Assuntos
Doença de Depósito de Glicogênio Tipo IIb/diagnóstico , Doença de Depósito de Glicogênio Tipo IIb/genética , Miocárdio Ventricular não Compactado Isolado/diagnóstico , Proteína 2 de Membrana Associada ao Lisossomo/genética , Mutação , Miotonia/diagnóstico , Doença de Depósito de Glicogênio Tipo IIb/complicações , Doença de Depósito de Glicogênio Tipo IIb/fisiopatologia , Humanos , Miocárdio Ventricular não Compactado Isolado/complicações , Miocárdio Ventricular não Compactado Isolado/genética , Masculino , Miotonia/complicações , Miotonia/genética , Linhagem , Fenótipo , Adulto JovemRESUMO
INTRODUCTION: Sleeve gastrectomy as an isolated procedure is a new option in bariatric surgery. The aim of this study was to evaluate its short and medium term (2 years) results in a multicenter setting. MATERIALS: This is a retrospective study including 446 patients undergoing surgery in 14 teaching, private, and public hospitals. RESULTS: The immediate post-operative course were uneventful in 83.3% of patients. Minor complications occurred in 10.9% of patients and major complications in 5.3%. Suture line leaks occurred in 4.3% of the whole series. There was no mortality and the rate of reoperation was 2%. Overall mean weight loss after two years was 32 kg and the mean excess weight loss was 62%. Weight loss was significantly greater in non super-obese patients (p=0.0003). CONCLUSIONS: This study confirms the feasibility of sleeve gastrectomy in a multicenter setting; it is efficacious at two years as an isolated bariatric procedure for non super-obese patients. It is possible that an additional second-stage procedure may be necessary for super-obese patients.
Assuntos
Cirurgia Bariátrica , Gastrectomia/métodos , Adulto , Estudos de Viabilidade , Feminino , Seguimentos , Humanos , Tempo de Internação , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Complicações Pós-Operatórias , Reoperação , Estudos Retrospectivos , Fatores de Tempo , Resultado do Tratamento , Redução de PesoRESUMO
Leucine aminopeptidase (LAP) mRNAs are induced in response to mechanical wounding, pathogen infection, and insect infestation (V. Pautot, F.M. Holzer, B. Reisch, L.L. Walling [1993] Proc Natl Acad Sci USA 90: 9906-9910). Polyclonal antibodies to a glutathione S-transferase-LAP fusion protein and affinity-purified antibodies recognizing LAP antigenic determinants detected four classes of polypeptides in tomato (Lycopersicon esculentum) leaves. All four classes had multiple polypeptides in two-dimensional polyacrylamide gel electrophoresis immunoblots. Although antigenically related to the wound-induced tomato LAP proteins, the 77- and 66-kD LAP-like proteins accumulated in both healthy and wounded leaves. Two classes of 55-kD polypeptides with distinctive isoelectric points were designated as plant LAPs; only the acidic LAP proteins accumulated to high levels after mechanical wounding or Pseudomonas syringae pv tomato infection of tomato leaves. The temporal accumulation of LAP mRNAs was correlated with the increase in acidic LAP protein subunits. A slow-migrating LAP activity was detected using a native gel assay after wounding. The molecular mass of the native wound-induced LAP enzyme was 353 kD. The 55-kD acidic LAP proteins were associated with induced LAP activity, whereas the neutral LAPs and the LAP-like proteins were not associated with this exopeptidase. A second, fast-migrating aminopeptidase was detected in both healthy and wounded tomato leaves. Cell fractionation experiments revealed that wound-induced LAP is a soluble enzyme.
RESUMO
LapA RNAs, proteins, and activities increased in response to systemin, methyl jasmonate, abscisic acid (ABA), ethylene, water deficit, and salinity in tomato (Lycopersicon esculentum). Salicylic acid inhibited wound-induced increases of LapA RNAs. Experiments using the ABA-deficient flacca mutant indicated that ABA was essential for wound and systemin induction of LapA, and ABA and systemin acted synergistically to induce LapA gene expression. In contrast, pin2 (proteinase inhibitor 2) was not dependent on exogenous ABA. Whereas both LapA and le4 (L. esculentum dehydrin) were up-regulated by increases in ABA, salinity, and water deficit, only LapA was regulated by octadecanoid pathway signals. Comparison of LapA expression with that of the PR-1 (pathogenesis-related 1) and GluB (basic beta-1,3-glucanase) genes indicated that these PR protein genes were modulated by a systemin-independent jasmonic acid-signaling pathway. These studies showed that at least four signaling pathways were utilized during tomato wound and defense responses. Analysis of the expression of a LapA1:GUS gene in transgenic plants indicated that the LapA1 promoter was active during floral and fruit development and was used during vegetative growth only in response to wounding, Pseudomonas syringae pv tomato infection, or wound signals. This comprehensive understanding of the regulation of LapA genes indicated that this regulatory program is distinct from the wound-induced pin2, ABA-responsive le4, and PR protein genes.
