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1.
Nat Plants ; 7(2): 123-128, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33558754

RESUMO

All crops are the product of a domestication process that started less than 12,000 years ago from one or more wild populations1,2. Farmers selected desirable phenotypic traits (such as improved energy accumulation, palatability of seeds and reduced natural shattering3) while leading domesticated populations through several more or less gradual demographic contractions2,4. As a consequence, the erosion of wild genetic variation5 is typical of modern cultivars, making them highly susceptible to pathogens, pests and environmental change6,7. The loss of genetic diversity hampers further crop improvement programmes to increase food production in a changing world, posing serious threats to food security8,9. Using both ancient and modern seeds, we analysed the temporal dynamics of genetic variation and selection during the domestication process of the common bean (Phaseolus vulgaris) in the southern Andes. Here, we show that most domestic traits were selected for before 2,500 years ago, with no or only minor loss of whole-genome heterozygosity. In fact, most of the changes at coding genes and linked regions that differentiate wild and domestic genomes are already present in the ancient genomes analysed here, and all ancient domestic genomes dated between 600 and 2,500 years ago are highly variable (at least as variable as modern genomes from the wild). Single seeds from modern cultivars show reduced variation when compared with ancient seeds, indicating that intensive selection within cultivars in the past few centuries probably partitioned ancestral variation within different genetically homogenous cultivars. When cultivars from different Andean regions are pooled, the genomic variation of the pool is higher than that observed in the pool of ancient seeds from north and central western Argentina. Considering that most desirable phenotypic traits are probably controlled by multiple polymorphic genes10, a plausible explanation of this decoupling of selection and genetic erosion is that early farmers applied a relatively weak selection pressure2 by using many phenotypically similar but genetically diverse individuals as parents. Our results imply that selection strategies during the past few centuries, as compared with earlier times, more intensively reduced genetic variation within cultivars and produced further improvements by focusing on a few plants carrying the traits of interest, at the cost of marked genetic erosion within Andean landraces.


Assuntos
Produtos Agrícolas/genética , Produtos Agrícolas/história , Domesticação , Fazendeiros/psicologia , Genoma de Planta , Phaseolus/genética , Argentina , Fazendeiros/estatística & dados numéricos , Variação Genética , Genótipo , História Antiga
2.
J Agric Food Chem ; 69(4): 1379-1390, 2021 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-33464885

RESUMO

A high content of potentially allergenic lectin in Phaseolus vulgaris L. beans is of increasing health concerns; however, understanding of the protein allergenicity mechanism on the molecular basis is scarce. In the present study, low-pH treatments were applied to modify black turtle bean lectin allergen, and a sensitization procedure was performed using the BALB/c mice for the allergenicity evaluation, while the conformational changes were monitored by the spectral analyses and the details were explored by the molecular dynamics simulation. Much milder anaphylactic responses were observed in BALB/c mice experiments. At the molecular level, the protein was unfolded in low acidic environments because of protonation, and α-helix was reduced with the exposure of trypsin cleavage sites, especially the improvement of protease accessibility for Lys121, 134, and 157 in the B cell epitope structural alterations. These results indicate that a low-pH treatment might be an efficient method to improve the safety of legume protein consumption.


Assuntos
Alérgenos/química , Lectinas/química , Phaseolus/imunologia , Alérgenos/imunologia , Animais , Linfócitos B/imunologia , Feminino , Manipulação de Alimentos , Hipersensibilidade Alimentar/imunologia , Humanos , Concentração de Íons de Hidrogênio , Lectinas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Phaseolus/genética , Sementes/química , Sementes/imunologia
3.
Nat Commun ; 12(1): 702, 2021 01 29.
Artigo em Inglês | MEDLINE | ID: mdl-33514713

RESUMO

Lima bean (Phaseolus lunatus L.), one of the five domesticated Phaseolus bean crops, shows a wide range of ecological adaptations along its distribution range from Mexico to Argentina. These adaptations make it a promising crop for improving food security under predicted scenarios of climate change in Latin America and elsewhere. In this work, we combine long and short read sequencing technologies with a dense genetic map from a biparental population to obtain the chromosome-level genome assembly for Lima bean. Annotation of 28,326 gene models show high diversity among 1917 genes with conserved domains related to disease resistance. Structural comparison across 22,180 orthologs with common bean reveals high genome synteny and five large intrachromosomal rearrangements. Population genomic analyses show that wild Lima bean is organized into six clusters with mostly non-overlapping distributions and that Mesomerican landraces can be further subdivided into three subclusters. RNA-seq data reveal 4275 differentially expressed genes, which can be related to pod dehiscence and seed development. We expect the resources presented here to serve as a solid basis to achieve a comprehensive view of the degree of convergent evolution of Phaseolus species under domestication and provide tools and information for breeding for climate change resiliency.


