Programa de Pós-graduação em Genética e Melhoramento de Plantas
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Item RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia(Universidade Federal de Goiás, 2017-12-18) Fagundes, Lucas França; Novaes, Evandro; http://lattes.cnpq.br/0568272239145336; Novaes, Evandro; Antunes, Adriana Maria; Moraes, Moemy Gomes deDue in most part to the rapid growth and adaptability of the Eucalyptus in Brazil, the country's forestry sector is one of the most competitive of the world. Drought is one of the main obstacles of the culture, which has been expanding to the country’s interior regions such as the Cerrado biome. The objective of this work was to identify genes and biological processes involved in the responses of the Eucalyptus roots to drought. The experiment was conducted in a completely randomized design with 2 treatments (control and deficit) x 3 root RNA gathering time (3, 6 and 12h after start of treatment) and three biological replicates (commercial clones 1528, I144, VM1). The plants grew in hydroponics for 50 days, before the experiment start by using PEG8000 to induce water deficit (osmotic potential of -0.9 MPa). The root RNA of the seedlings was extracted using the RNeasy Plant Mini® kit (Quiagen). RNA samples were sequenced on the HiSeq 2000 platform (Illumina), yielding approximately 626 million 50 bp sequences in single end mode. After quality control, the sequences were aligned in the reference genome of Eucalyptus grandis. After counting reads aligned on each gene, differential expression analyses were performed with edgeR package of R/Bioconductor. A total of 6169 differentially expressed genes (DEG) (FDR ≤ 5% and Log2 FoldChange > |2|) were identified. Of these, 3350 were up-regulated and 2819 were repressed under osmotic stress, considering all the time-series evaluations. At three hours after treatment the highest number of DEG was obtained, with 3150, followed by six hours (2494 DEG) and 12h (525 DEG). Functional enrichment analysis was performed with the Fisher exact test, by software Blast2GO, with the DEG associated with their respective Gene Ontology (GO) terms. As a total of 90 GO terms, enriched and non-redundant, was verified that our results indicate a rapid initial response of Eucalyptus roots induced by PEG 8000 in hydroponics. The most remarkable characteristics of this response under hydroponic conditions are: increased energy consumption via carbohydrate catabolism, activation of the glycolytic pathway, repression of the respiratory chain in the mitochondria, increase of the fermentation pathway, partial execution of the citric acid cycle and repression of the primary metabolism (DNA replication and RNA biosynthesis, development and growth). These enriched functional terms point to the predominance of hypoxia stress, induced by the high viscosity of PEG in hydroponics, as compared to the osmotic stress.Item Análise da diversidade e divergência genética em clones de Eucalyptus spp. potencialmente importantes para Goiás(Universidade Federal de Goiás, 2014-09-03) Maciel, Kelly de Jesus Silva; Novaes, Evandro; http://lattes.cnpq.br/0568272239145336; Novaes, Evandro; Coelho, Alexandre Siqueira Guedes; Vianello, Rosana PereiraThe success of the Brazilian forestry is due largely to the excellent adaptability of the Eucalyptus genus to our climate and soil conditions. The recent expansion of eucalypts to the North and Midwest regions of Brazil presents challenges such as the need for clones adapted to drought, high temperatures and nutrient deficient soils of Cerrado. Three clonal trials were installed in different regions of Goiás State to evaluate the adaptability and growth of 113 elite clones of Eucalyptus spp. The objective of this study was to estimate the genetic diversity and divergence among 90 of the clones used in three clonal trials installed in different regions of Goiás State. The clones were genotyped with nine microsatellite loci organized into four "multiplex" systems for PCR. The amplified fragments were separated on the ABI–3100 platform (Applied Biosystems). The genotyping was performed using the GeneMapper software (Applied Biosystems). Genetic diversity parameters were estimated using the GDA and Fstat programs. The parameters number of alleles (A), expected heterozygosity (He), observed heterozygosity (Ho), intrapopulation fixation index (f) and allelic richness were estimated for each microsatellite locus. The results showed that all loci used in this study were highly polymorphic, with an average of 16.78 alleles per locus. The EMBRA28 and EMBRA3 loci showed the highest number of alleles (24 and 22). In general, for most markers, the observed heterozygosity had similar estimates when compared to the expected heterozigosity under Hardy-Weinberg Equilibrium. As a result, the fixation index (f) did not differ significantly from zero. Analyses of genetic structure of the clones was performed using the software Structure (version 2.3.4), with K values ranging from 1 to 10, with 10 interactions each. Results indicated presence of three distinct genetic groups (K = 3). However, there was no clear relationship between the populations obtained and the different species of Eucalyptus used in the study. This result can be explained by the clone sample is originated from breeding programs where crosses may have admixed populations, disrupting some genetic structures. Most importantly, the molecular analyses indicate extraordinary genetic diversity within the clonal trials installed in Goiás. This genetic diversity can be exploited for breeding new genetic material adapted to the Cerrado conditions.Item Caracterização de microssatélites e desenvolvimento de marcadores SSR em khaya grandifoliola C. DC(Universidade Federal de Goiás, 2019-11-26) Sousa, Rodrigo Carlos Batista de; Soares, Thannya Nascimento; http://lattes.cnpq.br/5590256762396056; Novaes, Evandro; http://lattes.cnpq.br/0568272239145336; Novaes, Evandro; Coelho, Alexandre Siqueira Guedes; Soares, Thannya Nascimento; Guimaraes, Rejane AraujoAfrican mahogany (Khaya spp.) cultivation attracts increasing interest from producers, since their wood has similar characteristics to those of Brazilian mahogany with high commercial value in the European market. Despite its timber value throughout the world and its ecological importance in Africa, where it is classified as vulnerable to extinction, little is known about the species' levels of genetic diversity. Thus, this work aims to identify and characterize microsatellite regions in the transcriptome of Khaya grandifoliola, as well as to develop markers for studies of the genetic variability of the species. From the transcripts developed by Soares et al., (2019), microsatellite regions were identified and PCR primers were designed. Microsatellite sequences were classified according to the type and number of repetitions, as well as their location within transcripts. Using the microsatellite loci genetic parameters were estimated, such as the number of alleles per loci (K), expected and observed heterozygosity (He and Ho), intrapopulational fixation index (f), probability of exclusion of paternity and of identity (PE and PI). Within the African Mahogany transcripts, 37,925 microsatellite regions were detected, with 54% being mononucleotides, 29% dinucleotides and 15% trinucleotides. From 40 microsatellite loci, 12 were selected for labelling with fluorophores (6-FAM and HEX), forming two multiplex panels. The genotyping performed with the individuals identified an average of 3 alleles per locus. The expected (He = 0.53) and observed heterozygosity (Ho = 0.50) values used to estimate the genetic diversity in K. grandifoliola individuals were reasonable, given the low number of alleles per locus. The values found for PE (0.96) and PI (4.70x10-6) were similar to those found in studies of the same genus. Thus, these estimates indicate that the set of microsatellite markers is efficient in discriminating individuals. The SSR markers were also successfully transferred to other Khaya species. Therefore, these markers can be efficiently used in population studies with K. grandifoliola and other Khaya species.