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1.
Braz. j. biol ; 84: e256354, 2024.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1364518

RESUMO

Termites are known as social insects worldwide. Presently in China 473 species, 44 genera and 4 families of termites have been reported. Of them, 111 Reticulitermes species are widely spread in different zones of China. The dispersion flight season of these Chinese Reticulitermes species are usually started from February to June, but in some regions different species are distributed, sharing their boundaries and having overlapping flight seasons. These reasons become important sources of hybridization between two different heterospecific populations of termites. It was confirmed that the fertilized eggs and unfertilized eggs of some Reticulitermes termites have the capacity of cleavage. While the unfertilized eggs of R. aculabialis, R. chinensis and R. labralis cleaved normally and the only R. aculabialis unfertilized eggs develop in embryos. While, the R. flaviceps and R. chinensis were observed with their abnormal embryonic development, and not hatching of eggs parthenogenetically. They were reported more threatening to Chinese resources as they propagate with parthenogenesis, hybridization and sexual reproduction. Eggshell and macrophiles of eggs play important roles in species identification and control. Although, they are severe pests and cause a wide range of damages to wooden structures and products in homes, buildings, building materials, trees, crops, and forests in China's Mainland.


Os cupins são conhecidos como insetos sociais em todo o mundo. Atualmente na China foram relatadas 473 espécies, 44 gêneros e 4 famílias de cupins. Destas, 111 espécies de Reticulitermes estão amplamente distribuídas em diferentes zonas da China. A temporada de voo de dispersão dessas espécies chinesas de Reticulitermes geralmente começa de fevereiro a junho, mas em algumas regiões diferentes espécies são distribuídas, compartilhando seus limites e tendo temporadas de voo sobrepostas. Essas razões tornam-se importantes fontes de hibridização entre duas populações heteroespecíficas de cupins. Foi confirmado que os ovos fertilizados e não fertilizados de alguns cupins Reticulitermes possuem capacidade de clivagem. Já os ovos não fertilizados de R. aculabialis, R. chinensis e R. labralis clivaram normalmente, e os únicos ovos não fertilizados de R. aculabialis se desenvolvem em embriões. R. flaviceps e R. chinensis foram observados com desenvolvimento embrionário anormal, e não eclosão de ovos por partenogênese. Eles foram relatados como mais ameaçadores para os recursos chineses à medida que se propagam com partenogênese, hibridização e reprodução sexual. Casca de ovo e macrófilos de ovos desempenham papéis importantes na identificação e controle de espécies, embora sejam pragas graves e causem uma ampla gama de danos a estruturas e produtos de madeira em residências, edifícios, materiais de construção, árvores, plantações e florestas na China continental.


Assuntos
Animais , Partenogênese , Reprodução , Isópteros/crescimento & desenvolvimento , China , Hibridização Genética
2.
Plant Dis ; 2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36723957

