Water deficit is a major worldwide constraint to common bean(Phaseolus vulgaris L.)production,being photosynthesis one of the most affected physiological processes.To gain insights into the genetic basis of the photos...Water deficit is a major worldwide constraint to common bean(Phaseolus vulgaris L.)production,being photosynthesis one of the most affected physiological processes.To gain insights into the genetic basis of the photosynthetic response of common bean under water-limited conditions,a collection of 158 Portuguese accessions was grown under both well-watered and water-deficit regimes.Leaf gas-exchange parameters were measured and photosynthetic pigments quantified.The same collection was genotyped using SNP arrays,and SNP-trait associations tested considering a linear mixed model accounting for the genetic relatedness among accessions.A total of 133 SNP-trait associations were identified for net CO 2 assimilation rate,transpiration rate,stomatal conductance,and chlorophylls a and b,carotenes,and xanthophyll contents.Ninety of these associations were detected under water-deficit and 43 under well-watered conditions,with only two associations common to both treatments.Identified candidate genes revealed that stomatal regulation,protein translocation across membranes,redox mechanisms,hormone,and osmotic stress signaling were the most relevant processes involved in common bean response to water-limited conditions.These candidates are now preferential targets for common bean water-deficit-tolerance breeding.Additionally,new sources of water-deficit tolerance of Andean,Mesoamerican,and admixed origin were detected as accessions valuable for breeding,and not yet explored.展开更多
The knowledge on post-transcriptional regulation mechanisms implicated in seed development(SD)is still limited,particularly in one of the most consumed grain legumes,Phaseolus vulgaris L.We explore for the first time ...The knowledge on post-transcriptional regulation mechanisms implicated in seed development(SD)is still limited,particularly in one of the most consumed grain legumes,Phaseolus vulgaris L.We explore for the first time the miRNA expression dynamics in P.vulgaris developing seeds.Seventy-two known and 39 new miRNAs were found expressed in P.vulgaris developing seeds.Most of the miRNAs identified were more abundant at 10 and 40 days after anthesis,suggesting that late embryogenesis/early filling and desiccation were SD stages in which miRNA action is more pronounced.Degradome analysis and target prediction identified targets for 77 expressed miRNAs.While several known miRNAs were predicted to target HD-ZIP,ARF,SPL,and NF-Y transcription factors families,most of the predicted targets for new miRNAs encode for functional proteins.MiRNAs-targets expression profiles evidenced that these miRNAs could tune distinct seed developmental stages.MiRNAs more accumulated at early SD stages were implicated in regulating the end of embryogenesis,postponing the seed maturation program,storage compound synthesis and allocation.MiRNAs more accumulated at late SD stages could be implicated in seed quiescence,desiccation tolerance,and longevity with still uncovered roles in germination.The miRNAs herein described represent novel P.vulgaris resources with potential application in future biotechnological approaches to modulate the expression of genes implicated in legume seed traits with impact in horticultural production systems.展开更多
Quil-A and cholesterol can, under certain conditions, form nanoparticles, but the conditions for the formation of a homogeneous population of a particular kind of particles with the same morphology and size have remai...Quil-A and cholesterol can, under certain conditions, form nanoparticles, but the conditions for the formation of a homogeneous population of a particular kind of particles with the same morphology and size have remained elusive. However, a well-defined uniformity, as well as a high degree of batch-to-batch consistency, are prerequisites for adjuvant formulations to be used for practical vaccines. To accomplish the above stated tasks in the present study, we demonstrated that Quil-A and cholesterol form well-defined worm-like stable particles with a mean diameter of around 40 nm by dynamic light scattering (DLS) under carefully controlled thermodynamical conditions with little or no Quil-A degradation. The nanoparticles thus prepared possessed a significantly reduced hemolytic effect in comparison to unformulated free Quil-A. However, when the same conditions were applied to QS-21/cholesterol nanoparticle formation the morphology achieved was coil spring-like particles with a mean diameter of around 70 nm. This discrepancy in particle morphology and size was attributed to the differences in hydrophobicity of the Quil-A, being a heterogenic fraction of acylated as well as deacylated saponins, and the QS-21. With the process optimizations leading to a standardized particle size distribution and nanoparticle morphology presented here, NanoQuil F70 holds the potential as a well-tolerated vaccine adjuvant for veterinary use raising a Th1/Th2 balanced immune response.展开更多
