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au:"Machado, Marcos Antonio"

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Expression Quantitative Trait Loci (eQTL) mapping for callose synthases in intergeneric hybrids of Citrus challenged with the bacteria Candidatus Liberibacter asiaticus
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Curtolo, Maiara

; Granato, Laís Moreira

; Soratto, Tatiany Aparecida Teixeira

; Curtolo, Maisa

; Gazaffi, Rodrigo

; Takita, Marco Aurélio

; Cristofani-Yaly, Mariângela

; Machado, Marcos Antonio

Genetics and Molecular Biology

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2020, Volume 43 N. 2 elocation e20190133

Abstract: En | Text: En | PDF: En

Abstract Citrus plants have been extremely affected by Huanglongbing (HLB) worldwide, causing economic losses. HLB disease causes disorders in citrus plants, leading to callose deposition in the phloem vessel sieve plates. Callose is synthesized by callose synthases, which are encoded by 12 genes (calS1– calS12)in Arabidopsis thaliana. We evaluated the expression of eight callose synthase genes from Citrus in hybrids between Citrus sunki and Poncirus trifoliata infected with HLB. The objective of this work was to identify possible tolerance loci combining the expression quantitative trait loci (eQTL) of different callose synthases and genetic Single-Nucleotide Polymorphism (SNP) maps of C. sunki and P. trifoliata. The expression data from all CscalS ranged widely among the hybrids. Furthermore, the data allowed the detection of 18 eQTL in the C. sunki map and 34 eQTL in the P. trifoliata map. In both maps, some eQTL for different CscalS were overlapped; thus, a single region could be associated with the regulation of more than one CscalS. The regions identified in this work can be interesting targets for future studies of Citrus breeding programs to manipulate callose synthesis during HLB infection.

https://doi.org/10.1590/1678-4685-gmb-2019-0133 1108 downloads

Mutation in the xpsD gene of Xanthomonas axonopodis pv. citri affects cellulose degradation and virulence
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Baptista, Juliana Cristina

; Machado, Marcos Antonio

; Homem, Rafael Augusto

; Torres, Pablo Sebastián

; Vojnov, Adrian Alberto

; Amaral, Alexandre Morais do

Genetics and Molecular Biology

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2010, Volume 33 N. 1 Pages 146 - 153

Abstract: En | Text: En | PDF: En

The Gram-negative bacterium Xanthomonas axonopodis pv. citri, the causal agent of citrus canker, is a major threat to the citrus industry worldwide. Although this is a leaf spot pathogen, it bears genes highly related to degradation of plant cell walls, which are typically found in plant pathogens that cause symptoms of tissue maceration. Little is known on Xac capacity to cause disease and hydrolyze cellulose. We investigated the contribution of various open reading frames on degradation of a cellulose compound by means of a global mutational assay to selectively screen for a defect in carboxymethyl cellulase (CMCase) secretion in X. axonopodis pv. citri. Screening on CMC agar revealed one mutant clone defective in extracellular glycanase activity, out of nearly 3,000 clones. The insertion was located in the xpsD gene, a component of the type II secretion system (T2SS) showing an influence in the ability of Xac to colonize tissues and hydrolyze cellulose. In summary, these data show for the first time, that X. axonopodis pv. citri is capable of hydrolyzing cellulose in a T2SS-dependent process. Furthermore, it was demonstrated that the ability to degrade cellulose contributes to the infection process as a whole.

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Differential expression of pathogenicity- and virulence-related genes of Xanthomonas axonopodis pv. citri under copper stress
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Palmieri, Ana Carolina Basílio

; Amaral, Alexandre Morais do

; Homem, Rafael Augusto

; Machado, Marcos Antonio

Genetics and Molecular Biology

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2010, Volume 33 N. 2 Pages 348 - 353

Abstract: En | Text: En | PDF: En

In this study, we used real-time quantitative PCR (RT-qPCR) to evaluate the expression of 32 genes of Xanthomonas axonopodis pv. citri related to pathogenicity and virulence that are also involved in copper detoxification. Nearly all of the genes were up-regulated, including copA and copB. Two genes homologous to members of the type II secretion system (xcsH and xcsC) and two involved in the degradation of plant cell wall components (pglA and pel) were the most expressed in response to an elevated copper concentration. The type II secretion system (xcs operon) and a few homologues of proteins putatively secreted by this system showed enhanced expression when the bacteria were exposed to a high concentration of copper sulfate. The enhanced expression of the genes of secretion II system during copper stress suggests that this pathway may have an important role in the adaptative response of X. axonopodis pv. citri to toxic compounds. These findings highlight the potential role of these genes in attenuating the toxicity of certain metals and could represent an important means of bacterial resistance against chemicals used to control diseases.