RESUMO

Bacterial leaf streak (BLS) of barley is caused by the Gram-negative bacterial pathogen Xanthomonas translucens (Sapkota et al. 2020). In 2021, we observed multiple hill plots with BLS symptomatic plants in a barley stripe rust nursery in Vancouver, BC, Canada. We collected 29 leaf samples showing typical BLS symptoms (e.g. necrotic lesions; Fig. S1) and stored at 4 oC until bacterial isolation. Samples were surface-sterilized in 10% NaOCl for 20 sec and rinsed twice. About 1 cm2 of leaf tissue containing BLS characteristic lesions was macerated in 200 µL sterile H2O on a petri dish, incubated for 15 min, and 10 µl of the homogenates was streaked onto Wilbrink's - Boric Acid - Cephalexin (WBC) agar medium. Plates were incubated at 28-30 oC for 48 hrs. Four single colonies were obtained: BC10-1-2a (USask BC10-2a), BC10-1-2b (USask BC10-2b), UBC026 and UBC028. Colonies were grown in WBC broth and gDNA was extracted using E.Z.N.A. Bacterial DNA Kit (Omega Bio-Tek) or DNeasy Plant Pro Kit® (Qiagen) following manufacturer protocols. Genus-level identification was achieved using 16S rRNA sequencing with 27F/1492R primers (Lane 1991) of UBC026 (1,399 bp; NCBI # OP327375) and UBC028 (1,415 bp; NCBI #OP327376). Complete 16S rRNA sequences (1,533bp) of BC10-2a and BC10-2b (1,533 bp) were extracted from the draft whole-genome sequences (WGS) generated in this study. The 16S rRNA sequence homology values of 99.0-100% were recorded between the 4 strains. BLAST analyses of the 16S rRNA sequences to GenBank entries exhibited 99.5-100% similarity values (100% coverage) with the pathotype strains of Xtt DSM 18974T (LT604072) and X. translucens pv. undulosa (Xtu) CFBP 2055 (CP074361). Whole genomes of BC10-2a (JANUQY01) and BC10-2b (JANUQZ01) were sequenced (150-bp; reads 33.1 million; mean coverage 2125x) using NovaSeq Illumina, assembled (Unicycler v0.4.8; Wick et al. 2017) and analyzed to identify the strains to the species-level (Tambong et al. 2021). WGS of strains USask BC10-2a and USask BC10-2b exhibited genome-based DNA-DNA hybridization (dDDH; Meier-Kolthoff et al. 2013) and BLAST-based average nucleotide identity (ANIb; Richter et al. 2015) of 100%. The two strains also showed dDDH and ANIb of 90.4% (species-leel cut-off of 70%) and 98.780% and 98.80% (cut-off of 96%), respectively, with Xtt DSM 18974T (LT604072). In contrast, the WGS of BC10-2a and BC10-2b exhibited only 78.2% dDDH homology values with Xtu CFBP 2055T, suggesting that the strains are genetically more similar to Xtt. The assignment of these strains to Xtt is corroborated by phylogenomic analysis (Fig. S2; Meier-Kolthoff and Göker 2019) that showed the two strains clustering together (100% bootstrap) with the type strain DSM 18974T. These data suggest that these strains are taxonomically members of Xtt. Identification was also confirmed to the genus-level by LAMP assay using published X. translucens primers (Langlois et al. 2017). Pathovar-level identification was confirmed using a cbsA and S8.pep multiplex PCR diagnostic assay (Roman-Reyna et al. 2022). Koch's postulates were verified by greenhouse inoculation via leaf infiltration of UBC026 and UBC028 on 21-day old barley plants (line HB522) using an inoculum of 108 CFU ml-1 followed by re-isolation of the bacteria on WBC. The inoculated plants showed typical BLS symptoms similar to those observed in the field (Fig. S1). Water-inoculated plants had no symptoms. To our knowledge, this is the first published report of BLS of barley in British Columbia.

3.
Genetics ; 2023 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-36708142

RESUMO

The identification of individuals that have a recent hybrid ancestry (between populations or species) has been a goal of naturalists for centuries. Since the 1960s, codominant genetic markers have been used with statistical and computational methods to identify F1 hybrids and back crosses. Existing hybrid inference methods assume that alleles at different loci undergo independent assortment (are unlinked or in population linkage equilibrium). Genomic datasets include thousands of markers that are located on the same chromosome and are in population linkage disequilibrium which violate this assumption. Existing methods may therefore be viewed as composite likelihoods when applied to genomic datasets and their performance in identifying hybrid ancestry (which is a model-choice problem) is unknown. Here we develop a new program Mongrail that implements a full-likelihood Bayesian hybrid inference method that explicitly models linkage and recombination, generating the posterior probability of different F1 or F2 hybrid, or backcross, genealogical classes. We use simulations to compare the statistical performance of Mongrail with that of an existing composite likelihood method (NewHybrids) and apply the method to analyze genome sequence data for hybridizing species of barred and spotted owls.

4.
Methods Mol Biol ; 2626: 151-177, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36715904

RESUMO

Somatic follicle cells are critical support cells for Drosophila oogenesis, as they provide signals and molecules needed to produce a mature egg. Throughout this process, the follicle cells differentiate into multiple subpopulations and transition between three different cell cycle programs to support nurse cell and oocyte development. The follicle cells are mitotic in early egg chamber development, as they cover the germline cyst. In mid-oogenesis, follicle cells switch from mitosis to endocycling, increasing their ploidy from 2C to 16C. Finally, in late oogenesis, cells transition from endocycling to gene amplification, increasing the copy number of a small subset of genes, including the genes encoding proteins required for egg maturation. In order to explore the genetic regulation of these cell cycle switches and follicle cell development and specification, clonal analysis and the GAL4/UAS system are used frequently to reduce or increase expression of genes of interest. These genetic approaches combined with immunohistochemistry and in situ hybridization are powerful tools for characterizing the mechanisms regulating follicle cell development and the mitosis/endocycle and endocycle/gene amplification transitions. This chapter describes the genetic tools available to manipulate gene expression in follicle cells, as well as the methods and reagents that can be utilized to explore gene expression throughout follicle cell development.