Common bean(Phaseolus vulgaris L.),one of the most consumed food legumes worldwide,is threatened by two main constraints that are found frequently together in nature,water deficit(WD)and fusarium wilt(Fop).To understa...Common bean(Phaseolus vulgaris L.),one of the most consumed food legumes worldwide,is threatened by two main constraints that are found frequently together in nature,water deficit(WD)and fusarium wilt(Fop).To understand the shared and unique responses of common bean to Fop and WD,we analyzed the transcriptomic changes and phenotypic responses in two accessions,one resistant and one susceptible to both stresses,exposed to single and combined stresses.Physiological responses(photosynthetic performance and pigments quantification)and disease progression were also assessed.The combined FopWD imposition negatively affected the photosynthetic performance and increased the susceptible accession disease symptoms.The susceptible accession revealed a higher level of transcriptional changes than the resistant one,and WD single stress triggered the highest transcriptional changes.While 89 differentially expressed genes were identified exclusively in combined stresses for the susceptible accession,35 were identified in the resistant one.These genes belong mainly to“stress”,“signaling”,“cell wall”,“hormone metabolism”,and“secondary metabolism”functional categories.Among the up-regulated genes with higher expression in the resistant accession,the cysteine-rich secretory,antigen 5 and Pr-1(CAP)superfamily protein,a ribulose bisphosphate carboxylase family protein,and a chitinase A seem promising targets for multiple stress breeding.展开更多
Objective:To evaluate the effect of methanolic extract and ethyl acetate fraction of methanol extract prepared from the seeds of Blepharis(B.)persica on testosterone biosynthesis and also to elucidate the underlying m...Objective:To evaluate the effect of methanolic extract and ethyl acetate fraction of methanol extract prepared from the seeds of Blepharis(B.)persica on testosterone biosynthesis and also to elucidate the underlying mechanism.Methods:Forty-eight male Wistar rats were divided into eight groups(n=6 per group).GroupⅠreceived 0.3%w/w gum acacia suspension p.o.and served as the normal control group.GroupⅡwas administered testosterone propionate in arachis oil i.m.as the positive control group.GroupⅢtoⅣreceived B.persica methanolic extract p.o.at doses of 50,100 and 200 mg/kg body weight.GroupⅥtoⅦreceived B.persica ethyl acetate fraction p.o.at doses of 50,100 and 200 mg/kg body weight.The testis was used for biochemical estimation and histological studies.The effects of methanolic extract and ethyl acetate fraction of B.persica on testicular testosterone,mRNA expression corresponding to steroidogenic acute regulatory protein(StAR)and 3β-hydroxysteroid dehydrogenase(3β-HSD)along with 3β-HSD enzyme assay were evaluated in testicular tissues and sperm concentration.Ethyl acetate fraction of B.persica was subjected to column chromatography.Invitro studies were performed using TM3 cell line at three dose levels(50,100,200μg/mL),each for methanolic extract,ethyl acetate fraction and 2-benzoxazolinone for evaluation of their comparative effect on testosterone production.Results:Ethyl acetate fraction and methanolic extract of B.persica could elevate the testicular testosterone content compared to the normal control group.The treatment with methanolic extract and ethyl acetate fraction of B.persica increased the expression of mRNA corresponding to StAR by 6.7 fold and 10.6 fold,respectively,whereas the mRNA expression of 3β-HSD increased by 5.7 fold and 7.3 fold,respectively.Moreover,fraction and extract treatment exhibited increased 3β-HSD activity in the testicular tissues and were found to elevate sperm concentration in seminal fluid.The spermatogenic potential was further ensured by histological observations.2-benzoxazolinone was isolated from ethyl acetate fraction and identified using spectral studies.It showed the ability to increase the testosterone content in the TM3 Leydig cells.Conclusions:Methanolic extract and ethyl acetate fraction of B.persica are able to increase the testicular testosterone in rats by elevating mRNA expression of StAR and 3β-HSD in testicular tissues,leading to increase the sperm concentration.展开更多
The objective of this work was to study the microbial diversity of natural whey starters (NWS), with a special focus on Lactococcus strains, during 10 successive days of backslopping. Each day, whey samples were enume...The objective of this work was to study the microbial diversity of natural whey starters (NWS), with a special focus on Lactococcus strains, during 10 successive days of backslopping. Each day, whey samples were enumerated for their Lactococcus, Enterococcus, Leuconostoc and Lactobacillus levels. The diversity and dynamics of the Lactococcus population in whey were investigated using phenotypic methods such as acidifying aptitude and enzymatic activities. Molecular approach using REP-PCR, ERIC-PCR, plamid profiles, phages and prophages research and PFGE was also applied. Lactococci were the main population in whey. Strains from raw milk were sub-dominant. Based on PFGE and phenotypic results lactococci in whey displayed a more heterogeneous phenotype and pulsotype which may reflect greater variations than previously observed within starter. No phages and prophages were spotted on. Plasmids did not seem to be exchanged from strain to strain. The backslopping practice seemed to allow the strains of the starter to rapidly acquire a specificity of their own. The changes observed presumed a slow adaptation of the strains to the “back- slopping environment”. The study of NWS diversity constitutes an important step for the comprehension of acidification defects that recurrently occur in cheesemaking technologies using the backslopping practice.展开更多
