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1.
PLoS Negl Trop Dis ; 15(12): e0010062, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34941866

RESUMO

Schistosomes cause schistosomiasis, the world's second most important parasitic disease after malaria in terms of public health and social-economic impacts. A peculiar feature of these dioecious parasites is their ability to produce viable and fertile hybrid offspring. Originally only present in the tropics, schistosomiasis is now also endemic in southern Europe. Based on the analysis of two genetic markers the European schistosomes had previously been identified as hybrids between the livestock- and the human-infective species Schistosoma bovis and Schistosoma haematobium, respectively. Here, using PacBio long-read sequencing technology we performed genome assembly improvement and annotation of S. bovis, one of the parental species for which no satisfactory genome assembly was available. We then describe the whole genome introgression levels of the hybrid schistosomes, their morphometric parameters (eggs and adult worms) and their compatibility with two European snail strains used as vectors (Bulinus truncatus and Planorbarius metidjensis). Schistosome-snail compatibility is a key parameter for the parasites life cycle progression, and thus the capability of the parasite to establish in a given area. Our results show that this Schistosoma hybrid is strongly introgressed genetically, composed of 77% S. haematobium and 23% S. bovis origin. This genomic admixture suggests an ancient hybridization event and subsequent backcrosses with the human-specific species, S. haematobium, before its introduction in Corsica. We also show that egg morphology (commonly used as a species diagnostic) does not allow for accurate hybrid identification while genetic tests do.


Assuntos
Genoma Helmíntico , Hibridização Genética , Schistosoma haematobium/crescimento & desenvolvimento , Schistosoma haematobium/genética , Schistosoma/crescimento & desenvolvimento , Schistosoma/genética , Animais , Tamanho Corporal , Bulinus/parasitologia , Quimera/anatomia & histologia , Quimera/genética , Quimera/crescimento & desenvolvimento , Vetores de Doenças , Europa (Continente) , Feminino , Humanos , Masculino , Schistosoma/anatomia & histologia , Schistosoma haematobium/anatomia & histologia , Esquistossomose/parasitologia , Caramujos/parasitologia
2.
Sci Rep ; 11(1): 19345, 2021 09 29.
Artigo em Inglês | MEDLINE | ID: mdl-34588562

RESUMO

Abiotic stresses, including cold and drought, negatively affect maize (Zea mays L.) seed field emergence and later yield and quality. In order to reveal the molecular mechanism of maize seed resistance to abiotic stress at seed germination, the global transcriptome of high- vigour variety Zhongdi175 exposed to cold- and drought- stress was analyzed by RNA-seq. In the comparison between the control and different stressed sample, 12,299 differentially expressed genes (DEGs) were detected, of which 9605 and 7837 DEGs were identified under cold- and drought- stress, respectively. Functional annotation analysis suggested that stress response mediated by the pathways involving ribosome, phenylpropanoid biosynthesis and biosynthesis of secondary metabolites, among others. Of the obtained DEGs (12,299), 5,143 genes are common to cold- and drought- stress, at least 2248 TFs in 56 TF families were identified that are involved in cold and/or drought treatments during seed germination, including bHLH, NAC, MYB and WRKY families, which suggested that common mechanisms may be originated during maize seed germination in response to different abiotic stresses. This study will provide a better understanding of the molecular mechanism of response to abiotic stress during maize seed germination, and could be useful for cultivar improvement and breeding of high vigour maize cultivars.


Assuntos
Aclimatação/genética , Resposta ao Choque Frio/genética , Germinação , Melhoramento Vegetal/métodos , Zea mays/crescimento & desenvolvimento , Quimera/genética , Quimera/crescimento & desenvolvimento , Secas , Regulação da Expressão Gênica de Plantas , RNA-Seq , Sementes/crescimento & desenvolvimento , Transcriptoma , Zea mays/genética
3.
Transgenic Res ; 30(3): 251-261, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33786748

RESUMO

Histamine H2 receptor (HRH2) is closely associated with the development of cardiovascular and cerebrovascular diseases. However, systematic Hrh2 knockout mice did not exactly reflect the HRH2 function in specific cell or tissue types. To better understand the physiological and pathophysiological functions of endothelial HRH2, this study constructed a targeting vector that contained loxp sites flanking the ATG start codon located in Hrh2 exon 2 upstream and a neomycin (Neo) resistance gene flanked by self-deletion anchor sites within the mouse Hrh2 allele. The targeting vector was then electroporated into C57BL/6J embryonic stem (ES) cells, and positively targeted ES cell clones were micoinjected into C57BL/6J blastocysts, which were implanted into pseudopregnant females to obtain chimeric mice. The F1 generation of Hrh2flox/+ mice was generated via crossing chimeric mice with wild-type mice to excise Neo. We also successfully generated endothelial cell-specific knockout (ECKO) mice by crossing Hrh2flox/+ mice with Cdh5-Cre mice that specifically express Cre in endothelial cells and identified that Hrh2 deletion was only observed in endothelial cells. Hrh2flox/+ and Hrh2ECKO mice were normal, healthy and fertile and did not display any obvious abnormalities. These novel animal models will create new prospects for exploring roles of HRH2 during the development and treatment of related diseases.