Assuntos
Proteínas de Drosophila , Animais , Proteínas de Drosophila/metabolismo , Receptores Notch/metabolismo , Transdução de Sinais/fisiologia , Oogênese/genética , Drosophila/genética , Drosophila/metabolismo , Drosophila melanogaster/genética
5.
Diagnostics (Basel) ; 13(1)2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36611460

RESUMO

The expression of human epidermal growth factor receptor 2 (HER2) protein or gene transcripts is critical for therapeutic decision making in breast cancer. We examined the performance of a digitalized and artificial intelligence (AI)-assisted workflow for HER2 status determination in accordance with the American Society of Clinical Oncology (ASCO)/College of Pathologists (CAP) guidelines. Our preliminary cohort consisted of 495 primary breast carcinomas, and our study cohort included 67 primary breast carcinomas and 30 metastatic deposits, which were evaluated for HER2 status by immunohistochemistry (IHC) and in situ hybridization (ISH). Three practicing breast pathologists independently assessed and scored slides, building the ground truth. Following a washout period, pathologists were provided with the results of the AI digital image analysis (DIA) and asked to reassess the slides. Both rounds of assessment from the pathologists were compared to the AI results and ground truth for each slide. We observed an overall HER2 positivity rate of 15% in our study cohort. Moderate agreement (Cohen's κ 0.59) was observed between the ground truth and AI on IHC, with most discrepancies occurring between 0 and 1+ scores. Inter-observer agreement amongst pathologists was substantial (Fleiss´ κ 0.77) and pathologists' agreement with AI scores was 80.6%. Substantial agreement of the AI with the ground truth (Cohen´s κ 0.80) was detected on ISH-stained slides, and the accuracy of AI was similar for the primary and metastatic tumors. We demonstrated the feasibility of a combined HER2 IHC and ISH AI workflow, with a Cohen's κ of 0.94 when assessed in accordance with the ASCO/CAP recommendations.

6.
Anal Chim Acta ; 1239: 340630, 2023 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-36628741

RESUMO

BACKGROUND: As is known to all, pathogenic bacteria have a serious impact on human health. The development of sensitive, simple, rapid and low-cost bacterial detection method is necessary. Nowadays, some conventional methods (such as plate count, polymerase chain reaction (PCR) and immunological techniques) can not meet the above needs. This work was aimed at providing a new method for addressing these unmet needs. RESULT: This study proposed a novel PCR-free and label-free DNA sensor based on multiple linear hybridization chain reaction (ML-HCR) and cloth-based closed bipolar electrochemiluminescence for sensitive and rapid detection of Escherichia coli (E. coli). The target DNA can be obtained from the E. coli genomic DNA by using the restriction enzyme instead of PCR. The auxiliary probe-triggered ML-HCR is carried out with continuous hybridization of two hairpin DNA, and as a result the double stranded DNA is formed to provide a large number of binding sites for Ru(bpy)32+. The whole detection is PCR-free and label-free, and thus the detection procedure is easier and faster. Under optimized conditions, the linear detection range was from 102 to 107 CFU/mL, and the detection limit was low to 38 CFU/mL. In addition, the proposed DNA sensor has an acceptable selectivity, stability and reproducibility, and is successfully applied to detect E. coli in milk samples with the recoveries from 96.24% to 105.98%. SIGNIFICANCE: The proposed DNA sensor has broad application prospects in the fields of bacterial detection and gene diagnose. Further, this method has potential to be extended for establishing miniaturized, integrated, and automated detection system.


