Noélle Giacomini Lemos

Evaluation of environmental waters using the comet assay in Tilapia rendalli

Testing for environmental pollutants is an ever-growing concern. Various tests in organisms have been utilized for the detection and identification of toxic substances in the air, water and soil. In the present study, we utilized the comet assay in Tilapia rendalli to conduct an environmental assessment of Lake Igapó II, a lake located in the metropolitan area of Londrina, PR-Brazil. The results demonstrated that samples from Lake Igapó II had a significantly greater number of comets, mainly in classes 2 and 3. The results suggest a genotoxicity of the aquatic environment at Lake Igapó II and that the comet assay in T. rendalli provides adequate sensitivity to be utilized as a tool in the monitoring of water pollution and environmental risk assessment.

Development of classification criteria for resistance to soybean rust and differences in virulence among Japanese and Brazilian rust populations

Nos ultimos anos a ferrugem asiatica, causada pelo fungo Phakopsora pachyrhizi tornou-se uma das mais serias ameacas a producao de soja Brasileira. Linhagens melhoradas e variedades tem sido selecionadas para a resistencia a ferrugem da soja na Asia, entretanto para a selecao e utilizacao dessas fontes de resistencia, diferencas de virulencia entre populacoes Asiaticas e Brasileiras desse fungo devem ser consideradas. Neste trabalho sugerimos um criterio para se distinguir resistencia de susceptibilidade pela analise de quatro caracteres de resistencia: frequencia de lesoes contendo uredias, numero de uredias por lesao, frequencia de uredias abertas e nivel de esporulacao determinados pela utilizacao de 63 genotipos. Sob condicoes controladas em câmaras de crescimento, treze variedades de soja foram expostas a tres populacoes de fungos - uma populacao proveniente do Japao e duas populacoes provenientes do Brasil-e avaliadas quanto aos caracteres de resistencia mencionados acima. As populacoes Brasileiras diferiram entre si claramente quanto a virulencia e em relacao a populacao de isolados do Japao. Apenas dois genes de resistencia, Rpp4 presente na variedade PI459025 e Rpp5 presente na variedade Shiranui conferiram resistencia as tres populacoes da ferrugem. O numero de variedades ou genes resistentes uteis em ambos os paises parece ser limitado. Assim, um cultivar universalmente efetivo contra a ferrugem da soja deveria ser desenvolvido pela piramidacao de genes maiores de resistencia e pela introducao de resistencia horizontal.

Characterization of genes Rpp2, Rpp4, and Rpp5 for resistance to soybean rust

Asian rust, caused by the fungus Phakopsora pachyrhizi, is the most severe disease currently threatening soybean crops in Brazil. The development of resistant cultivars is a top priority. Genetic characterization of resistance genes is important for estimating the improvement when these genes are introduced into soybean plants and for planning breeding strategies against this disease. Here, we infected an F2 population of 140 plants derived from a cross between ‘An-76’, a line carrying two resistance genes (Rpp2 and Rpp4), and ‘Kinoshita’, a cultivar carrying Rpp5, with a Brazilian rust population. We scored six characters of rust resistance (lesion color [LC], frequency of lesions having uredinia [%LU], number of uredinia per lesion [NoU], frequency of open uredinia [%OU], sporulation level [SL], and incubation period [IP]) to identify the genetic contributions of the three genes to these characters. Furthermore, we selected genotypes carrying these three loci in homozygosis by marker-assisted selection and evaluated their genetic effect in comparison with their ancestors, An-76, PI230970, PI459025, Kinoshita and BRS184. All three genes contributed to the phenotypes of these characters in F2 population and when pyramided, they significantly contributed to increase the resistance in comparison to their ancestors. Rpp2, previously reported as being defeated by the same rust population, showed a large contribution to resistance, and its resistance allele seemed to be recessive. Rpp5 had the largest contribution among the three genes, especially to SL and NoU. Only Rpp5 showed a significant contribution to LC. No QTLs for IP were detected in the regions of the three genes. We consider that these genes could contribute differently to resistance to soybean rust, and that genetic background plays an important role in Rpp2 activity. All three loci together worked additively to increase resistance when they were pyramided in a single genotype indicating that the pyramiding strategy is one good breeding strategy to increase soybean rust resistance.