RESUMO
BACKGROUND: Somatosensory evoked potentials (SSEPs) are increasingly performed for the assessment of peripheral neuropathies, but no practical guidelines have yet been established in this specific application. STUDY AIM: To determine the relevant indication criteria and optimal technical parameters for SSEP recording in peripheral neuropathy investigation. METHODS: A survey was conducted among the French-speaking practitioners with experience of SSEP recording in the context of peripheral neuropathies. The results of the survey were analyzed and discussed to provide recommendations for practice. RESULTS: SSEPs appear to be a second-line test when electroneuromyographic investigation is not sufficiently conclusive, providing complementary and valuable information on central and proximal peripheral conduction in the somatosensory pathways. CONCLUSIONS: Guidelines for a standardized recording protocol, including the various parameters to be measured, are proposed. CLINICAL RELEVANCE: We hope that these proposals will help to recognize the value of this technique in peripheral neuropathy assessment in clinical practice.
Assuntos
Potenciais Somatossensoriais Evocados , Doenças do Sistema Nervoso Periférico/diagnóstico , Estimulação Elétrica/métodos , França , Humanos , Condução Nervosa , Guias de Prática Clínica como Assunto , Inquéritos e QuestionáriosRESUMO
Expression of proteinase inhibitor I and II genes was investigated during infection by Pseudomonas syringae pv. tomato, the causal agent of bacterial speck disease in tomato. Inoculation of leaves with P. s. pv. tomato of two inbred tomato lines that are resistant and susceptible to the pathogen resulted in the accumulation of proteinase inhibitor I and II mRNAs in this organ. Our data showed that in the lines used in this study, proteinase inhibitor II mRNAs accumulated in leaves to higher levels than proteinase inhibitor I mRNA in response to P. s. pv. tomato infection and wounding. Proteinase inhibitor II mRNAs accumulated more rapidly in disease-resistant than in disease-susceptible plants. Proteinase inhibitor I mRNAs were first detected in the disease-susceptible line during infection and wounding. In contrast to wounding, the systemic induction of these genes during pathogen ingression was limited. These data show that the plant proteinase inhibitors constitute one of the components of the plant defense system that are induced in response to bacterial pathogen invasion.
Assuntos
Regulação da Expressão Gênica , Proteínas de Plantas/genética , Inibidores de Proteases , Pseudomonas/fisiologia , Doenças das Plantas , Plantas/enzimologia , Plantas/genética , Plantas/microbiologia , RNA Mensageiro/metabolismoRESUMO
Tomato plants constitutively express a neutral leucine aminopeptidase (LAP-N) and an acidic LAP (LAP-A) during floral development and in leaves in response to insect infestation, wounding, and Pseudomonas syringae pv. tomato infection. To assess the physiological roles of LAP-A, a LapA-antisense construct (35S:asLapA1) was introduced into tomato. The 35S:asLapA1 plants had greatly reduced or showed undetectable levels of LAP-A and LAP-N proteins in healthy and wounded leaves and during floral development. Despite the loss of these aminopeptidases, no global changes in protein profiles were noted. The 35S:asLapA1 plants also exhibited no significant alteration in floral development and did not impact the growth and development of Manduca sexta and P. syringae pv. tomato growth rates during compatible or incompatible infections. To investigate the mechanism underlying the strong induction of LapA upon P. syringae pv. tomato infection, LapA expression was monitored after infection with coronatine-producing and -deficient P. syringae pv. tomato strains. LapA RNA and activity were detected only with the coronatine-producing P. syringae pv. tomato strain. Coronatine treatment of excised shoots caused increases in RNAs for jasmonic acid (JA)-regulated wound-response genes (LapA and pin2) but did not influence expression of a JA-regulated pathogenesis-related protein gene (PR-1). These results indicated that coronatine mimicked the wound response but was insufficient to activate JA-regulated PR genes.