Assuntos
Técnicas Biossensoriais , Escherichia coli , Humanos , Escherichia coli/genética , Reprodutibilidade dos Testes , DNA/genética , DNA/química , Hibridização de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase , Técnicas Biossensoriais/métodos , Limite de Detecção
7.
Anal Chim Acta ; 1239: 340738, 2023 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-36628775

RESUMO

A novel g-C3N4 nanosheets (g-C3N4 NSs)-assisted upconversion fluorescent aptasensor was proposed for Staphylococcus aureus (S. aureus) detection by adopting hybridization chain reaction (HCR) as a sensitizer. Two hairpin (H1 and H2) structured DNA probes were engineered predicated on the partial complementary sequence (cDNA) of S. aureus aptamer and modified on the exterior of the upconversion nanoparticles (UCNPs), respectively. The presence of S. aureus initiated the HCR system and activated H1 and H2 probes to form a double-helix away from the g-C3N4 NSs vicinity. This led to the decrease in peroxidase-like activity (PA) of the g-C3N4 NSs and corresponding fluorescence recovery proportional to the concentration of S. aureus (10-106 cfu mL-1). The method was applied to real food samples with acceptable recoveries (91.1-101.6%) and further validated by traditional plate counting method (p > 0.05).


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas , Staphylococcus aureus/genética , Hibridização de Ácido Nucleico , DNA Complementar , Técnicas Biossensoriais/métodos , Aptâmeros de Nucleotídeos/genética , Limite de Detecção
8.
Anticancer Res ; 43(2): 939-942, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36697075

RESUMO

BACKGROUND: The clear-cell variant of mucoepidermoid carcinoma (MEC) involving minor salivary glands is extremely rare in children. CASE REPORT: We report a case of clear-cell variant MEC in the minor salivary gland in a 10-year-old boy who presented with a mass of the right hard palate. Fine-needle aspiration showed features suggestive of clear-cell variant of MEC. Microscopically, the tumor cells showed predominant clear cells and scattered mucous cells. There was increased mitotic activity (6/mm2). No tumor necrosis or nuclear pleomorphism was identified. The tumor cells were positive for cytokeratin 7 (CK7), tumor protein p63, P40 (ΔNp63), CK5/6 and mucicarmine. Rearrangement of mastermind-like transcriptional coactivator 2 (MAML2) (11q21) gene was present in the tumor cells by fluorescence in situ hybridization, supporting the diagnosis of an intermediate-grade clear-cell variant of MEC. A right infrastructure maxillectomy for palate carcinoma with negative margins was performed. Grossly, the tumor was a 2.1 cm well-circumscribed, friable, pale tan mass with focal areas of cystic change. The final pathological diagnosis was clear-cell variant of MEC, intermediate grade, pT2. Post surgery, the patient recovered and was doing well, with no tumor recurrence or metastasis at the 6-month follow-up. CONCLUSION: To the best of our knowledge, this is the first documented case of clear-cell variant MEC in a child. Due to low to intermediate tumor grade, an overtly aggressive treatment should be avoided in a child.


Assuntos
Carcinoma Mucoepidermoide , Neoplasias das Glândulas Salivares , Masculino , Criança , Humanos , Proteínas de Ligação a DNA/genética , Transativadores/genética , Carcinoma Mucoepidermoide/diagnóstico , Carcinoma Mucoepidermoide/genética , Carcinoma Mucoepidermoide/cirurgia , Hibridização in Situ Fluorescente , Recidiva Local de Neoplasia , Fatores de Transcrição/genética , Palato/patologia , Neoplasias das Glândulas Salivares/genética , Neoplasias das Glândulas Salivares/cirurgia , Neoplasias das Glândulas Salivares/química
9.
Ecol Evol ; 13(1): e9673, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36699574

RESUMO

Obligate pollination mutualisms, in which plant and pollinator lineages depend on each other for reproduction, often exhibit high levels of species specificity. However, cases in which two or more pollinator species share a single host species (host sharing), or two or more host species share a single pollinator species (pollinator sharing), are known to occur in current ecological time. Further, evidence for host switching in evolutionary time is increasingly being recognized in these systems. The degree to which departures from strict specificity differentially affect the potential for hybridization and introgression in the associated host or pollinator is unclear. We addressed this question using genome-wide sequence data from five sympatric Panamanian free-standing fig species (Ficus subgenus Pharmacosycea, section Pharmacosycea) and their six associated fig-pollinator wasp species (Tetrapus). Two of the five fig species, F. glabrata and F. maxima, were found to regularly share pollinators. In these species, ongoing hybridization was demonstrated by the detection of several first-generation (F1) hybrid individuals, and historical introgression was indicated by phylogenetic network analysis. By contrast, although two of the pollinator species regularly share hosts, all six species were genetically distinct and deeply divergent, with no evidence for either hybridization or introgression. This pattern is consistent with results from other obligate pollination mutualisms, suggesting that, in contrast to their host plants, pollinators appear to be reproductively isolated, even when different species of pollinators mate in shared hosts.