Assuntos
Aclimatação/genética , Produtos Agrícolas/genética , Phaseolus/genética , Melhoramento Vegetal , Locos de Características Quantitativas , Argentina , Mapeamento Cromossômico , Mudança Climática , Domesticação , Genes de Plantas/genética , México , Dispersão Vegetal , RNA-Seq , Sementes , Sintenia
4.
Mol Genet Genomics ; 296(2): 379-390, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33449160

RESUMO

Common bean (Phaseolus vulgaris L.) is a short-day plant and its flowering time, and consequently, pod yield and quality is influenced by photoperiod. In this study, the photoperiodic-sensitive variety 'Hong jin gou', which flowers 31 days (d) earlier in short-day than in long-day, was used as the experimental material. Samples were collected to determine the growth and photosynthetic parameters in each daylength treatment, and transcriptome and metabolome data were conducted. We identified eight genes related to flowering by further screening for differentially expressed genes. These genes function to regulate the biological clock. The combination of differentially expressed genes and metabolites, together with the known regulation network of flowering time and the day-night expression pattern of related genes allow us to speculate on the regulation of flowering time in the common bean and conclude that TIMING OF CAB EXPRESSION1 (TOC1) plays a pivotal role in the network and its upregulation or downregulation causes corresponding changes in the expression of downstream genes. The regulatory network is also influenced by gibberellic acid (GA) and jasmonic acid (JA). These regulatory pathways jointly comprise the flowering regulatory network in common bean.


Assuntos
Perfilação da Expressão Gênica/métodos , Metabolômica/métodos , Phaseolus/fisiologia , Fatores de Transcrição/genética , Relógios Biológicos , Flores/fisiologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Redes Reguladoras de Genes/efeitos dos fármacos , Giberelinas/farmacologia , Ácidos Linoleicos/farmacologia , Phaseolus/genética , Phaseolus/metabolismo , Fotoperíodo , Proteínas de Plantas/genética , Análise de Sequência de RNA
5.
Food Chem ; 339: 127917, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32950898

RESUMO

Andean beans (Phaseolus vulgaris) chemical compositions and cooking characteristics contribute to a healthy diet. The objective of this study was to evaluate the influence of chemical composition on the cooking quality of 14 Andean beans genotypes with different seed coat colors. More specifically, water retention (WR), cooking time (CT), and solids released in the broth, were analysed. WR values ranged from 128.4% to 160.7% and CT ranged from 13.7 (BRS Embaixador) to 21.7 min (KID44). Andean beans showed variability in chemical composition, mainly starch content (39.43 g 100 g-1, BRS Realce to 51.92 g 100 g-1, LP15-04) and polymer composition. The profile of starch and interactions among minerals and chemical compounds influenced the cooking profiles than do the individual compounds. Andean beans traits of cooking, mainly CT, were influenced by their chemical composition; however they can be incorporated into diets without drastic changes in preparation methods.


Assuntos
Culinária/métodos , Phaseolus/química , Amido/análise , Amilopectina/análise , Amilose/análise , Genótipo , Minerais/análise , Análise Multivariada , Phaseolus/genética , Phaseolus/metabolismo , Sementes/química , Sementes/genética , Sementes/metabolismo
6.
PLoS One ; 15(12): e0243238, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33338076

RESUMO

Knowledge of genetic diversity in plant germplasm and the relationship between genetic factors and phenotypic expression is vital for crop improvement. This study's objectives were to understand the extent of genetic diversity and population structure in 60 common bean genotypes from East and Southern Africa. The common bean genotypes exhibited significant (p<0.05) levels of variability for traits such as days to flowering (DTF), days to maturity (DTM), number of pods per plant (NPP), number of seeds per pod (NSP), and grain yield per hectare in kilograms (GYD). About 47.82 per cent of the variation among the genotypes was explained by seven principal components (PC) associated with the following agronomic traits: NPP, NFF (nodes to first flower), DTF, GH (growth habit) and GYD. The SNP markers revealed mean gene diversity and polymorphic information content values of 0.38 and 0.25, respectively, which suggested the presence of considerable genetic variation among the assessed genotypes. Analysis of molecular variance showed that 51% of the genetic variation were between the gene pools, while 49% of the variation were within the gene pools. The genotypes were delineated into two distinct groups through the population structure, cluster and phylogenetic analyses. Genetically divergent genotypes such as DRK57, MW3915, NUA59, and VTTT924/4-4 with high yield and agronomic potential were identified, which may be useful for common bean improvement.


