role in ensuring food and nutritional security.However,its drought resistance mechanism is not fully understood.N6-methyladenosine(m^(6)A)modification of RNA,a prevalent epi-transcriptomic modification in eukaryotes,provides a binding site for m^(6)A readers and affects plant growth and stress responses by regulating RNA metabolism.In this study,we unveiled that the YT521-B homology(YTH)family gene SiYTH1 positively regulated the drought tolerance of foxtail millet.Notably,the siyth1 mutant exhibited reduced stomatal closure and augmented accumulation of excessive H_(2)O_(2)under drought stress.Further investigations demonstrated that SiYTH1 positively regulated the transcripts harboring m^(6)A modification related to stomatal closure and reactive oxygen species(ROS)scavenging under drought stress.SiYTH1 was uniformly distributed in the cytoplasm of SiYTH1-GFP transgenic foxtail millet.It formed dynamic liquid-like SiYTH1 cytosol condensates in response to drought stress.Moreover,the cytoplasmic protein SiYTH1 was identified as a distinct m^(6)A reader,facilitating the stabilization of its directly bound SiARDP and ROS scavenging-related transcripts under drought stress.Furthermore,natural variation analysis revealed SiYTH1AGTG as the dominant allele responsible for drought tolerance in foxtail millet.Collectively,this study provides novel insights into the intricate mechanism of m^(6)A reader-mediated drought tolerance and presents a valuable genetic resource for improving drought tolerance in foxtail millet breeding.展开更多
基金supported by the National Natural Science Foundation of China (32241042 and 31771807)the National Key R&D Program of China (2021YFF1000103)+1 种基金the China Agricultural Research System (CARS-06-04)the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences。
文摘Chlorophyll (Chl) content,especially Chl b content,and stomatal conductance (G_s) are the key factors affecting the net photosynthetic rate (P_n).Setaria italica,a diploid C_4 panicoid species with a simple genome and high transformation efficiency,has been widely accepted as a model in photosynthesis and drought-tolerance research.The current study characterized Chl content,G_s,and P_n of 48 Setaria mutants induced by ethyl methanesulfonate.A total of 24,34,and 35 mutants had significant variations in Chl content,G_s,and P_n,respectively.Correlation analysis showed a positive correlation between increased G_s and increased P_n,and a weak correlation between decreased Chl b content and decreased P_n was also found.Remarkably,two mutants behaved with significantly decreased Chl b content but increased P_n compared to Yugu 1.Seven mutants behaved with significantly decreased G_s but did not decrease P_(n )compared to Yugu 1.The current study thus identified various genetic lines,further exploration of which would be beneficial to elucidate the relationship between Chl content,G_s,and P_n and the mechanism underlying why C_4 species are efficient at photosynthesis and water saving.
文摘Background:The goal of the current study was to identify the potential therapeutic targets by analyzing the production of oxidants and their effects on the antioxidant system of Setaria digitata,a filarial worm that was collected from the peritoneal cavity of recently butchered calves.Methods:S.digitata was collected and the mitochondria-like particles(MLP),somatic isolates and the post-nuclear fraction were prepared for investigation.The radical intensity and the antioxidant enzyme activity in these fractions were evaluated.The effect of diethylcarbamazine(DEC),Q0,omeprazole,and L-methyl arginine on NOS production in the worm was also evaluated.Results:Experiments showed that coelomic fluid and whole worm showed significant levels of Nitric oxide compared to other fractions.The cytosol of the worm showed higher NOS activity than MLP.Superoxide dismutase and nitric oxide(NO)levels were found to be higher in MLP fraction of the worm.It was also found that DEC did not have any effect on the prevention of NO,while Q0,omeprazole,and L-methyl arginine treatment showed a decrease in NO production significantly.Conclusion:The results showed that by using the existing drugs we could reduce the propagation of S.digitata by inhibiting the production of NO.
基金supported by the Postdoctoral Management Committee,China(92948)the Natural Science Foundation of Shanxi Province,China(2012011032-1)the Chinese Agricultural Research System(CARS-07)
文摘Breeding of male-sterile lines has become the mainstream for the heterosis utilization in foxtail millet,but the genetic basis of most male-sterile lines used for the hybrid is still an area to be elucidated.In this study,a highly male-sterile line Gao146A was investigated.Genetic analysis indicated that the highly male-sterile phenotype was controlled by a single recessive gene a single recessive gene.Using F 2 population derived from cross Gao146A/K103,one gene controlling the highly male- sterility,tentatively named as ms1,which linked to SSR marker b234 with genetic distance of 16.7 cM,was mapped on the chromosome VI.These results not only laid the foundation for fine mapping of this highly male-sterile gene,but also helped to accelerate the improvement of highly male-sterile lines by using molecular marker assisted breeding method.
