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1.
Mol Genet Genomics ; 299(1): 28, 2024 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-38472470

RESUMO

Edaphic conditions of serpentine soils, naturally rich in heavy metals, act as a strong selection pressure that shapes specific metal-tolerant ecotypes. Medicago lupulina L. (black medick) is not only a widespread plant species that prefers calcareous and dry soil types but also grows at the borders of serpentine formations. It can also be found in waste and disturbed habitats. This is a species with reported phytoremediation potential, however, there is no published data regarding the impact of the environment on the genetic distribution of this species. The aim of our research was to explore how selection pressure of serpentine soils affects genetic diversity of M. lupulina and to test heavy-metal accumulation capacity of this species. Specimens of 11 M. lupulina populations were collected from serpentine outcrops located in Central and Eastern Bosnia as well as from non-serpentine sites. Soil and plant samples were analyzed for the total contents of heavy metals using air-acetylene flame atomic absorption spectroscopy. Genetic diversity was analyzed using AFLP (Amplified Fragment Length Polymorphism) markers. Serpentine soils showed high nickel, cobalt, chromium and iron concentrations. Nickel and manganese concentrations in soil samples and plant material showed statistically significant correlation. Although plants in two populations show the ability to extract Ni, M. lupulina does not show hyperaccumulating properties. Despite severe selective pressure, genetic diversity in serpentine populations is not reduced. Analyses of intrapopulation and interpopulation genetic diversity showed significant genetic differentiation among populations which is not related to their geographic distance. Population from non-metalliferous soil showed clear separation from all other populations. Diversity data suggest that serpentine populations maintain genetic diversity by undetected mechanisms and that edaphic factors rather than geography influence genetic structure analyzed M. lupulina populations.


Assuntos
Metais Pesados , Níquel , Níquel/análise , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Medicago , Metais Pesados/análise , Plantas , Variação Genética , Solo/química
2.
Heredity (Edinb) ; 132(2): 106-116, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38233486

RESUMO

Changes in epigenetic states can allow individuals to cope with environmental changes. If such changes are heritable, this may lead to epigenetic adaptation. Thus, it is likely that in sessile organisms such as plants, part of the spatial epigenetic variation found across individuals will reflect the environmental heterogeneity within populations. The departure of the spatial epigenetic structure from the baseline genetic variation can help in understanding the value of epigenetic regulation in species with different breadth of optimal environmental requirements. Here, we hypothesise that in plants with narrow environmental requirements, epigenetic variability should be less structured in space given the lower variability in suitable environmental conditions. We performed a multispecies study that considered seven pairs of congeneric plant species, each encompassing a narrow endemic with habitat specialisation and a widespread species. In three populations per species we used AFLP and methylation-sensitive AFLP markers to characterise the spatial genetic and epigenetic structures. Narrow endemics showed a significantly lower epigenetic than genetic differentiation between populations. Within populations, epigenetic variation was less spatially structured than genetic variation, mainly in narrow endemics. In these species, structural equation models revealed that such pattern was associated to a lack of correlation between epigenetic and genetic information. Altogether, these results show a greater decoupling of the spatial epigenetic variation from the baseline spatial genetic pattern in endemic species. These findings highlight the value of studying genetic and epigenetic spatial variation to better understand habitat specialisation in plants.


Assuntos
Epigênese Genética , Variação Genética , Humanos , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Metilação de DNA , Ecossistema
3.
Mol Ecol ; 33(6): e17283, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38288572

RESUMO

Avian embryos develop in an egg composition which reflects both maternal condition and the recent environment of their mother. In birds, yolk corticosterone (CORT) influences development by impacting pre- and postnatal growth, as well as nestling stress responses and development. One possible mechanism through which maternal CORT may affect offspring development is via changes to offspring DNA methylation. We sought to investigate this, for the first time in birds, by quantifying the impact of manipulations to maternal CORT on offspring DNA methylation. We non-invasively manipulated plasma CORT concentrations of egg-laying female zebra finches (Taeniopygia castanotis) with an acute dose of CORT administered around the time of ovulation and collected their eggs. We then assessed DNA methylation in the resulting embryonic tissue and in their associated vitelline membrane blood vessels, during early development (5 days after lay), using two established methods - liquid chromatography-mass spectrometry (LC-MS) and methylation-sensitive amplification fragment length polymorphism (MS-AFLP). LC-MS analysis showed that global DNA methylation was lower in embryos from CORT-treated mothers, compared to control embryos. In contrast, blood vessel DNA from eggs from CORT-treated mothers showed global methylation increases, compared to control samples. There was a higher proportion of global DNA methylation in the embryonic DNA of second clutches, compared to first clutches. Locus-specific analyses using MS-AFLP did not reveal a treatment effect. Our results indicate that an acute elevation of maternal CORT around ovulation impacts DNA methylation patterns in their offspring. This could provide a mechanistic understanding of how a mother's experience can affect her offspring's phenotype.


