Relata-se a ocorrência de Meloidogyne incognita (Est. I2) em pomar de videira na cidade de Petrolina, Pernambuco. Plantas da cultivar Festival enxertadas sobre 'IAC 766-Campinas', atacadas por Meloidogyne incognita, apresentavam sintomas de menor vigor, folhas amarelecidas e folhagem mais esparsa, e raízes com numerosas galhas em meio ao cultivo de feijão-de-porco (Canavalia ensiformis), também atacado pelo nematoide. Com o objetivo de avaliar a resistência de quatro porta-enxertos tropicais, oito de clima temperado, e as cultivares Niágara Rosada e Chardonnay quanto à reação a M. incognita e M. arenaria, em casa de vegetação, realizou-se este trabalho. Mudas das videiras, mantidas em vaso com solo esterilizado, foram inoculadas com 10.000 ovos + J2 de M. incognita ou M. arenaria por planta. Decorridos oito meses da inoculação, cada planta foi avaliada quanto ao número de galhas e número de ovos por planta, determinando-se, a seguir, o fator de reprodução de cada espécie do nematoide (FR=população inicial/população final) nos diferentes materiais testados. Entre os porta-enxertos avaliados, 'Harmony', 'Salt Creek', '1103 Paulsen', 'IAC 572-Jales', 'IAC 313-Tropical', 'K5BB Kober' e 'SO4', foram resistentes (FR<1,00) e 'Solferino', imune (FR=0,00) a M. incognita, sendo os demais suscetíveis ao nematoide. Quando se avaliou a reação da videira a M. arenaria, exceto '106-Traviú', '420 A', 'Rupestris du Lot' e '1103 Paulsen', os demais porta-enxertos foram resistentes (FR>1,00) ao nematoide. No entanto, ambas as cultivares copa foram suscetíveis às duas espécies de Meloidogyne testadas.

The occurrence of root-knot nematode Meloidogyne incognita (Est. I2) is reported in grapevine in Petrolina municipality, State of Pernambuco. Grape plants of Festival cultivar grafted on the 'IAC 766 - Campinas' rootstock attacked by M. incognita exhibited symptoms of less vigorous growth, yellowing and sparse leaves and multiple galls in the root system in an area cultivated with jack-bean (Canavalia ensiformis) also infected by the same nematode. In order to evaluate resistance of four tropical rootstocks, eight temperate and two cultivars Chardonnay and Niagara Rosada in reaction to M. incognita and M. arenaria, this study was carried out. Seedlings of the investigated grapevine genotypes were maintained in pots with sterilized soil and subsequently inoculated with 10,000 eggs + J2 of M. incognita or M. arenaria per plant in a six replicate design. Eight months after nematode inoculation, each plant was evaluated by counting gall and eggs. The final number of eggs (FP) of each Meloidogyne specie was calculated and the reproductive factor (RF = FP/IP), determined. Among the tested rootstocks, 'Harmony', 'Salt Creek', '1103 Paulsen', 'IAC 572-Jales', 'IAC 313-Tropical', 'K5BB Kober', 'SO4' , were resistant (RF<1.00) and, 'Solferino', immune (RF=0.00) to M. incognita, the remaining rootstocks were susceptible to the nematode. With exception of '106-Traviú', '420 A', 'Rupestris du Lot' and '1103 Paulsen', all tested rootstocks were resistant (RF>1.00) to M. arenaria. In contrast, both tested grapevine scion cultivars were susceptible to the investigated Meloidogyne species.

Simple Sequence Repeats (SSR) são marcadores moleculares adequados para estimar a variação genética de recursos genéticos; no entanto, o uso de SSR em escala requer a otimização de protocolos. Este trabalho objetivou identificar marcadores microssatélites, desenvolvidos para pera e outras espécies de frutas, para caracterizar coleções de germoplasma de pera e demonstrar seu uso em programas de melhoramento. De 62 marcadores investigados, 23 resultando em padrões reproduzíveis e polimórficos foram utilizados para genotipar uma amostra de 42 acessos do Banco Ativo de Germoplasma de Pera. Quando comparado com os 23 marcadores microssatélites, um subconjunto de 11 marcadores, selecionados com base em He, PIC e PId, foi usado para distinguir acessos e realizar a análise de agrupamento com eficácia similar, separando os acessos europeus, japoneses e chineses em grupos distintos. Esse subconjunto de marcadores constitui uma ferramenta útil para aplicações em manejo de recursos genéticos e melhoramento de pera.