Assuntos
Aminoácidos/fisiologia , Toxinas Bacterianas/metabolismo , Leucil Aminopeptidase/genética , Pseudomonas/patogenicidade , Solanum lycopersicum/genética , Sequência de Bases , Primers do DNA , Regulação para Baixo , Indenos , Solanum lycopersicum/enzimologia , Solanum lycopersicum/microbiologia , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/microbiologia , Pseudomonas/crescimento & desenvolvimento , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
Three gene constructions based on a mouse metallothionein I gene (mMT-I) were introduced into tobacco using a Ri plasmid vector system to test the effectiveness of animal gene regulatory signals in plant cells. No transcription from the native mouse gene was observed. In plant cells bearing chimeric mMT-I genes in which transcription was driven by the nopaline synthase promoter, neither polyadenylation nor splicing of mMT-I pre-mRNA was observed. Detailed comparisons of mMT-I sequences with those of known plant genes were carried out; slight differences in regions of known consensus sequences may be at least partly responsible for the non-recognition of mMT-I gene regulatory signals in plant cells, though other as yet unidentified, potentially necessary sequences may also be involved.
Assuntos
Regulação da Expressão Gênica , Metalotioneína/genética , Nicotiana/genética , Plantas Tóxicas , Animais , Sequência de Bases , Clonagem Molecular , DNA , DNA Recombinante , Éxons , Vetores Genéticos , Metalotioneína/biossíntese , Camundongos , Dados de Sequência Molecular , Regiões Promotoras Genéticas , Splicing de RNA , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , Transformação GenéticaRESUMO
This article describes several new expression vectors that capitalize on the ability of Agrobacterium rhizogenes to transfer DNA from its Ri plasmid to the plant nuclear genome. The intermediate vectors described include an expression cassette based on one of the three following promoters: the nopaline synthase promoter, or the cauliflower mosaic virus (CaMV) promoters responsible for transcription of either the 19S or 35S CaMV RNA. The termination and polyadenylation signals are either from the nopaline synthase gene or from CaMV. The expression micro-Ri plasmid described bears a selectable marker gene and an expression cassette cloned between the borders of the TL-region of the Ri plasmid of A. rhizogenes A4. Different strategies for using these vectors to introduce chimeric genes into plants are described, and the advantages and disadvantages of the two types of vectors are discussed.
Assuntos
Vetores Genéticos , Rhizobium/genética , Transformação Bacteriana , Cromossomos Bacterianos , Conjugação Genética , Meios de Cultura , DNA Bacteriano/análise , Regulação da Expressão Gênica , Plantas/microbiologia , PlasmídeosRESUMO
CONTEXT: Autopsy rates have been declining throughout the world, although preservation of the autopsy is considered a fundamental principle of medical care. In France, the 1994 bioethics law requires physicians to inform relatives before performing an autopsy. OBJECTIVE: To analyze the following factors that potentially influence hospital autopsy rates: legal constraints, autopsy reporting times, opinions of physicians requesting autopsies and pathologists regarding the usefulness of autopsy in patient care, and use of autopsy material in research publications. DESIGN: Record of the annual numbers of deaths and autopsies during a 10-year period (1988-1997). Record of the delays for transmission of final autopsy report to the requesting physician. Questionnaire analyzing the possible factors influencing autopsy rate. Categorization of articles published by pathologists according to the use of autopsy material. SETTING: A 1000-bed, university teaching hospital in the Paris, France, area. PARTICIPANTS: Questionnaire addressed to physicians, head nurses, and mortuary staff. RESULTS: A total of 1454 autopsies were reviewed. The autopsy rate declined from 15.4% in 1988 to 3.7% in 1997. This decline was marked after 1994 and tended to be slower for neurologic indications than for other indications. The final report had not been communicated within 180 days in 620 (42.6%) of 1454 autopsies. Fifty-five of 105 respondents considered that the bioethics law was one cause of the recent decrease of autopsy rate. Considering the contribution of autopsy to medical research, 94 (81%) of 116 articles dealing with central nervous system but only 28 (6%) of 464 articles dealing with other organs used autopsy-derived material. CONCLUSIONS: The 1994 bioethics law seems to contribute to the decline of autopsy. Inadequate delays for communicating autopsy results are frequent. Except for neuropathologists, autopsy is a minor source of research material.