10.
Evolution ; 77(1): 123-137, 2023 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-36625679

RESUMO

As anthropogenic activities are increasing the frequency and severity of droughts, understanding whether and how fast populations can adapt to sudden changes in their hydric environment is critically important. Here, we capitalize on the introduction of the Cuban brown anole lizard (Anolis sagrei) in North America to assess the contemporary evolution of a widespread terrestrial vertebrate to an abrupt climatic niche shift. We characterized hydric balance in 30 populations along a large climatic gradient. We found that while evaporative and cutaneous water loss varied widely, there was no climatic cline, as would be expected under adaptation. Furthermore, the skin of lizards from more arid environments was covered with smaller scales, a condition thought to limit water conservation and thus be maladaptive. In contrast to environmental conditions, genome-averaged ancestry was a significant predictor of water loss. This was reinforced by our genome-wide association analyses, which indicated a significant ancestry-specific effect for water loss at one locus. Thus, our study indicates that the water balance of invasive brown anoles is dictated by an environment-independent introduction and hybridization history and highlights genetic interactions or genetic correlations as factors that might forestall adaptation. Alternative water conservation strategies, including behavioral mitigation, may influence the brown anole invasion success and require future examination.


Assuntos
Lagartos , Animais , Lagartos/genética , Estudo de Associação Genômica Ampla , Aclimatação , Adaptação Fisiológica , Água
11.
Microorganisms ; 11(1)2023 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-36677460

RESUMO

We have previously reported the draft genome sequences of 59 endospore-forming Gram-positive bacterial strains isolated from Vietnamese crop plants due to their ability to suppress plant pathogens. Based on their draft genome sequence, eleven of them were assigned to the Brevibacillus and one to the Lysinibacillus genus. Further analysis including full genome sequencing revealed that several of these strains represent novel genomospecies. In vitro and in vivo assays demonstrated their ability to promote plant growth, as well as the strong biocontrol potential of Brevibacilli directed against phytopathogenic bacteria, fungi, and nematodes. Genome mining identified 157 natural product biosynthesis gene clusters (BGCs), including 36 novel BGCs not present in the MIBiG data bank. Our findings indicate that plant-associated Brevibacilli are a rich source of putative antimicrobial compounds and might serve as a valuable starting point for the development of novel biocontrol agents.

12.
ChemSusChem ; : e202300006, 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36601966

RESUMO

Creating new donor materials is crucial for further advancing organic solar cells. Random terpolymers have been adopted to overcome shortcomings of regular alternating donor-acceptor (D-A) polymers of which the performance is often susceptible to batch-to-batch variations. In general, the properties and performance of efficient D1 -A-D2 -A and D-A1 -D-A2 terpolymers are sensitive to the D1 /D2 or A1 /A2 monomer ratios. Side-chain hybridization is a strategy to address this problem. Here, six D1 -A-D2 -A-type random terpolymers comprising D1 and D2 monomers with the same π-conjugated D unit but with different side chains were synthesized. The side chains, containing either fluorine or trialkylsilyl substituents were chosen to provide near-identical optoelectronic properties but provide a tool to create a better-optimized film morphology when blended with a non-fullerene acceptor. This strategy allows improving the device performance to over 18 %, higher than that obtained with the corresponding D1 -A or D2 -A bipolymers (around 17 %). Hence, side-chain hybridization is a promising strategy to design efficient D1 -A-D2 -A terpolymer donors that are insensitive to the D1 /D2 monomer ratio, which is beneficial for the scaled-up synthesis of high-performance materials.

