Artículo de publicación ISIRoot hypoxia in fruit trees affects growth, vegetative development, and reproductive development, which is reflected in low productivity, poor fruit quality, and premature decay of trees. Using Illumina Hiseq2000, we performed transcriptome analysis of roots from two different rootstocks, ‘Mariana 2624’ and ‘Mazzard F12/1,’ which are tolerant and sensitive to hypoxia, respectively. Transcriptomes from control and hypoxia-stressed plants (6, 24, and 72 h) were compared, using Prunus persica (L.) as reference genome. Hypoxic conditions altered the transcription in both genotypes. There were a high number of common differentially expressed genes (DEG) between the two genotypes for each sampling time, but also exclusive DEG for each genotype, with a few DEG that presented opposite modes of regulations during the hypoxia treatment. An important group of DEGs exclusively upregulated in the tolerant genotype are associated to enzymes of posttranslational protein modifications, such as leucinerich repeat (LRR), kinases and ubiquitin-protein ligases, regulation of transcription, and process of oxide reduction. Singular enrichment analysis of gene ontology (GO), detected at least 115 GOs involved in the response to root hypoxia in the sensitive and/or tolerant genotypes. At least 25 GOs were identified as part of the baseline differences between the genotypes, most GO were disturbed in the sensitive genotype. The contribution from the baseline gene expression to the differential response between the Prunus genotypes is evidence that the resistant genotype is already Bprepared^ for a hypoxia event. An example are GO BP:0042221 of response to chemical stimulus; BP:0006979 of response to oxidative stress; MF:0016209 of antioxidant activity; MF:0016684 of oxidoreductase activity, acting on peroxide as acceptor; and MF:0004601 of peroxidase activity, which were disturbed only in the sensitive genotype, but not in the tolerant.FONDECYT (No. 1121117) and CEAF_R08I100

In sweet cherry trees, flowering is commercially important because the flowers, after fertilization, will generate the fruits. In P. avium, the flowering induction and flower organogensis are the first developmental steps towards flower formation and they occur within specialized organs known as floral buds during the summer, nine months before blooming. During this period the number of floral buds per tree and the bud fruitfulness (number of flowers per bud) are stablished affecting the potential yield of orchards and the plant architecture. The floral bud development is sensitive to any type of stress and the hotter and drier summers will interfere with this process and are calling for new adapted cultivars. A better understanding of the underlying molecular and hormonal mechanisms would be of help, but unlike the model plant Arabidopsis, very little is known about floral induction in sweet cherry. To explore the molecular mechanism of floral bud differentiation, high-throughput RNA sequencing was used to detect differences in the gene expression of P. avium floral buds at five differentiation stages. We found 2,982 differentially expressed genes during floral bud development. We identified genes associated with floral initiation or floral organ identity that appear to be useful biomarkers of floral development and several transcription factor families (ERF, MYB, bHLH, MADS-box and NAC gene family) with novel potential roles during floral transition in this species. We analyzed in deep the MADS-box gene family and we shed light about their key role during floral bud and organs development in P. avium. Furthermore, the hormonal-related signatures in the gene regulatory networks and the dynamic changes of absicic acid, zeatin and indolacetic acid contents in buds suggest an important role for these hormones during floral bud differentiation in sweet cherry. These data provide a rich source of novel informacion for functional and evolutionary studies about floral bud development in sweet cherry and new tools for biotechnology and breeding.

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Hexokinases (HXKs) and fructokinases (FRKs) are the only two families of enzymes in plants that have been identified as able to phosphorylate Glucose (Glc) and Fructose (Fru). Glc can only be phosphorylated in plants by HXKs, while Fru can be phosphorylated by either HXKs or FRKs. The various subcellular localizations of HXKs in plants indicate that they are involved in diverse functions, including anther dehiscence and pollen germination, stomatal closure in response to sugar levels, stomatal aperture and reducing transpiration. Its association with modulating programmed cell death, and responses to oxidative stress and pathogen infection (abiotic and biotic stresses) also have been reported. To extend our understanding about the function of HXK-like genes in the response of Prunus rootstocks to abiotic stress, we performed a detailed bioinformatic and functional analysis of hexokinase 3-like genes (HXK3s) from two Prunus rootstock genotypes, ‘M.2624’ (Prunus cerasifera Ehrh × P. munsoniana W.Wight & Hedrick) and ‘M.F12/1’ (P. avium L.), which are tolerant and sensitive to hypoxia stress, respectively. A previous large-scale transcriptome sequencing of roots of these rootstocks, showed that this HXK3-like gene that was highly induced in the tolerant genotype under hypoxia conditions. In silico analysis of gene promoters from M.2624 and M.F12/1 genotypes revealed regulatory elements that could explain differential transcriptional profiles of HXK3 genes. Subcellular localization was determinates by both bioinformatic prediction and expression of their protein fused to the green fluorescent protein (GFP) in protoplasts and transgenic plants of Arabidopsis. Both approaches showed that they are expressed in plastids. Metabolomics analysis of Arabidopsis plants ectopically expressing Prunus HXK3 genes revealed that content of several metabolites including phosphorylated sugars (G6P), starch and some metabolites associated with the TCA cycle were affected. These transgenic Arabidopsis plants showed improved tolerance to salt and drought stress under growth chamber conditions. Our results suggest that Prunus HXK3 is a potential candidate for enhancing tolerance to salt and drought stresses in stone fruit trees and other plants.