Assuntos
Atitude do Pessoal de Saúde , Autopsia/estatística & dados numéricos , Hospitais de Ensino/estatística & dados numéricos , Hospitais de Ensino/tendências , Serviço Hospitalar de Patologia/estatística & dados numéricos , Serviço Hospitalar de Patologia/tendências , Autopsia/legislação & jurisprudência , Ética Médica , França , Humanos , Consentimento Livre e Esclarecido/legislação & jurisprudência , Médicos , Inquéritos e Questionários , Doadores de Tecidos/estatística & dados numéricosRESUMO
BACKGROUND: The prevalence of fibromuscular dysplasia (FMD) in patients with cervical artery dissection (CAD) is unknown. Our objectives were to assess the risk of CAD recurring as a stroke or a transient ischemic attack and the association of these events with FMD. METHODS: We prospectively included and followed 103 consecutive patients who had been admitted for a CAD. The median follow-up was 4 years (range 4 months to 10 years). The main criteria for inclusion were a mural hematoma demonstrated by cervical magnetic resonance imaging and/or signs suggesting CAD on 2 other investigations. FMD was diagnosed on the so-called string of beads pattern by digital subtraction angiography. RESULTS: Five patients had CAD recurrence (60% occurred late). Four of these 5 patients had FMD. In 4 patients, CAD recurrence involved another cervical artery. CONCLUSION: The rate of symptomatic CAD recurrence was 1% per year and was often related to FMD.
Assuntos
Dissecação da Artéria Carótida Interna/complicações , Displasia Fibromuscular/complicações , Ataque Isquêmico Transitório/etiologia , Acidente Vascular Cerebral/etiologia , Dissecação da Artéria Vertebral/complicações , Adulto , Angiografia Digital , Anticoagulantes/uso terapêutico , Dissecação da Artéria Carótida Interna/tratamento farmacológico , Dissecação da Artéria Carótida Interna/epidemiologia , Dissecação da Artéria Carótida Interna/etiologia , Dissecação da Artéria Carótida Interna/patologia , Feminino , Displasia Fibromuscular/diagnóstico , Displasia Fibromuscular/epidemiologia , Seguimentos , Humanos , Incidência , Ataque Isquêmico Transitório/epidemiologia , Ataque Isquêmico Transitório/patologia , Angiografia por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Inibidores da Agregação Plaquetária/uso terapêutico , Prognóstico , Estudos Prospectivos , Recuperação de Função Fisiológica , Recidiva , Medição de Risco , Fatores de Risco , Acidente Vascular Cerebral/epidemiologia , Acidente Vascular Cerebral/patologia , Fatores de Tempo , Resultado do Tratamento , Ultrassonografia Doppler Dupla , Dissecação da Artéria Vertebral/tratamento farmacológico , Dissecação da Artéria Vertebral/epidemiologia , Dissecação da Artéria Vertebral/etiologia , Dissecação da Artéria Vertebral/patologiaRESUMO
UNLABELLED: Parkinson's disease (PD), a disorder of unknown etiology, is associated with the degeneration of dopaminergic neurons in nigro-striatal pathways. MPTP, a meperidine analog, causes parkinsonism in human and nonhuman primates. MPP+, the active metabolite of MPTP, inhibits the activity of respiratory chain complex I. In patients with PD, a reduced complex I activity was found in substantia nigra, skeletal muscle, and platelets. Because complex I is partially encoded by the mitochondrial genome, several studies have searched for mitochondrial (mt) DNA abnormalities in patients with PD. Our aim was to answer the following questions: (1) are there some abnormalities of mtDNA in PD? (2) if there are some, what are these abnormalities? and (3) what is the pathogenic role of these abnormalities? METHODS: The literature review was performed using Medline [National Library of Medicine, Washington] and Current Contents [Institute for Scientific Information, Philadelphia] databases. Periods screened were 1966-March, 1998 (Medline) and March 17, 1997-March 9, 1998 (Current Contents). Keywords were: "Parkinson" or "Parkinson's", and "mitochondrial DNA" or "mtDNA". We limited our research to articles in English and French. RESULTS: Medline search provided 59 articles. Current Contents search provided 22 articles. Twelve articles were found in both databases. Thirty-eight of the 69 articles were either reviews about mitochondrial diseases (19 articles) or original articles not related to mtDNA (19 articles). Our final selection included the remaining 31 articles.
