Gaspar Malone

IRAP and REMAP assessments of genetic similarity in rice.

Rice is a model genome for cereal research, providing important information about genome structure and evolution. Retrotransposons are common components of grass genomes, showing activity at transcription, translation and integration levels. Their abundance and ability to transpose make them good potential markers. In this study, we used 2 multilocus PCR-based techniques that detect retrotransposon integration events in the genome: IRAP (inter-retrotransposon amplified polymorphism) and REMAP (retrotransposon-microsatellite amplified polymorphism). Markers derived fromTos17, a copia-like endogenous retrotransposon of rice, were used to identify genetic similarity among 51 rice cultivars (Oryza sativa L.). Genetic similarity analysis was performed by means of the Dice coefficient, and dendrograms were developed by using the average linkage distance method. A cophenetic correlation coefficient was also calculated. The clustering techniques revealed a good adjustment between matrices, with correlation coefficients of 0.74 and 0.80, or lower (0.21) but still significant, between IRAP and REMAP-based techniques. Consistent clusters were found for Japanese genotypes, while a subgroup clustered the irrigated Brazilian genotypes.

Association between genetic distances in wheat (Triticum aestivum L.) as estimated by AFLP and morphological markers

Molecular and morphological data analyses matrices are very informative tools for the estimation of genetic distances. We used AFLP markers, morphological traits and combined analysis to estimate the genetic distances between wheat genotypes and ascertain any associations between the two techneques. Nineteen wheat (Triticum aestivum L.) genotypes were analyzed using amplified fragment length polymorphism (AFLP) markers and field evaluated for two years. The matrices obtained by morphological and molecular marker data analyses revealed a significant but moderate correlation (r = 0.47), indicating that such techniques sample distinct genome regions. The combined analysis was found to be highly correlated with AFLP (r = 0.97) and moderately correlated with morphological (r = 0.59) markers. A possible explanation for such results is a bias caused by the much higher number of AFLP (229) than morphological (17) markers. Thus, it is evident that the combined analysis is not efficient when a very dissimilar number of markers are used in each isolated technique. Therefore, to obtain a better knowledge of the degree of divergence among genotypes it is necessary to consider each analysis separately.

Dissimilaridade genética de linhagens de Trichogramma Westwood (Hymenoptera: Trichogrammatidae) através de marcadores moleculares ISSR

Demands for sustainable agricultural systems have forested the development of biological control techniques, such as the use of the egg parasitoid Trichogramma Westwood. However, the arduous identification of the parasitoid at the species level, due to the tiny size and the morphological similarities is an obstacle to increasing its use. Molecular markers are useful to reach the specimen genome and avoid environmental effects that could misguide identification. Several molecular markers techniques are available and the ISSR technique has been used to differentiate close individuals, due to its high polymorphism level, reproducibility and low cost. The objective of this study was to measure the level of genetic differentiation among five lines of Trichogramma, using ISSR markers: three belonging to the species T. pretiosum Riley, one to T. atopovirilia Oatman & Platner and one to T. bruni Nagaraja. Morphological identification of the parasitoids was conducted at ESALQ/USP - Piracicaba, SP. After DNA removal and standatization, PCR reactions were performed with 26 ISSR primers; 11 of them were selected because they presented greater polymorphism and consistency. Molecular data were converted into a binary matrix and analyzed (NTSYS v. 2.1). The 11 primers produced 172 polymorphic sections. Genetic similarity ranged from 19% to 96%, showing that the ISSR technique can efficiently identify DNA polymorphism in Trichogramma. Results also indicate important inter and intra-specific variations among the parasitoid lines.

Expressão diferencial de isoenzimas durante o processo de germinação de sementes de arroz em grandes profundidades de semeadura

The germination process consists of a complex and regulated group of physiologic and biochemical events that happen in the seed soon after beginning water absorption. The restart of the embryo growth due to water absorption involves the reactivation of many enzymes that will hydrolyze reserve substances, to supply redox potential and energy for germination. Plantlet emergence in monocotyledonous depends, principally, on the sowing depth and other factors such as genetic potential and seed vigor, because the embryo is nurtured by the seed stores along. In this study, isoenzimatic patterns of Esterase (EST), Acid Phosphatase (ACP), Malate Dehydrogenase (MDH), Alcohol Dehydrogenase (ADH) and Glutamate Oxalacetate Transaminase (GOT) were analyzed among germination processes at 20 cm in 34 red rice ecotypes and 5 commercial cultivars with the objective of identifying differential expression variations in the isoenzimatic systems analyzed. The five isoenzimatic systems analyzed showed variations in the enzymatic expression, mainly when compared with the patterns of the dry seed with the developing plantlets. From the results obtained it was concluded that ere is differential expression in the isozyme genes Esterase (EST), Acid Phosphatase (ACP), Malate Dehydrogenase (MDH), Alcohol Dehydrogenase (ADH) and Glutamate Oxalacetate Transaminase (GOT), showing participation in the germination process in rice.