基金funded by the National Key Project for Research on Transgenic Biology, China (2016ZX08002-002)the Innovation Project of Chinese Academy of Agricultural Sciences
文摘Foxtail millet（Setaria italica（L.）P.Beauv）is a naturally stress tolerant crop.Compared to other gramineous crops,it has relatively stronger drought and lower nutrition stress tolerance traits.To date,the scope of functional genomics research in foxtail millet（S.italic L.）has been quite limited.NAC（NAM,ATAF1/2 and CUC2）-like transcription factors are known to be involved in various biological processes,including abiotic stress responses.In our previous foxtail millet（S.italic L.）RNA seq analysis,we found that the expression of a NAC-like transcription factor,SiNAC110,could be induced by drought stress;additionally,other references have reported that SiNAC110 expression could be induced by abiotic stress.So,we here selected SiNAC110 for further characterization and functional analysis.First,the predicted SiNAC110 protein encoded indicated SiNAC110 has a conserved NAM（no apical meristem）domain between the 11–139 amino acid positions.Phylogenetic analysis then indicated that SiNAC110 belongs to subfamily III of the NAC gene family.Subcellular localization analysis revealed that the SiNAC110-GFP fusion protein was localized to the nucleus in Arabidopsis protoplasts.Gene expression profiling analysis indicated that expression of SiNAC110 was induced by dehydration,high salinity and other abiotic stresses.Gene functional analysis using SiNAC110 overexpressed Arabidopsis plants indicated that,under drought and high salt stress conditions,the seed germination rate,root length,root surface area,fresh weight,and dry weight of the SiNAC110 overexpressed lines were significantly higher than the wild type（WT）,suggesting that the SiNAC110 overexpressed lines had enhanced tolerance to drought and high salt stresses.However,overexpression of SiN AC110 did not affect the sensitivity of SiNAC110 overexpressed lines to abscisic acid（ABA）treatment.Expression analysis of genes involved in proline synthesis,Na＋/K＋transport,drought responses,and aqueous transport proteins were higher in the SiNAC110overexpressed lines than in the WT,whereas expression of ABA-dependent pathway genes did not change.These results indicated that overexpression of SiNAC110 conferred tolerance to drought and high salt stresses,likely through influencing the regulation of proline biosynthesis,ion homeostasis and osmotic balance.Therefore,SiNAC110 appears to function in the ABA-independent abiotic stress response pathway in plants.
基金supported by a grant(SIDA/2006/BT/04) awarded by National Science Foundation of Sri Lanka
文摘Objective:To clone,express and purify a putative parasitic nematode specific protein of Setaria digitata(S.digitata),filarial nematode that infects livestock and cause significant economic losses in Far East and Asia to he used for structural and functional analyses.Methods:To characterize uneharacterized gene of,S.digitata(SDUG),the herterologous expression of SDUG was carried out in the pET[cloned into pET45b(+)]expression system initially and co-expression of SDUC using chaperoiie plasmids pG-KJE8,pGro 7,pKJE7,pG-Tf2 and pTf16 containing chapcrone proteins of dnaK-dnaJ-grpE-groES-gro-E,groES-groEL,dnaK-dnaJ-grpE,groES-groEL-tig,and tig respectively,was carried out subsequently.Results:Expression of SDUG was seen when Escherichia coli strain BI.21(DE3)is used,while concentrating protein largely into the insoluble fraction.The co-expression of SDUG using chaperoiie plasmid mediated system indicated a significant increase of the protein in the soluble fraction.Of the chaperon plasniid sets,the highest amount of recombinant SDUP in the soluble fraction was seen when pGro7 was used in the presence of2 mg/mL L-arabinosc and 0.6M IPTG concentration in the culture medium and for 3 h of incubation at the temperature of 28℃.Recombinant SDUG was purified both from soluble and insoluble fractions using Ni affinity chromatography.SDS-PAGE and western blot analyses of these proteins revealed a single band having expected size of^24 kDa.Conclusions:SDUG seems to be more aggregate-prone and hydrophobic in nature and such protein can make soluble by correct selecting the inducer concentrations and induction temperature and its duration.
文摘Cell wall composition in monocotyledonous grasses has been identified as a key area of research for developing better feedstocks for forage and biofuel production.Setaria viridis and its close domesticated relative Setaria italica have been chosen as suitable monocotyledonous models for plants possessing the C4 pathway of photosynthesis including sorghum,maize,sugarcane,switchgrass and Miscanthus×giganteus.Accurate partial least squares regression（PLSR）models to predict S.italica stem composition have been generated,based upon Fourier transform mid-infrared（FTIR）spectra and calibrated with wet chemistry determinations of ground S.italica stem material measured using a modified version of the US National Renewable Energy Laboratory（NREL）acid hydrolysis protocol.The models facilitated a high-throughput screening analysis for glucan,xylan,Klason lignin and acid soluble lignin（ASL）in a collection of 183 natural S.italica variants and clustered them into classes,some possessing unique chemotypes.The predictive models provide a highly efficient screening tool for large scale breeding programs aimed at identifying lines or mutants possessing unique cell wall chemotypes.Genes encoding key catalytic enzymes of the lignin biosynthesis pathway exhibit a high level of conservation with matching expression profiles,measured by RT-q PCR,among accessions of S.italica,which closely mirror profiles observed in the different developmental regions of an elongating internode of S.viridis by RNASeq.