Assuntos
Grão Comestível/genética , Phaseolus/genética , Sementes/genética , África Oriental , África Austral , Grão Comestível/anatomia & histologia , Variação Genética , Genótipo , Phaseolus/anatomia & histologia , Fenótipo , Polimorfismo de Nucleotídeo Único , Sementes/anatomia & histologia
7.
PLoS One ; 15(10): e0239763, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33027258

RESUMO

Anthracnose, caused by the fungal pathogen Colletotrichum lindemuthianum, is one of the world's most destructive diseases of common bean. The use of resistant cultivars is the most cost-effective strategy to manage this disease; however, durable resistance is difficult to achieve due to the vast virulence diversity of the anthracnose pathogen. Finding new genes with broad-spectrum resistance increases the prospect of designing an effective anthracnose-management strategy. Genetic analysis confirmed the presence of a single, dominant anthracnose-resistance locus in AC, which we provisionally named Co-AC. Bulk segregant analysis and genetic mapping of two F2 populations from the crosses AC × PI207262 and AC × G 2333 were used to determine the position of the Co-AC locus in a 631 Kbp genomic region flanked by the SNP markers SS56 and SS92 on the lower arm of chromosome Pv01. By genotyping 77 F3 plants from the AC × PI207262 cross using nine additional markers, we fine-mapped the Co-AC locus to a significantly smaller genomic region (9.4 Kbp) flanked by the SNP markers SS102 and SS165. This 9.4 Kbp region harbors three predicted genes based on the common bean reference genome, notably including the gene model Phvul.001G244300, which encodes Clathrin heavy chain 1, a protein that supports specific stomatal regulation functions and might play a role in plant defense signaling. Because the Co-AC resistance locus is linked in cis, it can be selected with great efficiency using molecular markers. These results will be very useful for breeding programs aimed at developing bean cultivars with anthracnose resistance using marker-assisted selection. This study revealed the broad-spectrum resistance of AC to C. lindemuthianum and the existence of the Co-AC anthracnose-resistance locus. Fine mapping positioned this locus in a small genomic region on the lower end of chromosome Pv01 that contained three candidate genes for the Co-AC locus.


Assuntos
Resistência à Doença/genética , Phaseolus/genética , Cruzamento/métodos , Mapeamento Cromossômico/métodos , Colletotrichum/patogenicidade , Genes de Plantas/genética , Ligação Genética/genética , Marcadores Genéticos/genética , Genótipo , Phaseolus/microbiologia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Polimorfismo de Nucleotídeo Único/genética
8.
PLoS One ; 15(9): e0239263, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32986739

RESUMO

Mineral deficiency is worldwide one of the major problems associated with human health, and biofortification through breeding is considered an important strategy to improve the nutritional content of staple food in countries that face this problem. The assessment of genetic variability for seed nutrient contents is a first step in the development of a biofortified crop. From the germplasm bank IDR-IAPAR-EMATER, a set of 1,512 common bean accessions, consisting of local and commercial varieties and improved lines, was analyzed. High variability among the accessions was observed for all evaluated nutrient contents (P, K, Ca, Mg, Cu, Zn, Mn, Fe and S and protein). In the mean, the contents of the carioca and black market groups (Mesoamerican gene pool), were around 7% higher for the minerals Ca, Cu, Mn and Fe and between 2-4% higher for P, K, Mg and Zn than in the other groups with Mesoamerican and Andean common bean. Few differences were observed among the Mesoamerican accessions that belong to the carioca and black commercial groups. Wide variability was observed among the evaluated genotypes, and the concentrations of the best accessions exceeded the overall mean by 14-28%. Due to the high variability in the evaluated accessions, these results may contribute to the selection of promising parents for the establishment of mating blocks. The nutritional contents of many of the improved lines evaluated in this study were higher than those of the commercial cultivars, indicating the possibility of developing new biofortified cultivars.