Assuntos
Blastocisto/metabolismo , Quimera/genética , Células-Tronco Embrionárias/metabolismo , Receptores Histamínicos H2/genética , Animais , Antígenos CD/genética , Caderinas/genética , Quimera/crescimento & desenvolvimento , Códon de Iniciação/genética , Células Endoteliais/metabolismo , Éxons/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Integrases/genética , Camundongos , Camundongos Knockout , Neomicina/metabolismo
4.
BMC Plant Biol ; 21(1): 111, 2021 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-33627081

RESUMO

BACKGROUND: Over the life cycle of perennial trees, the dormant state enables the avoidance of abiotic stress conditions. The growth cycle can be partitioned into induction, maintenance and release and is controlled by complex interactions between many endogenous and environmental factors. While phytohormones have long been linked with dormancy, there is increasing evidence of regulation by DAM and CBF genes. To reveal whether the expression kinetics of CBFs and their target PtDAM1 is related to growth cessation and endodormancy induction in Populus, two hybrid poplar cultivars were studied which had known differential responses to dormancy inducing conditions. RESULTS: Growth cessation, dormancy status and expression of six PtCBFs and PtDAM1 were analyzed. The 'Okanese' hybrid cultivar ceased growth rapidly, was able to reach endodormancy, and exhibited a significant increase of several PtCBF transcripts in the buds on the 10th day. The 'Walker' cultivar had delayed growth cessation, was unable to enter endodormancy, and showed much lower CBF expression in buds. Expression of PtDAM1 peaked on the 10th day only in the buds of 'Okanese'. In addition, PtDAM1 was not expressed in the leaves of either cultivar while leaf CBFs expression pattern was several fold higher in 'Walker', peaking at day 1. Leaf phytohormones in both cultivars followed similar profiles during growth cessation but differentiated based on cytokinins which were largely reduced, while the Ox-IAA and iP7G increased in 'Okanese' compared to 'Walker'. Surprisingly, ABA concentration was reduced in leaves of both cultivars. However, the metabolic deactivation product of ABA, phaseic acid, exhibited an early peak on the first day in 'Okanese'. CONCLUSIONS: Our results indicate that PtCBFs and PtDAM1 have differential kinetics and spatial localization which may be related to early growth cessation and endodormancy induction under the regime of low night temperature and short photoperiod in poplar. Unlike buds, PtCBFs and PtDAM1 expression levels in leaves were not associated with early growth cessation and dormancy induction under these conditions. Our study provides new evidence that the degradation of auxin and cytokinins in leaves may be an important regulatory point in a CBF-DAM induced endodormancy. Further investigation of other PtDAMs in bud tissue and a study of both growth-inhibiting and the degradation of growth-promoting phytohormones is warranted.


Assuntos
Quimera/crescimento & desenvolvimento , Desenvolvimento Vegetal/genética , Dormência de Plantas/genética , Reguladores de Crescimento de Plantas/genética , Populus/crescimento & desenvolvimento , Populus/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Variação Genética
5.
PLoS One ; 15(12): e0243580, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33332448

RESUMO

Continued drought during the late growth stage of super hybrid rice (SHR) markedly reduces yield, and management practices that use water more efficiently can contribute greatly to high and stable yields from SHR. The absolute temperature differences (ATDs) between the rice plant and the atmosphere and between the soil and the atmosphere are believed to be important determinants of grain yield. However, it has not previously been determined whether these ATDs have any effect on SHR yields under water-saving cultivation. A two-year field experiment involving two SHR varieties, Liangyoupeijiu (LYPJ) and Y-Liangyou 9000 (YLY900), evaluated the effects of reducing water supply from mid-booting to maturity on grain yield, canopy relative humidity (CRH), leaf area index (LAI), and ATDs between the ambient temperature and the leaf surface, panicles, canopy, and soil. Grain yield increased significantly under shallow water irrigation (SW), by 8.84% (YLY900) and 12.26% (LYPJ), but decreased significantly under mild water stress (MS, -20 to -30 kPa), by 14.36% (YLY900) and 9.47% (LYPJ), as well as severe water stress (SS, -40 to -50 kPa), by 35.06% (YLY900) and 28.74% (LYPJ). As water supply decreased, so did the CRH and the ATDs, with significant decreases under MS and SS. The temperature differences were significantly and positively correlated with grain yield (P < 0.01) in both cultivars. LAI was increased under SW conditions, but was significantly decreased under MS and SS. Our study suggests that the dual goal of saving water while maintaining high yield can be achieved by applying SW irrigation from mid-booting to maturity and by adopting cultivation measures that maintain high CRH and high plant-atmosphere and soil-atmosphere ATDs in order to alleviate water stress. YLY900 has a higher yield potential than LYPJ under SW conditions, suggesting that its wide cultivation may help achieve this dual goal.


