RESUMO
In sweet cherry (Prunus avium), as in other temperate woody perennials, bud dormancy allows for survival in adverse environmental conditions during winter. During this process, environmental signals such as short days and/or low temperatures trigger internal signals that enable buds to become tolerant to the cold. The process involves tracking chilling units up to chilling the requirement fulfillment to resume growth, a transition involving transcriptional regulation, metabolic signaling, and epigenetic-related regulatory events. Massive sequencing of small RNAs was performed to identify miRNAs involved in sweet cherry dormancy by comparing their expression in field (regular seasonal) and controlled non-stop (continuous) chilling conditions. miRNAs highlighted by sequencing were validated using specific stem-loop PCR quantification, confirming expression patterns for known miRNAs such as miR156e, miR166c, miR172d, miR391, miR482c, and miR535b, as well as for newly proposed miRNAs. In silico prediction of the target genes was used to construct miRNA/target gene nodes. In particular, the involvement of the sweet cherry version for the miR156/SQUAMOSA PROMOTER-BINDING-LIKE PROTEIN genes whose expression was opposite in the two conditions suggests their involvement on dormancy regulation in sweet cherry. miRNA levels indicate that the regulation of stress-related genes and hormone synthesis modulates the expression of calcium metabolism and cell development-associated genes. Understanding the regulatory networks involved in sweet cherry dormancy, particularly in the context of miRNA involvement, represents the first step in the development of new agricultural strategies that may help overcome the increasing challenges presented by global climate change.
RESUMO
Temperate deciduous fruit tree species like sweet cherry (Prunus avium) require long periods of low temperatures to trigger dormancy release and flowering. In addition to sequence-based genetic diversity, epigenetic variation may contribute to different chilling requirements among varieties. For the low chill variety 'Royal Dawn' and high chill variety 'Kordia', we studied the methylome of floral buds during chilling accumulation using MethylC-seq to identify differentially methylated regions (DMRs) during chilling hours (CH) accumulation, followed by transcriptome analysis to correlate changes in gene expression with DNA methylation. We found that during chilling accumulation, DNA methylation increased from 173 CH in 'Royal Dawn' and 443 CH in 'Kordia' and was mostly associated with the CHH context. In addition, transcriptional changes were observed from 443 CH in 'Kordia' with 1,210 differentially expressed genes, increasing to 4,292 genes at 1,295 CH. While 'Royal Dawn' showed approximately 5,000 genes differentially expressed at 348 CH and 516 CH, showing a reprogramming that was specific for each genotype. From conserved upregulated genes that overlapped with hypomethylated regions and downregulated genes that overlapped with hypermethylated regions in both varieties, we identified genes related to cold-sensing, cold-signaling, oxidation-reduction process, metabolism of phenylpropanoids and lipids, and a MADS-box SVP-like gene. As a complementary analysis, we used conserved and non-conserved DEGs that presented a negative correlation between DNA methylations and mRNA levels across all chilling conditions, obtaining Gene Ontology (GO) categories related to abiotic stress, metabolism, and oxidative stress. Altogether, this data indicates that changes in DNA methylation precedes transcript changes and may occur as an early response to low temperatures to increase the cold tolerance in the endodormancy period, contributing with the first methylome information about the effect of environmental cues over two different genotypes of sweet cherry.
RESUMO
Chilling requirement (CR) for bud dormancy completion determines the time of bud break in apple (Malus × domestica Borkh.). The molecular control of bud dormancy is highly heritable, suggesting a strong genetic control of the trait. An available Infinium II SNP platform for genotyping containing 8,788 single nucleotide polymorphic markers was employed, and linkage maps were constructed in a F1 cross from the low CR M13/91 and the moderate CR cv. Fred Hough. These maps were used to identify quantitative trait loci (QTL) for bud break date as a trait related to dormancy release. A major QTL for bud break was detected at the beginning of linkage group 9 (LG9). This QTL remained stable during seven seasons in two different growing sites. To increase mapping efficiency in detecting contributing genes underlying this QTL, 182 additional SNP markers located at the locus for bud break were used. Combining linkage mapping and structural characterization of the region, the high proportion of the phenotypic variance in the trait explained by the QTL is related to the coincident positioning of Arabidopsis orthologs for ICE1, FLC, and PRE1 protein-coding genes. The proximity of these genes from the most explanatory markers of this QTL for bud break suggests potential genetic additive effects, reinforcing the hypothesis of inter-dependent mechanisms controlling dormancy induction and release in apple trees.