Assuntos
Biofortificação , Phaseolus/química , Banco de Sementes , Sementes/química , Brasil , Cruzamento , Genótipo , Humanos , Ferro/química , Minerais/química , Phaseolus/genética , Sementes/genética
9.
Mol Plant Microbe Interact ; 33(9): 1161-1175, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32633604

RESUMO

Halo blight disease of beans is caused by a gram-negative bacterium, Pseudomonas syringae pv. phaseolicola. The disease is prevalent in South America and Africa and causes crop loss for indigent people who rely on beans as a primary source of daily nutrition. In susceptible beans, P. syringae pv. phaseolicola causes water-soaking at the site of infection and produces phaseolotoxin, an inhibitor of bean arginine biosynthesis. In resistant beans, P. syringae pv. phaseolicola triggers a hypersensitive response that limits the spread of infection. Here, we used high-throughput mass spectrometry to interrogate the responses to two different P. syringae pv. phaseolicola isolates on a single line of common bean, Phaseolus vulgaris PI G19833, with a reference genome sequence. We obtained quantitative information for 4,135 bean proteins. A subset of 160 proteins with similar accumulation changes during both susceptible and resistant reactions included salicylic acid responders EDS1 and NDR1, ethylene and jasmonic acid biosynthesis enzymes, and proteins enabling vesicle secretion. These proteins revealed the activation of a basal defense involving hormonal responses and the mobilization of extracellular proteins. A subset of 29 proteins specific to hypersensitive immunity included SOBIR1, a G-type lectin receptor-like kinase, and enzymes needed for glucoside and phytoalexin production. Virus-induced gene silencing revealed that the G-type lectin receptor-like kinase suppresses bacterial infection. Together, the results define the proteomics of disease resistance to P. syringae pv. phaseolicola in beans and support a model whereby the induction of hypersensitive immunity reinstates defenses targeted by P. syringae pv. phaseolicola.


Assuntos
Resistência à Doença/genética , Phaseolus/genética , Doenças das Plantas/genética , Proteômica , Pseudomonas syringae/patogenicidade , Genoma de Planta , Phaseolus/microbiologia , Doenças das Plantas/microbiologia
10.
Mol Genet Genomics ; 295(6): 1325-1337, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32607601

RESUMO

The main edible organ of snap bean (Phaseolus vulgaris L.) is the pod, whose color is a main characteristic affecting its commercial use. Golden pods are popular with consumers; however, color instability affects their commercial exploitation and causes economic losses to the planters. In this study, we focused on the different pod color of two varieties of snap bean. The golden yellow color of snap bean pods is controlled by a single recessive nuclear gene located at 1-4.24 Mb of chromosome 2. To explore the physiological and molecular mechanism of the golden pod color, the golden bean line 'A18-1' and the green bean line 'Renaya' were selected as experimental materials. We analyzed the pigment contents, detected the intermediate products of chlorophyll biosynthesis, and identified differentially expressed genes using RNA-seq. The formation of golden bean pods reflects a chlorophyll deficiency, which was speculated to be caused by impairment of the Mg-protoporphyrin IX to chlorophyllide step. In 'A18-1' and 'Renaya' pods on 10, 14, and 18 days, five genes related to this step were differentially expressed, all of which were protochlorophyllide oxidoreductase (POR) genes. Among them, the expression changes of the Phvul. 004G112700, Phvul.007G157500, and Phvul. 004G112400 genes were consistent with the color change and physiological data during pod development in 'A18-1' and 'Renaya'. We speculated that the altered expression of these three POR genes might be related to changes in the chlorophyllide content. The results might provide insight into the understanding of chlorophyll biosynthesis and crop breeding for snap bean.


Assuntos
Clorofila/metabolismo , Regulação da Expressão Gênica de Plantas , Phaseolus/metabolismo , Proteínas de Plantas/metabolismo , Sementes/metabolismo , Vicia faba/metabolismo , Cor , Phaseolus/genética , Phaseolus/crescimento & desenvolvimento , Proteínas de Plantas/genética , Sementes/genética , Sementes/crescimento & desenvolvimento , Transcriptoma , Vicia faba/genética , Vicia faba/crescimento & desenvolvimento
11.
PLoS One ; 15(6): e0235215, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32598372

