RESUMO
The draft genome of the pear (Pyrus bretschneideri) using a combination of BAC-by-BAC and next-generation sequencing is reported. A 512.0-Mb sequence corresponding to 97.1% of the estimated genome size of this highly heterozygous species is assembled with 194× coverage. High-density genetic maps comprising 2005 SNP markers anchored 75.5% of the sequence to all 17 chromosomes. The pear genome encodes 42,812 protein-coding genes, and of these, ~28.5% encode multiple isoforms. Repetitive sequences of 271.9 Mb in length, accounting for 53.1% of the pear genome, are identified. Simulation of eudicots to the ancestor of Rosaceae has reconstructed nine ancestral chromosomes. Pear and apple diverged from each other ~5.4-21.5 million years ago, and a recent whole-genome duplication (WGD) event must have occurred 30-45 MYA prior to their divergence, but following divergence from strawberry. When compared with the apple genome sequence, size differences between the apple and pear genomes are confirmed mainly due to the presence of repetitive sequences predominantly contributed by transposable elements (TEs), while genic regions are similar in both species. Genes critical for self-incompatibility, lignified stone cells (a unique feature of pear fruit), sorbitol metabolism, and volatile compounds of fruit have also been identified. Multiple candidate SFB genes appear as tandem repeats in the S-locus region of pear; while lignin synthesis-related gene family expansion and highly expressed gene families of HCT, C3'H, and CCOMT contribute to high accumulation of both G-lignin and S-lignin. Moreover, alpha-linolenic acid metabolism is a key pathway for aroma in pear fruit.
Assuntos
Genoma de Planta , Pyrus/genética , Cromossomos de Plantas , Evolução Molecular , Frutas/genética , Duplicação Gênica , Genes de Plantas , Variação Genética , Genótipo , Anotação de Sequência Molecular , Dados de Sequência Molecular , Filogenia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Pyrus/imunologia , Sequências Repetitivas de Ácido Nucleico , Rosaceae/genética , Rosaceae/imunologia , Análise de Sequência de DNA , TranscriptomaRESUMO
BACKGROUND: Heat treatment (known as thermotherapy) together with in vitro culture of shoot meristem tips is a commonly used technology to obtain virus-free germplasm for the effective control of virus diseases in fruit trees. RNA silencing as an antiviral defense mechanism has been implicated in this process. To understand if high temperature-mediated acceleration of the host antiviral gene silencing system in the meristem tip facilitates virus-derived small interfering RNAs (vsiRNA) accumulation to reduce the viral RNA titer in the fruit tree meristem tip cells, we used the Apple stem grooving virus (ASGV)-Pyrus pyrifolia pathosystem to explore the possible roles of vsiRNA in thermotherapy. RESULTS: At first we determined the full-length genome sequence of the ASGV-Js2 isolate and then profiled vsiRNAs in the meristem tip of in vitro-grown pear (cv. 'Jinshui no. 2') shoots infected by ASGV-Js2 and cultured at 24 and 37 °C. A total of 7,495 and 7,949 small RNA reads were obtained from the tips of pear shoots cultured at 24 and 37 °C, respectively. Mapping of the vsiRNAs to the ASGV-Js2 genome revealed that they were unevenly distributed along the ASGV-Js2 genome, and that 21- and 22-nt vsiRNAs preferentially accumulated at both temperatures. The 5'-terminal nucleotides of ASGV-specific siRNAs in the tips cultured under different temperatures had a similar distribution pattern, and the nucleotide U was the most frequent. RT-qPCR analyses suggested that viral genome accumulation was drastically compromised at 37 °C compared to 24 °C, which was accompanied with the elevated levels of vsiRNAs at 37 °C. As plant Dicer-like proteins (DCLs), Argonaute proteins (AGOs), and RNA-dependent RNA polymerases (RDRs) are implicated in vsiRNA biogenesis, we also cloned the partial sequences of PpDCL2,4, PpAGO1,2,4 and PpRDR1 genes, and found their expression levels were up-regulated in the ASGV-infected pear shoots at 37 °C. CONCLUSIONS: Collectively, these results showed that upon high temperature treatment, the ASGV-infected meristem shoot tips up-regulated the expression of key genes in the RNA silencing pathway, induced the biogenesis of vsiRNAs and inhibited viral RNA accumulation. This study represents the first report on the characterization of the vsiRNA population in pear plants infected by ASGV-Js2, in response to high temperature treatment.
