Acidovorax citruli: genetic analyses and protocol for its detection in seeds

Bacterial fruit blotch of cucurbits (BFB), caused by the seed borne Gramnegative bacterium Acidovorax citrulli is a serious threat to cucurbit industry worldwide. Since late 1980`s after devastating outbreaks in watermelon fields in southern United States, BFB has spread worldwide and has been reported in other cucurbit crops such as melon, pumpkin, cucumber and squash. To date, there is evidence for the existence of at least two genetically and pathogenically distinct populations of A. citrulli. In Brazil, the first report of BFB was in 1991, in a watermelon field in São Paulo. Although widespread in the country, BFB has been a major problem to melon production. More precisely, BFB has caused significant yield losses to melon production in northeastern Brazil, which concentrates > 90% of the country`s melon production. Despite the management efforts and the recent advances in A. citrulli research, BFB is still a continuous threat to the cucurbit industry, including seed producers, growers and transplant nurseries. To better understand the population structure of A. citrulli strains in Brazil, and to provide a basis for the integrated management of BFB, we used pulsed-field gel electrophoresis (PFGE), multilocus sequence analysis (MLSA) of housekeeping and virulence-associated genes and pathogenicity tests on different cucurbit seedlings to characterize a Brazilian population of A. citrulli strains from different hosts and regions. Additionally, we conducted for the first time a comparative analysis of the A. citrulli group I and II population at genomic level and showed that these two groups differ on their genome sizes due to the presence of eight DNA segments, which are present in group II and absent in group I genomes. We also provide the first evidence to suggest that temperature might be a driver in the ecological adaptation of A. citrulli populations under nutrient-rich or -depleted conditions. Finally, in order to improve the routine detection of A. citrulli on melon seedlots, we designed a new primer set that is able to detect the different Brazilian haplotypes, thus minimizing the risk of false-negatives on PCR-based seed health testing.

Metodologias para a avaliação do potencial fisiológico de sementes de tabaco

Despite tobacco being a culture propagated by seeds, there is little information concerning tests that allow the distinction of similar germination lots in different levels of vigor. The diversity of cultivars available in the market, and a few peculiarities of the species, such as uneven maturation of the flowers, fruits and seeds, small size and seed dormancy, are considered obstacles for obtaining lots of tobacco of high physiological potential. Thus, this research was developed with the objective of adapting feasibility and vigor tests for evaluating the physiological potential of tobacco seed lots. We used nine lots of tobacco seeds of cultivar CSC 447 and nine lots of seeds of cultivar BAT 2101, belonging to variety groups Virginia and Burley, respectively. Initially, germination test was conducted to characterize the profile of the lots. For determining the feasibility and vigor of the tobacco seeds, germination tests were conducted in distinct temperatures, controlled emergence conditions, electric conductivity, artificial aging and in tetrazolium. For determining the isoenzymatic marker for seed quality, analyses were conducted with enzymes catalase, esterase, malate dehydrogenase and alcohol dehydrogenase. In conclusion, the emergence tests at 25oC and artificial aging at 41oC for 72 hours, are efficient in discriminating the lots of tobacco seeds in different levels of vigor. The electric conductivity and germination tests in different temperatures have distinct responses in relation to the genotype of the tobacco seeds. The tetrazolium test using the methodology with pre-conditioning in 3.5% sodium hypochlorite solution and subsequent emersion in 1.0% tetrazolium solution for 18 hours is efficient for the quick evaluation of the feasibility of tobacco seeds. The analysis of the profiles of enzymes catalase, esterase, malate dehydrogenase and alcohol dehydrogenase is efficient as markers for tobacco seed quality.