Sandra Patussi Brammer

Chromosome characterization in Thinopyrum ponticum (Triticeae, Poaceae) using in situ hybridization with different DNA sequences

Thinopyrum ponticum (2n = 10x = 70, JJJJsJs) belongs to the Triticeae tribe, and is currently used as a source of pathogen resistance genes in wheat breeding. In order to characterize its chromosomes, the number and position of 45S and 5S rDNA sites, as well as the distribution of the repetitive DNA sequences pAs1 and pSc119.2, were identified by fluorescent in situ hybridization. The number of nucleoli and NORs was also recorded after silver nitrate staining. Seventeen 45S and twenty 5S rDNA sites were observed on the short arms of 17 chromosomes, the 45S rDNA was always located terminally. On three other chromosomes, only the 5S rDNA site was observed. Silver staining revealed a high number of Ag-NORs (14 to 17) on metaphase chromosomes, whereas on interphase nuclei there was a large variation in number of nucleoli (one to 15), most of them (82.8%) ranging between four and nine. The pAs1 probe hybridized to the terminal region of both arms of all 70 chromosomes. In addition, a disperse labeling was observed throughout the chromosomes, except in centromeric and most pericentromeric regions. When the pSc119.2 sequence was used as a probe, terminal labeling was observed on the short arms of 17 chromosomes and on the long arms of five others. The relative position of 45S and 5S rDNA sites, together with the hybridization pattern of pAs1 and pSc119.2 probes, should allow whole chromosomes or chromosome segments of Th. ponticum to be identified in inbred lines of wheat x Th. ponticum.

Antioxidant responses of wheat plants under stress

Abstract Currently, food security depends on the increased production of cereals such as wheat (Triticum aestivum L.), which is an important source of calories and protein for humans. However, cells of the crop have suffered from the accumulation of reactive oxygen species (ROS), which can cause severe oxidative damage to the plants, due to environmental stresses. ROS are toxic molecules found in various subcellular compartments. The equilibrium between the production and detoxification of ROS is sustained by enzymatic and nonenzymatic antioxidants. In the present review, we offer a brief summary of antioxidant defense and hydrogen peroxide (H2O2) signaling in wheat plants. Wheat plants increase antioxidant defense mechanisms under abiotic stresses, such as drought, cold, heat, salinity and UV-B radiation, to alleviate oxidative damage. Moreover, H2O2 signaling is an important factor contributing to stress tolerance in cereals.

The importance for food security of maintaining rust resistance in wheat

Wheat is one of the main sources of calories and protein of the world’s population and therefore the pathogens that cause rust diseases of the crop are a real threat to food security. Besides the continuous evolution of rust pathogens which repeatedly results in overcoming the resistance of commercial varieties throughout the world, plant breeders are also now challenged by the impacts of global climatic changes. Agricultural practices will need to keep pace with the intensification of sustainable food production in order to face the challenge of feeding a world population estimated to reach about nine billion by 2050. Contemporary wheat breeding has increasingly focused on the future, culminating in the emergence of a global partnership for breeding new wheat varieties with resistance to rust pathogens. Plant breeding now employs a wide range of both long-established and frontier technologies aimed at achieving the United Nations Millennium Development Goals of ending hunger and extreme poverty (MDG1), while concurrently promoting environmental sustainability (MDG7) through global partnerships for development (MDG8).

Molecular cytogenetic characterization of parental genomes in the partial amphidiploid Triticum aestivum x Thinopyrum ponticum

The wheat line PF 839197 and six hybrid derivatives from a cross between PF 839197 and Thinopyrum ponticum were cytologically characterized by fluorescent in situ hybridization (FISH). Probes for the 5S and 45S rDNA genes (pTa794 and pTa71, respectively), a highly repetitive rye sequence (pSc119.2), the synthetic oligonucleotide (AAG)5, and total genomic DNA from Th. ponticum and rye were used. In the wheat line, a 1RS.1BL translocation was revealed by the labeling patterns produced with pSc119.2 and (AAG)5, and confirmed by genomic in situ hybridization (GISH) using rye genomic DNA as a probe. Analyses of partial amphiploids confirmed previous results indicating mitotic instability, with a tendency to stabilize at 2n = 42 or 56. GISH with Th. ponticum genomic DNA showed that in one hybrid derivative, with lower chromosome numbers (2n = 42-45), chromosomes were not labeled, whereas in the hybrids with 2n = 48-56 up to 14 chromosomes were labeled. These data suggest that the original chromosome set of these hybrids was 2n = 56, and that chromosomes from both genomes were lost by mitotic instability. FISH using the rDNA probes and GISH with Thinopyrum genomic DNA suggested that cells with 2n = 56 contained an entire wheat genome plus two monoploid chromosome sets of Th. ponticum.