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Ontogênese de caneluras em pedúnculo de flores de laranjeira doce infectados pelo vírus da tristeza dos citros estirpe "Capão Bonito"
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Tanaka, Francisco André Ossamu

; Kitajima, Elliot Watanabe

; Machado, Marcos Antonio

; Machado, Sílvia Rodrigues

Summa Phytopathologica

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dec 2009, Volume 35 N. 4 Pages 316 - 321

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Um estudo da ontogênese das caneluras induzidas em ramos de laranjeiras doces suscetíveis por isolados severos do vírus da tristeza dos citros (Citrus tristeza vírus - CTV) foi feito usando-se como modelo pedúnculos florais e de frutos. O menor calibre destes órgãos permite um melhor acompanhamento do processo. As observações foram feitas em laranjeira cv. Pêra infetada pelo isolado severo Capão Bonito do CTV. Cinco fases do processo de formação de caneluras puderam ser deduzidas pelas análises anatômicas. As primeiras alterações são representadas pelo aparecimento de células adensadas, hipertrofia e hiperplasia no parênquima e câmbio do floema e uma desorganização generalizada desta área. Segue-se uma atividade intensa do câmbio do floema adjacente e sua expansão em direção ao xilema. Esta invasão do xilema resulta na ruptura do anel do xilema pela massa celular do floema constituída de células recém formadas de parede celular delgada. Esta invasão do floema em direção ao xilema inicia um processo de degeneração dos vasos e parênquima do xilema. Finalmente há um colapso completo da região do xilema invadida, que é substituída pela massa do floema, resultando na canelura, notada ao se remover a casca.

An ontogenetic study of the stem pitting was carried out, using as a model system the peduncle of flowers and fruits of sweet orange cv. Pêra infected by a very severe isolate, Capão Bonito, of Citrus tristeza virus (CTV). Anatomical and ultrastructural analysis revealed five phases of this process. It starts with the localized appearance of chromatic cells in the phloem parenchyma, hypertrophy of companion and cambial cells and overall disorganization of the phloem and deposition of a dense material. The next step is characterized by the invasion of the xylem by an abnormal mass of phloem parenchymal cells formed by the disorganized proliferation. These changes lead to the third phase, where xylem ring is interrupted by the invading abnormal phloem proliferation, which produces undifferentiated cells with thin cell wall. The irregular activity of the vascular cambium continues followed by the complete degeneration of the xylem complemented by the appearance of dense phenolic deposits. Finally, there is the complete collapse of the xylem in the invaded region, and the corresponding space is occupied by the phloem mass which correspond to the stem pitting.

2685 downloads

Detection of the "Candidatus Liberibacter americanus" in phloem vessels of experimentally infected cataranthus roseus by scanning electron microscopy
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Tanaka, Francisco André O.

; Coletta-Filho, Helvécio Della

; Alves, Kely Cristina S.

; Spinelli, Myldred O.

; Machado, Marcos Antonio

; Kitajima, Elliot W.

Fitopatologia Brasileira

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dec 2007, Volume 32 N. 6 Pages 519 - 519

Abstract: En | Text: En | PDF: En

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Caracterização de um vírus baciliforme isolado de Solanum violaefolium transmitido pelos ácaros Brevipalpus phoenicis e Brevipalpus obovatus (Acari: Tenuipalpidae)
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Ferreira, Paulo de Tarso Oliveira

; Locali-Fabris, Eliane Cristina

; Freitas-Astúa, Juliana

; Antonioli-Luizon, Renata

; Gomes, Renata Takasugui

; Machado, Marcos Antonio

; Kitajima, Elliot Watanabe

Summa Phytopathologica

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sep 2007, Volume 33 N. 3 Pages 264 - 269