RESUMO
Epigenetic modifications can yield information about connections between genotype, phenotype variation and environmental conditions. Bud dormancy release in temperate perennial fruit trees depends on internal and environmental signals such as cold accumulation and photoperiod. Previous investigations have noted the participation of epigenetic mechanisms in the control of this physiological process. We examined whether epigenetic modifications were modulated in MADS-box genes, potential candidates for the regulation of bud dormancy and flowering in sweet cherry (Prunus avium L.). We identified and cloned two MADS-box genes homologous to the already-characterized dormancy regulators DORMANCY-ASSOCIATED MADS-box (DAM3 and DAM5) from Prunus persica (L.) Batsch. Bisulfite sequencing of the identified genes (PavMADS1 and PavMADS2), Methylated DNA Immunoprecipitation and small RNA deep sequencing were performed to analyze the presence of DNA methylations that could be guided by non-coding RNAs in the floral buds exposed to differential chilling hours. The results obtained reveal an increase in the level of DNA methylation and abundance of matching small interference RNAs (siRNAs) in the promoter of PavMADS1 when the chilling requirement is complete. For the first intron and 5' UTR of PavMADS1, de novo DNA methylation could be associated with the increase in the abundance of 24-nt siRNA matching the promoter area. Also, in the second large intron of PavMADS1, maintenance DNA methylation in all cytosine contexts is associated with the presence of homologous siRNAs in that zone. For PavMADS2, only maintenance methylation was present in the CG context, and no matching siRNAs were detected. Silencing of PavMADS1 and PavMADS2 coincided with an increase in Flowering Locus T expression during dormancy. In conclusion, DNA methylations and siRNAs appear to be involved in the silencing of PavMADS1 during cold accumulation and dormancy release in sweet cherry.
Assuntos
Prunus avium/genética , Prunus avium/metabolismo , Metilação de DNA/genética , Metilação de DNA/fisiologia , Epigênese Genética/genética , Epigênese Genética/fisiologia , Flores/genética , Flores/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismoRESUMO
Apple production depends on the fulfilment of a chilling requirement for bud dormancy release. Insufficient winter chilling results in irregular and suboptimal bud break in the spring, with negative impacts on apple yield. Trees from apple cultivars with contrasting chilling requirements for bud break were used to investigate the expression of the entire set of apple genes in response to chilling accumulation in the field and controlled conditions. Total RNA was analysed on the AryANE v.1.0 oligonucleotide microarray chip representing 57,000 apple genes. The data were tested for functional enrichment, and differential expression was confirmed by real-time PCR. The largest number of differentially expressed genes was found in samples treated with cold temperatures. Cold exposure mostly repressed expression of transcripts related to photosynthesis, and long-term cold exposure repressed flavonoid biosynthesis genes. Among the differentially expressed selected candidates, we identified genes whose annotations were related to the circadian clock, hormonal signalling, regulation of growth, and flower development. Two genes, annotated as FLOWERING LOCUS C-like and MADS AFFECTING FLOWERING, showed strong differential expression in several comparisons. One of these two genes was upregulated in most comparisons involving dormancy release, and this gene's chromosomal position co-localized with the confidence interval of a major quantitative trait locus for the timing of bud break. These results indicate that photosynthesis and auxin transport are major regulatory nodes of apple dormancy and unveil strong candidates for the control of bud dormancy.