Assuntos
Irrigação Agrícola , Oryza/crescimento & desenvolvimento , Irrigação Agrícola/métodos , Atmosfera , Quimera/crescimento & desenvolvimento , Grão Comestível/crescimento & desenvolvimento , Folhas de Planta/crescimento & desenvolvimento , Solo/química , Temperatura
6.
PLoS One ; 15(11): e0241914, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33206683

RESUMO

Alternative splicing (AS) is a post-transcriptional process common in plants and essential for regulation of environmental fitness of plants. In the present study, we focus on the AS events in poplar leaves to understand their effects on plant growth and development. The hybrid poplar (P.alba×P.glandulosa cv.84K) leaves were collected for RNA extraction. The extracted RNA was sequenced using on an Illumina HiSeq™ 2000 platform. Using the Populus trichocarpa genome as the reference, a total of 3810 AS genes were identified (9225 AS events), which accounted for 13.51% of all the expressed genes. Intron retention was the most common AS event, accounting for 43.86% of all the AS events, followed by alternative 3' splice sites (23.75%), alternative 5' splice sites (23.71%), and exon skipping (8.68%). Chromosomes 10 had the most condensed AS events (33.67 events/Mb) and chromosome 19 had the least (12.42 events/Mb). Association analysis showed that AS in the poplar leaves was positively correlated with intron length, exon number, exon length, and gene expression level, and was negatively correlated with GC content. AS genes in the poplar leaves were associated mainly with inositol phosphate metabolism and phosphatidylinositol signaling system pathways that would be significant on wooden plant production.


Assuntos
Processamento Alternativo , Quimera/genética , Proteínas de Plantas/genética , Populus/crescimento & desenvolvimento , Sequenciamento Completo do Genoma/métodos , Quimera/crescimento & desenvolvimento , Cromossomos de Plantas/genética , Éxons , Regulação da Expressão Gênica de Plantas , Sequenciamento de Nucleotídeos em Larga Escala , Íntrons , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Populus/genética , Sítios de Splice de RNA
7.
PLoS One ; 15(10): e0232071, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33052925

RESUMO

A PVL-positive, methicillin-susceptible Staphylococcus aureus was cultured from pus from cervical lymphadenitis of a patient of East-African origin. Microarray hybridisation assigned the isolate to clonal complex (CC) 80 but revealed unusual features, including the presence of the ORF-CM14 enterotoxin homologue and of an ACME-III element as well as the absence of etD and edinB. The isolate was subjected to both, Illumina and Nanopore sequencing allowing characterisation of deviating regions within the strain´s genome. Atypical features of this strain were attributable to the presence of two genomic regions that originated from other S. aureus lineages and that comprised, respectively, 3% and 1.4% of the genome. One deviating region extended from walJ to sirB. It comprised ORF-CM14 and the ACME-III element. A homologous but larger fragment was also found in an atypical S. aureus CC1/ST567 strain whose lineage might have served as donor of this genomic region. This region itself is a chimera comprising fragments from CC1 as well as fragments of unknown origin. The other deviating region comprised the region from htsB to ecfA2, i.e., another 3% of the genome. It was very similar to CC1 sequences. Either this suggests an incorporation of CC1 DNA into the study strain, or alternatively a recombination event affecting "canonical" CC80. Thus, the study strain bears witness of several recombination events affecting supposedly core genomic genes. Although the exact mechanism is not yet clear, such chimerism seems to be an additional pathway in the evolution of S. aureus. This could facilitate also a transmission of virulence and resistance factors and therefore offer an additional evolutionary advantage.


Assuntos
Proteínas de Bactérias/genética , Linfadenite/microbiologia , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Infecções Estafilocócicas/diagnóstico , Staphylococcus aureus/crescimento & desenvolvimento , Quimera/genética , Quimera/crescimento & desenvolvimento , Evolução Molecular , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Pessoa de Meia-Idade , Mutagênese Insercional , Recombinação Genética , Análise de Sequência de DNA , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificação , Supuração
8.
Genes (Basel) ; 11(10)2020 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-33023081