13.
Talanta ; 255: 124249, 2023 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-36610257

RESUMO

It is extremely necessary to establish a rapid and high-throughput method to detect mycotoxins in food, because grains and cereals are greatly vulnerable to mycotoxins before and after harvest. In this study, we developed a portable aptasensor based on streptavidin magnetic microspheres (MMPs) and hybridization chain reaction (HCR) to simultaneously detect T-2 toxin and zearalenone (ZEN) in corn and oat flour. The MMPs compete with the aptamer for binding, which releases more H0 and triggers HCR with the H1 intermediate modified using 6-FAM and BHQ-1 and the unmodified H2. Subsequently, placing the HCR system corresponding to T-2 and ZEN in a constant-temperature fluorescence detector resulted in well-recovered fluorescence of the HCR products. T-2 and ZEN exhibited good fluorescence response in the dynamic range of 0.001-10 ng mL-1 and 0.01-100 ng mL-1 with detection limits of 0.1 pg mL-1 and 1.2 pg mL-1, respectively. In addition, this strategy achieved the selective detection of T-2 and ZEN in the spiked corn and oat flour samples. The results are also in good agreement with those obtained using commercial ELISA kits. This developed aptasensor with the characteristics of simple operation and portability has the application potential of establishing sensitive and portable field detection of various mycotoxins.


Assuntos
Aptâmeros de Nucleotídeos , Micotoxinas , Toxina T-2 , Zearalenona , Zearalenona/análise , Toxina T-2/análise , Contaminação de Alimentos/análise , Micotoxinas/análise , Aptâmeros de Nucleotídeos/genética , Zea mays/metabolismo , Limite de Detecção
14.
Biosensors (Basel) ; 13(1)2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36671935

RESUMO

In the continuous combat against diseases, there is the need for tools that enable an improved diagnostic efficiency towards higher information density combined with reduced time-to-result and cost. Here, a novel fully integrated microfluidic platform, the Evalution™, is evaluated as a potential solution to this need. Encoded microparticles combined with channel-based microfluidics allow a fast, sensitive and simultaneous detection of several disease-related biomarkers. Since the binary code is represented by physically present holes, 210 different codes can be created that will not be altered by light or chemically induced degradation. Exploiting the unique features of this multiplex platform, hybridization chain reaction (HCR) is explored as a generic approach to reach the desired sensitivity. Compared to a non-amplified reference system, the sensitivity was drastically improved by a factor of 104, down to low fM LOD values. Depending on the HCR duration, the assay can be tuned for sensitivity or total assay time, as desired. The huge potential of this strategy was further demonstrated by the successful detection of a multiplex panel of six different nucleic acid targets including viruses and bacteria. The ability to not only discriminate these two categories but, with the same effort, also virus strains (human adenovirus and human bocavirus), virus subtypes (human adenovirus type B and D) and antibiotic-resistant bacteria (Streptococcus pneumonia), exemplifies the specificity of the developed approach. The effective, yet highly simplified, isothermal and protein-enzyme-free signal amplification tool reaches an LOD ranging from as low as 33 ± 4 to 151 ± 12 fM for the different targets. Moreover, direct detection in a clinically relevant sample matrix was verified, resulting in a detection limit of 309 ± 80 fM, approximating the low fM levels detectable with the gold standard analysis method, PCR, without the drawbacks related to protein enzymes, thermal cycling and elaborate sample preparation steps. The reported strategy can be directly transferred as a generic approach for the sensitive and specific detection of various target molecules in multiplex. In combination with the high-throughput capacity and reduced reagent consumption, the Evalution™ demonstrates immense potential in the next generation of diagnostic tools towards more personalized medicine.


Assuntos
Microfluídica , Técnicas de Amplificação de Ácido Nucleico , Humanos , Microfluídica/métodos , Hibridização de Ácido Nucleico , Reação em Cadeia da Polimerase , Técnicas de Amplificação de Ácido Nucleico/métodos , Bactérias/genética
15.
Int J Mol Sci ; 24(2)2023 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-36674808

