RESUMO
Commercial fig tree cultivation in Brazil involves a single cultivar, 'Roxo-de-Valinhos'. The use of a single cultivar results in serious diseases and related problems. The aim of this study was to characterize fig accessions by analyzing the natural root-knot nematode and leaf rust incidence in relation to the epigenomic profile of the plant, since epigenetic variations affect plant-pathogen interactions. All plants were attacked by nematodes, indicating susceptibility; Meloidogyne incognita was the root-knot nematode species involved. Joint analysis of data showed that methylation and leaf rust incidence were correlated when observed in the same phenological phase, presenting initial evidence of the same factorial pressure loads in genotypes, suggesting similar behavior within these genotypes.
O Brasil é o maior produtor de figos da América do Sul, porém o cultivo comercial brasileiro da figueira baseia-se na plantação de uma única cultivar, o Roxo-de-Valinhos, resultando em sérios problemas relativos a pragas e doenças. Uma vez que há variações epigenéticas na interação planta-patógeno, principalmente por meio da regulação gênica, o presente trabalho objetiva realizar a caracterização in vivo de acessos de figo, por meio da análise de incidência natural de nematoides formadores de galha e de incidência natural de ferrugem, correlacionada ao seu perfil epigenômico, a fim de subsidiar trabalhos de conservação, melhoramento genético e produção da cultura. A análise dos componentes principais dos dados da caracterização dos acessos foi realizada por meio da matriz de correlação residual obtida pela análise de variância conjunta utilizando o programa GENES. Pôde-se constatar que todas as plantas foram atacadas por este patógeno, evidenciando que são suscetíveis ao mesmo. Já a análise conjunta dos dados demonstrou que a metilação e a incidência de ferrugem em folhas, quando observadas na mesma fase fenológica da planta, se correlacionam, apresentando evidências iniciais de mesmas cargas fatoriais de pressão nos genótipos, com a premissa de comportamento semelhante nos mesmos, indicando que, além do fator genético, fatores abióticos também são responsáveis pelas alterações no fenótipo das plantas, evidenciando a plasticidade fenotípica das mesmas.
Assuntos
Ficus/parasitologia , Melhoramento Genético , BrasilRESUMO
Commercial fig tree cultivation in Brazil involves a single cultivar, 'Roxo-de-Valinhos'. The use of a single cultivar results in serious diseases and related problems. The aim of this study was to characterize fig accessions by analyzing the natural root-knot nematode and leaf rust incidence in relation to the epigenomic profile of the plant, since epigenetic variations affect plant-pathogen interactions. All plants were attacked by nematodes, indicating susceptibility; Meloidogyne incognita was the root-knot nematode species involved. Joint analysis of data showed that methylation and leaf rust incidence were correlated when observed in the same phenological phase, presenting initial evidence of the same factorial pressure loads in genotypes, suggesting similar behavior within these genotypes.
Assuntos
Ficus , Tylenchoidea , Animais , Incidência , Metilação , Raízes de Plantas , Plantas , Árvores/genéticaRESUMO
We assessed whether single nucleotide polymorphisms (SNPs) in the genes beta 1,4- galactosyltransferase (B4GALT1), luteinizing hormone receptor (LHR), follicle-stimulating hormone receptor (FSHR) and insulin-like growth factor 2 (IGF2) could be molecular markers for scrotal circumference (SC) in Nellore bulls. Animals with positive (+, n = 104) and negative (-, n = 74) expected progeny difference for scrotal circumference at 365 days (EPD SC 365) were selected and their SNPs were analyzed by restriction fragment length polymorphism (RFLP). The correlation between EPD SC 365 and expected progeny difference for age at first birth (EPD AFB) was also investigated. The SNPs in B4GALT1 and FSHR was not different between two groups analyzed. The CC genotype for LHR gene was most frequent in animals with EPD SC 365(+), whereas the TT was most frequent in the EPD SC 365(-). For IGF2 the CT and CC were the most frequent genotypes observed in animals with positive and negative EPD SC 365, respectively. The EPD SC 365 was negatively correlated with the EPD AFB (r = 0.23). We suggest that CC and TT genotypes for LHR and IGF2, respectively, could be possible molecular markers for SC selection in Nellore bulls, that can also predict for AFB.