Simple Sequence Repeats (SSR) are molecular markers suitable to assess the genetic variation of germplasm resources; however, large-scale SSR use requires protocol optimization. The present work aimed to identify SSR markers, developed for pear and other fruit species that are effective in characterizing pear germplasm collections and in demonstrating their use in providing support for genetic breeding programs. From a total of 62 SSR markers investigated, 23 yielding reproducible and polymorphic patterns were used to genotype a sample of 42 pear accessions of the Brazilian Pear Germplasm Bank (PGB). When compared to these 23 SSR markers, a subset of eleven markers, selected based on He, PIC and PId, was used to distinguish individual accessions and perform cluster analysis with similar efficacy. Genetic diversity analysis clustered the European, Japanese and Chinese accessions in distinct groups. This markers subset constitutes a valuable tool for several applications related to pear genetic resources management and breeding.

Os objetivos deste trabalho foram avaliar a diversidade genética de 79 acessos de soja de diferentes regiões do mundo, agrupá-los de acordo com a similaridade e testar a correlação entre os dois tipos de marcadores utilizados. Foram utilizados marcadores microssatélites genômicos (SSR) e funcionais (EST-SSR). Trinta pares de primers SSR foram selecionados (20 genômicos e 10 EST-SSR) de acordo com sua distribuição nos 20 grupos de ligação da soja, com sua unidade de repetição trinucleotídica e com seu conteúdo de informação polimórfica. Todos os lócus analisados foram polimórficos, e 259 alelos foram encontrados. O número de alelos por lócus variou entre 2-21, com média de 8,63. Os acessos possuem uma quantidade significativa de alelos raros, sendo os acessos 19, 35, 63 e 65 os que apresentaram maior número de alelos exclusivos. Os acessos 75 e 79 são os mais similares e os acessos 31 e 35, e 40 e 78 são os mais divergentes. Foi observada baixa correlação entre resultados de SSR e EST-SSR. Portanto, uma análise adequada de diversidade em soja deve ser feita utilizando-se tanto marcadores microssatélites genômicos como funcionais. A diversidade genética dos acessos selecionados é alta, tendo sido encontrados cinco grupos e vários subgrupos. Observou-se moderada relação entre divergência genética e origem geográfica dos acessos.

The objectives of this work were to investigate the genetic variation in 79 soybean (Glycine max) accessions from different regions of the world, to cluster the accessions based on their similarity, and to test the correlation between the two types of markers used. Simple sequence repeat markers present in genomic (SSR) and in expressed regions (EST-SSR) were used. Thirty SSR primer-pairs were selected (20 genomic and 10 EST-SSR) based on their distribution on the 20 genetic linkage groups of soybean, on their trinucleotide repetition unit and on their polymorphism information content. All analyzed loci were polymorphic, and 259 alleles were found. The number of alleles per locus varied from 2-21, with an average of 8.63. The accessions exhibit a significant number of rare alleles, with genotypes 19, 35, 63 and 65 carrying the greater number of exclusive alleles. Accessions 75 and 79 were the most similar and accessions 31 and 35, and 40 and 78, were the most divergent ones. A low correlation between SSR and EST-SSR data was observed, thus genomic and expressed microsatellite markers are required for an appropriate analysis of genetic diversity in soybean. The genetic diversity observed was high and allowed the formation of five groups and several subgroups. A moderate relationship between genetic divergence and geographic origin of accessions was observed.

A mutagênese tem sido tradicionalmente usada para gerar variabilidade genética em plantas ornamentais. Recentemente, tornou-se uma ferramenta poderosa na descoberta e análise da função gênica em genética reversa. Este trabalho objetivou comparar a eficiência da mutagênese física e química na geração de populações mutantes de petúnia. O dano genômico foi avaliado indiretamente por características de desenvolvimento de plântulas após o tratamento com doses de radiação gama de 0, 20, 40, 60, 80 e 100 Gy e do agente alquilante etil-metanossulfonato (EMS) de 0; 0,05; 0,1; 0,15; 0,2 e 0,25% (v/v). Radiação gama e EMS causaram danos ao desenvolvimento e reduziram a viabilidade das plântulas derivadas das sementes tratadas. As maiores doses de mutagênico diminuíram o número de plantas com folhas primárias aos 15 dias após a semeadura (DAS) em aproximadamente 44% e reduziram as taxas de sobrevivência a 55% (gama) e 28% (EMS) em relação aos controles. A redução na altura das plântulas foi proporcional ao aumento das dosagens de EMS, enquanto 40 e 60 Gy de radiação gama provocaram a redução mais significativa na altura de plantas. Abordagens de genética reversa requerem danos genômicos moderados, que permitam alta saturação de alelos mutantes com pequena redução no número de plantas viáveis, relacionados ao parâmetro biológico DL50, dosagem de mutagênico necessária para eliminar metade da população. Este valor correspondeu a 100 Gy de radiação gama e 0,1% de EMS. Os tratamentos foram empregados para a geração de populações mutantes de petúnia (M1 e M2) e novos mutantes morfológicos foram isolados.