基金Supported by Special Fund of Agricultural Science and Technology Achievement Transformation in Hebei Province(17826335D)Baoding Comprehensive Experimental Station of National Millet and Sorghum Industry Technology System(CARS-06-13.5-B2)
文摘In this study, the plant biomass production, biomass translocation rates across tissues and the lodging resistant-associated traits of millet （ Setaria italica L.） in North China were investigated. Among the four summer millet cultivars, Baogu 19 exhibited improved plant biomass （PB） production at flowering and maturity stages, biomass translocation amount （BTA） from vegetative tissues to seeds during filling period, and lodging resistant-associated （LRA） traits compared with other cultivars, including enhanced stem lignin contents, increased anti-broken resistance （ABR）, anti-puncturing resistance （APR）, and stem diameter （SD） of plants. Compared with treatment regular cultivation （RC）, high fertility treatment （HF） increased the plant BP, BTA from vegetative tissue to seed at filling stage, and the plant LRA traits; whereas high density treatment （HD） decreased the plant BP at plant level, plant BTA from vegetative tissues to seeds at filling stage, and the plant LRA traits. Correlation analysis revealed that stem ABR is significantly correlated with the plant lodging resistant-associated traits including APR and SD in the summer millet cultivars examined under various cultivation treatments. Our investigation indicates that cultivar Baogu 19 together with suitable fertilization and density can promote the plant biomass production, enhance vegetative tissue biomass translocation to seeds, and improve the lodging resistance of summer millet plants in North China.
基金supported by the National Natural Science Foundation of China(32001445 and 31871534)the Natural Science Foundation of Jiangsu Province(BK20200557)。
文摘Salinity,a major abiotic stress,reduces plant growth and severely limits agricultural productivity.Plants regulate salt uptake via calcineurin B-like proteins(CBLs).Although extensive studies of the functions of CBLs in response to salt stress have been conducted in Arabidopsis,their functions in Setaria italica are still poorly understood.The foxtail millet genome encodes seven CBLs,of which only SiCBL4 was shown to be involved in salt response.Overexpression of SiCBL5 in Arabidopsis thaliana sos3-1 mutant rescued its salt hypersensitivity phenotype,but that of other SiCBLs(SiCBL1,SiCBL2,SiCBL3,SiCBL6,and SiCBL7)did not rescue the salt hypersensitivity of the Atsos3-1 mutant.SiCBL5 harbors an N-myristoylation motif and is located in the plasma membrane.Overexpression of SiCBL5 in foxtail millet increased its salt tolerance,but its knockdown increased salt hypersensitivity.Yeast two-hybrid and firefly luciferase complementation imaging assays showed that SiCBL5 physically interacted with SiCIPK24 in vitro and in vivo.Cooverexpression of SiCBL5,SiCIPK24,and SiSOS1 in yeast conferred a high-salt-tolerance phenotype.Compared to wild-type plants under salt stress conditions,SiCBL5 overexpressors showed lower accumulations of Na^(+) and stronger Na^(+) efflux,whereas RNAi-SiCBL5 plants showed higher accumulations of Na^(+) and weaker Na^(+) efflux.These results indicate that SiCBL5 confers salt tolerance in foxtail millet by modulating Na^(+) homeostasis.
基金supported by the National Natural Science Foundation of China(31771807)the China Agriculture Research System(CARS06-13.5-A04)+1 种基金the National Key Research and Development Program of China(2018YFD1000700 and 2018YFD1000701)the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences。
文摘Male sterility is a common biological phenomenon in plant kingdom and has been used to generate male-sterile lines, which are important genetic resources for commercial hybrid seed production. Although increasing numbers of male-sterility genes have been identified in rice(Oryza sativa) and Arabidopsis(Arabidopsis thaliana), few male-sterility-related genes have been characterized in foxtail millet(Setaria italica). In this study, we isolated a male-sterile ethyl methanesulfonate-generated mutant in foxtail millet, no pollen 1(sinp1), which displayed abnormal Ubisch bodies, defective pollen exine and complete male sterility. Using bulk segregation analysis, we cloned SiNP1 and confirmed its function with CRISPR/Cas9 genome editing. SiNP1 encoded a putative glucose-methanol-choline oxidoreductase.Subcellular localization showed that the SiNP1 protein was preferentially localized to the endoplasmic reticulum and was predominantly expressed in panicle. Transcriptome analysis revealed that many genes were differentially expressed in the sinp1 mutant, some of which encoded proteins putatively involved in carbohydrate metabolism, fatty acid biosynthesis, and lipid transport and metabolism, which were closely associated with pollen wall development. Metabolome analysis revealed the disturbance of flavonoids metabolism and fatty acid biosynthesis in the mutant. In conclusion, identification of SiNP1 provides a candidate male-sterility gene for heterosis utilization in foxtail millet and gives further insight into the mechanism of pollen reproduction in plants.