Foram avaliados se polimorfismos de base única (SNPs) presentes nos genes beta-1,4- galactosiltransferase (B4GALT1), receptor de hormônio luteinizante (LHR), receptor de hormônio folículo estimulante (FSHR) e fator de crescimento semelhante à insulina 2 (IGF2) poderiam ser marcadores moleculares para o perímetro escrotal (PE) em touros da raça Nelore. Animais com diferença esperada de progênie positiva (+, n = 104) e negativa (-, n = 74) para PE aos 365 dias (DEP PE 365) foram selecionados e seus SNPs foram analisados utilizando a técnica de polimorfismo de comprimento de fragmentos de restrição (RFLP). A correlação entre DEP PE 365 e idade ao primeiro parto (DEP IPP) também foi investigada. Os SNPs dos genes B4GALT1 e FSHR não apresentaram diferença entre os dois grupos analisados. O genótipo CC para o gene LHR foi mais freqüente em animais com DEP PE 365 (+), enquanto o TT foi mais frequente no grupo com DEP PE 365 (-). Para o gene IGF2, os genótipos CT e CC foram mais freqüentes em animais com DEP PE 365 positiva e negativa, respectivamente. A DEP PE 365 foi negativamente correlacionada com a DEP IPP (r = -0,23). O genótipo CC para o gene LHR e genótipo TT para o gene IGF2 podem ser possíveis marcadores de PE para a seleção assistida em touros da raça Nelore, podendo ser ainda preditores para IPP.
Assuntos
Masculino , Animais , Bovinos , Bovinos/anatomia & histologia , Bovinos/genética , Fator de Crescimento Insulin-Like II/análise , Polimorfismo Genético/genética , Receptores do FSH/análise , Receptores do LH/análise , /análiseRESUMO
We assessed whether single nucleotide polymorphisms (SNPs) in the genes beta 1,4- galactosyltransferase (B4GALT1), luteinizing hormone receptor (LHR), follicle-stimulating hormone receptor (FSHR) and insulin-like growth factor 2 (IGF2) could be molecular markers for scrotal circumference (SC) in Nellore bulls. Animals with positive (+, n = 104) and negative (-, n = 74) expected progeny difference for scrotal circumference at 365 days (EPD SC 365) were selected and their SNPs were analyzed by restriction fragment length polymorphism (RFLP). The correlation between EPD SC 365 and expected progeny difference for age at first birth (EPD AFB) was also investigated. The SNPs in B4GALT1 and FSHR was not different between two groups analyzed. The CC genotype for LHR gene was most frequent in animals with EPD SC 365(+), whereas the TT was most frequent in the EPD SC 365(-). For IGF2 the CT and CC were the most frequent genotypes observed in animals with positive and negative EPD SC 365, respectively. The EPD SC 365 was negatively correlated with the EPD AFB (r = 0.23). We suggest that CC and TT genotypes for LHR and IGF2, respectively, could be possible molecular markers for SC selection in Nellore bulls, that can also predict for AFB.(AU)
Foram avaliados se polimorfismos de base única (SNPs) presentes nos genes beta-1,4- galactosiltransferase (B4GALT1), receptor de hormônio luteinizante (LHR), receptor de hormônio folículo estimulante (FSHR) e fator de crescimento semelhante à insulina 2 (IGF2) poderiam ser marcadores moleculares para o perímetro escrotal (PE) em touros da raça Nelore. Animais com diferença esperada de progênie positiva (+, n = 104) e negativa (-, n = 74) para PE aos 365 dias (DEP PE 365) foram selecionados e seus SNPs foram analisados utilizando a técnica de polimorfismo de comprimento de fragmentos de restrição (RFLP). A correlação entre DEP PE 365 e idade ao primeiro parto (DEP IPP) também foi investigada. Os SNPs dos genes B4GALT1 e FSHR não apresentaram diferença entre os dois grupos analisados. O genótipo CC para o gene LHR foi mais freqüente em animais com DEP PE 365 (+), enquanto o TT foi mais frequente no grupo com DEP PE 365 (-). Para o gene IGF2, os genótipos CT e CC foram mais freqüentes em animais com DEP PE 365 positiva e negativa, respectivamente. A DEP PE 365 foi negativamente correlacionada com a DEP IPP (r = -0,23). O genótipo CC para o gene LHR e genótipo TT para o gene IGF2 podem ser possíveis marcadores de PE para a seleção assistida em touros da raça Nelore, podendo ser ainda preditores para IPP.(AU)
Assuntos
Animais , Masculino , Bovinos , Bovinos/anatomia & histologia , Bovinos/genética , Polimorfismo Genético/genética , beta-N-Acetilglucosaminilglicopeptídeo beta-1,4-Galactosiltransferase/análise , Receptores do LH/análise , Receptores do FSH/análise , Fator de Crescimento Insulin-Like II/análiseRESUMO