RESUMO

The present study aimed to investigate and compare fitness-related traits and ploidy levels of purebreds and hybrids produced from sturgeon broodstock with both normal and abnormal ploidy levels. We used diploid Acipenser ruthenus and tetraploid A. baerii males and females to produce purebreds and reciprocal hybrids of normal ploidy levels. Likewise, we used diploid A. ruthenus and tetraploid A. baerii females mated to pentaploid and hexaploid A. baerii males to produce hybrids of abnormal ploidy levels. Fertilization of ova of A. ruthenus and A. baerii of normal ploidy with the sperm of pentaploid and hexaploid A. baerii produced fully viable progeny with ploidy levels that were intermediate between those of the parents as was also found in crosses of purebreds and reciprocal hybrids of normal ploidy levels. The A. ruthenus × pentaploid A. baerii and A. ruthenus × hexaploid A. baerii hybrids did not survive after 22 days post-hatch (dph). Mean body weight and cumulative survival were periodically checked at seven-time intervals. The recorded values of mean body weight were significantly higher in A. baerii × pentaploid A. baerii hybrids than other groups at three sampling points (160, 252 and 330 dph). In contrast, the highest cumulative survival was observed in A. baerii × A. ruthenus hybrids at all sampling points (14.47 ± 5.70 at 497 dph). Overall, most of the studied sturgeon hybrids displayed higher mean BW and cumulative survival compared to the purebreds. The utilization of sturgeon hybrids should be restricted to aquaculture purposes because they can pose a significant genetic threat to native populations through ecological interactions.


Assuntos
Quimera/genética , Evolução Molecular , Peixes/genética , Oócitos/fisiologia , Ploidias , Espermatozoides/fisiologia , Animais , Quimera/crescimento & desenvolvimento , Cromossomos , Feminino , Peixes/crescimento & desenvolvimento , Genoma , Masculino , Oócitos/citologia , Espermatozoides/citologia
9.
Cell Rep ; 31(2): 107507, 2020 04 14.
Artigo em Inglês | MEDLINE | ID: mdl-32294444

RESUMO

Protozoan parasites in the genus Leishmania produce a broad spectrum of diseases in their human hosts. The strain and species-specific genes controlling these diverse clinical outcomes have remained poorly tractable using reverse genetics approaches. A cryptic sexual cycle involving a meiotic-like process has been described in Leishmania but is so far confined to parasites growing in the sand fly vector. Here, we describe the reproducible in vitro generation of hybrid clones using axenic culture forms of Leishmania tropica promastigotes. Analysis of SNPs marker inheritance and whole-genome sequencing data indicate that the progeny clones are full genomic hybrids. The demonstration that mating-competent forms arise in culture should facilitate experimental study of the mating biology of Leishmania and the generation of large numbers of recombinant parasites for positional cloning of important genes.


Assuntos
Técnicas de Cultura de Células/métodos , Leishmania/crescimento & desenvolvimento , Reprodução/fisiologia , Animais , Quimera/genética , Quimera/crescimento & desenvolvimento , Marcadores Genéticos/genética , Insetos Vetores/parasitologia , Leishmania/genética , Leishmania tropica/genética , Leishmania tropica/crescimento & desenvolvimento , Psychodidae/parasitologia , Sequenciamento Completo do Genoma/métodos
10.
Genes (Basel) ; 11(2)2020 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-32075264

RESUMO

Interspecific hybridization creates genetic variation useful for crop improvement. However, whether pollen from a different genus affects the genomic stability and/or transcriptome of the recipient species during intergeneric pollination has not been investigated. Here, we crossed japonica rice cv. Z12 with the maize accession B73 (pollen donor) and obtained a morphologically stable line, MU1, exhibiting moderate dwarfism, higher tiller number, and increased grain weight compared with Z12. To reveal the genetic basis of these morphological changes in MU1, we performed whole-genome resequencing of MU1 and Z12. Compared with Z12, MU1 showed 107,250 single nucleotide polymorphisms (SNPs) and 23,278 insertion/deletions (InDels). Additionally, 5'-upstream regulatory regions (5'UTRs) of 429 and 309 differentially expressed genes (DEGs) in MU1 contained SNPs and InDels, respectively, suggesting that a subset of these DEGs account for the variation in 5'UTRs. Transcriptome analysis revealed 2190 DEGs in MU1 compared with Z12. Genes up-regulated in MU1 were mainly involved in photosynthesis, generation of precursor metabolites, and energy and cellular biosynthetic processes; whereas those down-regulated in MU1 were involved in plant hormone signal transduction pathway and response to stimuli and stress processes. Quantitative PCR (qPCR) further identified the expression levels of the up- or down-regulated gene in plant hormone signal transduction pathway. The expression level changes of plant hormone signal transduction pathway may be significant for plant growth and development. These findings suggest that mutations caused by intergeneric pollination could be the important reason for changes of MU1 in agronomic traits.