RESUMO

Nitrogen-doped multi-walled carbon nanotubes (N-MWCNTs) are widely used for drug delivery. One of the main challenges is to clarify their interaction with hypoxia-inducible factor 1 alpha (HIF-1α), the lack of which leads to oncological and cardiovascular diseases. In the presented study, N-MWCNTs were synthesized by catalytic chemical vapor deposition and irradiated with argon ions. Their chemical state, local structure, interfaces, Stone-Wales defects, and doping with nitrogen were analyzed by high resolution transmission electron microscopy (HRTEM), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and near-edge X-ray absorption fine structure (NEXAFS) spectroscopy. Using experimental data, supercells of functionalized N-MWCNTs with an oxygen content of 2.7, 4 and 6 at. % in carboxyl groups were built by quantum chemical methods. Our analysis by the self-consistent charge density functional tight-binding (SCC DFTB) method shows that a key role in the functionalization of CNTs with carboxyl groups belongs to Stone-Wales defects. The results of research in the decoration of CNTs with HIF-1α demonstrate the possibility of wave-diffusion drug delivery. The nature of hybridization and relaxation determines the mechanism of oxygen regulation with HIF-1α molecules, namely, by OH-(OH-C) and OH-(O=C) chemical bonds. The concentration dependence of drug release in the diffusion mode suggests that the best pattern for drug delivery is provided by the tube with a carboxylic oxygen content of 6 at. %.


Assuntos
Nanotubos de Carbono , Nanotubos de Carbono/química , Microscopia Eletrônica de Transmissão , Nitrogênio , Oxigênio
16.
Int J Mol Sci ; 24(2)2023 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-36675124

RESUMO

The halophytic wild relatives within Triticeae might provide valuable sources of salt tolerance for wheat breeding, and attempts to use these sources of tolerance have been made for improving salt tolerance in wheat by distant hybridization. A novel wheat substitution line of K17-1078-3 was developed using common wheat varieties of Chuannong16 (CN16), Zhengmai9023 (ZM9023), and partial amphidiploid Trititrigia8801 (8801) as parents, and identified as the 3E(3D) substitution line. The substitution line was compared with their parents for salt tolerance in hydroponic culture to assess their growth. The results showed that less Na+ accumulation and lower Na+/K+ ratio in both shoots and roots were achieved in K17-1078-3 under salinity compared to its wheat parents. The root growth and development of K17-1078-3 was less responsive to salinity. When exposed to high salt treatment, K17-1078-3 had a higher photosynthesis rate, more efficient water use efficiency, and greater antioxidant capacity and stronger osmotic adjustment ability than its wheat parents. In conclusion, a variety of physiological responses and root system adaptations were involved in enhancing salt tolerance in K17-1078-3, which indicated that chromosome 3E possessed the salt tolerance locus. It is possible to increase substantially the salt tolerance of wheat by the introduction of chromosome 3E into wheat genetic background.


Assuntos
Plântula , Triticum , Plântula/genética , Tetraploidia , Melhoramento Vegetal , Poaceae/genética , Tolerância ao Sal/genética , Cromossomos de Plantas/genética
17.
Plants (Basel) ; 12(2)2023 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-36678948

RESUMO

Trifolium medium L. is a wild polyploid relative of the agriculturally important red clover that possesses traits promising for breeding purposes. To date, T. medium also remains the only clover species with which agriculturally important red clover has successfully been hybridized. Even though allopolyploid origin has previously been suggested, little has in fact been known about the T. medium karyotype and its origin. We researched T. medium and related karyotypes using comparative cytogenomic methods, such as fluorescent in situ hybridization (FISH) and RepeatExplorer cluster analysis. The results indicate an exceptional karyotype diversity regarding numbers and mutual positions of 5S and 26S rDNA loci and centromeric repeats in populations of T. medium ecotypes and varieties. The observed variability among T. medium ecotypes and varieties suggests current karyotype instability that can be attributed to ever-ongoing battle between satellite DNA together with genomic changes and rearrangements enhanced by post-hybridization events. Comparative cytogenomic analyses of a T. medium hexaploid variety and diploid relatives revealed stable karyotypes with a possible case of chromosomal rearrangement. Moreover, the results provided evidence of T. medium having autopolyploid origin.