Soybean physiology and gene expression during drought.

Soybean genotypes MG/BR46 (Conquista) and BR16, drought-tolerant and -sensitive, respectively, were compared in terms of morphophysiological and gene-expression responses to water stress during two stages of development. Gene-expression analysis showed differential responses in Gmdreb1a and Gmpip1b mRNA expression within 30 days of water-deficit initiation in MG/BR46 (Conquista) plants. Within 45 days of initiating stress, Gmp5cs and Gmpip1b had relatively higher expression. Initially, BR16 showed increased expression only for Gmdreb1a, and later (45 days) for Gmp5cs, Gmdefensin and Gmpip1b. Only BR16 presented down-regulated expression of genes, such as Gmp5cs and Gmpip1b, 30 days after the onset of moisture stress, and Gmgols after 45 days of stress. The faster perception of water stress in MG/BR46 (Conquista) and the better maintenance of up-regulated gene expression than in the sensitive BR16 genotype imply mechanisms by which the former is better adapted to tolerate moisture deficiency.

Resistance to Asian soybean rust in soybean lines with the pyramided three Rpp genes

In this study, the influence of genetic background on the resistance level of a soybean line carrying Rpp2, Rpp4, and Rpp5 was evaluated by backcrossing it with a susceptible variety. It was also evaluated eight lines which carry these Rpp genes against five Asian soybean rust (ASR) isolates, in order to determine the likely range of resistance against ASR isolates differing in pathogenicity. The results indicated that a high level of resistance against various ASR isolates could be retained in lines carrying the three Rpp genes in susceptible genetic backgrounds, although minor influences of plant genetic background and ASR pathogenicity to the ASR resistance could occur. Thus, lines with the pyramided three Rpp genes should be effective against a complex pathogen population consisting of diverse Phakopsora pachyrhizi isolates.

Cloning and quantitative expression analysis of drought-induced genes in soybean.

We determined the expression levels of DREB transcription factor (Gmdreb1) and of the genes Gmgols, Gmpip1b, Gmereb, and Gmdefensin in drought-tolerant (MG/BR46-Conquista) and drought-sensitive (BR16) genotypes of soybean, during drought. The trial was carried out in a controlled-environment chamber, set up to provide drought conditions. Sequences of Arabidopsis thaliana DREB-family proteins were used to build a phylogenetic tree through the alignment of the conserved regions near the AP2 domain. We found that Gmdreb1 is similar to Atrap2.1, which is located near the AtDREB1 and AtDREB2 families. The amplified fragment was cloned and sequenced; alignment with the sequence available at Genbank showed total similarity. Expression analysis showed that under drought: a) Gmdreb1 expression increased in leaves and roots of both genotypes and expression level changes occurred that were correlated with the length of the water-deficit period; b) there were increased expression levels of Gmdefensin in roots of MG/BR46; c) expression of Gmgols increased in leaves and roots of the two genotypes; d) Gmpip1b expression generally increased, except in roots of BR16, and e) the same was found for Gmereb, except in roots of MG/BR46.

Morpho-anatomical and micromorphometrical evaluations in soybean genotypes during water stress.

In a greenhouse experiment, morpho-anatomical and micromorphometrical analyses of two soybean cultivars, MG/BR46 (Conquista) and BR16-tolerant and sensitive to drought, respectively-were used to study their water-deficit-tolerance strategies. Drought treatments were applied at reproductive stages from R2 to R7, where evaluations were conducted at 30 days and 45 days after stress started, respectively. The total length of Conquista plants (shoot + root) was greater than of BR16 plants. Pod dry weight was adversely affected due to the lack of moisture, decreasing productivity even of Conquista plants. Both the cultivars had normal development of root hairs; however, it was observed a decrease in the cortex:central cylinder ratio in BR16 stressed for 30 days, and they also showed similar leaflet thickness and stomata distribution. Differences in drought tolerance observed between the two cultivars seemed to be related to factors other than morphological traits since this species has a short lifecycle.