Caracterização e comparação de genes expressos em ápices meristemáticos de cana-de-açúcar cultivados em SP e RN

In this work, we used sugarcane as a model due to its importance for sugar and ethanol production. Unlike the current plant models, sugarcane presents a complex genetics and an enormous allelic variation. Here, we report the analysis of SAGE libraries produced using the shoot apical meristem from contrasted genotypes by flowering induction (non-flowering vs. early-flowering varieties) grown under São Paulo state conditions. The expression pattern was analyzed using samples from São Paulo (SP) and Rio Grande do Norte (RN) states. These results showed that cDNAs identified by SAGE libraries had differential expression only in São Paulo state samples. Furthermore, the cDNA identified CYP (Citocrome P450) was chosen for in silico and genome characterization because it was found in SAGE libraries and subtractive libraries from samples from RN. Phylogenetic trees showed the relationship for these sequences. Furthermore, the qRT-PCR for CYP showed a potential role as flowering indutor for RN samples considering different isophorms. Considering the results present here, it can be consider that CYP gene may be used as molecular marker

Germinação de sementes e otimização de técnicas de micropropagação de umbuzeiro (spondias tuberosa, arr.) anacardiaceae

The brazilian-plum (Spondias tuberosa, His) is a tropical fruit tree that has been consolidated in the market for agribusiness processing, due to its characteristic flavor of fruit. Accordingly, studies to optimize the propagation of plants are necessary for production of seedlings with agronomic and quality assurance measures. This study aimed at determining the efficient techniques for uniform seed germination, as brazilian-plum seed present mechanical dormancy, and establish optimal culture media for multiplication of shoots from the in vitro micropropagation. Firstly, in a greenhouse at the Universidade Federal do Rio Grande do Norte, was evaluated the influence of different methods of breaking dormancy in the emergence of seedlings of brazilian-plum and speed of germination (IVG) of seeds. After 60 days of cultivation, it was found that splay in the distal portion of the seed was the best treatment, with rates of 85.33% in germinability and 3.415 of IVG, compared with the treatment of seed-soaking in water for 12h + humus and the control group. Subsequently, new sources of seedling explants were obtained in studies of tissue culture. Laboratory of Plant Biotechnology that the university, was used stem apex, nodal segments and internodes in search of decontamination with various concentrations of calcium hypochlorite [Ca(OCl)2] and micropropagation, inoculating them in half WPM (1980) with various concentrations of 6-benzylaminopurine (BAP). We used 10 sample units with three replications for different concentrations of [Ca(OCl)2], BAP and explants type. After thirty days, which was observed for the control of contamination, during the establishment in vitro, concentrations of [Ca(OCl)2] between 0.5% and 2.0% were effective in combating exogenous contamination of the apex. In nodal segments and internodes, concentrations of [Ca(OCl)2] between 1.0% and 2.0% and 1.5% and 2.0% were respectively, sufficient to reduce the percentage of losses in these infestations explants. For micropropagation, the culture medium supplemented with 0.1 mg.L-1 BAP promotes better development of multiple shoots per explants from nodal segment. However, success does not get to shoot training in internodal segment

Análise de cDNAs identificados em bibliotecas substrativas de cDNA para floração de variedades de cana-de-açúcar cultivadas no Rio Grande do Norte(RN): fator de transição NAC, Calmodulina e Fosfatidiltransferase

The flowering is a physiological process that it is vital for plants. This physiological process has been well studied in the plant model Arabidopsis, but in sugarcane this process is not well known. The transition of the shoot apical meristem from vegetative to flowering is a critical factor for plant development. At Brazil northeastern region, the transition to flowering in sugarcane has an important effect as it may reduce up to 60% its production. This is a consequence of the sugar translocation from stalks to the shoot apical meristem which is necessary during the flowering process. Therefore, the aim of this work was to explore and analyze cDNAs previously identified using subtractive cDNA libraries. The results showed that these cDNAs showed differential expression profile in varieties of sugarcane (early x late flowering). The in silico analysis suggested that these cDNAs had homology to calmodulin, NAC transcription factor and phosphatidylinositol, a SEC14, which were described in the literature as having a role in the process of floral development. To better understand the role of the cDNA homologous to calmodulin, tobacco plants were transformed with overexpression cassettes in sense and antissense orientation. Plants overexpressing the cassette in sense orientation did not flowered, while plants overexpressing the cassette in the antissense orientation produced flowers. The data obtained in this study suggested the possible role from CAM sequence, SEC14 and NAC in the induction/floral development pathway in sugarcane, this is the first study in order to analyze these genes in the sugarcane flowering process.

