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Mol Genet Genomics ; 295(1): 107-120, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31506717


The oriental gall wasp Dryocosmus kuriphilus represents a limiting pest for the European Chestnut (Castanea sativa, Fagaceae) as it creates severe yield losses. The European Chestnut is a deciduous tree, having major social, economic and environmental importance in Southern Europe, covering an area of 2.53 million hectares, including 75,000 ha devoted to fruit production. Cultivars show different susceptibility and very few are resistant to gall wasp. To deeply investigate the plant response and understand which factors can lead the plant to develop or not the gall, the study of transcriptome is basic (fundamental). To date, little transcriptomic information are available for C. sativa species. Hence, we present a de novo assembly of the chestnut transcriptome of the resistant Euro-Japanese hybrid 'Bouche de Bétizac' (BB) and the susceptible cultivar 'Madonna' (M), collecting RNA from buds at different stages of budburst. The two transcriptomes were assembled into 34,081 (BB) and 30,605 (M) unigenes, respectively. The former was used as a reference sequence for further characterization analyses, highlighting the presence of 1444 putative resistance gene analogs (RGAs) and about 1135 unigenes, as putative MiRNA targets. A global quantitative transcriptome profiling comparing the resistant and the susceptible cultivars, in the presence or not of the gall wasp, revealed some GO enrichments as "response to stimulus" (GO:0050896), and "developmental processes" (e.g., post-embryonic development, GO:0009791). Many up-regulated genes appeared to be transcription factors (e.g., RAV1, AP2/ERF, WRKY33) or protein regulators (e.g., RAPTOR1B) and storage proteins (e.g., LEA D29) involved in "post-embryonic development". Our analysis was able to provide a large amount of information, including 7k simple sequence repeat (SSR) and 335k single-nucleotide polymorphism (SNP)/INDEL markers, and generated the first reference unigene catalog for the European Chestnut. The transcriptome data for C. sativa will contribute to understand the genetic basis of the resistance to gall wasp and will provide useful information for next molecular genetic studies of this species and its relatives.

Fagaceae/genética , Transcriptoma/genética , Vespas/patogenicidade , Animais , Europa (Continente) , Fagaceae/parasitologia , Perfilação da Expressão Gênica/métodos , Repetições de Microssatélites/genética , Anotação de Sequência Molecular/métodos , Doenças das Plantas/parasitologia , Polimorfismo de Nucleotídeo Único/genética , Fatores de Transcrição/genética , Árvores/genética , Árvores/parasitologia , Regulação para Cima/genética
Foods ; 8(12)2019 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-31766769


Single-cultivar juices may be a valuable way to introduce different versions of a product to the market and obtain price discrimination. To communicate a product's value, complex characteristics incorporated by each cultivar must be identified. New sensory methods rely on the assessor's ability to recall attributes; however, the use of objective vocabularies may improve the sensory profiling. This work aimed to profile monovarietal apple juices by using projective mapping (PM) combined with ultra-flash profiling (UFP) supported by a sensory wheel built with a text-mining tool. Samples were also analyzed for physicochemical parameters to provide more information to the assessment. The assessor coordinates from PM were used in multiple factor analysis with confidence ellipses to assess differences among samples. A goodness-of-fit test was applied to select the most meaningful descriptors generated through the UFP test by calculating the expected frequency of choosing a descriptor from the sensory wheel and comparing it with the observed values. The methodology provided a more accurate sensory profile compared to previous research on fresh apples and juices. Elstar, Jonagold, and Pinova were considered as sweet juices, and Gravensteiner was described as sour and astringent, with green-apple notes. Rubinette was described as having a strong taste and cloudy aspect.

PLoS One ; 13(4): e0195408, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29608620


The growing area of European hazelnut (Corylus avellana L.) is increasing, as well as the number of producing countries, and there is a pressing need for new improved cultivars. Hazelnut conventional breeding process is slow, due to the length of juvenile phase and the high heterozygosity level. The development of genetic linkage maps and the identification of molecular markers tightly linked to QTL (quantitative trait loci) of agronomic interest are essential tools for speeding up the selection of seedlings carrying desired traits through marker-assisted selection. The objectives of this study were to enrich a previous linkage map and confirm QTL related to time of leaf budburst, using an F1 population obtained by crossing Tonda Gentile delle Langhe with Merveille de Bollwiller. Genotyping-by-Sequencing was used to identify a total of 9,999 single nucleotide polymorphism markers. Well saturated linkage maps were constructed for each parent using the double pseudo-testcross mapping strategy. A reciprocal translocation was detected in Tonda Gentile delle Langhe between two non-homologous chromosomes. Applying a bioinformatic approach, we were able to disentangle 'pseudo-linkage' between markers, removing markers around the translocation breakpoints and obtain a linear order of the markers for the two chromosomes arms, for each linkage group involved in the translocation. Twenty-nine QTL for time of leaf budburst were identified, including a stably expressed region on LG_02 of the Tonda Gentile delle Langhe map. The stability of these QTL and their coding sequence content indicates promise for the identification of specific chromosomal regions carrying key genes involved in leaf budburst.