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Solano-violeta (Solanum violaefolium) é uma planta ornamental rasteira usada para cobrir solos de áreas sombreadas. Um vírus que induz manchas anelares nas folhas desta planta, tentativamente designado Solanum violaefolium ringspot virus - SvRSV, transmitido pelo ácaro Brevipalpus phoenicis (Acari: Tenuipalpidae) foi encontrado em Piracicaba, SP. Trata-se de um vírus baciliforme que se assemelha a outros vírus do tipo citoplasmático transmitidos por Brevipalpus sp. Este trabalho teve como objetivo relatar propriedades biológicas e estabelecer uma caracterização molecular parcial do SvRSV. O vírus pode ser transmitido mecanicamente a várias outras espécies botânicas, causando lesões localizadas. Entre as espécies avaliadas, Datura stramonium mostrou-se a melhor hospedeira experimental. Observou-se também a manifestação de sintomas nestas plantas após infestação das mesmas por B. obovatus previamente alimentado em lesões de SvRSV, confirmando esta outra espécie de ácaro como vetor do vírus. Suas propriedades físicas in vitro foram: temperatura de inativação 40-45 ºC; ponto final de diluição 10-3-10-4; longevidade in vitro 12 dias. Em secções ultrafinas, as partículas do SvRSV mostraram-se levemente mais delgadas e mais longas que as de outros vírus do mesmo grupo. A partir do dsRNA do SvRSV foi construída uma biblioteca de cDNA e foram identificadas duas possíveis regiões codificadoras das proteínas de movimento e replicase viral. Baseado nestas regiões foram desenhados "primers" para amplificação do RNA do SvRSV por RT-PCR. Sondas baseadas nas seqüências obtidas hibridizaram com ss- e dsRNA de D. stramonium infectadas pelo vírus. Ensaios preliminares de RT-PCR e hibridização não resultaram em reação com o vírus da leprose dos citros, tipo citoplasmático (CiLV-C).

Solanum violaefolium is an ornamental plant, with prostrate, trailing growth habit and is cultivated in shaded areas. A virus that causes ringspot symptoms on its leaves, tentatively named as Solanum violaefolium ringspot virus (SvRSV) and transmitted by Brevipalpus phoenicis (Acari: Tenuipalpidae) was found in Piracicaba city, São Paulo State. It is a bacilliform virus that resembles other cytoplasmic viruses transmitted by Brevipalpus sp. The objective of this work is to describe the biological properties and establish partial molecular characterization of the SvRSV. The SvRSV can be transmitted mechanically to several plant species causing local lesions. Among the tested species, Datura stramonium was proved to be the best experimental host. It was observed that S. violaefolium plants were infested by B. obovatus that also transmitted SvRSV in preliminary assays. Its in vitro physical properties were: thermal inactivation point: 40-45 ºC; dilution end point: 10-3-10-4 and in vitro longevity: 12 days. Ultrathin sections revealed that SvRSV particles are slightly thinner and longer than other cytoplasmic-type viruses transmitted by Brevipalpus sp. A cDNA library from dsRNA molecules extracted from SvRSV was constructed and two regions, which putatively code for the replicase and movement proteins were identified. Based on these sequences, primer pairs were designed for SvRSV´s RNA amplification by RT-PCR. Indeed, probes based on such sequences hybridized with ss- and dsRNA from infected D. stramonium plants. Preliminary molecular assays using primers and probes of SvRSV did not detect Citrus leprosis virus, cytoplasmic type (CiLV-C), another cytoplasmic type Brevipalpus-transmitted viruses.

3825 downloads

Expression profile of oxidative and antioxidative stress enzymes based on ESTs approach of citrus
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Peroni, Luis Antonio

; Ferreira, Renato Rodrigues

; Figueira, Antonio

; Machado, Marcos Antonio

; Stach-Machado, Dagmar Ruth

Genetics and Molecular Biology

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2007, Volume 30 N. 3 Pages 872 - 880

Abstract: En | Text: En | PDF: En

Plants not only evolve but also reduce oxygen in photosynthesis. An inevitable consequence of this normal process is the production of reactive oxygen species (ROS). Plants are adequately protected by the presence of multiple antioxidative enzymes in the cytosol and also in the different cell organelles such as chloroplasts, mitochondria, and peroxisomes. Traditionally, ROS were considered to be only a toxic byproduct of aerobic metabolism. However, recently it has become apparent that plants actively produce these molecules which may control many different physiological processes such as abiotic and biotic stress response, pathogen defense and systemic signaling. The search results using the Citrus Genome Program in Brazil (CitEST) for oxidative stress and the antioxidant enzyme system in Citrus Sinensis variety ‘Pera IAC’ indicated that the multiple ROS-scavenging enzymes were expressed throughout all citrus tissues. The analyses demonstrated the ubiquitous expression of metallothioneins, probably indicating a constitutive expression pattern. Oxalate oxidase has been identified as the most abundant expressed gene in developing fruits, which suggests a specific function in the ripening of citrus fruit. Moreover, infected leaves with Xylella fastidiosa and Leprosis citri showed a massive change in their ROS gene expression profile which may indicate that the suppression of ROS detoxifying mechanisms may be involved in the induction of the diseases.