Assuntos
DNA Mitocondrial/genética , Doença de Parkinson/genética , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina , Animais , Encéfalo/patologia , Humanos , MEDLINE , Mutação , National Library of Medicine (U.S.) , Doença de Parkinson/patologia , Doença de Parkinson Secundária/induzido quimicamente , Doença de Parkinson Secundária/genética , Primatas , Estados UnidosRESUMO
Leaf discs of C. intybus were inoculated with an Agrobacterium tumefaciens strain harboring a neomycin phosphotransferase (neo) gene for kanamycin resistance and a mutant acetolactate synthase gene (csr1-1) from Arabidopsis thaliana conferring resistance to sulfonylurea herbicides. A regeneration medium was optimized which permitted an efficient shoot regeneration from leaf discs. Transgenic shoots were selected on rooting medium containing 100 mg/l kanamycin sulfate. Integration of the csr1-1 gene into genomic DNA of kanamycin resistant chicory plants was confirmed by Southern blot hybridizations. Analysis of the selfed progenies (S1 and S2) of two independent transformed clones showed that kanamycin and chlorsulfuron resistances were inherited as dominant Mendelian traits. The method described here for producing transformed plants will allow new opportunities for chicory breeding.
RESUMO
The maize autonomous transposable element Ac was introduced into haploid Nicotiana plumbaginifolia via Agrobacterium tumefaciens transformation of leaf disks. All the regenerated transformants (R0) were diploid and either homozygous or heterozygous for the hygromycin resistance gene used to select primary transformants. The Ac excision frequency was determined using the phenotypic assay of restoration of neomycin phosphotransferase activity and expression of kanamycin resistance among progeny seedlings. Some of the R0 plants segregated kanamycin-resistant seedlings in selfed progeny at a high frequency (34 to 100%) and contained one or more transposed Ac elements. In the primary transformants Ac transposition probably occurred during plant regeneration or early development. Other R0 transformants segregated kanamycin-resistant plants at a low frequency (< or = 4%). Two transformants of this latter class, containing a unique unexcised Ac element, were chosen for further study in the expectation that their kanamycin resistant progeny would result from independent germinal transposition events. Southern blot analysis of 32 kanamycin-resistant plants (R1 or R2), selected after respectively one or two selfings of these primary transformants, showed that 27 had a transposed Ac at a new location and 5 did not have any Ac element. Transposed Ac copy number varied from one to six and almost all transposition events were independent. Southern analysis of the R2 and R3 progeny of these kanamycin-resistant plants showed that Ac continued to transpose during four generations, and its activity increased with its copy number. The frequency of Ac transposition, from different loci, remained low (< or = 7%) from R0 to R3 generations when only one Ac copy was present.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Agrobacterium tumefaciens/genética , Elementos de DNA Transponíveis , Nicotiana/genética , Plantas Tóxicas , Zea mays/genética , Sequência de Bases , Southern Blotting , Clonagem Molecular/métodos , DNA Bacteriano/genética , Dados de Sequência Molecular , Mutagênese Insercional , Oligodesoxirribonucleotídeos , Plantas Geneticamente Modificadas , Reação em Cadeia da Polimerase/métodos , Mapeamento por Restrição , Homologia de Sequência do Ácido Nucleico , Transformação GenéticaRESUMO
The KNAT2 (for KNOTTED-like from Arabidopsis thaliana 2) homeobox gene is expressed in the vegetative apical meristem. It is also active during flower development, suggesting a function in the structuring of flowers. To investigate its role, we used a DEXAMETHASONE (DEX)-inducible system to generate transgenic plants that overexpressed a fusion of KNAT2 with the hormone binding domain of the glucocorticoid receptor. DEX-induced plants were similar to plants overexpressing the closely related KNAT1 gene, indicating overlapping functions, although we observed differences as well. In particular, KNAT2-GR activation induced ectopic carpel features. First, KNAT2 induced the homeotic conversion of nucellus into carpel-like structures. Second, KNAT2 induced stigmatic papillae on rosette leaves in the ap2-5 background. Third, ectopic expression of the carpel identity gene AGAMOUS (AG) was observed in carpels and ovules. Interestingly, the homeotic conversion was not dependent on AG activity, because it was maintained in the ag-1 ap2-5 double mutant. Therefore, our data indicate that KNAT2 also must activate other carpel regulators. Together, these results suggest that KNAT2 plays a role in carpel development.