Assuntos
Corticosterona , Passeriformes , Animais , Feminino , Corticosterona/farmacologia , Corticosterona/análise , Metilação de DNA , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , DNA
4.
Plant Physiol ; 194(2): 787-804, 2024 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-37815230

RESUMO

Root development influences plant responses to environmental conditions, and well-developed rooting enhances plant survival under abiotic stress. However, the molecular and genetic mechanisms underlying root development and abiotic stress tolerance in plants remain unclear. In this study, we identified the MYB transcription factor-encoding gene IbMYB73 by cDNA-amplified fragment length polymorphism and RNA-seq analyses. IbMYB73 expression was greatly suppressed under abiotic stress in the roots of the salt-tolerant sweet potato (Ipomoea batatas) line ND98, and its promoter activity in roots was significantly reduced by abscisic acid (ABA), NaCl, and mannitol treatments. Overexpression of IbMYB73 significantly inhibited adventitious root growth and abiotic stress tolerance, whereas IbMYB73-RNAi plants displayed the opposite pattern. IbMYB73 influenced the transcription of genes involved in the ABA pathway. Furthermore, IbMYB73 formed homodimers and activated the transcription of ABA-responsive protein IbGER5 by binding to an MYB binding sites I motif in its promoter. IbGER5 overexpression significantly inhibited adventitious root growth and abiotic stress tolerance concomitantly with a reduction in ABA content, while IbGER5-RNAi plants showed the opposite effect. Collectively, our results demonstrated that the IbMYB73-IbGER5 module regulates ABA-dependent adventitious root growth and abiotic stress tolerance in sweet potato, which provides candidate genes for the development of elite crop varieties with well-developed root-mediated abiotic stress tolerance.


Assuntos
Ácido Abscísico , Ipomoea batatas , Ácido Abscísico/farmacologia , Ácido Abscísico/metabolismo , Ipomoea batatas/genética , Ipomoea batatas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Estresse Fisiológico/fisiologia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
5.
Comp Immunol Microbiol Infect Dis ; 104: 102100, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38043450

RESUMO

Microsporum canis is considered the common dermatophyte agent associated with ringworm in felines and canines. In the present study, we sampled n = 548 felines and canines for the probable isolation of M. canis. The rate of isolation from the cats and dogs was 70.27 % (52/74) and 1.68 % (8/474), respectively and Persian cats were found to be highly susceptible to M. canis infection. The strains were evaluated for their production of phospholipase, lipase, catalase, and hemolysis and their ability to grow at 35 â„ƒ. All the strains were identified as low producers of catalase and n = 17 strains exhibited high thermotolerance ability. Terbinafine was found to be the most effective antifungal drug and fluconazole was the least effective, in vitro. AFLP analysis revealed three genotypes of M. canis with 15 sub-clusters showing ≥ 90 % similarity and 7 sub-clusters exhibiting 100 % similarity. However, the phenotypic characters cannot be attributed based on the AFLP profiles.


Assuntos
Doenças do Gato , Dermatomicoses , Doenças do Cão , Animais , Gatos , Cães , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Catalase/farmacologia , Dermatomicoses/tratamento farmacológico , Dermatomicoses/microbiologia , Dermatomicoses/veterinária , Impressões Digitais de DNA/veterinária , Doenças do Gato/microbiologia , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados/veterinária , Doenças do Cão/microbiologia , Microsporum/genética
6.
J Clin Microbiol ; 61(11): e0080623, 2023 11 21.
Artigo em Inglês | MEDLINE | ID: mdl-37877725