Assuntos
Flexiviridae/crescimento & desenvolvimento , Temperatura Alta , Brotos de Planta/virologia , Pyrus/virologia , RNA Interferente Pequeno/genética , Flexiviridae/genética , Flexiviridae/efeitos da radiação , Inativação Gênica , Brotos de Planta/imunologia , Brotos de Planta/efeitos da radiação , Pyrus/imunologia , Pyrus/efeitos da radiação , RNA Interferente Pequeno/metabolismo , RNA Viral/antagonistas & inibidoresRESUMO
Erwinia amylovora is a necrogenic bacterium, causing the fire blight disease on many rosaceous plants. Triggering oxidative burst by E. amylovora is a key response by which host plants try to restrain pathogen spread. Electron transport chain (ETC) of chloroplasts is known as an inducible source of reactive oxygen species generation in various stresses. This research was performed to assess the role of this ETC in E. amylovora-host interaction using several inhibitors of this chain in susceptible and resistant apple and pear genotypes. All ETC inhibitors delayed appearance of disease necrosis, but the effects of methyl viologen, glutaraldehyde, and DCMU were more significant. In the absence of inhibitors, resistant genotypes showed an earlier and severe H2O2 generation and early suppression of redox dependent, psbA gene. The effects of inhibitors were corresponding to the redox potential of ETC inhibitory sites. In addition, delayed necrosis appearance was associated with the decreased disease severity and delayed H2O2 generation. These results provide evidences for the involvement of this ETC in host oxidative burst and suggest that chloroplast ETC has significant role in E. amylovora-host interaction.
Assuntos
Resistência à Doença , Erwinia amylovora/fisiologia , Malus/fisiologia , Doenças das Plantas/imunologia , Pyrus/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Cloroplastos/metabolismo , Transporte de Elétrons , Genótipo , Interações Hospedeiro-Patógeno , Peróxido de Hidrogênio/metabolismo , Malus/imunologia , Malus/metabolismo , Malus/microbiologia , Doenças das Plantas/microbiologia , Brotos de Planta/imunologia , Brotos de Planta/microbiologia , Brotos de Planta/fisiologia , Pyrus/imunologia , Pyrus/metabolismo , Pyrus/microbiologia , Explosão RespiratóriaRESUMO
Approximately 70% of birch pollen allergic patients in Europe experience hypersensitivity reactions to Immunoglobulin E (IgE) cross-reactive food sources. This so-called pollen-food syndrome (PFS) is defined by allergic symptoms elicited promptly by the ingestion of fruits, nuts, or vegetables in these patients. So far, in the literature, less attention has been given to Bet v 1 cross-reactive symptoms caused by pear (Pyrus communis). In the Netherlands, pears are widely consumed. The primary objective of this study was to measure the type and severity of allergic symptoms during pear challenges in birch pollen allergic patients, with a positive history of pear allergy, using two different pear varieties. Fifteen patients were included, skin prick test (SPT), prick-to-prick test (PTP), specific Immunoglobulin E (sIgE), and single-blind oral challenges were performed with two pear (Pyrus communis) varieties: the 'Cepuna' (brand name Migo®) and the 'Conference' pears. All patients were sensitized to one or both pear varieties. A total of 12 out of 15 participants developed symptoms during the 'Cepuna' food challenge and 14/15 reacted during the 'Conference' challenge. Challenges with the 'Cepuna' pears resulted in less objective symptoms (n = 2) in comparison with challenges with 'Conference' pears (n = 7). Although we did not find significance between both varieties in our study, we found a high likelihood of fewer and less severe symptoms during the 'Cepuna' challenges. Consequently selected pear sensitized patients can try to consume small doses of the 'Cepuna' pear outside the birch pollen season.
Assuntos
Alérgenos/imunologia , Betula/imunologia , Hipersensibilidade Alimentar/diagnóstico , Pólen/imunologia , Pyrus/imunologia , Adulto , Reações Cruzadas , Feminino , Hipersensibilidade Alimentar/imunologia , Frutas/imunologia , Humanos , Imunoglobulina E/imunologia , Masculino , Pessoa de Meia-Idade , Países Baixos , Método Simples-Cego , Testes Cutâneos , Adulto JovemRESUMO
Occupational allergies such as pollinosis are reported in several agricultural works in Japan. Many pollens and spores were observed in Japanese pear orchard during the artificial pollination season. By the study on daily symptoms in an allergic farmer, we confirmed that the pollinosis symptoms were most common and most severe during the artificial pollination. These results suggest that the exposure to allergenic pollens and spores induces allergic symptoms. Thus, caution should be paid for the avoidance of the exposure to these allergenic pollens and spores to prevent the allergy.
Assuntos
Doenças dos Trabalhadores Agrícolas/etiologia , Pólen/efeitos adversos , Pyrus/efeitos adversos , Rinite Alérgica Sazonal/etiologia , Doenças dos Trabalhadores Agrícolas/imunologia , Feminino , Humanos , Imunoglobulina E/sangue , Japão , Lycopodium/efeitos adversos , Lycopodium/imunologia , Exposição Ocupacional , Pólen/imunologia , Pyrus/imunologia , Rinite Alérgica Sazonal/imunologia , Esporos/imunologiaRESUMO
In a district of Japanese pear cultivators, a questionnaire survey and an IgE antibody survey were conducted on the pollinosis. A high prevalence of 36.3 percent of the farmers complained of pollinosis symptoms. By the IgE antibody survey, the symptoms were found to be related to the airborne pollens in the orchard.