Assuntos
Temperatura Baixa , Genes de Plantas , Malus/genética , Relógios Circadianos , Análise por Conglomerados , Flavonoides/biossíntese , Flores/genética , Flores/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Malus/crescimento & desenvolvimento , Análise de Sequência com Séries de Oligonucleotídeos , Locos de Características Quantitativas , Estações do Ano , Transdução de SinaisRESUMO
The search for low chill cultivars for peach production in areas with low chill accumulation (temperatures below 7.2°C) has stimulated studies about the higher temperatures effects on low chill cultivars. The objective of this research was to evaluate the influence of mild temperatures on flowering and leafing of 'Turmalina' and 'Ágata' cultivars with low and medium low chilling requirements, respectively. Plants obtained by grafting flower twigs in the autumn, after the grafting, were submitted to chilling treatments (150, 250, 350, 500 and 672 hours) at three temperature levels (5°C, 10°C and 15°C). After the end of the treatments, the plants had been taken to greenhouse (20°C±5°C) and evaluated weekly. The results showed that satisfactory flower and leaf development occur even at temperatures as high as 15°C for a period of 150 hours, in 'Turmalina' peach. Temperatures of 10°C for a period equal or superior to 350 hours promote satisfactory leafing in 'Ágata' peach. Temperatures of 10°C and 15°C cause earlier development of flowers and leaves in both cultivars when compared to 5°C.
A busca por cultivares pessegueiro com baixa necessidade em frio para regiões com pouco acúmulo de frio (temperaturas abaixo de 7,2°C) tem estimulado estudos sobre o efeito de altas temperaturas sobre cultivares de baixa necessidade de frio. O objetivo desta pesquisa foi avaliar a influência de temperaturas amenas na floração e brotação da 'Turmalina' e 'Ágata', duas cultivares com baixa e média necessidade de frio, respectivamente. Plantas obtidas por enxertia no outono foram submetidas a tratamentos de frio (150, 250, 350, 500 e 672 horas) em três níveis de temperatura (5°C, 10°C e 15°C). Após os tratamentos, estas plantas foram colocadas em casa de vegetação a temperatura de 20°C±5°C e avaliadas semanalmente. Conclui-se que temperatura de até 15°C por um período de 150 horas é capaz de resultar em floração e brotação satisfatórias para a cultivar 'Turmalina'. Temperaturas de 10°C por um período igual ou superior a 350 horas promovem brotação satisfatória na cultivar 'Ágata'. Temperaturas de 10°C e 15°C comparadas a 5°C causam antecipação da floração e brotação e ambas cultivares.
RESUMO
The search for low chill cultivars for peach production in areas with low chill accumulation (temperatures below 7.2°C) has stimulated studies about the higher temperatures effects on low chill cultivars. The objective of this research was to evaluate the influence of mild temperatures on flowering and leafing of 'Turmalina' and 'Ágata' cultivars with low and medium low chilling requirements, respectively. Plants obtained by grafting flower twigs in the autumn, after the grafting, were submitted to chilling treatments (150, 250, 350, 500 and 672 hours) at three temperature levels (5°C, 10°C and 15°C). After the end of the treatments, the plants had been taken to greenhouse (20°C±5°C) and evaluated weekly. The results showed that satisfactory flower and leaf development occur even at temperatures as high as 15°C for a period of 150 hours, in 'Turmalina' peach. Temperatures of 10°C for a period equal or superior to 350 hours promote satisfactory leafing in 'Ágata' peach. Temperatures of 10°C and 15°C cause earlier development of flowers and leaves in both cultivars when compared to 5°C.