RESUMO

The infection proportion of Candida orthopsilosis, a member of the C. parapsilosis complex, has increased globally in recent years, and nosocomial outbreaks have been reported in several countries. This study aimed to establish microsatellite loci-based typing method that was able to effectively distinguish among C. orthopsilosis isolates. Three reference C. orthopsilosis genome sequences were analyzed to identify repeat loci. DNA sequences containing over eight bi- or more nucleotide repeats were selected. A total of 51 loci were initially identified, and locus-specific primers were designed and tested with 20 epidemiologically unrelated isolates. Four loci with excellent reproducibility, specificity, and resolution for molecular typing purposes were identified, and the combined discriminatory power (DP, based on 20 epidemiologically unrelated isolates) of these four loci was 1.0. Reproducibility was demonstrated by consistently testing three strains each in triplicate, and stability, demonstrated by testing 10 successive passages. Then, we collected 48 C. orthopsilosis non-duplicate clinical isolates from the China Hospital Invasive Fungal Surveillance Net study to compare the DP of the microsatellite-based typing with internal transcribed spacer (ITS) and amplified fragment length polymorphism (AFLP) typing analyses, using ATCC 96139 as a reference strain. These 49 isolates were subdivided into 12 microsatellite types (COMT1-12), six AFLP types, and three ITS types, while all the isolates with the same COMT belonged to consistent AFLP and ITS type, demonstrating the high DP of our microsatellite-type method. According to our results, COMT12 was found to be the predominant type in China, and COMT5 was the second largest and responsible for causing a nosocomial outbreak. This microsatellite-type method is a valuable tool for the differentiation of C. orthopsilosis and could be vital for epidemiological studies to determine strain relatedness and monitor transmission.


Assuntos
Candidíase , Infecção Hospitalar , Humanos , Candida parapsilosis , Candida/genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Candidíase/diagnóstico , Candidíase/epidemiologia , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Reprodutibilidade dos Testes , Hospitais , Surtos de Doenças , Genótipo , Repetições de Microssatélites , Técnicas de Tipagem Micológica/métodos
7.
Sci Total Environ ; 905: 167214, 2023 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-37730049

RESUMO

Carpinus betulus L., the hornbeam, is a component of lowland and highland forests in Europe. By examining the postglacial migratory history of thermophilic tree species, the study aimed to unravel their putative glacial microrefugia in the Carpathian region. The present study points to the two distinct genetic AFLP groups of C. betulus in the Carpathian region that represent different genetic lineages based on Bayesian analysis. They differed in Nei's gene diversity index h, and the analysis of molecular variance AMOVA showed a percentage variation of the populations between the groups of 13.74 %. Principal coordinate analysis (PCoA) of 368 AFLP tree samples confirmed the presence of two genetic groups. Ninety-five populations underwent principal component analysis (PCA) to show the main correlations between genetic diversity indices and bioclimatic/climate variables (WorldClim and Carpatclim). The generalized logistic model (GLM) showed the significance of Nei's genetic index h in delimiting genetic groups. The results of population-genetic and multivariate analyses determined that the two genetic groups nowadays are spatially diffused and do not show a clear geographic pattern, pointing to a genetic melting pot. We found ecological links between genetic diversity and bioclimatic characteristics, especially the precipitation in the coldest quarter - Bio19. The refugial Maxent model indicates a significant contribution of the Bio7 variable (both linked with a continental type of climate) to the occurrence of the species during the LGM in Europe. We suggest the relict character of hornbeam populations in a specific climatic-terrain niche in the northern part of the Carpathian Basin.


Assuntos
Variação Genética , Refúgio de Vida Selvagem , Teorema de Bayes , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Filogeografia , Betulaceae , Filogenia
8.
J Genet ; 1022023.
Artigo em Inglês | MEDLINE | ID: mdl-37537853

RESUMO

Investigating the population genetic structure of parasites and their host plants can provide valuable insights into their coevolutionary processes. In this study, we assessed and compared the population genetic diversity and structure of 12 Rhus gall aphid (Schlechtendalia chinensis) populations and their respective host plant (Rhus chinensis) using amplified fragment length polymorphism (AFLP) markers. Analysis of molecular variance (AMOVA) revealed that both the aphid and its host plant exhibited higher genetic variance within populations than among them, indicating that their coevolutionary history may have produced analogous patterns of population genetic structure. Additionally, we considered alternative factors that could contribute to this outcome, such as intraspecific gene flow, hybridization, or environmental influences. Our analysis did not reveal a significant correlation between genetic and geographic distances of either the aphid or host plant populations, leading us to reject the isolation-by-distance model as a plausible explanation for the demographic histories of these two species.