Assuntos
Quimera/crescimento & desenvolvimento , Produtos Agrícolas/crescimento & desenvolvimento , Perfilação da Expressão Gênica/métodos , Oryza/genética , Sequenciamento Completo do Genoma/métodos , Zea mays/genética , Quimera/genética , Produtos Agrícolas/genética , Flores/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Hibridização Genética , Mutação INDEL , Oryza/crescimento & desenvolvimento , Melhoramento Vegetal , Proteínas de Plantas/genética , Polinização , Polimorfismo de Nucleotídeo Único , Zea mays/crescimento & desenvolvimento
11.
Br J Nutr ; 123(2): 121-134, 2020 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-31637992

RESUMO

The experiment was conducted to investigate the effects of dietary threonine (Thr) on growth performance and muscle growth, protein synthesis and antioxidant-related signalling pathways of hybrid catfish Pelteobagrus vachelli♀ × Leiocassis longirostris♂. A total of 1200 fish (14·19 (se 0·13) g) were randomly distributed into six groups with four replicates each, fed six diets with graded level of Thr (9·5, 11·5, 13·5, 15·4, 17·4 and 19·3 g/kg diets) for 56 d. Results showed (P < 0·05) that dietary Thr (1) increased percentage weight gain, specific growth rate, feed efficiency and protein efficiency ratio; (2) up-regulated growth hormone (GH), insulin-like growth factor 1 (IGF-1), proliferating cell nuclear antigen, myogenic regulation factors (MyoD, Myf5, MyoG and Mrf4) and myosin heavy chain (MyHC) mRNA levels; (3) increased muscle protein content via regulating the protein kinase B/target of rapamycin signalling pathway and (4) decreased malondialdehyde and protein carbonyl contents, increased catalase, glutathione-S-transferase, glutathione reductase and GSH activities, up-regulated mRNA levels of antioxidant enzymes related to NFE2-related factor 2 and γ-glutamylcysteine ligase catalytic subunit. These results suggest that Thr has a potential role to improve muscle growth and protein synthesis, which might be due to the regulation of GH-IGF system, muscle growth-related gene, antioxidative capacity and protein synthesis-related signalling pathways. Based on the quadratic regression analysis of specific growth rate, the Thr requirement of hybrid catfish (14·19-25·77 g) was estimated to be 13·77 g/kg of the diet (33·40 g/kg of dietary protein).


Assuntos
Ração Animal/análise , Peixes-Gato/crescimento & desenvolvimento , Peixes-Gato/metabolismo , Músculos/efeitos dos fármacos , Treonina/administração & dosagem , Animais , Elementos de Resposta Antioxidante/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Peixes-Gato/genética , Quimera/crescimento & desenvolvimento , Quimera/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Feminino , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Hormônio do Crescimento/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Masculino , Músculos/metabolismo , Biossíntese de Proteínas/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos
12.
Virol J ; 16(1): 145, 2019 11 27.
Artigo em Inglês | MEDLINE | ID: mdl-31771593

RESUMO

BACKGROUND: Rice stripe mosaic virus (RSMV) is a tentative new Cytorhabdovirus species in family Rhabdoviridae transmitted by the leafhopper Recilia dorsalis. Although the virus was first detected in southern China in 2015, few studies have investigated rice symptoms and yield losses caused by RSMV infection. METHODS: In this study, we observed and systematically compared symptoms of three virally infected, representative varieties of indica, hybrid and japonica rice and determined the yield parameters of the artificially inoculated plants. RESULTS: The three RSMV-infected cultivated rice varieties exhibited slight dwarfing, striped mosaicism, stiff, crinkled or even twisted leaves, an increased number of tillers, delayed heading, cluster-shaped shortening of panicles and mostly unfilled grains. Slight differences in symptom occurrence time were observed under different environmental conditions. For example, mosaic symptoms appeared earlier and crinkling symptoms appeared later, with both symptoms later receding in some infected plants. Yield losses due to RSMV also differed among varieties. The most serious yield reduction was experienced by indica rice (cv. Meixiangzhan), followed by hybrid indica rice (cv. Wuyou 1179) and then japonica (cv. Nipponbare). Single panicle weight, seed setting rate and 1000-kernel weight were reduced in the three infected varieties compared with healthy plants-by 85.42, 94.85 and 31.56% in Meixiangzhan; 52.43, 53.06 and 25.65% in Wuyou 1179 and 25.53, 49.32 and 23.86% in Nipponbare, respectively. CONCLUSIONS: Our findings contribute basic data for field investigations, formulation of prevention and control strategies and further study of the pathogenesis of RSMV.


Assuntos
Oryza/crescimento & desenvolvimento , Oryza/virologia , Doenças das Plantas/virologia , Infecções por Rhabdoviridae/virologia , Rhabdoviridae/crescimento & desenvolvimento , Quimera/anatomia & histologia , Quimera/crescimento & desenvolvimento , Quimera/virologia , China , Oryza/anatomia & histologia
13.
Sci Rep ; 9(1): 14284, 2019 10 03.
Artigo em Inglês | MEDLINE | ID: mdl-31582777