Genetic analysis of adult-plant resistance to leaf rust in a double haploid wheat (Triticum aestivum L. em Thell) population

A genetic analysis of adult plant resistance to leaf rust (Puccinia triticina) was performed in in vitro obtained double haploid progenies from a cross between the Brazilian wheat cultivar Trigo BR 35, which, under the high inoculum pressure of the southern region, has been resistant to leaf rust for more than 12 years, and the susceptible cultivar IAC 13-Lorena. Haplodiploidization via in vitro gimnogenesis was done by somatic elimination of the pollen donor genome after maize pollination of the F1 plants. The advantages and usefulness of double haploids (DH) for genetic analysis of complex inherited traits like durable adult-plant resistance to wheat leaf rust were evident: it was possible to analyze inheritance patterns in this cross by using only the 35 DH homozygous segregant lines obtained by in vitro embryo culture of F1 flowers pollinated by maize, this number being equivalent to 1,225 conventional F2 lines because of lack of heterozygosity. After being infected with MCG and LPG races, the results indicated that Trigo BR 35 has two resistance genes. One of the genes expressed resistance only after the intermediate stage of plant development (5-6 leaves).

Viabilidade polínica como seleção assistida no programa de melhoramento genético de triticale

The objective of this study was to analyze the pollen grain viability of 52 triticale genotypes from the 2005 crossing block of the Embrapa Wheat breeding program. The spikes were collected prior to anthesis and cytological analyses were performed upon fixing the anthers from the basal, medium, and apical regions of the spike and replicated three times for each sample. For each slide, about 200 pollen grains were analyzed and they were classified into two different groups: 1) normal or viable and 2) non-viable or abnormal. The obtained data were analyzed and the means were compared by the Duncan test at 5% propability. The results of analysis of variance demonstrated that the differences among the genotypes were highly significant. However, most of the genotypes studied showed a high percentage of normal pollens. Cytological analysis of the pollen grain is potentially useful as assisted selection in a breeding program, especially to decrease the possibility of obtaining sterile plants or plants with less grain production.

Mitotic instability in wheat×Thinopyrum ponticum derivatives revealed by chromosome counting, nuclear DNA content and histone H3 phosphorylation pattern

To evaluate the mitotic stability of Triticum aestivum×Thinopyrum ponticum derivatives (BC2F7 and BC2F5 doubled haploids), chromosome counting by both conventional and immunostaining techniques, and measurement of DNA content were performed. The wheat progenitor line, PF 839197, the wheat recurrent parent CEP 19 and the control Chinese Spring were also investigated. In the hybrid derivatives, chromosome number ranged from 2n=36 to 60, with a predominance of chromosome numbers higher than 2n=42, that was confirmed by determination of nuclear DNA content. Chinese Spring’ and PF 839197 were stable, but CEP 19 showed chromosome number variation (20%). Analyses of non-pretreated cells revealed the presence of anaphase bridges, lagging chromatids, chromosome fragments and micronuclei. Immunostaining with an antibody recognizing histone H3 phosphorylated showed dicentric chromatids forming anaphase bridges and pericentromeric phosphorylation at centric chromosome fragments but not at lagging chromatids. The possible causes of the observed mitotic instability are discussed.

Reactive Oxygen Species and Antioxidant Enzymes Involved in Plant Tolerance to Stress

Plants are continuously exposed to several stress factors in field, which affect their production. These environmental adversities generally induce the accumulation of reactive oxygen species (ROS), which can cause severe oxidative damage to plants. ROS are toxic molecules found in various subcellular compartments. The equilibrium between the production and detoxification of ROS is sustained by enzymatic and nonenzymatic antioxidants. Due to advances in molecular approaches during the last decades, nowadays it is possible to develop economically important transgenic crops that have increased tolerance to stresses. This chapter discusses the oxidative stress and damage to plants. In addition, it reports the involvement of antioxidant enzymes in the tolerance of plants to various stresses.

Identificação de fontes de resistência à ferrugem da folha do trigo em acessos de Aegilops tauschii

In Brazil, losses caused by wheat leaf rust (Puccinia triticina) occur annually. The widespread incidence in the different producing areas varies in intensity, depending on climatic conditions, level of genetic resistance of the cultivars and efficacy of chemical control. The use of resistant cultivars is the most efficient control method, and the genes conferring resistance to leaf rust fungus are denominated Lr. Several of these genes have already been identified and mapped. Some of them were mapped directly in hexaploid wheats, while others were initially found in related species, with lower ploidy level and later transferred to cultivated wheat. Among related species, Aegilops tauschii, the diploid species donor of the D genome of cultivated wheat, is indicated as an important source of resistance genes to leaf rust. The objective of this work was to evaluate the resistance of A. tauschii accessions from the Active Germplasm Bank of Embrapa Trigo (BAG - Passo Fundo, RS) to leaf rust, in order to incorporate this resistance in commercial varieties. The reaction to race SPJ-RS of Puccinia triticina was evaluated in forty accessions of A. tauschii and 25% (10 accessions) were resistant. The data obtained in this study can help in choosing resistant accessions to be used as parents in breeding programs in order to increase resistance to the pathogen.

Genomic in situ Hybridization in Triticeae: A Methodological Approach

In several plant groups, especially those with polyploid complexes as Triticum (the wheat genus, Poaceae), related species can be used as important sources of genes. In the tribe Triti‐ ceae as a whole, which comprises other important cereals as barley (Hordeum vulgare) and rye (Secale cereale), there are high rates of successful interspecific hybridization [1-2]. Due to the ease in obtaining these hybrids, plus the high amount of available information on the genomes of the species, the interspecific hybrids are potentially useful for the genetic im‐ provement of these crops [3-4]. Thus, the hybrids and their derivatives from breeding pro‐ grams can be analyzed by means of different approaches, aiming the full knowledge on the phenotypic constitution of the plant material for its subsequent utilization.