Assuntos
Afídeos , Rhus , Animais , Afídeos/genética , Rhus/genética , Rhus/parasitologia , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Variação Genética
9.
Genes (Basel) ; 14(7)2023 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-37510311

RESUMO

(1) Introduction: Pathogenic variants in the CFTR (Cystic Fibrosis Transmembrane conductance Regulator, OMIM: 602421) gene cause Cystic Fibrosis (CF, OMIM: 219700) and CF-related disorders (CF-RD), often accompanied by obstructive azoospermia due to congenital bilateral aplasia of vas deferens (CBAVD, OMIM: 277180) in male patients. The L138ins (c.413_415dup; p. (Leu138dup)) is a mild variant in the CFTR gene that is relatively common among CF-patients in Slavic populations. The frequency of this variant in Russian infertile men has not been sufficiently studied; (2) Materials and Methods: The sample consisted of 6033 Russian infertile men. The patients were tested for 22 common in Russian populations pathogenic variants of the CFTR gene and the IVS9Tn-polymorphic locus of the intron 9. Molecular-genetic studies were performed using amplified fragment length polymorphism (AFLP-PCR), multiplex ligation-dependent probe amplification (MLPA), and nested PCR (for analysis of the IVS9Tn-polymorphic locus); (3) Results: Pathogenic variants in the CFTR were detected in 3.9% of patients. The most frequent variants were F508del and CFTRdele2.3(21kb), accounted for 61.0% and 7.1% of detected variants, respectively. The L138ins variant was detected in 17 (0.28%) individuals: one of them was homozygous, 10 patients were heterozygous, and 6 patients were compound-heterozygous (F508del/L138ins, n = 4; L138ins/N1303K, n = 1; L138ins/5T, n = 1). Two pathogenic CF-causing variants in the CFTR gene were detected in 8 patients, including 7 compound heterozygous (F508del/L138ins, n = 4; F508del/N1303K, n = 1; 2184insA/E92K, n = 1; 3849+10kbC>T/E92K, n = 1) and one homozygous (L138ins/L138ins). The L138ins variant was found in 7 out of 16 (43.75%) chromosomes in six of these patients. The most common pathogenic variant, F508del, was identified in five out of them, in 5 of 16 (31.25%) chromosomes. The allele frequency (AF) of the L138ins variant in the sample has been found to be 0.0014.; (4) Conclusions: The L138ins variant of the CFTR gene is the third most common variant after F508del and CFTRdele2.3(kb) among Russian infertile men.


Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística , Infertilidade Masculina , Humanos , Masculino , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Frequência do Gene , Mutação , Federação Russa/epidemiologia , Infertilidade Masculina/genética
10.
Sci Rep ; 13(1): 9030, 2023 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-37270656

RESUMO

Understanding the genetic variability of hybrids provides information on their current and future evolutionary role. In this paper, we focus on the interspecific hybrid Ranunculus circinatus × R. fluitans that forms spontaneously within the group Ranuculus L. sect. Batrachium DC. (Ranunculaceae Juss.). Genome-wide DNA fingerprinting using amplified fragment length polymorphisms (AFLP) was employed to determine the genetic variation among 36 riverine populations of the hybrid and their parental species. The results demonstrate a strong genetic structure of R. circinatus × R. fluitans within Poland (Central Europe), which is attributed to independent hybridization events, sterility of hybrid individuals, vegetative propagation, and isolation through geographical distance within populations. The hybrid R. circinatus × R. fluitans is a sterile triploid, but, as we have shown in this study, it may participate in subsequent hybridization events, resulting in a ploidy change that can lead to spontaneous fertility recovery. The ability to produce unreduced female gametes of the hybrid R. circinatus × R. fluitans and the parental species R. fluitans is an important evolutionary mechanism in Ranunculus sect. Batrachium that could give rise to new taxa.