RESUMO

In the last decade, avian gene preservation research has focused on the use of the early precursors of the reproductive cells, the primordial germ cells (PGCs). This is because avian PGCs have a unique migration route through the vascular system which offers easy accessibility. Furthermore, culturing of the cells in vitro, freezing/thawing, reintegration into a recipient embryo and the development of the germ cells can be carried out in well-defined laboratory circumstances. The efficient recovery of the donor genotype and the frequency of germline transmission from the surrogate host animals are still areas which need further development. Thus, the aim of the present study was to investigate an infertile interspecific hybrid (recipient) as an appropriate host for primordial germ cells from native poultry breeds. Guinea fowl × chicken hybrids were produced, the crossing was repeated inversely. The phenotype, the hatching time, the hatching rate, the sex ratio, the presence of own germ cells, the fertility and the phenotype of viable hybrids and the incidence of chromosomal abnormalities of dead hybrid embryos were described. 6.65% viable offspring was obtained with crossing of Guinea fowl females with domestic fowl males. Crossing of domestic fowl hens with Guinea fowl male resulted in lower fertility, 0.14% viable offspring. Based on the investigations, the observed offspring from the successful crossing were sterile male hybrids, thus an extreme form of Haldane's rule was manifested. The sterile hybrid male embryos were tested by injecting fluorescently labeled chicken PGCs. The integration rate of labeled PGCs was measured in 7.5-day, 14.5-day and 18.5-day old embryonic gonads. 50%, 5.3% and 2.4% of the injected hybrid embryos survived and 40%, 5.3% and 2.4% of the examined gonads contained fluorescent labeled donor PGCs. Therefore, these sterile hybrid males may be suitable recipients for male PGCs and possibly for female PGCs although with lower efficiency. This research work shows that the sterility of hybrids can be used in gene conservation to be a universal host for PGCs of different avian species.


Assuntos
Criopreservação , Espécies em Perigo de Extinção , Galliformes/genética , Animais , Cruzamento/métodos , Galinhas/genética , Galinhas/crescimento & desenvolvimento , Quimera/genética , Quimera/crescimento & desenvolvimento , Criopreservação/métodos , Feminino , Galliformes/crescimento & desenvolvimento , Células Germinativas/citologia , Células Germinativas/crescimento & desenvolvimento , Células Germinativas/metabolismo , Hibridização Genética , Masculino
14.
PLoS One ; 14(5): e0216939, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31141543

RESUMO

In the present two-year study, an attempt was made to estimate the grain yield, grain nutrient uptake, and oil quality of three commonly grown maize (Zea mays L.) hybrids fertilized with varied levels of nitrogen (N), phosphorus (P) and potassium (K). Results obtained from both the experimental years indicated that application of 125% of recommended dose of fertilizer (RDF) recorded maximum grain yield (10.37 t ha-1; 124% higher than control). When compared with 100% RDF, grain yield reduction with nutrient omission was 44% for N omission, 17% for P omission, and 27% for K omission. Nitrogen uptake was increased with increasing NPK levels up to 150% RDF that was statistically at par (p ≥ 0.01) with 125% RDF. Increasing trend in P and K uptake was observed with successive increase in NPK levels up to 125% RDF, above which it declined. The protein content was significantly higher in grains of var. P 3396 with 125% RDF. Nutrient management has significant (p ≤ 0.01) role in the grain oil content. Saturated fatty acids (palmitic, stearic and arachidic acid) content decreased, and unsaturated fatty acid (oleic, linoleic and linolenic acid) increased with increasing NPK levels. The average oleic acid desaturation and linoleic acid desaturation ratios were increased with increasing NPK levels up to 100 and 125% RDF, respectively. However, average monounsaturated fatty acids (MUFA): poly-unsaturated fatty acids (PUFA), saturated: unsaturated as well as linoleic: linolenic acid ratios were increased on receiving 75% RDF, and beyond that it showed decreasing trend. The omission of K had the highest inhibitory effect on corn oil quality followed by N and P omission.


Assuntos
Óleo de Milho/química , Grão Comestível/efeitos dos fármacos , Nitrogênio/farmacologia , Fósforo/farmacologia , Potássio/farmacologia , Zea mays/efeitos dos fármacos , Quimera/crescimento & desenvolvimento , Quimera/metabolismo , Óleo de Milho/metabolismo , Grão Comestível/crescimento & desenvolvimento , Grão Comestível/metabolismo , Ácidos Graxos/biossíntese , Ácidos Graxos/classificação , Ácidos Graxos/isolamento & purificação , Ácidos Graxos Insaturados/biossíntese , Ácidos Graxos Insaturados/classificação , Ácidos Graxos Insaturados/isolamento & purificação , Fertilizantes/análise , Humanos , Nitrogênio/metabolismo , Nutrientes/metabolismo , Nutrientes/farmacologia , Fósforo/metabolismo , Proteínas de Plantas/biossíntese , Proteínas de Plantas/isolamento & purificação , Potássio/metabolismo , Zea mays/crescimento & desenvolvimento , Zea mays/metabolismo
15.
Proc Natl Acad Sci U S A ; 116(22): 10852-10857, 2019 05 28.
Artigo em Inglês | MEDLINE | ID: mdl-31085653