5744 downloads

CitEST libraries
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Targon, Maria Luísa P. Natividade

; Takita, Marco Aurélio

; Amaral, Alexandre M. do

; Souza, Alessandra A. de

; Locali-Fabris, Eliane Cristina

; Dorta, Sílvia de Oliveira

; Borges, Kleber Martins

; Souza, Juliana Mendonça de

; Rodrigues, Carolina Munari

; Lucheta, Adriano Reis

; Freitas-Astúa, Juliana

; Machado, Marcos Antonio

Genetics and Molecular Biology

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2007, Volume 30 N. 3 Pages 1019 - 1023

Abstract: En | Text: En | PDF: En

In order to obtain a better understanding of what is citrus, 33 cDNA libraries were constructed from different citrus species and genera. Total RNA was extracted from fruits, leaves, flowers, bark, seeds and roots, and subjected or not to different biotic and abiotic stresses (pathogens and drought) and at several developmental stages. To identify putative promoter sequences, as well as molecular markers that could be useful for breeding programs, one shotgun library was prepared from sweet orange (Citrus sinensis var. Olimpia). In addition, EST libraries were also constructed for a citrus pathogen, the oomycete Phythophthora parasitica in either virulent or avirulent form. A total of 286,559 cDNA clones from citrus were sequenced from their 5’ end, generating 242,790 valid reads of citrus. A total of 9,504 sequences were produced in the shotgun library and the valid reads were assembled using CAP3. In this procedure, we obtained 1,131 contigs and 4,083 singletons. A total of 19,200 cDNA clones from P. parasitica were sequenced, resulting in 16,400 valid reads. The number of ESTs generated in this project is, to our knowledge, the largest citrus sequence database in the world.

2947 downloads

Frequency and distribution of microsatellites from ESTs of citrus
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Palmieri, Darío Abel

; Novelli, Valdenice Moreira

; Bastianel, Marinês

; Cristofani-Yaly, Mariângela

; Astúa-Monge, Gustavo

; Carlos, Eduardo Fermino

; Oliveira, Antonio Carlos de

; Machado, Marcos Antonio

Genetics and Molecular Biology

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2007, Volume 30 N. 3 Pages 1009 - 1018

Abstract: En | Text: En | PDF: En

Nearly 65,000 citrus EST (Expressed Sequence Tags) have been investigated using the CitEST project database. Microsatellites were investigated in the unigene sequences from Citrus spp. and Poncirus trifoliata. From these sequences, approximately 35% of the non-redundant ESTs contained SSRs. The frequencies of different SSR motifs were similar between Citrus spp and trifoliate orange. In general, mononucleotide repeats appeared to be the most abundant SSRs in the CitEST database, but we also identify di-, tri-, tetra-, penta- and hexanucleotide repeats. The AG/CT and AAG/CTT were the most common dinucleotide and trinucleotide motifs, with frequencies of 54.4% and 25.2%, respectively. Primer sequences flanking SSR motifs were successfully designed and synthesized. After in silico polymorphism analysis, a subset of sixty-eight primers was validated in different Citrus spp. and Poncirus trifoliata. PCR-amplification revealed polymorphism in citrus with all tested primer pairs and showed the potential of these markers for linkage mapping. Our study showed that the CitEST database can be exploited for the development of SSR markers that can amplify Citrus spp. and related genus for comparative mapping and other genetic analyses.

3514 downloads Cited 3 times in SciELO

10.

Characterization of Citrus tristeza virus isolates from grapefruit (Citrus paradisi Macf.) accessions of Citrus Active Germplasm Bank
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Corazza-Nunes, Maria Júlia

; Machado, Marcos Antonio

; Stach-Machado, Dagmar Ruth

; Nunes, William Mário Carvalho

; Carvalho, Sérgio Alves de

; Müller, Gerd Walter

Summa Phytopathologica

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sep 2006, Volume 32 N. 4 Pages 322 - 327