Assuntos
Ranunculus , Humanos , Feminino , Ranunculus/genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Evolução Biológica , Ploidias , Hibridização Genética , Genoma
11.
J Appl Microbiol ; 134(5)2023 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-37188640

RESUMO

AIMS: Develop quantitative assays (qPCR) to determine the wheat rhizosphere competence of inoculant strains Bacillus amyloliquefaciens W10 and Pseudomonas protegens FD6, and their suppressive efficacies against the sharp eyespot pathogen Rhizoctonia cerealis. METHODS AND RESULTS: Antimicrobial metabolites of strains W10 and FD6 decreased in vitro growth of R. cerealis. A qPCR assay for strain W10 was designed from a diagnostic AFLP fragment and the rhizosphere dynamics of both strains in wheat seedlings were compared by culture-dependent (CFU) and qPCR assays. The qPCR minimum detection limits for strains W10 and FD6 were log 3.04 and log 4.03 genome (cell) equivalents g-1 soil, respectively. Inoculant soil and rhizosphere abundance determined by CFU and qPCR were highly correlated (r > 0.91). In wheat bioassays, rhizosphere abundance of strain FD6 was up to 80-fold greater (P < 0.001) than strain W10 at 14 and 28 days postinoculation. Both inoculants reduced (P < 0.05) rhizosphere soil and root abundance of R. cerealis by up to 3-fold. CONCLUSIONS: Strain FD6 exhibited greater abundance in wheat roots and rhizosphere soil than strain W10 and both inoculants decreased the rhizosphere abundance of R. cerealis.


Assuntos
Bacillus amyloliquefaciens , Bacillus amyloliquefaciens/genética , Triticum , Rizosfera , Solo , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Rhizoctonia , Doenças das Plantas/prevenção & controle
12.
Mycoses ; 66(7): 585-593, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-36971239

RESUMO

Sporotrichosis is an implantation mycosis caused by the dimorphic fungus Sporothrix and mostly involves cutaneous and subcutaneous tissues and the lymphatic vessels. Among more than 50 different species, only Sporothrix schenckii, Sporothrix globosa and Sporothrix brasiliensis are frequently reported to cause infections in humans. Sporothrix brasiliensis is remarkably virulent and has been spreading rapidly in Brazil and other Latin American countries. In this study, we aimed to determine the genetic relatedness and antifungal susceptibility of Sporothrix strains by analysing 89 isolates from humans and cats in Curitiba, Southern Brazil. Calmodulin sequencing identified 81 S. brasiliensis and seven S. schenckii isolates. Amplified fragment length polymorphism genotyping analysis showed feline and human isolates clustering together. In vitro susceptibility testing with seven antifungals demonstrated a broad activity against all tested S. brasiliensis isolates, with no significant differences in minimal inhibitory concentration (MIC) values between feline and human isolates. Resistance was solely observed in one human isolate against itraconazole and posaconazole, with MICs of ≥16 µg/mL against both antifungals. Whole genome sequencing (WGS) analysis on this isolate and two related susceptible isolates did not reveal any unique substitutions in resistance-associated genes, including cyp51, hmg and erg6, when compared to two related susceptible isolates. The novel antifungal olorofim exhibited excellent activity against this large isolate collection, with all isolates considered as susceptible. Altogether, we indicate zoonotic transmission based on genotyping and revealed a broad activity of seven common antifungals, including olorofim, against a large S. brasiliensis isolate collection.


Assuntos
Sporothrix , Esporotricose , Humanos , Animais , Gatos , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Genótipo , Brasil , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Esporotricose/microbiologia , Testes de Sensibilidade Microbiana
13.
Chromosome Res ; 31(1): 3, 2023 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-36692656

RESUMO

Like other cecidomyiid Diptera, Hessian fly has stable S chromosomes and dispensable E chromosomes that are retained only in the germ line. Amplified fragment length polymorphisms (AFLP), suppressive subtractive hybridization (SSH), fluorescent in-situ hybridization (FISH), and sequencing were used to investigate similarities and differences between S and E chromosomes. More than 99.9% of AFLP bands were identical between separated ovary and somatic tissue, but one band was unique to ovary and resembled Worf, a non-LTR retrotransposon. Arrayed clones, derived by SSH of somatic from ovarian DNA, showed no clones that were unique to ovary. FISH with BAC clones revealed a diagnostic banding pattern of BAC positions on both autosomes and both sex chromosomes, and each E chromosome shared a pattern with one of the S chromosomes. Sequencing analysis showed that E chromosomes are nearly identical to S chromosomes, since no sequence could be confirmed to belong only to E chromosomes. There were a few questionably E-specific sequences that are candidates for further investigation. Thus, the E chromosomes appear to be derived from S chromosomes by the acquisition or conversion of sequences that produce the negatively heteropycnotic region around the centromere.