RESUMO

In perennial plants, seasonal shifts provide cues that control adaptive growth patterns of the shoot apex. However, where these seasonal cues are sensed and communicated to the shoot apex remains unknown. We demonstrate that systemic signals from leaves play key roles in seasonal control of shoot growth in model tree hybrid aspen. Grafting experiments reveal that the tree ortholog of Arabidopsis flowering time regulator FLOWERING LOCUS T (FT) and the plant hormone gibberellic acid (GA) systemically convey seasonal cues to the shoot apex. GA (unlike FT) also acts locally in shoot apex, downstream of FT in seasonal growth control. At the shoot apex, antagonistic factors-LAP1, a target of FT and the FT antagonist TERMINAL FLOWER 1 (TFL1)-act locally to promote and suppress seasonal growth, respectively. These data reveal seasonal changes perceived in leaves that are communicated to the shoot apex by systemic signals that, in concert with locally acting components, control adaptive growth patterns.


Assuntos
Reguladores de Crescimento de Plantas/metabolismo , Brotos de Planta/crescimento & desenvolvimento , Transdução de Sinais/fisiologia , Arabidopsis/fisiologia , Proteínas de Arabidopsis/metabolismo , Quimera/crescimento & desenvolvimento , Giberelinas/metabolismo , Fotoperíodo , Fenômenos Fisiológicos Vegetais , Estações do Ano
16.
Elife ; 82019 05 16.
Artigo em Inglês | MEDLINE | ID: mdl-31090541

RESUMO

While lower vertebrates contain adult stem cells (aSCs) that maintain homeostasis and drive un-exhaustive organismal growth, mammalian aSCs display mainly the homeostatic function. Here, we use lineage analysis in the medaka fish gill to address aSCs and report separate stem cell populations for homeostasis and growth. These aSCs are fate-restricted during the entire post-embryonic life and even during re-generation paradigms. We use chimeric animals to demonstrate that p53 mediates growth coordination among fate-restricted aSCs, suggesting a hierarchical organisation among lineages in composite organs like the fish gill. Homeostatic and growth aSCs are clonal but differ in their topology; modifications in tissue architecture can convert the homeostatic zone into a growth zone, indicating a leading role for the physical niche defining stem cell output. We hypothesise that physical niches are main players to restrict aSCs to a homeostatic function in animals with fixed adult size.


Assuntos
Tecido Adiposo/crescimento & desenvolvimento , Células-Tronco Adultas/metabolismo , Brânquias/crescimento & desenvolvimento , Oryzias/crescimento & desenvolvimento , Tecido Adiposo/metabolismo , Animais , Diferenciação Celular/genética , Quimera/genética , Quimera/crescimento & desenvolvimento , Genes p53/genética , Brânquias/metabolismo , Homeostase/genética , Humanos , Oryzias/metabolismo , Nicho de Células-Tronco/genética
17.
Heredity (Edinb) ; 123(3): 318-336, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30911141

RESUMO

Heterosis refers to the superior performance of F1 hybrids over their respective parental inbred lines. Although the genetic and expression basis of heterosis have been previously investigated, the metabolic basis for this phenomenon is poorly understood. In a preliminary morphological study in Brassica juncea, we observed significant heterosis at the 50% flowering stage, wherein both the growth and reproduction of F1 reciprocal hybrids were greater than that of their parents. To identify the possible metabolic causes or consequences of this heterosis, we carried out targeted LC-MS analysis of 48 primary (amino acids and sugars) and secondary metabolites (phytohormones, glucosinolates, flavonoids, and phenolic esters) in five developmental tissues at 50% flowering in hybrids and inbred parents. Principal component analysis (PCA) of metabolites clearly separated inbred lines from their hybrids, particularly in the bud tissues. In general, secondary metabolites displayed more negative heterosis values in comparison to primary metabolites. The tested primary and secondary metabolites displayed both additive and non-additive modes of inheritance in F1 hybrids, wherein the number of metabolites showing an additive mode of inheritance were higher in buds and siliques (52.77-97.14%) compared to leaf tissues (47.37-80%). Partial least regression (PLS) analysis further showed that primary metabolites, in general, displayed higher association with morphological parameters in F1 hybrids. Overall, our results are consistent with a resource-cost model for heterosis in B. juncea, where metabolite allocation in hybrids appears to favor growth, at the expense of secondary metabolism.