Mobilização de reservas e partição de metabólitos durante a germinação da semente e o estabelecimento da plântula em moringa

Seed germination and seedling establishment are critical processes for commercial plantation and depend directly on reserve mobilization as a source of cellular fuels and biosynthetic precursors. In this way, we investigated the coordination among reserve mobilization, metabolite partitioning, and mobilizing enzyme activities in Moringa oleifera Lam (moringa) an oil-seeded species employed in biofuel production. Seeds were germinated under controlled conditions and seedlings were grown hydroponically at a greenhouse. Samples were harvested at 0, 4, 8, 10, 12, 16, and 20 days after imbibition (DAI). The contents of dry mass (DM), neutral lipids (NL), soluble proteins (SP), starch, total soluble sugars (TSS), non-reducing sugars (NRS), and total free amino acids (TFAA) as the activity of isocitrate lyase (ICL), acid proteases, and amylases were determined. The mobilization of storage proteins was initiated during seed germination whereas the mobilization of storage lipids and starch was triggered throughout seedling establishment although all reserves have been depleted until 20 DAI. The partitioning of DM and metabolites to the roots and the shoots was uneven during seedling establishment. Low shoot/root ratio on the basis of DM could be related to the natural occurrence of moringa in drought climates. In the roots, TSS, NRS, and TFAA were accumulated from 12 to 16 DAI and then were consumed until the end of the experiment. In the shoots, TSS and TFAA were consumed in parallel with NRS accumulation from 12 to 20 DAI. The activity of ICL, acid proteases, and amylases was coordinated with the mobilization of lipids, proteins and starch respectively. Thus, we propose that the patterns of reserve mobilization and metabolite partitioning verified in moringa seem distinct from those found to other tree species and may be involved in metabolic strategies to enable environment colonization

Caracterização ecofisiológica de sementes de espécies lenhosas da Caatinga

Dormancy is an inherent property of the seeds that define the environmental conditions in which they are able to germinate and their presence is an adaptive trait common in species inhabiting semiarid regions. Moreover, the ability of seedling establishment in these environments has been related to the size, strength and chemical characteristics of the seeds. This study investigated patterns of dormancy and germination speed in tree species of the Caatinga, exploring how the seed size influence the processes of germination, seedling size and biomass allocation. In addition, we aim to investigate the chemical characteristics of the reserves, to verify a possible relationship between nutritional content and the process of seed germination. Therefore, seeds were collected from ten species of woody Caatinga for tests of breaking dormancy, germination and biochemical characterization. Overall, the results show that the scarification treatments mechanical and chemical, and thermal shock influenced the percentage and speed of germination in 50 % of the species, suggesting that they have some level of physical dormancy in the seeds. Biochemical characterization showed the existence of large amounts of carbohydrates in the seeds of all species, low proportion of protein and low amounts of neutral lipids. Using linear regression, we demonstrated the existence of a significant relationship between seed size and the ratio of root/shoot where the largest seeds invested a greater amount of resources for shoot growth. The relationship between germination speed and non-reducing sugar content was also significant, so these compounds is related to the maintenance of physiological seed quality. These results confirm some relationships discussed in the literature for cultivated species, but can be applied to the species native to the Caatinga

Prospecção de genes associados ao processo de floração em tomateiro.

Flowering is a process marked by switch of shoot apical meristem to floral meristem, and it involves a complex regulation by endogenous and environmental factors. Analyses of key flowering genes have been carried out primarily in Arabidopsis thaliana and have provided a foundation for understanding the underlying molecular genetic mechanisms controlling different aspects of floral development. Several homologous have been found in other species, but for crops species such as tomatoes this process is not well known. The aim of this work was to use the genetic natural variation associated to the flowering process and use molecular tools such as subtractive libraries and real time PCR in order to identify and analyze the expression from genes that may be associated to flowering in these two species: L. esculentum cv Micro-Tom and L. pimpinellifolium. Our results showed there were identified many genes related to vegetative and possibly to the flowering process. There were also identified many sequences that were unknown. We ve chosen three genes to analyze the expression by real time PCR. The histone H2A gene gave an expression higher in L. pimpinellifolium, due to this the expression of this gene may be associated to flowering in this specie. It was also analyzed the expression of an unknown gene that might be a key factor of the transition to flowering, also in L. pimpinellifolium. For the elongation factor 1-α expression, the expression results were not informative, so this gene may have a constitutive expression in vegetative and flowering state. The results observed allowed us to identify possible genes that may be related to the flowering process. For further results it will be necessary a better characterization of them.