Assuntos
Dípteros , Animais , Feminino , Dípteros/genética , Sequência de Bases , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Cromossomos Sexuais/genética , DNA/genética
14.
PeerJ ; 10: e14512, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36545382

RESUMO

Apple scab is a serious disease that restricts the growth of cultivated apples. The objective of this study is to investigate the genetic variations and genetic structure of Venturia inaequalis in Gansu Province, China. 108 isolates of the pathogen V. inaequalis from the Jingning, Lingtai, Jingchuan, Xifeng, Ning and Maiji regions were collected, and their genetic diversity was analyzed using AFLP molecular marker technique. The results showed that genetic diversity was present among the isolates but was not statistically significant. Genetic distance values ranged from 0.0095 to 0.0762. Cluster analysis results showed that the 108 isolates could be divided into two clusters using a similarity coefficient of 0.69. A total of 104 isolates were contained in cluster I while four isolates were contained in Cluster II. From the AMOVA analysis, 98% of variations were observed within the same region, while 2% were observed across different regions. The analysis of population structure showed that 108 isolates had two common ancestors, with the Jingning isolates mainly being derived from the red ancestor. PCoA analysis showed that the Jingning isolates were independent to a certain extent. The different geographical location caused the genetic difference of the isolates. The genetic diversity of apple scab in Gansu Province is greatly aided by this work, which also offers a theoretical foundation for the use of molecular markers in assisted breeding to create novel resistant types.


Assuntos
Ascomicetos , Malus , Malus/genética , Ascomicetos/genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Melhoramento Vegetal , Variação Genética/genética
15.
J Microbiol Methods ; 203: 106605, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36341783

RESUMO

In fields such as the food industry, it is very important to identify target bacteria at the species level or lower for optimal product quality control. Bacteria identification at the subspecies or lower level requires time-consuming and high-cost analyses such as multi-locus sequence typing and amplified fragment length polymorphism analyses. Herein, we developed a primer design algorithm for precisely identifying bacteria based on a whole genome DNA sequence that is easy to apply. The algorithm designs primer sets that produce fragments from all input sequences and maximizes the differences in the amplicon size or amplicon sequence among input sequences. We demonstrate that the primer sets designed by the algorithm clearly classified six subspecies of Lactobacillus delbrueckii, and we observed that the resolution of the method is equal to that of a multi-locus sequence analysis. The algorithm allows the easy but precise identification of bacteria within a short time. (SHRS is available freely from PyPI under the MIT license.).


Assuntos
Bactérias , Lactobacillus delbrueckii , Tipagem de Sequências Multilocus/métodos , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Bactérias/genética , Lactobacillus delbrueckii/genética , Algoritmos
16.
Sci Rep ; 12(1): 19765, 2022 11 17.
Artigo em Inglês | MEDLINE | ID: mdl-36396718

RESUMO

Understanding how environmental factors shape patterns of genetic and phenotypic variations in a species is necessary for conservation and plant breeding. However, these factors have not yet been completely understood in tuberous orchid species used to make 'Salep', an important ingredient in traditional medicine and beverages in middle eastern countries and India. In many areas, increasing demand has pushed species to the brink of extinction. In this study, 198 genotypes from 18 populations of the endangered species Orchis mascula L. spanning a large-scale climatic gradient in northern Iran were used to investigate patterns of genetic diversity and plant functional traits. Populations were sampled from three land cover types (woodland, shrubland, and pastureland/grassland). Plant height, stem length, number of flowers, bulb fresh and dry weight, glucomannan, and starch concentrations showed high variation among populations and were significantly related to land cover type. In general, genetic diversity was high, particularly in those from eastern Hyrcanian; additionally, populations showed a high level of genetic differentiation (G'st = 0.35) with low gene flow (Nm = 0.46). The majority of genetic differentiation occurred within populations (49%) and land cover types (20%). The population structural analysis using the AFLP marker data in K = 4 showed a high geographical affinity for 198 O. mascula genotypes, with some genotypes having mixed ancestry. Temperature and precipitation were found to shape genetic and phenotypic variation profoundly. Significant isolation by the environment was observed, confirming the strong effect of environmental variables on phenotypic and genetic variation. Marker-trait association studies based on MLM1 and MLM2 models revealed significant associations of P-TGG + M-CTT-33 and E-AGG + M-CGT-22 markers with plant height and glucomannan content. Overall, a combination of large-scale climatic variables and land cover types significantly shaped genetic diversity and functional trait variation in O. mascula populations.