A busca por cultivares pessegueiro com baixa necessidade em frio para regiões com pouco acúmulo de frio (temperaturas abaixo de 7,2°C) tem estimulado estudos sobre o efeito de altas temperaturas sobre cultivares de baixa necessidade de frio. O objetivo desta pesquisa foi avaliar a influência de temperaturas amenas na floração e brotação da 'Turmalina' e 'Ágata', duas cultivares com baixa e média necessidade de frio, respectivamente. Plantas obtidas por enxertia no outono foram submetidas a tratamentos de frio (150, 250, 350, 500 e 672 horas) em três níveis de temperatura (5°C, 10°C e 15°C). Após os tratamentos, estas plantas foram colocadas em casa de vegetação a temperatura de 20°C±5°C e avaliadas semanalmente. Conclui-se que temperatura de até 15°C por um período de 150 horas é capaz de resultar em floração e brotação satisfatórias para a cultivar 'Turmalina'. Temperaturas de 10°C por um período igual ou superior a 350 horas promovem brotação satisfatória na cultivar 'Ágata'. Temperaturas de 10°C e 15°C comparadas a 5°C causam antecipação da floração e brotação e ambas cultivares.
RESUMO
In Santa Fe, the central-east area of Argentina, the expansion of fruit tree crops requires the introduction and evaluation of low chilling varieties. The aims of this study was to characterize the phenological behaviour and the reproductive traits of peach (Prunus persica L. Batsch) and nectarine varieties with chilling requirements ranging from 150 to 650 chilling hours (CH). Low chilling varieties ( 450 CH) had higher flower density (22.0-56.5 flowers m-1), fruit set (26.0-56.7%), fruit yield (9.3-36.6 kg per plant), and low coefficient of fruit yield variability than varieties with a higher chilling requirement (>500 CH). The phenology of high chilling requirement varieties had the advantage that blooming and fruit set happened after the period of late frost occurrence. However, they did not satisfy their chilling requirement, showing an inadequate vegetative and reproductive behaviour. Low chilling varieties began to sprout during July, and the period of full bloom occurred from July 14th to August 4th. Fruit harvest was initiated on October 19th with the cv. 'Flordastar' and it extended for 58 days. However, on low chilling varieties the harvest time was reduced in ten days. Flower density, fruit set and fruit yield showed a negative relationship with the chilling requirement of the variety.
Na região centro-leste do estado de Santa Fé, Argentina, para a expansão dos frutais é preciso introduzir variedades de baixo requerimento de frio. O objetivo deste estudo foi caracterizar o comportamento durante as fases de crescimento e frutificação de variedades de pêssego (Prunus persica L. Batsch) e nectarina com requerimentos de frio que variam entre 150-650 horas de frio (CH). As variedades de pêssego de baixa necessidade de frio ( 450 CH) mostraram maior densidade de flores (22.0-56.5 flores m-1), de estabelecimento de frutos (26.0-56.7%), e rendimento de fruta (9.3-36.6 kg por planta) comparado com variedades de pêssego com necessidade de frio maior (>500 CH). As variedades de maior necessidade de frio tiveram a vantagem da floração, e o estabelecimento dos frutos aconteceu depois do período de risco de geadas. Porém, essas variedades não têm satisfeita sua necessidade de frio, apresentando comportamento vegetativo e reprodutivo inadequado. As variedades de baixa necessidade de frio brotaram entre 4 a 31 de julho e o período de plena floração aconteceu entre 14 de julho a 4 de agosto. A colheita de fruta começou em 19 de outubro com o cv. 'Flordastar', e sua duração foi de 58 dias. Para as variedades de baixa necessidade de frio o período de colheita foi dez dias mais curto.
RESUMO
The search for low chill cultivars for peach production in areas with low chill accumulation (temperatures below 7.2°C) has stimulated studies about the higher temperatures effects on low chill cultivars. The objective of this research was to evaluate the influence of mild temperatures on flowering and leafing of 'Turmalina' and 'Ágata' cultivars with low and medium low chilling requirements, respectively. Plants obtained by grafting flower twigs in the autumn, after the grafting, were submitted to chilling treatments (150, 250, 350, 500 and 672 hours) at three temperature levels (5°C, 10°C and 15°C). After the end of the treatments, the plants had been taken to greenhouse (20°C±5°C) and evaluated weekly. The results showed that satisfactory flower and leaf development occur even at temperatures as high as 15°C for a period of 150 hours, in 'Turmalina' peach. Temperatures of 10°C for a period equal or superior to 350 hours promote satisfactory leafing in 'Ágata' peach. Temperatures of 10°C and 15°C cause earlier development of flowers and leaves in both cultivars when compared to 5°C.