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Isolados do vírus da tristeza dos citros (CTV) de 35 acessos de pomelos que fazem parte do Banco Ativo de Germoplasma de Citros, localizado no Centro APTA Citros Sylvio Moreira, Cordeirópolis, São Paulo, Brasil, pertencente ao Instituto Agronômico de Campinas (IAC), foram caracterizados através dos sintomas observados nas árvores, indexação biológica, diagnóstico imunológico e análise SSCP (polimorfismo de conformação de fita simples) do gene da proteína do capsídeo. O grupo de plantas que, em geral, apresentou menor volume de copa e severo sintoma de canelura diferenciou-se significativamente do grupo com maior desenvolvimento vegetativo e fraco a moderado sintoma de canelura. No entanto, a maioria dos isolados de CTV das plantas de ambos os grupos induziu fraca reação em limão galego e nenhuma relação entre títulos do vírus nos tecidos e sintomatologia foi observada na avaliação sorológica conduzida por DAS-ELISA. A reação com diferentes anticorpos monoclonais e os distintos padrões eletroforéticos obtidos por SSCP demonstraram que há uma grande diversidade entre os isolados de CTV que infectam os acessos de pomelos. Alta complexidade de bandas dentro de um mesmo isolado foi também observada nos perfis SSCP, demonstrando que cada isolado é constituído por uma mistura de diferentes haplótipos de CTV. Padrões SSCP semelhantes foram observados entre algumas plantas com fortes sintomas de caneluras e entre algumas plantas com sintomas fracos ou moderados de caneluras.

Citrus tristeza virus (CTV) isolates from 35 grapefruit accessions belonging to Citrus Active Germplasm Bank of the "Instituto Agronômico de Campinas" located at the "Centro APTA Citros Sylvio Moreira", Cordeirópolis, São Paulo state, Brazil, were characterized and evaluated through symptoms in the trees, biological indexing, immunological diagnosis with different monoclonal antibodies and SSCP analysis (single-strand conformation polymorphism) of the coat protein gene. Symptomatology indicated that, in general, the group of plants with smaller canopy volume and severe stem pitting differed significantly from the group that presented greater vegetative development and mild to moderate stem pitting. However, the isolates from most of the accessions induced mild reaction on Mexican lime. The serological evaluation through the DAS-ELISA using monoclonal antibodies did not reveal any association between virus titer in the plant tissue and symptoms. The reaction with different monoclonal antibodies and the distinct electrophoresis patterns obtained through SSCP showed that there is a high degree of diversity among the isolates that infect these grapefruit accessions. High complexity within the same isolate was also observed in the SSCP profiles. This finding indicates that the CTV isolates from these plants are a complex mixture of CTV haplotypes. Similar SSCP banding patterns were observed among some plants with strong stem pitting symptoms, and among some plants with weak or moderate stem pitting symptoms.

2335 downloads Cited 1 time in SciELO

11.

The citrus leprosis pathosystem
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Bastianel, Marinês

; Freitas-Astúa, Juliana

; Kitajima, Elliot Watanabe

; Machado, Marcos Antonio

Summa Phytopathologica

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sep 2006, Volume 32 N. 3 Pages 211 - 220

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

A leprose dos citros é considerada a principal virose na citricultura brasileira, com maior destaque no Estado de São Paulo, principalmente pelos altos custos demandados para o controle químico do vetor, o ácaro Brevipalpus phoenicis. Além da relevância dessa virose para a citricultura local, sua importância mundial vem sendo ampliada consideravelmente nos últimos anos, principalmente com a disseminação do vírus em novos países da América do Sul e Central. No Brasil, apesar da sua importância econômica e ocorrência por mais de sete décadas, os mais importantes avanços no entendimento das interações do patossistema leprose têm sido obtidos apenas nos últimos dez anos. Essa revisão aborda os diferentes aspectos dessa doença, trazendo um breve histórico da doença, principais características da leprose, alternativas de controle, sua crescente importância econômica na cadeia citrícola nacional, os mais recentes relatos de sua ocorrência em outros países e os novos resultados obtidos pela pesquisa buscando um melhor entendimento das interações entre ácaro, vírus e hospedeiro vegetal.

Citrus leprosis is considered the main viral disease for the Brazilian citrus production, particularly for the State of São Paulo, due to the high costs spent for the chemical control of its vector, the tenuipalpid mite Brevipalpus phoenicis. In addition, its global importance has significantly increased in the last years, with the dissemination of the virus to new countries in South and Central America. In Brazil, despite its economical importance and occurrence for more than seven decades, the most significant advances towards understanding the pathosystem interactions have been obtained only in the last ten years. This review focuses on various aspects of the disease, beginning with a historical view, its main characteristics, alternatives for its control, its increasing economical importance in Brazil and abroad, and the new data on the search for understanding the interactions amongst the mite vector, the virus, and the plant host.

5337 downloads Cited 11 times in SciELO

12.