Assuntos
Variação Genética , Orchidaceae , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Melhoramento Vegetal , Orchidaceae/genética , Fenótipo
17.
Ann Bot ; 130(7): 1041-1056, 2022 12 31.
Artigo em Inglês | MEDLINE | ID: mdl-36413156

RESUMO

BACKGROUND AND AIMS: Invasive plants often colonize wide-ranging geographical areas with various local microenvironments. The specific roles of epigenetic and genetic variation during such expansion are still unclear. Chenopodium ambrosioides is a well-known invasive alien species in China that can thrive in metalliferous habitats. This study aims to comprehensively understand the effects of genetic and epigenetic variation on the successful invasion of C. ambrosioides. METHODS: We sampled 367 individuals from 21 heavy metal-contaminated and uncontaminated sites with a wide geographical distribution in regions of China. We obtained environmental factors of these sampling sites, including 13 meteorological factors and the contents of four heavy metals in soils. Microsatellite markers were used to investigate the demographic history of C. ambrosioides populations in China. We also analysed the effect of epigenetic variation on metalliferous microhabitat adaptation using methylation-sensitive amplified polymorphism (MSAP) markers. A common garden experiment was conducted to compare heritable phenotypic variations among populations. KEY RESULTS: Two distinct genetic clusters that diverged thousands of years ago were identified, suggesting that the eastern and south-western C. ambrosioides populations in China may have originated from independent introduction events without recombination. Genetic variation was shown to be a dominant determinant of phenotypic differentiation relative to epigenetic variation, and further affected the geographical distribution pattern of invasive C. ambrosioides. The global DNA unmethylation level was reduced in metalliferous habitats. Dozens of methylated loci were significantly associated with the heavy metal accumulation trait of C. ambrosioides and may contribute to coping with metalliferous microenvironments. CONCLUSIONS: Our study of C. ambrosioides highlighted the dominant roles of genetic variation in large geographical range expansion and epigenetic variation in local metalliferous habitat adaptation.


Assuntos
Chenopodium ambrosioides , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Ecossistema , Espécies Introduzidas , Epigênese Genética , Variação Genética
18.
Genes (Basel) ; 13(11)2022 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-36360327

RESUMO

Abiotic stresses cause a significant decrease in productivity and growth in agricultural products, especially barley. Breeding has been considered to create resistance against abiotic stresses. Pyramiding genes for tolerance to abiotic stresses through selection based on molecular markers connected to Mega MQTLs of abiotic tolerance can be one of the ways to reach Golden Barley. In this study, 1162 original QTLs controlling 116 traits tolerant to abiotic stresses were gathered from previous research and mapped from various populations. A consensus genetic map was made, including AFLP, SSR, RFLP, RAPD, SAP, DArT, EST, CAPS, STS, RGA, IFLP, and SNP markers based on two genetic linkage maps and 26 individual linkage maps. Individual genetic maps were created by integrating individual QTL studies into the pre-consensus map. The consensus map covered a total length of 2124.43 cM with an average distance of 0.25 cM between markers. In this study, 585 QTLs and 191 effective genes related to tolerance to abiotic stresses were identified in MQTLs. The most overlapping QTLs related to tolerance to abiotic stresses were observed in MQTL6.3. Furthermore, three MegaMQTL were identified, which explained more than 30% of the phenotypic variation. MQTLs, candidate genes, and linked molecular markers identified are essential in barley breeding and breeding programs to develop produce cultivars resistant to abiotic stresses.