Assuntos
Doenças dos Trabalhadores Agrícolas/etiologia , Pyrus/efeitos adversos , Rinite Alérgica Sazonal/etiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Doenças dos Trabalhadores Agrícolas/epidemiologia , Doenças dos Trabalhadores Agrícolas/imunologia , Feminino , Humanos , Imunoglobulina E/sangue , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Ácidos Oleicos , Pólen/efeitos adversos , Pólen/imunologia , Pyrus/imunologia , Rinite Alérgica Sazonal/epidemiologia , Rinite Alérgica Sazonal/imunologiaRESUMO
The European pear psylla, Cacopsylla pyri L. (Hemiptera: Psyllidae), is one of the most serious arthropod pests of pear. Since proper control of this pest is essential, better understanding of the complex plant-pest relationship is mandatory. This research deals with constitutive polyphenolic profiles in leaves of 22 pear cultivars of diverse origin (P. communis, P. pyrifolia, and P. pyrifolia × P. communis) and different resistance to psylla. The study was designed to show which differences in the polyphenolic profile of leaves from resistant and susceptible pear cultivars could be utilized as information in subsequent breeding programs. The results demonstrated that the leaves of Oriental pear cultivars contained much higher amounts of p-hydroxybenzoic acid, ferulic acid, aesculin, and naringin, that, together with detected 3-O-(6â³-O-p-coumaroyl)-hexoside, apigenin, apigenin 7-O-rutinoside, and hispidulin, indicated a clear difference between the species and might represent phenolics responsible for psylla resistance.
Assuntos
Hemípteros/fisiologia , Doenças das Plantas/parasitologia , Folhas de Planta/química , Polifenóis/química , Pyrus/parasitologia , Animais , Resistência à Doença , Doenças das Plantas/imunologia , Folhas de Planta/imunologia , Folhas de Planta/parasitologia , Polifenóis/imunologia , Pyrus/química , Pyrus/imunologiaRESUMO
Orchard workers in north China are highly exposed to orchard pollens, especially peach and other Rosaceae family pollens during pollination season. The aim of this study was to investigate whether occupational allergy to peach tree pollen as a member of Rosaceae family is IgE-mediated and to evaluate the cross-reactivity among Rosaceae family pollens. Allergen skin test and conjunctival challenge test were performed; enzyme linked immune-sorbent assay (ELISA), inhibiting ELISA, western immunoblotting and inhibiting western immunoblotting were done with Rosaceae family orchard pollens, including peach, apricot, cherry, apple and pear tree pollens. Mass spectrometry was also performed to probe the main allergen component and cross-reactive protein. Sensitizations to peach pollen were found in both skin test and conjunctival challenge in the patients. Serum specific IgE to three pollens (peach, apricot and cherry) were detected through ELISA. When peach pollen used as solid phase, ELISA inhibition revealed other four kinds of pollens capable of inducing partial to strong inhibitions (45% to 87%), with the strongest inhibition belonging to apricot pollen (87%). Western blotting showed predominant IgE binding to a 20 KD protein among these pollens, which appeared to be a cross-reactive allergen component through western blotting inhibition. It was recognized as a protein homologous to glutathione s-transferase 16 from Arabidopsis thaliana. Peach and other Rosaceae family tree pollen may serve as a potential cause of IgE mediated occupational respiratory disease in orchard workers in north China.
Assuntos
Asma Ocupacional/imunologia , Glutationa Transferase/imunologia , Imunoglobulina E/imunologia , Melhoramento Vegetal , Prunus persica/imunologia , Rinite Alérgica Sazonal/imunologia , Adulto , Western Blotting , Reações Cruzadas , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Malus/imunologia , Espectrometria de Massas , Prunus/imunologia , Prunus armeniaca/imunologia , Pyrus/imunologia , Rosaceae/imunologia , Testes CutâneosRESUMO
BACKGROUND: The pear psylla, Cacopsylla bidens (Sulc), is one of the most damaging pests of commercial pear orchards in Israel. Psylla control is a major obstacle to efficient integrated pest management, necessitating research on cultivars with natural resistance to pear psylla. Recently, two pear accessions (Py.760-261 and Py.701-202) from the local Newe Ya'ar fruit tree live collection were identified as having apparent resistance to pear psylla. Our goal was to evaluate the resistance of these two accessions relative to the commercial cultivar Spadona Estiva, and to identify whether the resistance mechanisms in the former interfere with insect colonisation of the plant (antixenosis) or inhibit insect growth, development, reproduction and survival (antibiosis). RESULTS: Settlement and development of C. bidens was evaluated under natural conditions (pear orchard), semi-natural conditions (potted plants), and on detached branches and leaves (laboratory). Our results indicate that the selection Py.760-261 is 10 times more resistant than Spadona while Py.701-202 is five times more resistant. CONCLUSIONS: The resistance mechanism in both accessions appears to be antibiosis affecting nymph survival. These resistant accessions may be used as rootstock or as a source of resistant genes in breeding programmes.