RESUMO
The advance of Next Generation Sequencing (NGS) technologies allows high-throughput genotyping at a reasonable cost, although, in the case of peach, this technology has been scarcely developed. To date, only a standard Genotyping by Sequencing approach (GBS), based on a single restriction with ApeKI to reduce genome complexity, has been applied in peach. In this work, we assessed the performance of the double-digest RADseq approach (ddRADseq), by testing 6 double restrictions with the restriction profile generated with ApeKI. The enzyme pair PstI/MboI retained the highest number of loci in concordance with the in silico analysis. Under this condition, the analysis of a diverse germplasm collection (191 peach genotypes) yielded 200,759,000 paired-end (2 × 250 bp) reads that allowed the identification of 113,411 SNP, 13,661 InDel and 2133 SSR. We take advantage of a wide sample set to describe technical scope of the platform. The novel platform presented here represents a useful tool for genomic-based breeding for peach.
Assuntos
Genoma de Planta , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Prunus persica/enzimologia , Prunus persica/genética , Análise de Sequência de DNA/métodos , Biologia Computacional/métodos , DNA de Plantas/genética , DNA de Plantas/isolamento & purificação , Desoxirribonucleases de Sítio Específico do Tipo II/genética , Loci Gênicos , Técnicas de Genotipagem/métodos , Melhoramento Vegetal , Polimorfismo de Nucleotídeo ÚnicoRESUMO
O presente trabalho teve como objetivo caracterizar nutricionalmente plantas de pessegueiro na região produtora de Pelotas, RS. Foram amostrados 65 pomares de pessegueiros das cultivares Maciel, Granada, Esmeralda e Sensação nos municípios de Pelotas, Canguçu e Morro Redondo no ciclo produtivo de 2018. As amostras foliares, após secas e moídas, foram analisadas quanto aos seus teores de nitrogênio (N), fósforo (P), potássio (K), cálcio (Ca), magnésio (Mg), boro (B), cobre (Cu), ferro (Fe), manganês (Mn) e zinco (Zn) e os resultados foram apresentados em distribuições de frequência nas classes de suficiência indicadas para a cultura na região além da correlação entre os nutrientes. Observou-se maiores deficiências nos teores de nitrogênio, cobre, ferro e zinco. Entretanto, para a maioria das amostras, os teores de potássio encontraram-se na classe acima do normal, encontrou-se correlações entre P/Ca, P/Cu, Ca/P, Ca/Mg e Ca/B. É possível constatar a carência do uso de ferramentas para avaliar o estado nutricional dos pomares, principalmente a diagnose foliar, o que levou ao uso inadequado de fertilizantes e consequentemente um desbalanço nutricional das plantas.(AU)
This study set out to assess the nutritional status of peach trees in the commercial peach-growing region of Pelotas, Rio Grande do Sul. The sample includes 65 peach orchards of the Maciel, Granada, Esmeralda, and Sensação cultivars in the municipalities of Pelotas, Canguçu, and Morro Redondo during the 2018 agricultural cycle. After drying and grinding the leaf samples for an analysis of their nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), boron (B), copper (Cu), iron (Fe), manganese (Mn), and zinc (Zn) content, the results were presented in frequency distributions divided into classes based on the nutrient requirements indicated for the crop in the region, in addition to the correlations between the nutrients. Greater deficiencies were observed in nitrogen, copper, iron, and zinc content. However, potassium content was found in the above normal class for most of the samples, and there were correlations between P/Ca, P/Cu, Ca/P, Ca/Mg, and Ca/B. An insufficient use of the tools to assess the nutritional status of the orchards was observed, especially the leaf diagnosis, which led to the inappropriate use of fertilizers and the consequent nutrient imbalance in the plants.(AU)
Assuntos
Prunus persica/química , Prunus persica/enzimologia , Prunus persica/fisiologia , Valor Nutritivo , EstercoRESUMO
O presente trabalho teve como objetivo caracterizar nutricionalmente plantas de pessegueiro na região produtora de Pelotas, RS. Foram amostrados 65 pomares de pessegueiros das cultivares Maciel, Granada, Esmeralda e Sensação nos municípios de Pelotas, Canguçu e Morro Redondo no ciclo produtivo de 2018. As amostras foliares, após secas e moídas, foram analisadas quanto aos seus teores de nitrogênio (N), fósforo (P), potássio (K), cálcio (Ca), magnésio (Mg), boro (B), cobre (Cu), ferro (Fe), manganês (Mn) e zinco (Zn) e os resultados foram apresentados em distribuições de frequência nas classes de suficiência indicadas para a cultura na região além da correlação entre os nutrientes. Observou-se maiores deficiências nos teores de nitrogênio, cobre, ferro e zinco. Entretanto, para a maioria das amostras, os teores de potássio encontraram-se na classe acima do normal, encontrou-se correlações entre P/Ca, P/Cu, Ca/P, Ca/Mg e Ca/B. É possível constatar a carência do uso de ferramentas para avaliar o estado nutricional dos pomares, principalmente a diagnose foliar, o que levou ao uso inadequado de fertilizantes e consequentemente um desbalanço nutricional das plantas.
This study set out to assess the nutritional status of peach trees in the commercial peach-growing region of Pelotas, Rio Grande do Sul. The sample includes 65 peach orchards of the Maciel, Granada, Esmeralda, and Sensação cultivars in the municipalities of Pelotas, Canguçu, and Morro Redondo during the 2018 agricultural cycle. After drying and grinding the leaf samples for an analysis of their nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), boron (B), copper (Cu), iron (Fe), manganese (Mn), and zinc (Zn) content, the results were presented in frequency distributions divided into classes based on the nutrient requirements indicated for the crop in the region, in addition to the correlations between the nutrients. Greater deficiencies were observed in nitrogen, copper, iron, and zinc content. However, potassium content was found in the above normal class for most of the samples, and there were correlations between P/Ca, P/Cu, Ca/P, Ca/Mg, and Ca/B. An insufficient use of the tools to assess the nutritional status of the orchards was observed, especially the leaf diagnosis, which led to the inappropriate use of fertilizers and the consequent nutrient imbalance in the plants.
Assuntos
Esterco , Prunus persica/enzimologia , Prunus persica/fisiologia , Prunus persica/química , Valor NutritivoRESUMO
Most oxidoreductases that use NAD+ or NADP+ to transfer electrons in redox reactions display a strong preference for the cofactor. The catalytic efficiency of peach glucitol dehydrogenase (GolDHase) for NAD+ is 1800-fold higher than that for NADP+. Herein, we combined structural and kinetic data to reverse the cofactor specificity of this enzyme. Using site-saturation mutagenesis, we obtained the D216A mutant, which uses both NAD+ and NADP+, although with different catalytic efficiencies (1000 ± 200 and 170 ± 30 M-1 s-1, respectively). This mutant was used as a template to introduce further mutations by site-directed mutagenesis, using information from the fruit fly NADP-dependent GolDHase. The D216A/V217R/D218S triple mutant displayed a 2-fold higher catalytic efficiency with NADP+ than with NAD+. Overall, our results indicate that the triple mutant has the potential to be used for metabolic and cellular engineering and for cofactor recycling in industrial processes.