18.
Plants (Basel) ; 12(2)2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36678992

RESUMO

Among cereals, triticale (×Trititcoseale Wittmack ex A. Camus) represents a number of advantages such as high grain yield even in marginal environments, tolerance to drought, cold and acid soils, as well as lower production costs. Together with high biomass of grain and straw, triticale is also considered as an industrial energy crop. As an artificial hybrid, it has not evolved naturally, which is reflected in narrow genetic diversity causing a resistance collapse in recent years. Here, we describe a novel, synthetic tetraploid triticale, which was developed by the crossing of rye (Secale cereale L.) with einkorn wheat (Triticum monococcum spp. monococcum), which possess Sr35 stem rust resistance gene. Three subsequent generations of alloploids were obtained by chromosome doubling followed by self-pollination. The cytogenetic analyses revealed that the amphiploids possess a set of 28 chromosomes (14 of Am-genome and 14 of R-genome). The values of the most important yield-shaping traits for these tetraploid triticale form, including thousand-grain weight, plant height and stem length were higher compared to parental genotypes, as well as standard hexaploid triticale cultivars. This study shows that this tetraploid triticale genetic stock can be an interesting pre-breeding germplasm for triticale improvement or can be developed as a new alternative crop.

19.
Plants (Basel) ; 12(2)2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36678995

RESUMO

The Cattleya (Orchidaceae-Laeliinae subtribe) intergeneric hybrids, such as Brassolaeliocattleya (Blc.), have great ornamental value, due to their compact-size, with large and high color diversity of flowers. Artificial induction of polyploidy brings agronomic, ornamental and genetic benefits to plants. Polyploidization efficiency depends on factors, such as the type of antimitotic, polyploidization method, concentrations, exposure times and type of explant. This study aimed to develop a protocol to polyploidize Blc. orchids, by testing two types of explants (seeds and protocorms), concentrations and exposure times to colchicine. The effects of colchicine on the in vitro development of explants were also investigated. The responses of explants to colchicine depended on the concentrations, exposure time and the interaction of these factors. Flow cytometric analysis evidenced high endopolyploidy and allowed the separation of polyploidized (4C, 8C and 16C peaks) from non-polyploidized (only 2C and 4C peaks) plants. The highest percentage of polyploid plants was regenerated from protocorms (16.4%) treated with colchicine instead of seeds (3.2%). Protocorms treated with colchicine at 500-750 µM for 18 h resulted in the best percentage of polyploidization. Additionally, in vitro natural polyploidization using protocorms was reported (11.5%). Cytological analyses allowed an estimation of the number of chromosomes of the parents (≡70), polyploidized (≡140) and non-polyploidized progeny (≡70).

20.
Plants (Basel) ; 12(2)2023 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-36679086

RESUMO

The remarkable species diversity of the genus Sorbus is a result of polyploidization and frequent hybridization between interacting species of different cytotypes. Moreover, hybridization is possible between several parental taxa. Gametophytic apomixis, which is common among polyploid Sorbus taxa, indicates the role of clonal reproduction in the evolutionary stabilization of hybridogeneous genotypes. The precise determination of the origin of seeds and their quantitative evaluation may elucidate inter-cytotype interactions, the potential role of mixed-cytotype populations in evolutionary success, and the long-term survival of some hybrid species. We investigated the reproduction modes of selected species of Sorbus in mixed-cytotype populations in eastern Slovakia, Central Europe. We determined the pollen quality, seed production rate, and the ploidy level of mature trees, as well as the origin of the embryo and endosperm in seeds of the stenoendemics S. amici-petri, S. dolomiticola, and S. hornadensis. The tetraploids S. amici-petri and S. hornadensis are characterized by regular and highly stainable pollen grains and reproduce predominantly via pseudogamous apomixis. In contrast, triploid S. dolomiticola usually has oval, heterogenous, and weakly stainable pollen grains, suggesting male meiotic irregularities. Although seeds originate via pseudogamous apomixis in S. dolomiticola as well, the ploidy level of sperm cells participating in the fertilization of central cells is usually determined by co-occurring species of different cytotypes. This suggests that maintaining mating partners is necessary for the long-term survival of a triploid species. We documented rare BIII hybrids and the residual sexuality in tetraploids. The distribution of seeds of meiotic and apomeiotic origins in S. amici-petri shows bimodal characteristics; however, genotypes with predominantly sexual seed types are rare. Reproduction modes documented in polyploid stenoendemics of Sorbus and inferred microevolutionary intercytotype relationships highlight the mixed-cytotype populations as the source of biodiversity in apomictic plant complexes. We suggest that conservation efforts should focus on maintaining the species and cytotypic diversity of Sorbus populations, especially when it comes to the conservation of triploid species.

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