A busca por cultivares pessegueiro com baixa necessidade em frio para regiões com pouco acúmulo de frio (temperaturas abaixo de 7,2°C) tem estimulado estudos sobre o efeito de altas temperaturas sobre cultivares de baixa necessidade de frio. O objetivo desta pesquisa foi avaliar a influência de temperaturas amenas na floração e brotação da 'Turmalina' e 'Ágata', duas cultivares com baixa e média necessidade de frio, respectivamente. Plantas obtidas por enxertia no outono foram submetidas a tratamentos de frio (150, 250, 350, 500 e 672 horas) em três níveis de temperatura (5°C, 10°C e 15°C). Após os tratamentos, estas plantas foram colocadas em casa de vegetação a temperatura de 20°C±5°C e avaliadas semanalmente. Conclui-se que temperatura de até 15°C por um período de 150 horas é capaz de resultar em floração e brotação satisfatórias para a cultivar 'Turmalina'. Temperaturas de 10°C por um período igual ou superior a 350 horas promovem brotação satisfatória na cultivar 'Ágata'. Temperaturas de 10°C e 15°C comparadas a 5°C causam antecipação da floração e brotação e ambas cultivares.
RESUMO
In Santa Fe, the central-east area of Argentina, the expansion of fruit tree crops requires the introduction and evaluation of low chilling varieties. The aims of this study was to characterize the phenological behaviour and the reproductive traits of peach (Prunus persica L. Batsch) and nectarine varieties with chilling requirements ranging from 150 to 650 chilling hours (CH). Low chilling varieties ( 450 CH) had higher flower density (22.0-56.5 flowers m-1), fruit set (26.0-56.7%), fruit yield (9.3-36.6 kg per plant), and low coefficient of fruit yield variability than varieties with a higher chilling requirement (>500 CH). The phenology of high chilling requirement varieties had the advantage that blooming and fruit set happened after the period of late frost occurrence. However, they did not satisfy their chilling requirement, showing an inadequate vegetative and reproductive behaviour. Low chilling varieties began to sprout during July, and the period of full bloom occurred from July 14th to August 4th. Fruit harvest was initiated on October 19th with the cv. 'Flordastar' and it extended for 58 days. However, on low chilling varieties the harvest time was reduced in ten days. Flower density, fruit set and fruit yield showed a negative relationship with the chilling requirement of the variety.
Na região centro-leste do estado de Santa Fé, Argentina, para a expansão dos frutais é preciso introduzir variedades de baixo requerimento de frio. O objetivo deste estudo foi caracterizar o comportamento durante as fases de crescimento e frutificação de variedades de pêssego (Prunus persica L. Batsch) e nectarina com requerimentos de frio que variam entre 150-650 horas de frio (CH). As variedades de pêssego de baixa necessidade de frio ( 450 CH) mostraram maior densidade de flores (22.0-56.5 flores m-1), de estabelecimento de frutos (26.0-56.7%), e rendimento de fruta (9.3-36.6 kg por planta) comparado com variedades de pêssego com necessidade de frio maior (>500 CH). As variedades de maior necessidade de frio tiveram a vantagem da floração, e o estabelecimento dos frutos aconteceu depois do período de risco de geadas. Porém, essas variedades não têm satisfeita sua necessidade de frio, apresentando comportamento vegetativo e reprodutivo inadequado. As variedades de baixa necessidade de frio brotaram entre 4 a 31 de julho e o período de plena floração aconteceu entre 14 de julho a 4 de agosto. A colheita de fruta começou em 19 de outubro com o cv. 'Flordastar', e sua duração foi de 58 dias. Para as variedades de baixa necessidade de frio o período de colheita foi dez dias mais curto.