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.
A new plant commensal Pseudomonas veronii isolate (strain R4) was identified from a Xiphinema index biocontrol screen. Isolated from grapevine roots from vineyards in central Chile, the strain R4 exhibited a slower yet equivalently effective nematicide activity as the well-characterized P. protegens CHA0. Whole genome sequencing of strain R4 and comparative analysis among the available Pseudomonas spp. genomes allowed for the identification of gene clusters that encode putative extracellular proteases and lipase synthesis and secretion systems, which are proposed to mediate—at least in part—the observed nematicidal activity. In addition, R4 strain presented relevant gene clusters related to metal tolerance, which is typical in P. veronii. Bioinformatics analyses also showed gene clusters associated with plant growth promoting activity, such as indole-3-acetic acid synthesis. In addition, the strain R4 genome presented a metabolic gene clusters associated with phosphate and ammonia biotransformation from soil, which could improve their availability for plants.Electronic supplementary materialThe online version of this article (doi:10.1186/s40793-016-0198-y) contains supplementary material, which is available to authorized users.
The fungi Botrytis cinerea and Erysiphe necator are responsible for gray mold and powdery mildew diseases, respectively, which are among the most devastating diseases of grapes. Two endochitinase (ech42 and ech33) genes and one N-acetyl-β-D-hexosaminidase (nag70) gene from biocontrol agents related to Trichoderma spp. were used to develop a set of 103 genetically modified (GM) 'Thompson Seedless' lines (568 plants) that were established in open field in 2004 and evaluated for fungal tolerance starting in 2006. Statistical analyses were carried out considering transgene, explant origin, and plant response to both fungi in the field and in detached leaf assays. The results allowed for the selection of the 19 consistently most tolerant lines through two consecutive years (2007-2008 and 2008-2009 seasons). Plants from these lines were grafted onto the rootstock Harmony and established in the field in 2009 for further characterization. Transgene status was shown in most of these lines by Southern blot, real-time PCR, ELISA, and immunostrips; the most tolerant candidates expressed the ech42-nag70 double gene construct and the ech33 gene from a local Hypocrea virens isolate. B. cinerea growth assays in Petri dishes supplemented with berry juices extracted from the most tolerant individuals of the selected population was inhibited. These results demonstrate that improved fungal tolerance can be attributed to transgene expression and support the iterative molecular and physiological phenotyping in order to define selected individuals from a population of GM grapevines.
AbstractPrecambrian rocks comprise nearly one-quarter of the surface of Brazil and range from Paleoarchean (ca. 3.6 Ga) to the latest Ediacaran (0.542 Ga) in age. Except for controversial phosphatized ‘embryo-like’ microfossils like those from the lower Ediacaran Doushantuo Formation, China and complex rangeomorphs, Brazilian research has revealed all major categories of Precambrian life forms described elsewhere – microbialites, biomarkers, silicified microfossils, palynomorphs, vase-shaped microfossils, macroalgae, metazoans, vendobionts and ichnofossils – but the paleobiological significance of this record has been little explored. At least four occurrences of these fossils offer promise for increased understanding of the following aspects of Precambrian biospheric evolution: (i) the relationship of microbialites in 2.1–2.4 Ga old carbonates of the Minas Supergroup in the Quadrilátero Ferrífero, Minas Gerais (the oldest Brazilian fossils) to the development of the early oxygenic atmosphere and penecontemporaneous global tectonic and climatic events; (ii) the evolutionary and biostratigraphic significance of Mesoproterozoic to Ediacaran organic-walled microfossils in central–western Brazil; (iii) diversity and paleoecological significance of vase-shaped heterotrophic protistan microfossils in the Urucum Formation (Jacadigo Group) and possibly the Bocaina Formation (Corumbá Group), of Mato Grosso do Sul; and (iv) insights into the record of skeletogenesis and paleoecology of latest Ediacaran metazoans as represented by the abundant organic carapaces ofCorumbellaand calcareous shells of the index fossilCloudina, of the Corumbá Group, Mato Grosso do Sul. Analysis of the Brazilian Precambrian fossil record thus holds great potential for augmenting paleobiological knowledge of this crucial period on Earth and for developing more robust hypotheses regarding possible origins and evolutionary pathways of biospheres on other planets.
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