Diversidade de Phytophthora parasitica isolados de Citrus usando seqüências de nucleotídeos da região ITS-5.8S rDNA
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Rosa, Daniel Dias

; Machado, Marcos Antonio

; Targon, Maria Luisa Penteado Natividade

; Furtado, Edson Luiz

Summa Phytopathologica

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jun 2006, Volume 32 N. 2 Pages 188 - 191

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Realizou-se estudo para caracterização e verificação da diversidade genética de Phytophthora parasitica, agente causador da gomose dos citros. Quatorze isolados de Phytophthora parasitica, provenientes do Estado de São Paulo, foram seqüenciados a partir das regiões internas transcritas (ITS1 e ITS2) do gene 5.8S. Obtiveram-se seqüências de 812 pb a 860 pb que foram comparadas com seqüências de outras espécies de Phytophthora spp depositadas no NCBI. Foram feitos estudos filogenéticos, utilizando-se o método "neighbor-joining" com 1000 "bootstrap" e construído o dendrograma mais representativo. Obtiveram-se os resultados de 98,88% a 100% de similaridade genética entre os 14 isolados paulistas, e 99,5% a 98,8% entre estes e a seqüência de P. nicotianae (gi| 8927482) obtida do GenBank NCBI.

Phytophthoraparasitica is been considered one of the most prevalent citrus soilborn pathogens in Brazil. Fourteen isolates were collected in different region of São Paulo state. Amplification of the ITS region using primers ITS1 and ITS2 produced DNA fragments of 812 to 860 bp, that were compared with other sequences obtained to Gene Bank (NCBI). Neighbor-joining analysis, with 1000 bootstrap revealed an average similarity of 99,3 to 100% between the isolates, and 98,1 to 99,9% between S. Paulo state isolates and P. nicotianae (= Phytophthoraparasitica)obtained from the Gene Bank (gi|8927482). The ITS1 was more conserved than to the ITS2. This is a good tool for quick diagnosis of Phytophthora citrus diseases.

2205 downloads Cited 1 time in SciELO

13.

Brazilian coffee genome project: an EST-based genomic resource
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Vieira, Luiz Gonzaga Esteves

; Andrade, Alan Carvalho

; Colombo, Carlos Augusto

; Moraes, Ana Heloneida de Araújo

; Metha, Ângela

; Oliveira, Angélica Carvalho de

; Labate, Carlos Alberto

; Marino, Celso Luis

; Monteiro-Vitorello, Claúdia de Barros

; Monte, Damares de Castro

; Giglioti, Éder

; Kimura, Edna Teruko

; Romano, Eduardo

; Kuramae, Eiko Eurya

; Lemos, Eliana Gertrudes Macedo

; Almeida, Elionor Rita Pereira de

; Jorge, Érika C.

; Albuquerque, Érika V. S.

; Silva, Felipe Rodrigues da

; Vinecky, Felipe

; Sawazaki, Haiko Enok

; Dorry, Hamza Fahmi A.

; Carrer, Helaine

; Abreu, Ilka Nacif

; Batista, João A. N.

; Teixeira, João Batista

; Kitajima, João Paulo

; Xavier, Karem Guimarães

; Lima, Liziane Maria de

; Camargo, Luis Eduardo Aranha de

; Pereira, Luiz Filipe Protasio

; Coutinho, Luiz Lehmann

; Lemos, Manoel Victor Franco

; Romano, Marcelo Ribeiro

; Machado, Marcos Antonio

; Costa, Marcos Mota do Carmo

; Sá, Maria Fátima Grossi de

; Goldman, Maria Helena S.

; Ferro, Maria Inês T.

; Tinoco, Maria Laine Penha

; Oliveira, Mariana C.

; Van Sluys, Marie-Anne

; Shimizu, Milton Massao

; Maluf, Mirian Perez

; Eira, Mirian Therezinha Souza da

; Guerreiro Filho, Oliveiro

; Arruda, Paulo

; Mazzafera, Paulo

; Mariani, Pilar Drummond Sampaio Correa

; Oliveira, Regina L.B.C. de

; Harakava, Ricardo

; Balbao, Silvia Filippi

; Tsai, Siu Mui

; Mauro, Sonia Marli Zingaretti di

; Santos, Suzana Neiva

; Siqueira, Walter José

; Costa, Gustavo Gilson Lacerda

; Formighieri, Eduardo Fernandes

; Carazzolle, Marcelo Falsarella

; Pereira, Gonçalo Amarante Guimarães

Brazilian Journal of Plant Physiology

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mar 2006, Volume 18 N. 1 Pages 95 - 108