Qualidade fisiológica de sementes de genótipos de algodoeiro sob estresse salino

The germination of cotton seeds and the seedlings emergency are generally delayed and reduced by the salinity. Although the cotton is considered a tolerant culture, it can suffer substantial reductions in regarding its growth and production when exposed to salinity condition. The aims of this study went evaluate the effect of the saline stress in the germination phase to four cotton genotypes (BRS Rubi, BRS Safira, BRS 201 and CNPA 187 8H), using different osmotic potentials generated with increment of sodium chloride (NaCl). The saline stress was simulated using NaCl aqueous solutions in the potentials: 0.0 (Control); -0.2; -0.4; -0.6; -0.8 and -1.0 MPa. The treatments were monitored by means of tests for analysis of seeds, germination, first counting, speed germination index, length of shoot, radicle length, dry weigth of embrionic axis and shoot/radicle ratio. The tests for germination, first counting and index of germination speed were accomplished using 50 seeds for repetition and for the study of length of shoot, radicle length, dry weigth of embrionic axis and shoot/radicle ratio were used 20 seeds by repetition. For both tests four repetitions were accomplished by genotype for each one of the potentials. The seeds of each repetition were involved in papers Germitest humidified with NaCl solution corresponding to the potential. The repetitions of both tests were maintained in a germinator with saturated humidity. The analysis were initiate four days after the induction of the saline stress. The evaluations of the first three variables analyzed were accomplished daily; the seeds were remove and counted when its germinated. For the length tests just the repetitions corresponding to the potential of NaCl 0,0 MPa were analysis 4 days after the beginning of the induction of the saline stress. The analysis of the repetitions of the potentials -0,2 and -0,4 and of the potentials -0,6, -0,8 and -1,0 MPa they were accomplished with 12 and 20 days, respectively. For accomplishment of the analisis of this test the shoot of the 20 plantules of each repetition was separate from the radicle and both parts were measured. The statistical analyses were performed using the GENMOD and GLM procedures of the SAS. For the variable germination, the cultivates CNPA 187 8H and BRS Safira stood out for the potential -0.8 MPa, with averages of 89% and 81%, respectively. The test of speed germination index to cultivate BRS Safira presented the largest averages for the two higher saline potentials. It was observed that the increase of the saline potential reduces the germination percentage and speed germination index. For each day of evaluation it was verified that the increase of the saline potential causes a reduction of the length both of the shoot and of the radicle. The radicle tends to grow more than the shoot until the potential -0,4 MPa

Conservação de espécies nativas em um assentamento rural do município de Apodi-RN: impactos socioambientais e econômicos e o uso da biotecnologia como alternativa promissora

The Caatinga biome has a high diversity of potential and their conservation constitutes one of the greatest challenges of Brazilian science. The sustainable management of the Caatinga emerges as an alternative that through the formation of systems agrossilvipastoris, enables the use of forest resources sustainably, ensuring their conservation, regeneration and recovery. In RN this technique has been developed mainly in settlements of Agrarian Reform, such as P. A. Moaci Lucena, and their impacts go beyond the environmental aspect and reverberate socially and economically on the quality of life of family farmers. Despite the efficiency of the Sustainable Management of the Caatinga in the conservation of native species, many forests species of this biome faces serious problems of propagation and for this reason have become vulnerable to extinction, as is the case of Mimosa caesalpiniifolia Benth . Thus , it is evident the need to use sustainable alternatives to overcome the difficulties of propagation of this species and enable their replacement in areas where their existence is threatened. The Plant Biotechnology is considered as a promising alternative in this sense, considering that by micropropagation enables the large-scale production of seedlings with high health genetics status. This work has the following objectives: evaluate the perception of family farmers of P. A. Moaci Lucena in relation to social, environmental and economic impacts of the Sustainable Management of the Caatinga and check the conditions of germination and in vitro propagation of Mimosa caesalpiniifolia Benth that enabling the production of seedlings of this specie on a large scale. To achieve the first objective, semi-structured interviews showed that in the perception of farmers PA Moaci Lucena, the Sustainable Management of the Caatinga was responsible for generating many social, environmental and economic impacts that affected directly in the improvement in the quality of life of the families of the Settlement Project Moaci Lucena. Have to achieve the second objective, were investigated the influence of different substrates and concentrations of growth regulator BAP in the germination and shoot induction in vitro of Mimosa caesalpiniifolia Benth. The vermiculite was presented as the most suitable substrate for germination of this species, because it provided a more rapid germination, higher growth rates and higher dry matter accumulation. Regarding micropropagation, the concentration of 17.76 μmol/L of BAP presented a more responsive in relation to multiplication rate and the number of shoots in M. caesalpiniifolia, thus constituting the most suitable concentration for the in vitro propagation of this specie