RESUMO

Anthracnose (ANT) and angular leaf spot (ALS) caused by Colletotrichum lindemuthianum and Pseudocercospora griseola, respectively, are devastating diseases of common bean around the world. Therefore, breeders are constantly searching for new genes with broad-spectrum resistance against ANT and ALS. This study aimed to characterize the genetic resistance of California Dark Red Kidney (CDRK) to C. lindemuthianum races 73, 2047, and 3481 and P. griseola race 63-39 through inheritance, allelism testing, and molecular analyses. Genetic analysis of response to ANT and ALS in recombinant inbred lines (RILs) from a CDRK × Yolano cross (CY) showed that the resistance of CDRK cultivar is conferred by a single dominant loci, which we named CoPv01CDRK/PhgPv01CDRK. Allelism tests performed with race 3481showed that the resistance gene in CDRK is independent of the Co-1 and Co-AC. We conducted co-segregation analysis in genotypes of 110 CY RILs and phenotypes of the RILs in response to different races of the ANT and ALS pathogens. The results revealed that CoPv01CDRK and PhgPv01CDRK are coinherited, conferring resistance to all races. Genetic mapping of the CY population placed the CoPv01CDRK/PhgPv01CDRK loci in a 245 Kb genomic region at the end of Pv01. By genotyping 19 RILs from the CY population using three additional markers, we fine-mapped the CoPv01CDRK/PhgPv01CDRK loci to a smaller genomic region of 33 Kb. This 33 Kb region harbors five predicted genes based on the common bean reference genome. These results can be applied in breeding programs to develop bean cultivars with ANT and ALS resistance using marker-assisted selection.


Assuntos
Colletotrichum/fisiologia , Resistência à Doença/genética , Genes de Plantas , Ligação Genética , Marcadores Genéticos , Phaseolus/genética , Doenças das Plantas/genética , California , Mapeamento Cromossômico , Genótipo , Phaseolus/microbiologia , Fenótipo , Doenças das Plantas/microbiologia
12.
Phytopathology ; 110(8): 1428-1436, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32301679

RESUMO

Common bean (Phaseolus vulgaris) is one of the most consumed agricultural products in the world. Its production is affected by common bacterial blight (CBB) caused by Xanthomonas citri pv. fuscans and X. phaseoli pv. phaseoli. In this work, we investigated the spectrum, genetics, and inheritance of common bean resistance to X. citri pv. fuscans. Inoculation of nine selected cultivars with an X. citri pv. fuscans strain showed that BRS Radiante and IAPAR 16 were resistant. These two cultivars were also resistant to six X. phaseoli pv. phaseoli strains of different geographic origins, demonstrating their broad-spectrum resistances. BRS Radiante sustained smaller X. citri pv. fuscans populations than two susceptible cultivars. Stomatal densities of IAPAR 16 and BRS Radiante were significantly higher than or not different from susceptible cultivars. BRS Radiante showed the lowest general combining ability values and the combination BRS Radiante × Carioca MG the lowest specific combining ability (SCA) values, revealing the capacity of BRS Radiante to increase resistance to X. citri pv. fuscans. Positive and negative parental SCA values indicated dominant and recessive genes involved in X. citri pv. fuscans resistance. Resistance of the BRS Radiante × Carioca MG cross segregated in a 9:7 ratio in the F2 population, indicating that it is governed by two complementary dominant genes. Maximum likelihood analysis showed that the resistance of BRS Radiante to X. citri pv. fuscans is conferred by a gene of major effect with contribution of additional polygenes. This study contributes with important knowledge on the resistance against CBB in Brazilian common bean cultivars as well as with molecular tools for confirmation of common bean hybrids.


Assuntos
Phaseolus/genética , Xanthomonas/genética , Brasil , DNA Bacteriano , Doenças das Plantas
13.
Food Chem ; 321: 126680, 2020 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-32247181

RESUMO

Seed phytic acid reduces mineral bioavailability by chelating minerals. Consumption of common bean seeds with the low phytic acid 1 (lpa1) mutation improved iron status in human trials but caused adverse gastrointestinal effects, presumably due to increased stability of lectin phytohemagglutinin L (PHA-L) compared to the wild type (wt). A hard-to-cook (HTC) defect observed in lpa1 seeds intensified this problem. We quantified the HTC phenotype of lpa1 common beans with three genetic backgrounds. The HTC phenotype in the lpa1 black bean line correlated with the redistribution of calcium particularly in the cell walls, providing support for the "phytase-phytate-pectin" theory of the HTC mechanism. Furthermore, the excess of free cations in the lpa1 mutation in combination with different PHA alleles affected the stability of PHA-L lectin.