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

O café é um dos principais produtos agrícolas, sendo considerado o segundo item em importância do comércio internacional de "commodities". O gênero Coffea pertence à família Rubiaceae que também inclui outras plantas importantes. Este gênero contém aproximadamente 100 espécies, mas a produção comercial é baseada somente em duas espécies, Coffea arabica e Coffea canephora, que representam aproximadamente 70 % e 30 % do mercado total de café, respectivamente. O Projeto Genoma Café Brasileiro foi desenvolvido com o objetivo de disponibilizar os modernos recursos da genômica à comunidade científica e aos diferentes segmentos da cadeia produtiva do café. Para isso, foram seqüenciados 214.964 clones escolhidos aleatoriamente de 37 bibliotecas de cDNA de C. arabica, C. canephora e C. racemosa representando estádios específicos do desenvolvimento de células e de tecidos do cafeeiro, resultando em 130.792, 12.381 e 10.566 seqüências de cada espécie, respectivamente, após processo de trimagem. Os ESTs foram agrupados em 17.982 contigs e em 32.155 singletons. A comparação destas seqüências pelo programa BLAST revelou que 22 % não tiveram nenhuma similaridade significativa às seqüências no banco de dados do National Center for Biotechnology Information (de função conhecida ou desconhecida). A base de dados de ESTs do cafeeiro resultou na identificação de cerca de 33.000 unigenes diferentes. Os resultados de anotação das seqüências foram armazenados em base de dados "online" em <A HREF="http://www.lge.ibi.unicamp.br/cafe">http://www.lge.ibi.unicamp.br/cafe</A>. Os recursos desenvolvidos por este projeto disponibilizam ferramentas genéticas e genômicas que podem ser decisivas para a sustentabilidade, a competitividade e a futura viabilidade da agroindústria cafeeira nos mercados interno e externo.

Coffee is one of the most valuable agricultural commodities and ranks second on international trade exchanges. The genus Coffea belongs to the Rubiaceae family which includes other important plants. The genus contains about 100 species but commercial production is based only on two species, Coffea arabica and Coffea canephora that represent about 70 % and 30 % of the total coffee market, respectively. The Brazilian Coffee Genome Project was designed with the objective of making modern genomics resources available to the coffee scientific community, working on different aspects of the coffee production chain. We have single-pass sequenced a total of 214,964 randomly picked clones from 37 cDNA libraries of C. arabica, C. canephora and C. racemosa, representing specific stages of cells and plant development that after trimming resulted in 130,792, 12,381 and 10,566 sequences for each species, respectively. The ESTs clustered into 17,982 clusters and 32,155 singletons. Blast analysis of these sequences revealed that 22 % had no significant matches to sequences in the National Center for Biotechnology Information database (of known or unknown function). The generated coffee EST database resulted in the identification of close to 33,000 different unigenes. Annotated sequencing results have been stored in an online database at <A HREF="http://www.lge.ibi.unicamp.br/cafe">http://www.lge.ibi.unicamp.br/cafe</A>. Resources developed in this project provide genetic and genomic tools that may hold the key to the sustainability, competitiveness and future viability of the coffee industry in local and international markets.

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14.

Caracterização de genótipos de Citrus spp. através de marcadores RAPD
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Bastianel, Marinês

; Dornelles, Ana Lúcia Cunha

; Machado, Marcos Antonio

; Wickert, Ester

; Maraschin, Simone de Farias

; Della Coletta Filho, Helvécio

; Schäfer, Gilmar

Ciência Rural

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oct 2001, Volume 31 N. 5 Pages 763 - 768

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Em programas de melhoramento de citros, a caracterização adequada dos recursos genéticos disponíveis é de grande importância, principalmente devido às características biológicas da cultura, como a heterozigosidade, a embrionia nucelar e o longo ciclo reprodutivo. A facilidade com que ocorrem hibridações (interespecíficas e intergenéricas) e a embrionia nucelar favoreceram a formação e a preservação de novas combinações, classificadas como espécies. Neste estudo, marcadores RAPDs foram utilizados para analisar 15 acessos de Citrus spp., sendo quatro variedades de laranjeiras doce (C. sinensis Osbeck), quatro tangerineiras (C. reticulata Blanco, C. nobilis Loureiro, C. sunki Loureiro e C. deliciosa Tenore), uma laranjeira azeda (C. aurantium L.), um pomeleiro (C. paradisi Macf.), uma torangeira (C. grandis Osbeck), uma cidreira (C. medica L.), uma limeira ácida (C. latifolia) e dois híbridos (Citrus clementina T. x (C. tangerina T. x C. paradisi Macf.)). Doze sequências iniciadoras aleatórias foram utilizadas para estudar os 15 genótipos, encontrando-se um grau de similaridade mínimio de 0,81 ("Simple Matching") entre as tangerineiras. Os menores graus de similaridade foram encontrados entre as espécies de Citrus menos aparentadas (C. medica, C. grandis e C. latifolia). As quatro cultivares de laranjeiras doces não puderam ser diferenciadas pelos marcadores RAPD utilizados, apresentando similaridade máxima.