Assuntos
Quimera/metabolismo , Vigor Híbrido , Padrões de Herança , Metaboloma , Mostardeira/metabolismo , Metabolismo Secundário/genética , Quimera/genética , Quimera/crescimento & desenvolvimento , Produtos Agrícolas , Flavonoides/biossíntese , Flavonoides/química , Flores/genética , Flores/crescimento & desenvolvimento , Flores/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Glucosinolatos/biossíntese , Glucosinolatos/química , Mostardeira/genética , Mostardeira/crescimento & desenvolvimento , Fenóis/química , Fenóis/metabolismo , Melhoramento Vegetal , Reguladores de Crescimento de Plantas/biossíntese , Reguladores de Crescimento de Plantas/química , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Óleos de Plantas/metabolismo , Análise de Componente Principal
18.
Methods Mol Biol ; 1874: 403-430, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30353528

RESUMO

The unique characteristics of the avian embryo, with its large opaque yolk, have necessitated the development of different approaches to transgenesis from those that have been successful in mammalian species. Genetic modification of birds was greatly advanced by the ability to grow long-term cultures of primordial germ cells (PGCs). These cells are obtained from embryos, established in culture, and can be propagated without losing the ability to contribute to the germline when reintroduced into a host animal. PGCs can be genetically modified in culture using traditional transfection and selection techniques, including gene targeting and site-specific nuclease approaches. Here, we describe our methods for deriving cell lines, long-term culture, genetic modification, production of germline chimeras and obtaining fully transgenic birds with the desired genetic modifications.


Assuntos
Animais Geneticamente Modificados/crescimento & desenvolvimento , Galinhas/genética , Quimera/crescimento & desenvolvimento , Células Germinativas/citologia , Animais , Linhagem Celular , Células Cultivadas , Galinhas/crescimento & desenvolvimento , Técnicas de Cocultura , Feminino , Técnicas de Transferência de Genes , Células Germinativas/metabolismo , Masculino , Ratos
19.
Theor Med Bioeth ; 39(4): 301-319, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-30167942

RESUMO

This paper presents an account of how human spontaneous embryonic chimeras are formed. On the prevalent view in the philosophical literature, it is said that chimeras are the product of two embryos that fuse to form a new third embryo. We call this version of fusion synthesis. In contrast to synthesis, we present an alternative mechanism for chimera formation called incorporation, wherein one embryo incorporates the cells of a second embryo into its body. We argue that the incorporation thesis explains other types of chimera formation, which are better understood, and is more consistent than synthesis with what is known about embryological development. Incorporation also has different implications than synthesis and so avoids the philosophical puzzles that are often said to accompany embryonic chimera formation-puzzles which pose problems to the human embryo's persistence from fertilization to the fetal stage of human development.


Assuntos
Quimera/crescimento & desenvolvimento , Gêmeos/genética , Quimera/metabolismo , Humanos
20.
Microbiol Res ; 214: 101-113, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30031472

RESUMO

Maize, a crop cultivated worldwide, was investigated for plant tissue and crop stage specific colonization of endophytic bacteria. Such bacterial interactions have high potential to enhance maize grain yield by means of biological nitrogen fixation and/or plant growth promoting activities. In this study endophytic bacteria were isolated from a hybrid PEEHM-5 and composite PC-4 maize varieties using root, stem and leaf tissues of plants at vegetative, flowering and maturity stages of growth. PEEHM-5 harbored higher endophytic bacterial population than PC-4 at all growth stages, with highest in roots and at flowering stage. Morphologically 188 different endophytic isolates (82 from PEEHM-5, 106 from PC-4) were screened for plant growth promoting attributes viz. P, K, Zn solubilization, production of hormones, siderophore, ACC deaminase, HCN, biological nitrogen fixation and biocontrol of two maize fungal pathogens. Thirty one potential PGP isolates on RFLP analysis of their amplified 16S rRNA gene, were clustered in 13 phylogenetic groups. On sequencing and blasting of amplified 16S rRNA gene of representative isolates from each group identified PC-4 endophytic bacterial isolates as Bacillus aryabhattai, Pantoea cypripedii, Bacillus licheniformis, Klebsiella sp., Pantoea dispersa, Klebsiella variicola, Pantoea sp., Agrobacterium larrymoorei and PEEHM-5 endophytic bacterial isolates as Bacillus sp., Bacillus amyloliquefaciens, Lactococcus lactis, Bacillus cereus and Staphylococcus hominis. In planta evaluation of potential isolates at variable chemical fertilizer input indicated their potential in compensating nearly 25% of the fertilizer input as observed on their improvement of shoot and root parameters. Lactococcus lactis inoculation influenced maximum followed by Pantoea and Klebsiella isolates.


Assuntos
Bactérias/classificação , Bactérias/isolamento & purificação , Biodiversidade , Endófitos/classificação , Endófitos/isolamento & purificação , Zea mays/crescimento & desenvolvimento , Zea mays/microbiologia , Bactérias/genética , Bactérias/metabolismo , Quimera/crescimento & desenvolvimento , Quimera/microbiologia , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Endófitos/genética , Filogenia , Reguladores de Crescimento de Plantas/metabolismo , Folhas de Planta/microbiologia , Raízes de Plantas/microbiologia , Caules de Planta/microbiologia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
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