Assuntos
Cálcio/química , Lectinas/química , Phaseolus/química , Ácido Fítico/química , Fito-Hemaglutininas/química , Culinária , Dureza , Temperatura Alta , Mutação , Phaseolus/genética , Sementes/química , Sementes/genética
14.
Plant Sci ; 294: 110445, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32234228

RESUMO

Phosphate acquisition by plants is an essential process that is directly implicated in the optimization of crop yields. Purple acid phosphatases (PAPs) are ubiquitous metalloenzymes, which catalyze the hydrolysis of a wide range of phosphate esters and anhydrides. While some plant PAPs display a preference for ATP as the substrate, others are efficient in hydrolyzing phytate or 2-phosphoenolpyruvate (PEP). PAP from red kidney bean (rkbPAP) is an efficient ATP- and ADPase, but has no activity towards phytate. Crystal structures of this enzyme in complex with ATP analogues (to 2.20 and 2.60 Å resolution, respectively) complement the recent structure of rkbPAP with a bound ADP analogue (ChemBioChem 20 (2019) 1536). Together these complexes provide the first structural insight of a PAP in complex with molecules that mimic biologically relevant substrates. Homology modeling was used to generate three-dimensional structures for the active sites of PAPs from tobacco (NtPAP) and thale cress (AtPAP26) that are efficient in hydrolyzing phytate and PEP as preferred substrates, respectively. The combining of crystallographic data, substrate docking simulations and a phylogenetic analysis of 49 plant PAP sequences (including the first PAP sequences reported from Eucalyptus) resulted in the identification of several active site residues that are important in defining the substrate specificities of plant PAPs; of particular relevance is the identification of a motif ("REKA") that is characteristic for plant PAPs that possess phytase activity. These results may inform bioengineering studies aimed at identifying and incorporating suitable plant PAP genes into crops to improve phosphorus acquisition and use efficiency. Organic phosphorus sources increasingly supplement or replace inorganic fertilizer, and efficient phosphorus use of crops will lower the environmental footprint of agriculture while enhancing food production.


Assuntos
Fosfatase Ácida/metabolismo , Fosfatase Ácida/genética , Bioengenharia/métodos , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Phaseolus/genética , Phaseolus/metabolismo , Especificidade por Substrato
15.
Plant Physiol Biochem ; 151: 77-87, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32200193

RESUMO

Molybdenum is considered one of the most important micronutrients applied as a foliar fertilizer for common dry bean. In this study, molybdenum oxide nanoparticles (MoO3-NPs) were applied in different concentrations (0, 10, 20, 30 and 40 ppm) over two sequent seasons, 2018 and 2019, to investigate their effect on the plant morphological criteria, yield, and the genomic stability of DNA. The results showed that the application of 40 ppm MoO3-NPs as a foliar fertilizer showed preferable values of plant morphological criteria, such as the number of leaves and branches per plant, as well as the fresh and dry weight with regard to the common bean plant. In addition, the seed yield increased by 82.4% and 84.1% with 40 ppm, while the shoot residue increased by 32.2% and 32.1% with 20 ppm of MoO3-NPs during two seasons, 2018 and 2019, respectively. Furthermore, the common bean treated with 20 and 40 ppm MoO3-NPs had positive unique bands with ISSR primer 848 at 1400 bp (Rf 0.519) and with primer ISSR2M at 200 bp (Rf 0.729), respectively. In addition, SDS-PAGE reveald some proteins in seedlings which were absent in the flowering stage at 154, 102, 64, 37 and 34 KDa, which may be due to differences in plant proteins required for metabolic processes in each stage. In conclusion, the application of 40 ppm MoO3-NPs was more effective on the productivity of the common bean plants.


Assuntos
Fertilizantes , Instabilidade Genômica , Molibdênio/administração & dosagem , Phaseolus/crescimento & desenvolvimento , Phaseolus/genética , DNA de Plantas/genética , Nanopartículas Metálicas , Óxidos/administração & dosagem , Folhas de Planta , Plântula
16.
Int J Mol Sci ; 21(6)2020 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-32183068