Assuntos
Proteínas de Arabidopsis , Arabidopsis/genética , Proteínas de Homeodomínio/genética , Proteínas de Plantas/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/ultraestrutura , Dexametasona/farmacologia , Regulação da Expressão Gênica de Plantas , Microscopia Confocal , Microscopia Eletrônica de Varredura , Dados de Sequência Molecular , Estruturas Vegetais/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/ultraestrutura , beta-Galactosidase/genéticaRESUMO
The wound-induced leucine aminopeptidase (EC 3.4.11.1) genes, LapA1 and LapA2, from tomato (Lycopersicon esculentum Mill.) were isolated and characterized. The genes were organized in a tandem array with approximately 6 kb separating their coding regions. Quantitation of LapA mRNA levels in conjunction with nuclear run-on experiments indicated that LapA genes were primarily under transcriptional control after wounding and infection with Pseudomonas syringae pv. tomato. In contrast, actin genes were down-regulated after pathogen infection. The sequences of the LapA1 and LapA2 5'-flanking regions were determined and several potential regulatory motifs were identified. Ribonuclease protection studies revealed that LapA1 and LapA2 had short 18-bp 5'-untranslated regions (UTR), both genes were expressed after wounding, and LapA1 mRNAs were 3.3-fold more abundant than LapA2 transcripts. While the region surrounding LapA1 was conserved, the 3'-UTRs and 3'-flanking regions of LapA2 had diverged in two inbred tomato lines. The accumulation of LapA mRNAs and of LAP-A (acidic pI), LAP-N (neutral pI) and LAP-related proteins were examined in two monocot and five dicot species. The LAP-N and 66- and 77-kDa LAP-related proteins were detected in healthy and wounded leaves of all plants examined. The LAP-A proteins were only detected in nightshade and their accumulation was distinct from that observed in tomato.
Assuntos
Leucil Aminopeptidase/genética , Proteínas de Plantas/genética , Solanum lycopersicum/genética , Regiões 3' não Traduzidas , Motivos de Aminoácidos , Sequência de Bases , Leucil Aminopeptidase/isolamento & purificação , Leucil Aminopeptidase/metabolismo , Solanum lycopersicum/enzimologia , Dados de Sequência Molecular , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , RNA de Plantas/análise , Alinhamento de SequênciaRESUMO
The spatial and temporal activity of the entire and individual promoter domains of the rolA gene of Agrobacterium rhizogenes was investigated and correlated with the distinctive features of the phenotypes of transgenic tobacco plants. The GUS assay was performed in the presence of an oxidative catalyst during the development of transgenic plants expressing chimeric genes containing the beta-glucuronidase coding sequence under the control of the different promoter domains. In situ hybridization was also used on transgenic plants harbouring rolA under the control of the entire or deleted promoter. This paper demonstrates for the first time that the entire rolA promoter, composed of domains, A, B and C, is silent in seeds, then activated at the onset of germination in the cotyledons and in the elongation zone of the radicle and is finally expressed throughout the vegetative and floral phases. Domains B + C, which were sufficient to induce wrinkled leaves and short internodes, were active in all the stem tissues, but only in the companion cells of the phloem strands of the leaves. Domain C, which specified a dwarf phenotype with normal leaves, was weakly expressed in the stem vascular bundles and in the leaf internal phloem. These results indicate that the vascular bundles are the primary targets for the generation of the short internode phenotype. Furthermore, the local expression of rolA in the stem vascular bundles induced a size reduction of the surrounding parenchyma cells, suggesting the existence of some diffusible factor(s) associated with the expression of the rolA gene.
Assuntos
Proteínas de Bactérias/genética , Regulação da Expressão Gênica , Genes Bacterianos , Nicotiana/crescimento & desenvolvimento , Plantas Tóxicas , Rhizobium/genética , Técnicas de Transferência de Genes , Glucuronidase , Hibridização In Situ , Morfogênese , Fenótipo , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Proteínas Recombinantes de Fusão , Distribuição Tecidual , Nicotiana/anatomia & histologia , Nicotiana/genéticaRESUMO
A leucine aminopeptidase (EC 3.4.11.1) cDNA clone (DR57) that was induced in response to Pseudomonas syringae pv. tomato (P.s. tomato) infection was isolated using a subtractive hybridization-enriched cDNA probe. Genomic DNA blot analysis showed that the tomato genome had two leucine aminopeptidase genes. The levels of DR57 mRNAs after P.s. tomato infection and mechanical wounding were determined in two inbred tomato lines that exhibit susceptibility and resistance to P.s. tomato. DR57 mRNAs were detected 12 hours after infection and 4 hours after wounding. Furthermore, DR57 mRNAs were systemically induced in response to wounding. DR57 mRNAs were induced in leaves after Spodoptera littoralis feeding but were not detected in detached leaf controls. Possible roles for the DR57 leucine aminopeptidase in the defense reactions are discussed.