Traditionally, mutagenesis has been used to introduce novel genetic variability in ornamental crops. More recently, it has become a powerful tool in gene discovery and functional analyses in reverse genetics approaches. The present work aimed to compare the efficiency of physical and chemical agents in generating mutant populations of petunia. We have indirectly evaluated the genomic damage by analyzing developmental characteristics of the plantlets derived from treated seeds employing gamma radiation at 0, 20, 40, 60, 80 and 100 Gy and the alkylating agent ethyl-methanesulfonate (EMS) at 0, 0.05, 0.1, 0.15, 0.2 and 0.25% (v/v). Gamma rays and EMS caused developmental defects and decreased seedling viability in plants obtained from the mutagenized seeds. High mutagen doses reduced in approximately 44% the number of plants with primary leaves at 15 days after sowing (DAS) and decreased seedling survival rates to 55% (gamma) and 28% (EMS), in comparison to untreated controls. Seedling height decrease was proportional to increasing EMS dosage, whereas 40 and 60 Gy of gamma irradiation caused the most significant reduction in height. Moderate DNA damage allowing a high saturation of mutant alleles in the genome and the generation of viable plants for reverse genetics studies was correlated to the biological parameter LD50, the dose required to kill half of the tested population. It corresponded to 100 Gy for gamma radiation and 0.1% for EMS treatment. The optimized mutagen treatments were used to develop petunia mutant populations (M1 and M2) and novel morphological mutants were identified.

The endogenous time-keeping mechanism is responsible for organizing plant physiology and metabolism according to periodic environmental changes, such as diurnal cycles of light and dark and seasonal progression throughout the year. In plants, circadian rhythms control gene expression, stomatal opening, and the timing component of the photoperiodic responses, leading to enhanced fitness due to increased photosynthetic rates and biomass production. We have investigated the citrus genome databases of expressed sequence tags (EST) in order to identify genes coding for functionally characterized proteins involved in the endogenous time-keeping mechanism in Arabidopsis thaliana. Approximately 180,000 EST sequences from 53 libraries were investigated and 81 orthologs of clock components were identified. We found that the vast majority of Arabidopsis circadian clock genes are present in citrus species, although some important components are absent such as SRR1 and PRR5. Based on the identified transcripts, a model for the endogenous oscillatory mechanism of citrus is proposed. These results demonstrate the power of comparative genomics between model systems and economically important crop species to elucidate several aspects of plant physiology and metabolism.

Water deficit is one of the most critical environmental stresses to which plants are submitted during their life cycle. The evolutionary and economic performance of the plant is affected directly by reducing its survival in the natural environment and its productivity in agriculture. Plants respond to water stress with biochemical and physiological modifications that may be involved in tolerance or adaptation mechanisms. A great number of genes have been identified as transcriptionally regulated for water deficit. EST sequencing projects provide a significant contribution to the discovery of expressed genes. The identification and determination of gene expression patterns is important not only to understand the molecular bases of plant responses but also to improve water stress tolerance. In our citrus transcriptome survey we have attempted to identify homologs to genes known to be induced and regulated under water stress conditions. We have identified 89 transcripts whose deduced amino acid sequences share similarities with proteins involved in uptake and transport of water and ion, 34 similar to components of the osmolyte metabolism, 67 involved in processes of membranes and proteins protection and 115 homologs of reactive oxygen species scavenger. Many drought-inducible genes identified are known to be regulated by development, salt, osmotic and low temperature. Their possible roles in specific or general mechanisms of water stress citrus responses are discussed.

Studies employing model species have elucidated several aspects of photoperception and light signal transduction that control plant development. However, the information available for economically important crops is scarce. Citrus genome databases of expressed sequence tags (EST) were investigated in order to identify genes coding for functionally characterized proteins responsible for light-regulated developmental control in model plants. Approximately 176,200 EST sequences from 53 libraries were queried and all bona fide and putative photoreceptor gene families were found in citrus species. We have identified 53 orthologs for several families of transcriptional regulators and cytoplasmic proteins mediating photoreceptor-induced responses although some important Arabidopsis phytochrome- and cryptochrome-signaling components are absent from citrus sequence databases. The main gene families responsible for phototropin-mediated signal transduction were present in citrus transcriptome, including general regulatory factors (14-3-3 proteins), scaffolding elements and auxin-responsive transcription factors and transporters. A working model of light perception, signal transduction and response-eliciting in citrus is proposed based on the identified key components. These results demonstrate the power of comparative genomics between model systems and economically important crop species to elucidate several aspects of plant physiology and metabolism.