RESUMO

Actin plays a critical role in the rhizobium-legume symbiosis. Cytoskeletal rearrangements and changes in actin occur in response to Nod factors secreted by rhizobia during symbiotic interactions with legumes. These cytoskeletal rearrangements are mediated by diverse actin-binding proteins, such as actin depolymerization factors (ADFs). We examined the function of an ADF in the Phaseolus vulgaris-rhizobia symbiotic interaction (PvADFE). PvADFE was preferentially expressed in rhizobia-inoculated roots and nodules. PvADFE promoter activity was associated with root hairs harbouring growing infection threads, cortical cell divisions beneath root hairs, and vascular bundles in mature nodules. Silencing of PvADFE using RNA interference increased the number of infection threads in the transgenic roots, resulting in increased nodule number, nitrogen fixation activity, and average nodule diameter. Conversely, overexpression of PvADFE reduced the nodule number, nitrogen fixation activity, average nodule diameter, as well as NODULE INCEPTION (NIN) and EARLY NODULIN2 (ENOD2) transcript accumulation. Hence, changes in ADFE transcript levels affect rhizobial infection and nodulation, suggesting that ADFE is fine-tuning these processes.


Assuntos
Fatores de Despolimerização de Actina/metabolismo , Phaseolus/metabolismo , Proteínas de Plantas/metabolismo , Rhizobium/metabolismo , Fatores de Despolimerização de Actina/genética , Fixação de Nitrogênio , Phaseolus/genética , Phaseolus/microbiologia , Proteínas de Plantas/genética , Rhizobium/genética
17.
PLoS One ; 15(3): e0229909, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32134988

RESUMO

Stable transformation of common bean (Phaseolus vulgaris L.) has been successful, to date, only using biolistic-mediated transformation and shoot regeneration from meristem-containing embryo axes. In this study, using precultured embryo axes, and optimal co-cultivation conditions resulted in a successful transformation of the common bean cultivar Olathe using Agrobacterium tumefaciens strain EHA105. Plant regeneration through somatic embryogenesis was attained through the preculture of embryo axes for 12 weeks using induced competent cells for A. tumefaciens-mediated gene delivery. Using A. tumefaciens at a low optical density (OD) of 0.1 at a wavelength of 600 nm for infection and 4-day co-cultivation, compared to OD600 of 0.5, increased the survival rate of the inoculated explants from 23% to 45%. Selection using 0.5 mg L-1 glufosinate (GS) was effective to identify transformed cells when the bialaphos resistance (bar) gene under the constitutive 35S promoter was used as a selectable marker. After an 18-week selection period, 1.5% -2.5% inoculated explants, in three experiments with a total of 600 explants, produced GS-resistant plants through somatic embryogenesis. The expression of bar was confirmed in first- and second-generation seedlings of the two lines through reverse polymerase chain reaction. Presence of the bar gene was verified through genome sequencing of two selected transgenic lines. The induction of regenerable, competent cells is key for the successful transformation, and the protocols described may be useful for future transformation of additional Phaseolus germplasm.


Assuntos
Agrobacterium tumefaciens/genética , Phaseolus/genética , Melhoramento Vegetal/métodos , Plantas Geneticamente Modificadas/genética , Transformação Genética , Agrobacterium tumefaciens/efeitos dos fármacos , Aminobutiratos/farmacologia , DNA de Plantas/genética , Farmacorresistência Bacteriana/genética , Vetores Genéticos , Herbicidas/farmacologia , Compostos Organofosforados/farmacologia , Fenótipo , RNA de Plantas/genética
18.
Sci Rep ; 10(1): 1680, 2020 02 03.
Artigo em Inglês | MEDLINE | ID: mdl-32015448

RESUMO

The investigation of substrate spectrum towards five racemic (rac-) aryl glycidyl ethers (1a-5a) indicated that E. coli/pveh3, an E. coli BL21(DE3) transformant harboring a PvEH3-encoding gene pveh3, showed the highest EH activity and enantiomeric ratio (E) towards rac-3a. For efficiently catalyzing the kinetic resolution of rac-3a, the activity and E value of PvEH3 were further improved by site-directed mutagenesis of selected residues. Based on the semi-rational design of an NC-loop in PvEH3, four single-site variants of pveh3 were amplified by PCR, and intracellularly expressed in E. coli BL21(DE3), respectively. E. coli/pveh3E134K and /pveh3T137P had the enhanced EH activities of 15.3 ± 0.4 and 16.1 ± 0.5 U/g wet cell as well as E values of 21.7 ± 1.0 and 21.2 ± 1.1 towards rac-3a. Subsequently, E. coli/pveh3E134K/T137P harboring a double-site variant gene was also constructed, having the highest EH activity of 22.4 ± 0.6 U/g wet cell and E value of 24.1 ± 1.2. The specific activity of the purified PvEH3E134K/T137P (14.5 ± 0.5 U/mg protein) towards rac-3a and its catalytic efficiency (kcat/Km of 5.67 mM-1 s-1) for (S)-3a were 1.7- and 3.54-fold those (8.4 ± 0.3 U/mg and 1.60 mM-1 s-1) of PvEH3. The gram-scale kinetic resolution of rac-3a using whole wet cells of E. coli/pveh3E134K/T137P was performed at 20 °C for 7.0 h, producing (R)-3a with 99.4% ees and 38.5 ± 1.2% yield. Additionally, the mechanism of PvEH3E134K/T137P with remarkably improved E value was analyzed by molecular docking simulation.