Assuntos
Hordeum , Hordeum/genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Técnica de Amplificação ao Acaso de DNA Polimórfico , Melhoramento Vegetal , Estresse Fisiológico/genética
19.
Am J Bot ; 109(10): 1641-1651, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36112611

RESUMO

PREMISE: Parthenogenesis is the capacity of organisms to develop embryos from unfertilized eggs. When parthenogenesis is coupled with unreduced gamete formation (apomeiosis), genetically maternal progeny result. Genetic elucidation of this form of reproduction in plants, apomixis, has important agronomic implications. However, genetic characterization of apomeiosis and parthenogenesis has been problematic in part because the traits usually co-occur and are restricted to polyploids. In this work, the inheritance of parthenogenetic embryo development, by itself, was studied at the diploid level. METHODS: Progeny resulting from a cross between a diploid (2n = 18), heterozygous, parthenogenetic pollen donor, and a diploid, wildtype, sexual seed parent were evaluated. Paternity was tested with conserved orthologous sequence (COS) markers, reproductive development of F1s was evaluated with microscopy of cleared ovules, and an amplified fragment length polymorphism (AFLP) marker (Eagc × Macg.615) co-segregating with parthenogenesis was characterized at the sequence level. RESULTS: Of 102 diploid biparental progeny, 47 exhibited parthenogenetic embryo and endosperm development, and 55 lacked development of the egg and central cell. This result is consistent with Mendelian inheritance for a single locus (P = 0.43). Isolation and sequencing of the AFLP marker indicates that it is likely a portion of a Ty-Gypsy retrotransposon. Attempts to develop a sequence-characterized amplified region marker from the AFLP were unsuccessful. CONCLUSIONS: This work shows that parthenogenesis can be transmitted simply at the diploid level. This advance is key in the development of a tractable system in Erigeron aimed at the identification of the parthenogenesis locus using genetic mapping strategies.


Assuntos
Erigeron , Magnoliopsida , Diploide , Erigeron/genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Magnoliopsida/genética , Partenogênese/genética , Desenvolvimento Embrionário
20.
Front Cell Infect Microbiol ; 12: 953750, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36118044

RESUMO

Introduction: Recently, India witnessed an unprecedented surge of coronavirus disease 2019 (COVID-19)-associated mucormycosis (CAM) cases. In addition to patient management issues, environmental Mucorales contamination possibly contributed to the outbreak. A recent study evaluated environment contamination by Mucorales in the hospital setting. However, a considerable number of CAM patients were never admitted to a hospital before the development of the disease. The present study, therefore, planned to evaluate Mucorales contamination of patients' residences. Methods: The residential environment of 25 patients with CAM living in north India was surveyed. Air samples were collected from indoor and immediate outdoor vicinity of the patients' residence and cultured on Dichloran Rose-Bengal Chloramphenicol (DRBC) agar with benomyl for selective isolation of Mucorales. Surface swab samples were also collected from the air coolers fitted in those residences and cultured on DRBC agar. The isolates were identified by phenotypic and genotypic methods. Amplified fragment length polymorphism (AFLP) was employed to evaluate the genetic relatedness of the environmental and patients' clinical isolates. Results: The median spore count (mean ± SD, cfu/m3) of Mucorales in the air of patients' bedrooms was significantly higher than in the air in other rooms in those residences (3.55 versus 1.5, p = 0.003) or the air collected directly from the front of the air cooler (p < 0.0001). The Mucorales spore count in the environment did not correlate with either ventilation of the room or hygiene level of the patients' residences. Rhizopus arrhizus was isolated from the environment of all patients' residences (n = 25); other Mucorales species isolated were Cunninghamella bertholletiae (n = 14), Rhizopus microsporus (n = 6), Rhizopus delemar (n = 6), Syncephalastrum racemosum (n = 1), Lichtheimia corymbifera (n = 1), and Mucor racemosus (n = 1). Genetic relatedness was observed between 11 environmental isolates from the patients' bedrooms and respective clinical isolates from patients. Discussion: The study supported the view that the patients might have acquired Mucorales from the home environment during the post-COVID-19 convalescence period. Universal masking at home during patients' convalescence period and environmental decontamination could minimize exposure in those susceptible patients.


Assuntos
COVID-19 , Mucorales , Mucormicose , Ágar , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Benomilo , Cloranfenicol , Convalescença , Humanos , Mucorales/genética , Mucormicose/epidemiologia
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