Plant hormones play a crucial role in integrating endogenous and exogenous signals and in determining developmental responses to form the plant body throughout its life cycle. In citrus species, several economically important processes are controlled by phytohormones, including seed germination, secondary growth, fruit abscission and ripening. Integrative genomics is a powerful tool for linking newly researched organisms, such as tropical woody species, to functional studies already carried out on established model organisms. Based on gene orthology analyses and expression patterns, we searched the Citrus Genome Sequencing Consortium (CitEST) database for Expressed Sequence Tags (EST) consensus sequences sharing similarity to known components of hormone metabolism and signaling pathways in model species. More than 600 homologs of functionally characterized hormone metabolism and signal transduction members from model species were identified in citrus, allowing us to propose a framework for phytohormone signaling mechanisms in citrus. A number of components from hormone-related metabolic pathways were absent in citrus, suggesting the presence of distinct metabolic pathways. Our results demonstrated the power of comparative genomics between model systems and economically important crop species to elucidate several aspects of plant physiology and metabolism.

A transformação genética através da eletroporação de protoplastos foi estabelecida para espécies vegetais comercialmente importantes. Neste trabalho, fontes de explante, intensidades de campo elétrico, soluções de eletroporação, configuração da molécula de DNA e pré-tratamentos osmóticos foram avaliados para otimizar a expressão transiente do gene repórter em tecidos eletroporados de Stylosanthes guianensis, uma leguminosa forrageira tropical. Taxas elevadas de transformação foram obtidas empregando-se explantes cotiledonares e 250 V cm-1 de intensidade de campo elétrico. DNA plasmidial linear, solução de eletroporação livre de cloro e pré-tratamento osmótico com 1,6 mol L-1 de manitol favorecerem a expressão transiente do gene repórter, porém não significativamente. A amplificação por PCR específica do transgene foi usada para demonstrar a ocorrência de transformação nos tecidos.

Genetic transformation though protoplast electroporation has been established for commercially important plant species. In this work, explant sources, electric field strengths, electroporation buffers, DNA forms and osmotic pretreatment were assayed in order to optimize transient reporter gene expression in electroporated tissues of Stylosanthes guianensis, a tropical forage legume. Higher transformation rates were obtained employing cotyledonary explants and an electric field strength of 250 V cm-1. Linear plasmid DNA, chloridefree electroporation buffer and osmotic pretreatment with 1.6 mol L-1 mannitol also improved transient transformation but non-significantly. Transgene specific PCR amplification was employed to prove the transformed status of the tissues.

A transformação genética é um valioso recurso para o melhoramento e para a pesquisa em Genética, Fisiologia e Bioquímica. Plantas transgênicas são usadas para complementar ou substituir mutantes na pesquisa fundamental e para auxiliar em estudos de fenômenos biológicos complexos como patogenicidade, organização do genoma, captação de luz e transdução de sinais. Nesta revisão, são discutidas abordagens recentes visando a introdução, screening e seleção de transformantes, estudos sobre expressão do transgene e uso de plantas geneticamente modificadas. Ensaios de campo e culturas transgênicas comerciais são listadas assim como as principais espécies e características modificadas. O potencial das plantas transgênicas para a produção de compostos animais, remediação biológica e síntese de polímeros é igualmente apresentado.

Genetic transformation is a powerful tool for plant breeding and genetical, physiological or biochemical research, consequently it is an extremely dynamic field. Transgenic plants are commonly used to complete or substitute mutants in basic research, helping the studies of complex biological situations such as pathogenesis process, genome organization, light reception and signal transduction. In this review, recent approaches for foreign gene introduction (e.g. Agrobiolistics, whole tissue electroporation, in planta Agrobacterium transformation), screening (reporter gene possibilities and performance) and transformant selection (ipt selective marker) are discussed. Transgene expression and mechanisms underlying (trans)gene inactivation are presented. Practical applications of genetically modified plants, field tests and commercial transgenic crops worldwide and in Brazil are listed, as well as the main traits and species modified. Potential uses of transgenic plants for animal compound production, biological remediation and synthetic polymer assembly are also shown.