IDENTIFICAÇÃO DO ESTÁDIO ADEQUADO PARA REALIZAÇÃO DE ANÁLISES ISOENZIMÁTICAS NA CARACTERIZAÇÃO DE CULTIVARES DE ARROZ 1

Enzymatic systems commonly used in cultivar characterization are products of gene expression and are, therefore, highly influenced by development stages, the organ from which they are collected and the environment. Usually, these factors are not considered when several enzymatic complexes are used from a single protein extraction, resulting in an inefficient reading and interpretation of the results. This study was carried out to evaluate the adequate stage for protein extraction for each enzymatic complex, when there is a maximum phenotypic expression, to be used in rice isoenzymatic characterization. Two lots, one of high and one of low physiologic quality, for each of the rice varieties El Paso L144, IRGA 417 and EEA 406, were analyzed using the Esterase, Acid Phosphatase, Glutamate Dehydrogenase, Glutamate Oxalacetate Transaminase, and Malate Dehydrogenase analyses. Six development stages (0, 2, 4, 6, 8 and 10 days) were used for protein extraction. Results suggested that each isoenzymatic complex requires an adequate moment for protein extraction; it was not possible to identify a development stage where the maximum phenotypic expression of the Esterase, Acid Phosphatase, Glutamate Dehydrogenase, Glutamate Oxalacetate Transaminase, and Malate Dehydrogenase isoenzymatics complexes presented the same profile; it is not recommended to analyze several isoenzymatics complexes from a single protein extraction; the physiologic quality of EEA 406 seeds affected the Esterase and Glutamate Oxalacetate Transaminase isoenzymatic profiles.

Emprego de modelos gráficos na seleção de genitores de milho para hibridização e mapeamento genético

Associating phenotypic to molecular data can be a powerful tool for the selection of parental genotypes for breeding and mapping purposes. Thus, the objectives of the study were: i) to estimate the genetic dissimilarity among 30 maize inbred lines (15 tolerant and 15 sensitive to flooding); ii) to select potential parents for mapping and breeding; iii) to compare the efficiency of different graphical models in displaying the calculated distances. A total of 21 RAPD primers were used for the estimation of genetic dissimilarity. The genetic dissimilarity was obtained according to the complement of Dice similarity coefficient, clustering procedure was performed by the average linkage method and the cophenetic coefficient was obtained. The complement of Dice similarity coefficient was subjected to principal components and multidimensional scale analyses, and the output efficiency was tested by the correlation between the original distances and those presented in the graphs. The clustering techniques did not reveal a perfect agreement with the original matrix, with correlations of 0.70, 0.53 and 0.75 for the dendrogram, principal components and multidimensional scale analyses, respectively. Among the tested techniques employed, multidimensional scale analyses gave more precise outputs, since this technique showed higher agreement with the original distance matrix, and preserved distances between all genotype pairs. Besides, this technique is the most indicated when the objective is to plan crosses, since it displays the distances between genotype pairs.

Variabilidade genética interespecífica em formigas cortadeiras do gênero Acromyrmex que ocorrem no Estado do Rio Grande do Sul

At present, a few number of taxonomists work on identification of leaf cutting ants. Morphological-based methods do not always produce a reliable identification, due to the strong polymorphism observed even in the same colony. This fact leads to hypothesize that, besides those variations, others may also occur related to geographic distribution of the ants. Six Acromyrmex species were sampled at four locations in the State of Rio Grande do Sul and analyzed by RAPD and AFLP. Fifty UBC primers, originated from the University of British Columbia, were evaluated to select a set of primers that could be useful for species identification and genetic variability studies. Only 13 primers, which amplified fragments of all species, were selected. Although the selected primers produced fragments that allowed various ways for identification of A. heyeri, A. ambiguus, A. crassispinus, A. striatus, A. laticeps and A. aspersus, only UBC 354, UBC348 and UBC356 primers allowed a reliable identification showing the most visible fragments. Results by dendogram and morphological-based identification, showed that A. striatus and A. laticeps are less related to the other species. However A. ambiguus e A. crassispinus are the most genetically related ants in the State of Rio Grande do Sul. The results indicate that a reliable identification of Acromyrmex can be carried out by RAPD and AFLP, yet allowing verification of genetic distance between species.