Assuntos
Cresóis/metabolismo , Epóxido Hidrolases/metabolismo , Compostos de Epóxi/metabolismo , Phaseolus/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Catálise , Epóxido Hidrolases/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Cinética , Simulação de Acoplamento Molecular/métodos , Mutagênese Sítio-Dirigida/métodos , Phaseolus/genética , Estereoisomerismo , Especificidade por Substrato/fisiologia
19.
PLoS One ; 15(2): e0228680, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32017794

RESUMO

Common bean (Phaseolus vulgaris L.) is a major source of proteins and one of the most important edible foods for more than three hundred million people in the world. The common bean plants are frequently attacked by spider mite (Tetranychus urticae Koch), leading to a significant decrease in plant growth and economic performance. The use of resistant cultivars and the identification of the genes involved in plant-mite resistance are practical solutions to this problem. Hence, a comprehensive study of the molecular interactions between resistant and susceptible common bean cultivars and spider mite can shed light into the understanding of mechanisms and biological pathways of resistance. In this study, one resistant (Naz) and one susceptible (Akhtar) cultivars were selected for a transcriptome comparison at different time points (0, 1 and 5 days) after spider mite feeding. The comparison of cultivars in different time points revealed several key genes, which showed a change increase in transcript abundance via spider mite infestation. These included genes involved in flavonoid biosynthesis process; a conserved MYB-bHLH-WD40 (MBW) regulatory complex; transcription factors (TFs) TT2, TT8, TCP, Cys2/His2-type and C2H2-type zinc finger proteins; the ethylene response factors (ERFs) ERF1 and ERF9; genes related to metabolism of auxin and jasmonic acid (JA); pathogenesis-related (PR) proteins and heat shock proteins.


Assuntos
Ácaros/patogenicidade , Phaseolus/imunologia , Transcriptoma , Animais , Perfilação da Expressão Gênica , Genes de Plantas , Interações Hospedeiro-Patógeno , Infestações por Ácaros , Phaseolus/genética , Phaseolus/parasitologia , Fatores de Tempo
20.
Sci Rep ; 10(1): 3628, 2020 02 27.
Artigo em Inglês | MEDLINE | ID: mdl-32107403

RESUMO

Common bean variety choice by farmers in Uganda is driven by seed yield plus end-use quality traits like market class and cooking time. Limited genotype by environment information is available for traits valued by consumers. This research evaluated yield, seed size, hydration properties, and cooking time of 15 common bean genotypes within market classes recognized by consumers along with three farmers' checks at nine on-farm locations in Uganda for two seasons. Yield ranged from 71 to 3,216 kg ha-1 and was largely controlled by location (21.5% of Total Sums of Squares [TSS]), plus the interaction between location and season (48.6% of TSS). Cooking time varied from 19 to 271 minutes with the genotypes Cebo Cela and Ervilha consistently cooking fastest in 24 and 27 minutes respectively. Comparatively, the local checks (NABE-4, NABE-15, and Masindi yellow) took 35 to 45 minutes to cook. Cooking time was largely controlled by genotype (40.6% of TSS). A GGE biplot analysis uncovered the presence of two mega-environments for yield and one mega-environment for cooking time. Identification of mega-environments for these traits will help expedite common bean breeding, evaluation, and variety selection through reduction of number of test environments needed for phenotype evaluations. The high yielding and fast cooking genotypes from this study can be targeted as parental materials to improve existing common bean germplasm for these important traits.


Assuntos
Phaseolus/genética , Sementes/química , Cruzamento , Culinária , Fazendas , Interação Gene-Ambiente , Genótipo , Phaseolus/química , Phaseolus/classificação , Phaseolus/crescimento & desenvolvimento , Fenótipo , Sementes/classificação , Sementes/genética , Sementes/crescimento & desenvolvimento , Uganda
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