Corylus/crescimento & desenvolvimento , Corylus/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/genética , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Agricultura , Mapeamento Cromossômico , Cromossomos de Plantas , Ligação Genética , Marcadores Genéticos , Fenótipo , Melhoramento Vegetal , Análise de Sequência de DNA
Mol Biol Rep ; 39(4): 4997-5008, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22362313


Hazelnut is a monoecious species characterized by mid-winter blooming and sporophytic incompatibility. The molecular mechanisms at the basis of the female flower development and of the pollen-stigma interaction are little known, although pollination in this species is a critical factor to ensure good yield. Differential display technique was used to study genes expressed during the female flower development, comparing styles before emergence from the bud and styles at full bloom. The full-length cDNA clone, designated CavPrx (Corylus avellana peroxidase) and isolated in mature styles, was characterized as a sequence encoding for a 330 amino acids protein, containing all the conserved features of class III peroxidases. CavPrx resulted expressed only in styles, with a peak in mature styles pollinated with compatible pollen. Class III peroxidases are expressed in several different plant tissue types and are involved in a broad spectrum of physiological processes. Until now, four peroxidases expressed in the stigma were identified in Arabidopsis thaliana and Senecio squalidus: they were assumed to be possibly involved in pollen-pistil interaction, pollen tube penetration/growth and/or in defence against pathogens. CavPrx is the first gene for a floral peroxidase isolated in hazelnut and its expression pattern suggests a possible role in the pollination process.

Corylus/enzimologia , Corylus/genética , Flores/enzimologia , Flores/genética , Genes de Plantas/genética , Peroxidase/genética , Proteínas de Plantas/genética , Sequência de Aminoácidos , Sequência de Bases , Corylus/crescimento & desenvolvimento , DNA Complementar/genética , DNA Complementar/isolamento & purificação , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Peroxidase/química , Peroxidase/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Padrões de Referência
Genome ; 53(5): 384-99, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20616869


There is a lack of published microsatellite data that characterizes Camellia spp. To address this, an initial study of sequence tagged microsatellite site (STMS) variation was undertaken with 132 accessions of Camellia spp., which included 24 accessions representing 22 different species or varieties as well as 63 cultivars of C. japonica, 33 cultivars of C. sasanqua, 7 cultivars of C. x vernalis, 3 cultivars of C. x hiemalis, and 2 cultivars of C. hybrida. The four primer sets used (MSCJAF37, MSCJAH46, MSCJAF25, and MSCJAH38) successfully amplified polymorphic alleles in all the species analysed, showing cross-transferability. Overall, 96 alleles were scored. MSCJAH38 primers produced the highest number of bands (30), while MSCJAH46 primers yielded the lowest number (15). The genetic distance between pairs of accessions was estimated on the basis of the Nei coefficient and a principal coordinate analysis was performed. The plot revealed a main differentiation between the C. japonica cultivars and the winter camellias. The distribution of the genetic variation, attributed by AMOVA, particularly highlighted genetic overlap among C. sasanqua cultivars and the cultivars belonging to C. x vernalis, C. x hiemalis, and C. hybrida. In conclusion, this study demonstrated that STMS markers offer a suitable method for detection of genetic variability and molecular study of camellia genotypes.

Camellia/genética , DNA de Plantas/genética , Variação Genética , Repetições de Microssatélites/genética , Alelos , Camellia/classificação , Primers do DNA/genética , Frequência do Gene , Filogenia , Polimorfismo Genético , Sitios de Sequências Rotuladas , Especificidade da Espécie
Transgenic Res ; 19(1): 17-27, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19507046


Eight transgenic grapevine lines transformed with the coat protein gene of Grapevine fanleaf virus (GFLV-CP) were analyzed for a correlation between transgene expression, siRNAs production and DNA methylation. Bisulphite genome sequencing was used for a comprehensive analysis of DNA methylation. Methylated cytosine residues of CpG and CpNpG sites were detected in the GFLV-CP transgene, in the T7 terminator and in the 35S promoter of three grapevines without transgene expression, but no detectable level of siRNAs was recorded in these lines. The detailed analysis of 8 lines revealed the complex arrangements of T-DNA and integrated binary vector sequences as crucial factors that influence transgene expression. After inoculation with GFLV, no change in the levels of cytosine methylation was observed, but transgenic and untransformed plants produced short siRNAs (21-22 nt) indicating that the grapevine plants responded to GFLV infection by activating a post-transcriptional gene silencing mechanism.

Citosina/metabolismo , Inativação Gênica/fisiologia , Imunidade Inata/genética , Vírus de Plantas/imunologia , RNA Interferente Pequeno/biossíntese , Transgenes/fisiologia , Vitis/genética , Metilação de DNA/genética , DNA-Citosina Metilases/metabolismo , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Doenças das Plantas/virologia , Plantas Geneticamente Modificadas , Transformação Genética/fisiologia , Transgenes/genética , Vitis/imunologia , Vitis/metabolismo , Vitis/virologia