Nuclear reprogramming mediated by somatic cell nuclear transfer (SCNT) has many applications in medicine. However, animal clones show increased rates of abortion and reduced neonatal viability. Herein, we used exosomal-miRNA profiles as a non-invasive biomarker to identify pathological pregnancies. MiRNAs play important roles in cellular proliferation and differentiation during early mammalian development. Thus, the aim of this study was to identify exosomal-miRNAs in maternal blood at 21 days of gestation that could be used for diagnosis and prognosis during early clone pregnancies in cattle. Out of 40 bovine-specific miRNAs, 27 (67.5%) were with low abundance in the C-EPL (Clone - Early pregnancy loss) group compared with the C-LTP (Clone - Late pregnancy) and AI-LTP (Artificial Insemination - Late pregnancy) groups, which had similar miRNAs levels. Bioinformatics analysis of the predicted target genes demonstrated signaling pathways and functional annotation clusters associated with critical biological processes including cell proliferation, differentiation, apoptosis, angiogenesis and embryonic development. In conclusion, our results demonstrate decreased exosomal-miRNAs in maternal blood at 21 days of gestation in cloned cattle pregnancies that failed to reach term. Furthermore, the predicted target genes regulated by these 27 miRNAs are strongly associated with pregnancy establishment and in utero embryonic development.
Assuntos
Aborto Espontâneo/genética , Ácidos Nucleicos Livres/metabolismo , Exossomos/metabolismo , MicroRNAs/metabolismo , Animais , Bovinos , Diferenciação Celular , Proliferação de Células/genética , Ácidos Nucleicos Livres/genética , Reprogramação Celular , Clonagem de Organismos , Biologia Computacional , Desenvolvimento Embrionário , Feminino , Perfilação da Expressão Gênica , Inseminação Artificial , MicroRNAs/genética , Anotação de Sequência Molecular , Mães , Técnicas de Transferência Nuclear , Gravidez , Transdução de SinaisRESUMO
The chicken (Gallus gallus) embryo has been used as a classic model system for developmental studies because of its easy accessibility for surgical manipulation during embryonic development. Sex determination in birds is chromosomally based (ZZ for males and ZW for females); however, the basic mechanism of sex determination is still unknown. Here, the dynamics of expression of candidate genes implicated in vertebrate sex determination and differentiation were studied during embryonic chicken gonadal development. Gene expression profiles were obtained before, during, and after gonadal sex differentiation in females and males for DMRT1, SOX3, SOX9, DAX1, SCII, HINTZ, HINTW, and the male hypermethylated (MHM) region. Transcripts for the HINTZ, DMRT1, DAX1, SCII, and SOX9 genes were observed in both sexes, but expression was higher in male gonads and may be correlated with testicular differentiation. The expression patterns of HINTW, SOX3, and MHM suggest that they may act in ovary development and may be involved in meiosis entry. MHM was upregulated and DMRT1 was downregulated in females at the same developmental stage. This may indicate a regulation of DMRT1 by MHM ncRNA. Similar dynamics were observed between HINTW and HINTZ. This study reports on the MHM expression profile during gonadal development and its correlation with the expression of genes involved in vertebrate sex determination.
Assuntos
Gônadas/crescimento & desenvolvimento , Processos de Determinação Sexual , Diferenciação Sexual/genética , Animais , Embrião de Galinha , Metilação de DNA/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Masculino , Cromossomos Sexuais/genética , Testículo/crescimento & desenvolvimento , Testículo/metabolismoRESUMO
In addition to methylated cytosines (5-mCs), hydroxymethylcytosines (5-hmCs) are present in CpG dinucleotide-enriched regions and some transcription regulator binding sites. Unlike methylation, hydroxymethylation does not result in silencing of gene expression, and the most commonly used methods to study methylation, such as techniques based on restriction enzymatic digestion and/or bisulfite modification, are unable to distinguish between them. Genomic imprinting is a process of gene regulation where only one member of an allelic pair is expressed depending on the parental origin. Chromosome 11p15.5 has an imprinting control region (ICR2) that includes a differentially methylated region (KvDMR1) that guarantees parent-specific gene expression. The objective of the present study was to determine the presence of 5-hmC at the KvDMR1 in human placentas. We analyzed 16 third-trimester normal human placentas (chorionic villi). We compared two different methods based on real-time PCR after enzymatic digestion. The first method distinguished methylation from hydroxymethylation, while the other method did not. Unlike other methylation studies, subtle variations of methylation in ICRs could represent a drastic deregulation of the expression of imprinted genes, leading to important phenotypic consequences, and the presence of hydroxymethylation could interfere with the results of many studies. We observed agreement between the results of both methods, indicating the absence of hydroxymethylation at the KvDMR1 in third-trimester placentas. To the best of our knowledge, this is the first study describing the investigation of hydroxymethylation in human placenta using a genomic imprinting model.
