Análise da expressão dos genes da subfamília DREB em resposta à seca em plantas de Coffea canephora Conilon

Coffea canephora is one of the most economically important coffee species and in Brazil, Conilon is the most widely cultivated plant of this species. Abiotic stresses such as temperature variations and drought periods are factors that significantly affect their production and tend to worsen with globally recognized climate changes. In an attempt to understand the molecular responses of coffee plants in water deficit conditions, recent studies have identified candidate genes (CGs) as CcDREB1D. This gene showed increased expression in response to drought in the leaves of clone 14 (drought tolerant) in relation to the clone 22 (sensitive to drought) of C. canephora Conilon. Based on these results, the identification of DREB genes and their subgroups (SGs) of C. canephora, the objective is to analyze in silico and also in vivo these genes expression in leaf and root of tolerant (14, 73 and 120) and sensitive clones (22) of C. canephora Conilon submitted or not to a water deficit. In silico expressions of all DREB genes were analyzed from the Coffee Genome Hub Database and in vivo expression was performed by the technique "reverse transcription-quantitative PCR" (RT-qPCR). In silico gene expression analysis was possible to identify DREB genes with potential responses to abiotic stresses, corroborating some validated in vivo results. In this analysis, several genes showed differential expression in response to drought among the SGs (IIV), the tolerant and sensitive clones and the leaf and root. These differentially expressed genes were identified as potential CGs and among them, it was found that most tolerant clones showed increased expression in relation to sensitive in response to drought, with higher expression levels for clones 14 and 73. These highest levels were observed in leaves compared to the roots and SG-I stood at greater number of genes expressed in response to drought. These results suggest that DREB CGs, as Cc05_g06840, Cc02_g03420 e Cc08_g13960, play an important role in the regulatory mechanisms of response to drought in C. canephora Conilon.

Avaliação do potencial de uso agrícola da fluorita com óxido silício (AgroSiCa), derivado da fabricação de fertilizantes fosfatados

Phosphate fertilizers are critical for crop production in tropical soils, which are known for having high phosphate-fixing capacity and aluminium saturation, as well as low pH and calcium contents. Fluorine is a component of many phosphate rocks used to make phosphate fertilizers, via a process that generates hexafluorosilicic acid (H2SiF6). While many treatment technologies have been proposed for removal of fluorine in industrial facilities, little attention has been given to a process of neutralizing H2SiF6 with calcium oxide aiming to find out an alternative and sustainable use of a by-product with a great potential for beneficial use in tropical agriculture. This study evaluated the effect of a by-product of phosphoric acid production (fluorite with silicon oxide, hereafter called AgroSiCa) in levels of phosphorus (P), calcium (Ca), silicon (Si), aluminum (Al) and fluorine (F) and some others parameters in soils as on growth of soybean and corn. Experiments were conducted in a greenhouse condition at the Federal University of Lavras (UFLA), Lavras, Minas Gerais, using different types of soils in tropical regions and different doses of AgroSiCa. The application of AgroSiCa resulted in a slight increase in soil pH and significant increases in calcium, phosphorus and silicon in the soil solution and the shoots of corn and soybeans. We also found very low levels of fluoride in all soil leachates. A significant reduction of labile aluminum levels found in all soils after the cultivation of corn and soybeans. In sum, AgroSiCa improved soil properties and contributed to better growth of both cultures. In sum, AgroSiCa improved soil properties and contributed to a better growth of both crops. Our results show that reacting H2SiF6 derived from the wet-process phosphoric acid production with calcium oxide leads to a by-product with potential for agricultural use, especially when applied in highly-weathered soils. Besides providing calcium and silicon to plants, the use of such by-product in soils with high phosphate-fixing capacity and high aluminium saturation delivers additional benefits, since fluoride and silicon can play an important role in improving soil conditions due to the formation of less plant-toxic forms of aluminium, as well as upon decreasing phosphate fixation, thus improving root development and making fertilizer-derived phosphate more available for plant growth.

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.

Influência do etileno na regulação do florescimento de Coffea arabica L.

Coffee is one of the most valuable exported agricultural product worldwide as well as the second most traded commodity after oil. This crop represents a source of employment and considerably accounts for the revenue of the country, besides its undeniable importance for local and world socioeconomical scenery. The quality of the coffee drink is directly related to the fruit ripening stage, which is found in different stages at harvesting due to the sequential flowering, increasing costs with the production and generating a lower-quality beverage. The asynchronous flowering of the coffee tree comes from the uneven development of buds, which can be influenced by environmental factors, taking into account that the anthesis takes place after a water deficit period, followed by precipitation. Changes in the hormone balance have been observed after water deficit and rehydration cycles, and such oscillation on the water status of the plant increased the level of ethylene in some species. This finding led to the association between the requirements at the flowering stage of the coffee tree and the involvement of the ethylene in promoting anthesis. The application of 1- MCP (ethylene action inhibitor) triggered the anthesis in coffee trees. And, in coffee seedlings in greenhouse conditions after a period of water shortage followed by irrigation, the profile of gene expression of the biosynthesis route and signaling of ethylene are changed in leaves and roots. The increased levels of the ethylene precursor (ACC) in roots throughout the dry season and the transportation of this compound into shoots may be the signal to trigger the anthesis in coffee tree after rehydration. Thus, in order to better understand the role of the ethylene in regulation of flowering, we analyzed the effects of the exogenous application of 1-MCP in different physiological characteristics and in the expression of genes related to the ethylene biosynthesis and signaling pathways in coffee leaves and bud from plants under field conditions. The evaluations and tissue sampling were carried out in different times treatments implementation: T1 – control, T2 – 1-MCP+Break-Thru, and T3 – Break-Thru. There was influence of 1-MCP and Break-Thru in gas exchange parameters and of 1-MCP contributed to the increase in relative water content. RT-qPCR analyses showed a different behavior in relation to the profile of gene expression in leaves and buds analyzed in the present study (CaACS1 – like, CaACO1 – like, CaACO4 – like, CaETR4 – like) and the expression levels were changed a few hours (2h) after applying the products. However, there was flowering only in plants treated with 1-MCP, in the absence of rain and irrigation.