Caracterização funcional do transcriptoma de amendoim forrageiro.

O uso do amendoim forrageiro em consórcios com gramíneas nas pastagens e como cobertura verde, consorciado com culturas comerciais, tem crescido nos últimos anos. A análise do genoma funcional permite a identificação de genes de interesse agronômico. Assim, o objetivo deste trabalho foi realizar a anotação funcional de genes do transcriptoma de folhas de Arachis pintoi. Dos 98.432 transcritos analisados, 69% apresentaram correspondências com o banco de dados de proteínas do National Center of Biotechnology Information. As classes função molecular (36%) e processo biológico (35,8%) representaram a maioria dos termos de Gene Ontology atribuídos, enquanto o componente celular (28,2%) apresentou menor número. A análise de expressão diferencial identificou 1.550 e 1.357 genes com maior nível de expressão nas cultivares Amarillo e Belomonte, respectivamente. A análise de enriquecimento dos genes mostrou que 55,63% pertencem à classe componente celular, seguida por função molecular (26,48%) e processo biológico (20,89%). Esses resultados são o primeiro relato de anotação funcional de A. pintoi que irá fornecer uma importante fonte de informação para avanços nos estudos de expressão, silenciamento e edição gênica nos programas de melhoramento de Arachis.Editores técnicos: Rodrigo Souza Santos; Fabiano Marçal Estanislau

Efeito da fermentação natural e do envelhecimento sobre a qualidade química e sensorial de cachaça orgânica Brasileira.

This study verified the effect of the spontaneous fermentation/natural ferment (NF) on the chemo-sensory quality of cachaça, comparing to the commercial ferment (CF). The effect of ageing (maturation) was also analysed in the beverage. Microbiological analysis (plating on selective media for total/wild yeast and bacteria counting) and physico-chemical analysis (pH, acidity and soluble solids) were performed in the samples of the must and the ferment collected during three cycles of fermentation in a semi-industrial scale. Samples of cachaça were stored in 5-L oak containers for 45 days, subsequently analyzing the physico-chemical characteristics (pH, acidity, alcohol content, copper and secondary compounds) and sensory acceptability (aroma, flavour, colour, body and global impression). The fermentation with NF showed higher number of wild yeasts; however there was no difference in the number of bacteria comparing to CF. An intense acidification occurred during the preparation of NF, which was also observed in the initial cycles of fermentation, but decreased afterwards. Greater alterations in cachaça composition were found to be more exclusively related to the maturation than to the type of ferment, except for the acidity. However, there was a significant loss of aroma, flavour and global impression after maturation but only in cachaça produced with the CF. The results revealed a strong interaction between ferment and maturation of the beverage, suggesting that substances produced by the microorganisms from different inocula during fermentation reacted differently with the wood components of the barrels influencing the sensory attributes.Este trabalho teve por objetivo avaliar o efeito da fermentação espontânea/fermento natural (NF) sobre a qualidade fisico-química e aceitabilidade da cachaça, comparando-se com o fermento comercial (CF). O efeito do envelhecimento (maturação) da bebida foi também avaliado. Análises microbiológicas (plaqueamento em meios seletivos para contagem de leveduras totais/selvagens e bactérias) e físico-químicas (pH, acidez e sólidos solúveis) foram conduzidas nas amostras do mosto e do fermento coletadas durante três ciclos fermentativos (escala semi-industrial). Amostras de cachaça foram armazenadas em barris de carvalho de 5 litros por 45 dias, seguindo-se a análise físico-química (pH, acidez, teor alcoolico, cobre e compostos secundários) e de aceitabilidade sensorial (aroma, sabor, cor, corpo e impressão global). A fermentação com NF mostrou maior número de leveduras selvagens; no entanto, não houve diferença significativa no número de bactérias quando comparada com CF. Uma intensa acidificação foi observada durante o preparo de NF, a qual foi verificada nos ciclos iniciais da fermentação, decrescendo a seguir. As alterações na composição da cachaça foram mais significativas em relação à maturação do que ao tipo de fermento, com exceção da acidez. No entanto, houve uma perda significativa de aroma, sabor e impressão global após a maturação na cachaça produzida com CF. Os resultados revelaram uma forte interação entre fermento e envelhecimento (maturação) da bebida, sugerindo que substâncias produzidas pelos microrganismos a partir de inóculos diferentes reagiram diferentemente com componentes da madeira dos barris, influenciando nos atributos sensoriais de aceitabilidade

Linkage disequilibrium and population structure in wild and cultivated populations of rubber tree (Hevea brasiliensis).

Abstract: Among rubber tree species, which belong to the Hevea genus of the Euphorbiaceae family, Hevea brasiliensis (Willd. ex Adr.de Juss.) Muell. Arg. is the main commercial source of natural rubber production worldwide. Knowledge of the population structure and linkage disequilibrium (LD) of this species is essential for the efficient organization and exploitation of genetic resources. Here, we obtained single-nucleotide polymorphisms (SNPs) using a genotyping-by-sequencing (GBS) approach and then employed the SNPs for the following objectives: (i) to identify the positions of SNPs on a genetic map of a segregating mapping population, (ii) to evaluate the population structure of a germplasm collection, and (iii) to detect patterns of LD decay among chromosomes for future genetic association studies in rubber tree. A total of 626 genotypes, including both germplasm accessions (368) and individuals from a genetic mapping population (254), were genotyped. A total of 77,660 and 21,283 SNPs were detected by GBS in the germplasm and mapping populations, respectively. The mapping population, which was previously mapped, was constructed with 1,062 markers, among which only 576 SNPs came from GBS, reducing the average interval between two adjacent markers to 4.4 cM. SNPs from GBS genotyping were used for the analysis of genetic structure and LD estimation in the germplasm accessions. Two groups, which largely corresponded to the cultivated and wild populations, were detected using STRUCTURE and via principal coordinate analysis. LD analysis, also using the mapped SNPs, revealed that non-random associations varied along chromosomes, with regions of high LD interspersed with regions of low LD. Considering the length of the genetic map (4,693 cM) and the mean LD (0.49 for cultivated and 0.02 for wild populations), a large number of evenly spaced SNPs would be needed to perform genome-wide association studies in rubber tree, and the wilder the genotypes used, the more difficult the mapping saturation