RESUMO
In addition to methylated cytosines (5-mCs), hydroxymethylcytosines (5-hmCs) are present in CpG dinucleotide-enriched regions and some transcription regulator binding sites. Unlike methylation, hydroxymethylation does not result in silencing of gene expression, and the most commonly used methods to study methylation, such as techniques based on restriction enzymatic digestion and/or bisulfite modification, are unable to distinguish between them. Genomic imprinting is a process of gene regulation where only one member of an allelic pair is expressed depending on the parental origin. Chromosome 11p15.5 has an imprinting control region (ICR2) that includes a differentially methylated region (KvDMR1) that guarantees parent-specific gene expression. The objective of the present study was to determine the presence of 5-hmC at the KvDMR1 in human placentas. We analyzed 16 third-trimester normal human placentas (chorionic villi). We compared two different methods based on real-time PCR after enzymatic digestion. The first method distinguished methylation from hydroxymethylation, while the other method did not. Unlike other methylation studies, subtle variations of methylation in ICRs could represent a drastic deregulation of the expression of imprinted genes, leading to important phenotypic consequences, and the presence of hydroxymethylation could interfere with the results of many studies. We observed agreement between the results of both methods, indicating the absence of hydroxymethylation at the KvDMR1 in third-trimester placentas. To the best of our knowledge, this is the first study describing the investigation of hydroxymethylation in human placenta using a genomic imprinting model.
RESUMO
We analyzed two single nucleotide polymorphisms (SNPs) of the IGF2 and CYP21 genes in Nellore cattle participating in the Brazilian Animal Breeding Program. The SNPs were found in exon 6 of the IGF2 (insulin-like growth factor 2) gene (RFLP/MboII) as well as in the promoter region of the CYP21 (steroid 21-hydroxylase) gene (RFLP/HpaII) of these animals. The TC heterozygotes were significantly more frequent than CC and TT homozygotes in the RFLP/MboII polymorphism. The T allele was significantly more frequent than the C allele in RFLP/HpaII polymorphism. This population was found to be in Hardy-Weinberg equilibrium for these SNPs. Association of these polymorphisms with expected progeny differences of reproductive and productive traits was investigated, but proved to be significant only for DP550 (expected progeny differenced for weight at 365 days - IGF2 - RFLP/MboII) and DP450 (expected progeny differenced for weight at 450 days - CYP21 - RFLP/HpaII). This is the first study on the occurrence of these two polymorphisms in this Zebu breed of cattle. A total of 147 Nellore animals participating in the Breeding Program of the Nellore Breed (PMGRN) under the management of the National Association of Breeders and Researchers (ANCP) in the city of Ribeirão Preto were analyzed.
Assuntos
Composição Corporal/genética , Bovinos/genética , Fator de Crescimento Insulin-Like II/genética , Esteroide 21-Hidroxilase/genética , Agricultura , Alelos , Criação de Animais Domésticos , Animais , Peso Corporal/genética , Cruzamento , Frequência do Gene , Marcadores Genéticos , Fenótipo , Polimorfismo de Nucleotídeo ÚnicoRESUMO
The testis-specific protein Y-encoded gene (TSPY) is a Y-specific gene present in variable copy number in many mammalian species, including cattle. We tested the applicability of the TSPY gene as a Y-specific marker to predict preimplantation embryo sex in Nelore (Bos indicus) cattle. Two blastomeres were removed from each embryo. A total of 36 single blastomeres and the remaining cells of their 18 matched in vitro conceived embryos were screened for TSPY amplification by nested-PCR. The results obtained from a single blastomere and the remaining cells of the same embryo were concordant in all cases. All blastomeres (16/16) from eight embryos produced with sexed sperm (specific for production of male embryos) were TSPY-positive. We conclude that TSPY is a good male-specific marker, the usefulness of which is probably enhanced by the high copy number. Other methods that are less time-consuming, such as real-time PCR, could be improved with the use of the TSPY gene sequences to generate primers and/or probes. This is the first report to demonstrate the applicability of the TSPY gene for sexing single cells in cattle.