文摘Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important cereal crops in China. Evaluation of germplasm and genetic diversity of foxtail millet is still in its infancy, but prolamin could play an important role as a protein marker. To investigate the genetic diversity and population structure of foxtail millet from different ecological zones of China, 90 accessions of foxtail millet were collected from three major ecological areas: North, Northwest, and Northeast China. The prolamin contents were examined by acid polyacrylamide gel electrophoresis (acid-PAGE). Five to twenty-two prolamin bands appeared in tested varieties, of which were polymorphic, so prolamin patterns of foxtail millet varieties can be used in variety identification and evaluation. Structure analysis identified six groups, which matches their pedigree information but not their geographic origins. This indicated a high degree (87.78%) of consistency with a phylogenetic classification based on SSR. The results showed prolamin banding patterns were an effective method for analyzing foxtail millet genetic variability.
基金Supported by the Deanship of Scientific Research(DSR),King Abdulaziz University,Jeddah(Grant No.407/142/1434)
文摘Objective: To analyse the efficacy of diethylcarbamazine(DEC), tetramisole and chlorpromazine on the longevity and activity of glucose-6-phosphatase and succinate dehydrogenase in the microi lariae recovered from the peripheral circulation of the rats before and after the treatment.Methods: Setaria cer vi worms were implanted in white rats via laparotomy and microfilaraemic rats were divided into 4 groups. Groups 1, 2 and 3 were treated with DEC, tetramisole and chlorpromazine respectively, while Group 4 served as infected control. Longevity of microi lariae and dif erential leucocyte counts were recorded till the disappearance of microi lariae from peripheral blood. Glucose-6-phosphatase and succinate dehydrogenase enzymes were localized in the microi lariae recovered from normal and treated rats.Results: The microi lariae survived for 48 days in untreated rats while survival was reduced to 15, 21 and 27 days after treatment with DEC, tetramisole and chlorpromazine, respectively. Eosinophils and neutrophils increased during 2nd and 3rd weeks, whereas the lymphocytes increased during 4-7 weeks. DEC treatment resulted in slight decrease in the localization of succinate dehydrogenase but not in glucose-6-phosphatase. Tetramisole and chlorpromazine treatment did not show any appreciable change in the localization of both the above enzymes. Conclusions: DEC proved the most ef ective drug which cleared the microi laraemia within 15 days and reduced the activity of succinate dehydrogenase to some extent followed by tetramisole and chlorpromazine which took more time for the clearance of microi lariae and had no ef ect on the localization of both glucose-6-phosphatase and succinate dehydrogenase.
基金This work was supported by the National Key R&D Program of China(2023YFD1200700/2023YFD1200702/2018YFD1000700/2018YFD1000704).
文摘Foxtail millet(Setaria italica),a vital drought-resistant crop,plays a significant role in ensuring food and nutritional security.However,its drought resistance mechanism is not fully understood.N6-methyladenosine(m^(6)A)modification of RNA,a prevalent epi-transcriptomic modification in eukaryotes,provides a binding site for m^(6)A readers and affects plant growth and stress responses by regulating RNA metabolism.In this study,we unveiled that the YT521-B homology(YTH)family gene SiYTH1 positively regulated the drought tolerance of foxtail millet.Notably,the siyth1 mutant exhibited reduced stomatal closure and augmented accumulation of excessive H_(2)O_(2)under drought stress.Further investigations demonstrated that SiYTH1 positively regulated the transcripts harboring m^(6)A modification related to stomatal closure and reactive oxygen species(ROS)scavenging under drought stress.SiYTH1 was uniformly distributed in the cytoplasm of SiYTH1-GFP transgenic foxtail millet.It formed dynamic liquid-like SiYTH1 cytosol condensates in response to drought stress.Moreover,the cytoplasmic protein SiYTH1 was identified as a distinct m^(6)A reader,facilitating the stabilization of its directly bound SiARDP and ROS scavenging-related transcripts under drought stress.Furthermore,natural variation analysis revealed SiYTH1AGTG as the dominant allele responsible for drought tolerance in foxtail millet.Collectively,this study provides novel insights into the intricate mechanism of m^(6)A reader-mediated drought tolerance and presents a valuable genetic resource for improving drought tolerance in foxtail millet breeding.