文摘Water deficit is a major worldwide constraint to common bean(Phaseolus vulgaris L.)production,being photosynthesis one of the most affected physiological processes.To gain insights into the genetic basis of the photosynthetic response of common bean under water-limited conditions,a collection of 158 Portuguese accessions was grown under both well-watered and water-deficit regimes.Leaf gas-exchange parameters were measured and photosynthetic pigments quantified.The same collection was genotyped using SNP arrays,and SNP-trait associations tested considering a linear mixed model accounting for the genetic relatedness among accessions.A total of 133 SNP-trait associations were identified for net CO 2 assimilation rate,transpiration rate,stomatal conductance,and chlorophylls a and b,carotenes,and xanthophyll contents.Ninety of these associations were detected under water-deficit and 43 under well-watered conditions,with only two associations common to both treatments.Identified candidate genes revealed that stomatal regulation,protein translocation across membranes,redox mechanisms,hormone,and osmotic stress signaling were the most relevant processes involved in common bean response to water-limited conditions.These candidates are now preferential targets for common bean water-deficit-tolerance breeding.Additionally,new sources of water-deficit tolerance of Andean,Mesoamerican,and admixed origin were detected as accessions valuable for breeding,and not yet explored.
文摘The knowledge on post-transcriptional regulation mechanisms implicated in seed development(SD)is still limited,particularly in one of the most consumed grain legumes,Phaseolus vulgaris L.We explore for the first time the miRNA expression dynamics in P.vulgaris developing seeds.Seventy-two known and 39 new miRNAs were found expressed in P.vulgaris developing seeds.Most of the miRNAs identified were more abundant at 10 and 40 days after anthesis,suggesting that late embryogenesis/early filling and desiccation were SD stages in which miRNA action is more pronounced.Degradome analysis and target prediction identified targets for 77 expressed miRNAs.While several known miRNAs were predicted to target HD-ZIP,ARF,SPL,and NF-Y transcription factors families,most of the predicted targets for new miRNAs encode for functional proteins.MiRNAs-targets expression profiles evidenced that these miRNAs could tune distinct seed developmental stages.MiRNAs more accumulated at early SD stages were implicated in regulating the end of embryogenesis,postponing the seed maturation program,storage compound synthesis and allocation.MiRNAs more accumulated at late SD stages could be implicated in seed quiescence,desiccation tolerance,and longevity with still uncovered roles in germination.The miRNAs herein described represent novel P.vulgaris resources with potential application in future biotechnological approaches to modulate the expression of genes implicated in legume seed traits with impact in horticultural production systems.
文摘Quil-A and cholesterol can, under certain conditions, form nanoparticles, but the conditions for the formation of a homogeneous population of a particular kind of particles with the same morphology and size have remained elusive. However, a well-defined uniformity, as well as a high degree of batch-to-batch consistency, are prerequisites for adjuvant formulations to be used for practical vaccines. To accomplish the above stated tasks in the present study, we demonstrated that Quil-A and cholesterol form well-defined worm-like stable particles with a mean diameter of around 40 nm by dynamic light scattering (DLS) under carefully controlled thermodynamical conditions with little or no Quil-A degradation. The nanoparticles thus prepared possessed a significantly reduced hemolytic effect in comparison to unformulated free Quil-A. However, when the same conditions were applied to QS-21/cholesterol nanoparticle formation the morphology achieved was coil spring-like particles with a mean diameter of around 70 nm. This discrepancy in particle morphology and size was attributed to the differences in hydrophobicity of the Quil-A, being a heterogenic fraction of acylated as well as deacylated saponins, and the QS-21. With the process optimizations leading to a standardized particle size distribution and nanoparticle morphology presented here, NanoQuil F70 holds the potential as a well-tolerated vaccine adjuvant for veterinary use raising a Th1/Th2 balanced immune response.