In citrus improvement programs the characterization of the available genetic resources is of great importance, mainly concerning biological characteristics of the culture, as the heterozigosity, nucellar the embriony and long reproductive cycle. Favored by nucellar embriony interespecific and intergeneric hybridizations and genotypes preservation happen easily. RAPDs markers were used to analyze 15 Citrus spp., four sweet orange (C. sinensis Osbeck), (C. medica, C. grandis e C. latifolia), four mandarins (C. reticulata Blanco, C. nobilis Loureiro, C. sunki Loureiro e C. deliciosa Tenore), a sour orange (C. aurantium L.), a grapefruit (C. paradisi Marcf.), a pummelo (C. grandis Osbeck), a cidra (C. medica L.), a lime (C. latifolia) and two hybrids (Citrus clementina T. x (C. tangerina T. x C. paradisi Macf.)). Genetic similarities of 15 Citrus genotypes obtained with twelve random primers, indicated a minimum similarity degree of 0.81 (simple matching) among the mandarins. Lower similarity degrees were obtained among less related Citrus species (C. medica, C. grandis e C. latifolia). The four varieties of sweet oranges (C. sinensis Osbeck) could not be differentiated by RAPD markers, showing maximum similarity.

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15.

MUTANTES DE LARANJA - 'PÊRA' COM NÚMERO REDUZIDO DE SEMENTES, OBTIDOS ATRAVÉS DE MUTAÇÕES INDUZIDAS
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LATADO, RODRIGO ROCHA

; TULMANN NETO, AUGUSTO

; ANDO, AKIHIKO

; IEMMA, ANTONIO FRANCISCO

; POMPEU JUNIOR, JORGINO

; FIGUEIREDO, JOSÉ ORLANDO

; PIO, ROSE MARY

; MACHADO, MARCOS ANTONIO

; NAMEKATA, TAKAO

; CERAVOLO, LEONARDO

; ROSSI, ANTONIO CARLOS

Revista Brasileira de Fruticultura

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aug 2001, Volume 23 N. 2 Pages 339 - 344

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

A obtenção de cultivares de citros com pequeno número de sementes é importante quando o objetivo é a produção de frutas para o consumo in natura. O objetivo deste trabalho foi o de avaliar, por três anos (1997, 1998 e 1999), o número médio de sementes e 15 outras características agronômicas, de 127 mutantes putativos de laranja-'Pêra', selecionados após irradiação de borbulhas com raios-gama e 8 plantas selecionadas do controle não irradiado. Os mutantes foram divididos em 16 grupos baseados nos tipos de mutações observados e em cada grupo, como controle comum, foram incluídas plantas de laranja-'Pêra' comercial. Observou-se que 46 mutantes putativos e uma planta do controle não irradiado (PSC) apresentaram redução significativa no número de sementes por fruto, nos três anos consecutivos, sendo que 15 mutantes apresentaram frutos com média entre uma e duas sementes e 9 mutantes, frutos com média inferior a uma semente. Dentre estes 9 mutantes, os de número 27, 28 e 58, que apresentaram também alterações significativas no diâmetro de copa ou na altura de planta, além do 59 e 101, que não apresentaram alterações nas outras 15 características avaliadas, possuem um maior potencial para serem lançados como novos mutantes de laranja-'Pêra' com pequeno número de sementes.

Citrus varieties with low number of seeds are important for in natura fruit market. The objective of the present work was to evaluate, for three years (1997, 1998 and 1999), the average number of seeds per fruit and 15 other agronomic characteristics of 127 putative mutated clones of sweet orange 'Pêra', selected after gamma-irradiation of budwood and 8 non irradiated control clones. Mutants were divided in 16 groups based on the type of mutation observed 'Pêra' commercial control plants (PCC) were included. It was observed that 46 putative mutants and one non-irradiated plant (PSC) showed significant reduction in seed number per fruit. Fifteen mutants had 1 -- 2 seeds on an average and 9 mutants less than one seed per fruit. Among them, the mutants 27, 28 and 58 showed other significant changes in the tree crown diameter or plant height. The mutants 59 and 101 showed no significant differences in the other 15 characteristics evaluated, indicating that they have a good potential to be released as new sweet orange 'Pêra' mutants with low number of seeds.

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