Mapping the sex determination locus in the hāpuku (Polyprion oxygeneios) using ddRAD sequencing

Background  Hāpuku (Polyprion oxygeneios) is a member of the wreckfish family (Polyprionidae) and is highly regarded as a food fish. Although adults grow relatively slowly, juveniles exhibit low feed conversion ratios and can reach market size in 1–2 years, makingP. oxygeneiosa strong candidate for aquaculture. However, they can take over 5years to reach sexual maturity in captivity and are not externally sexually dimorphic, complicating many aspects of broodstock management. Understanding the sex determination system ofP. oxygeneiosand developing accurate assays to assign genetic sex will contribute significantly towards its full-scale commercialisation.  Results  DNA from parents and sexed offspring (n = 57) from a single family of captive bredP. oxygeneioswas used as a template for double digestion Restriction-site Associated DNA (ddRAD) sequencing. Two libraries were constructed usingSbfI–SphI andSbfI –NcoI restriction enzyme combinations, respectively. Two runs on an Illumina MiSeq platform generated 70,266,464 raw reads, identifying 19,669 RAD loci. A combined sex linkage map (1367cM) was constructed based on 1575 Single Nucleotide Polymorphism (SNP) markers that resolved into 35 linkage groups. Sex-specific linkage maps were of similar size (1132 and 1168cM for male and female maps respectively). A single major sex-determining locus, found to be heterogametic in males, was mapped to linkage group 14. Several markers were found to be in strong linkage disequilibrium with the sex-determining locus. Allele-specific PCR assays were developed for two of these markers, SphI6331 and SphI8298, and demonstrated to accurately differentiate sex in progeny within the same pedigree. Comparative genomic analyses indicated that many of the linkage groups within theP. oxygeneiosmap share a relatively high degree of homology with those published for the European seabass (Dicentrarchus labrax).  Conclusion  P. oxygeneioshas an XX/XY sex determination system. Evaluation of allele-specific PCR assays, based on the two SNP markers most closely associated with phenotypic sex, indicates that a simple molecular assay for sexingP. oxygeneiosshould be readily attainable. The high degree of synteny observed withD. labraxshould aid further molecular genetic study and exploitation of hāpuku as a food fish

Assembly of the 373k gene space of the polyploid sugarcane genome reveals reservoirs of functional diversity in the world's leading biomass crop

BACKGROUND: Sugarcane cultivars are polyploid interspecific hybrids of giant genomes, typically with 10-13 sets of chromosomes from 2 Saccharum species. The ploidy, hybridity, and size of the genome, estimated to have >10 Gb, pose a challenge for sequencing. RESULTS: Here we present a gene space assembly of SP80-3280, including 373,869 putative genes and their potential regulatory regions. The alignment of single-copy genes in diploid grasses to the putative genes indicates that we could resolve 2-6 (up to 15) putative homo(eo)logs that are 99.1% identical within their coding sequences. Dissimilarities increase in their regulatory regions, and gene promoter analysis shows differences in regulatory elements within gene families that are expressed in a species-specific manner. We exemplify these differences for sucrose synthase (SuSy) and phenylalanine ammonia-lyase (PAL), 2 gene families central to carbon partitioning. SP80-3280 has particular regulatory elements involved in sucrose synthesis not found in the ancestor Saccharum spontaneum. PAL regulatory elements are found in co-expressed genes related to fiber synthesis within gene networks defined during plant growth and maturation. Comparison with sorghum reveals predominantly bi-allelic variations in sugarcane, consistent with the formation of 2 "subgenomes" after their divergence approximately 3.8-4.6 million years ago and reveals single-nucleotide variants that may underlie their differences. CONCLUSIONS: This assembly represents a large step towards a whole-genome assembly of a commercial sugarcane cultivar. It includes a rich diversity of genes and homo(eo)logous resolution for a representative fraction of the gene space, relevant to improve biomass and food production

Genome of Herbaspirillum seropedicae Strain SmR1, a Specialized Diazotrophic Endophyte of Tropical Grasses

The molecular mechanisms of plant recognition, colonization, and nutrient exchange between diazotrophic endophytes and plants are scarcely known. Herbaspirillum seropedicae is an endophytic bacterium capable of colonizing intercellular spaces of grasses such as rice and sugar cane. The genome of H. seropedicae strain SmR1 was sequenced and annotated by The Paraná State Genome Programme—GENOPAR. The genome is composed of a circular chromosome of 5,513,887 bp and contains a total of 4,804 genes. The genome sequence revealed that H. seropedicae is a highly versatile microorganism with capacity to metabolize a wide range of carbon and nitrogen sources and with possession of four distinct terminal oxidases. The genome contains a multitude of protein secretion systems, including type I, type II, type III, type V, and type VI secretion systems, and type IV pili, suggesting a high potential to interact with host plants. H. seropedicae is able to synthesize indole acetic acid as reflected by the four IAA biosynthetic pathways present. A gene coding for ACC deaminase, which may be involved in modulating the associated plant ethylene-signaling pathway, is also present. Genes for hemagglutinins/hemolysins/adhesins were found and may play a role in plant cell surface adhesion. These features may endow H. seropedicae with the ability to establish an endophytic life-style in a large number of plant species