文摘Common bean(Phaseolus vulgaris L.),one of the most consumed food legumes worldwide,is threatened by two main constraints that are found frequently together in nature,water deficit(WD)and fusarium wilt(Fop).To understand the shared and unique responses of common bean to Fop and WD,we analyzed the transcriptomic changes and phenotypic responses in two accessions,one resistant and one susceptible to both stresses,exposed to single and combined stresses.Physiological responses(photosynthetic performance and pigments quantification)and disease progression were also assessed.The combined FopWD imposition negatively affected the photosynthetic performance and increased the susceptible accession disease symptoms.The susceptible accession revealed a higher level of transcriptional changes than the resistant one,and WD single stress triggered the highest transcriptional changes.While 89 differentially expressed genes were identified exclusively in combined stresses for the susceptible accession,35 were identified in the resistant one.These genes belong mainly to“stress”,“signaling”,“cell wall”,“hormone metabolism”,and“secondary metabolism”functional categories.Among the up-regulated genes with higher expression in the resistant accession,the cysteine-rich secretory,antigen 5 and Pr-1(CAP)superfamily protein,a ribulose bisphosphate carboxylase family protein,and a chitinase A seem promising targets for multiple stress breeding.
基金supported by Charotar University of Science and Technology through CHARUSAT Research Grant sanctioned to Dr.Manan Raval[CHARUSAT SEED RESEARCH GRANT/RPCP/MAR/12].
文摘Objective:To evaluate the effect of methanolic extract and ethyl acetate fraction of methanol extract prepared from the seeds of Blepharis(B.)persica on testosterone biosynthesis and also to elucidate the underlying mechanism.Methods:Forty-eight male Wistar rats were divided into eight groups(n=6 per group).GroupⅠreceived 0.3%w/w gum acacia suspension p.o.and served as the normal control group.GroupⅡwas administered testosterone propionate in arachis oil i.m.as the positive control group.GroupⅢtoⅣreceived B.persica methanolic extract p.o.at doses of 50,100 and 200 mg/kg body weight.GroupⅥtoⅦreceived B.persica ethyl acetate fraction p.o.at doses of 50,100 and 200 mg/kg body weight.The testis was used for biochemical estimation and histological studies.The effects of methanolic extract and ethyl acetate fraction of B.persica on testicular testosterone,mRNA expression corresponding to steroidogenic acute regulatory protein(StAR)and 3β-hydroxysteroid dehydrogenase(3β-HSD)along with 3β-HSD enzyme assay were evaluated in testicular tissues and sperm concentration.Ethyl acetate fraction of B.persica was subjected to column chromatography.Invitro studies were performed using TM3 cell line at three dose levels(50,100,200μg/mL),each for methanolic extract,ethyl acetate fraction and 2-benzoxazolinone for evaluation of their comparative effect on testosterone production.Results:Ethyl acetate fraction and methanolic extract of B.persica could elevate the testicular testosterone content compared to the normal control group.The treatment with methanolic extract and ethyl acetate fraction of B.persica increased the expression of mRNA corresponding to StAR by 6.7 fold and 10.6 fold,respectively,whereas the mRNA expression of 3β-HSD increased by 5.7 fold and 7.3 fold,respectively.Moreover,fraction and extract treatment exhibited increased 3β-HSD activity in the testicular tissues and were found to elevate sperm concentration in seminal fluid.The spermatogenic potential was further ensured by histological observations.2-benzoxazolinone was isolated from ethyl acetate fraction and identified using spectral studies.It showed the ability to increase the testosterone content in the TM3 Leydig cells.Conclusions:Methanolic extract and ethyl acetate fraction of B.persica are able to increase the testicular testosterone in rats by elevating mRNA expression of StAR and 3β-HSD in testicular tissues,leading to increase the sperm concentration.
文摘The objective of this work was to study the microbial diversity of natural whey starters (NWS), with a special focus on Lactococcus strains, during 10 successive days of backslopping. Each day, whey samples were enumerated for their Lactococcus, Enterococcus, Leuconostoc and Lactobacillus levels. The diversity and dynamics of the Lactococcus population in whey were investigated using phenotypic methods such as acidifying aptitude and enzymatic activities. Molecular approach using REP-PCR, ERIC-PCR, plamid profiles, phages and prophages research and PFGE was also applied. Lactococci were the main population in whey. Strains from raw milk were sub-dominant. Based on PFGE and phenotypic results lactococci in whey displayed a more heterogeneous phenotype and pulsotype which may reflect greater variations than previously observed within starter. No phages and prophages were spotted on. Plasmids did not seem to be exchanged from strain to strain. The backslopping practice seemed to allow the strains of the starter to rapidly acquire a specificity of their own. The changes observed presumed a slow adaptation of the strains to the “back- slopping environment”. The study of NWS diversity constitutes an important step for the comprehension of acidification defects that recurrently occur in cheesemaking technologies using the backslopping practice.
