Sample and Library Preparation for PacBio Long-Read Sequencing in Grapevine.

PacBio long-read sequencing is a third-generation technology that generates long reads up to 20 kilobases (kb), unlike short-read sequencing instruments that produce up to 600 bases. Long-read sequencing is particularly advantageous in higher organisms, such as humans and plants, where repetitive regions in the genome are more abundant. The PacBio long-read sequencing uses a single molecule, real-time approach where the SMRT cells contain several zero-mode waveguides (ZMWs). Each ZMW contains a single DNA molecule bound by a DNA polymerase. All ZMWs are flushed with deoxy nucleotides with a fluorophore specific to each nucleotide. As the sequencing proceeds, the detector detects the wavelength of the fluorescence and the nucleotides are read in real-time. This chapter describes the sample and library preparation for PacBio long-read sequencing for grapevine.

A Review of Ampelometry: Morphometric Characterization of the Grape (Vitis spp.) Leaf.

Grape (Vitis spp.) is one of the most important horticultural crops, cultivated worldwide on more than 7.3 million hectares for various purposes such as winemaking, fresh fruit consumption, rootstock, and ornamental plants. Based on the inter- and intraspecific morphological variability, several descriptor lists, manuals and ampelographic studies are available for identification. Among the organs, leaves have the most traits, while the young shoot, bunch and berry are also important in the characterization of the genotypes. Vitis species and cultivars are described by leaf morphological characterization developed in many ways for the identification of genotypes, to clarify synonymies and distinct clones or evaluate the diversity of wild Vitis taxa. Morphometric-also known as ampelometric-evaluation has an extensive background in the literature. However, for some reasons, only a part of the literature is cited, despite its significant scientific value. In this paper, we summarize the efforts of metric characterization of the grapevine leaf with the introduction of the scientific objectives and reviewing the studies showing the innovations in phenotyping during the past 120 years.

Gene expression profiling identifies pathways involved in seed maturation of Jatropha curcas.

BACKGROUND: Jatropha curcas, a tropical shrub, is a promising biofuel crop, which produces seeds with high content of oil and protein. To better understand the maturation process of J. curcas seeds and to improve its agronomic performance, a two-step approach was performed in six different maturation stages of seeds: 1) generation of the entire transcriptome of J. curcas seeds using 454-Roche sequencing of a cDNA library, 2) comparison of transcriptional expression levels using a custom Agilent 8x60K oligonucleotide microarray. RESULTS: A total of 793,875 high-quality reads were assembled into 19,382 unique full-length contigs, of which 13,507 could be annotated with Gene Ontology (GO) terms. Microarray data analysis identified 9111 probes (out of 57,842 probes), which were differentially expressed between the six maturation stages. The expression results were validated for 75 selected transcripts based on expression levels, predicted function, pathway, and length. Result from cluster analyses showed that transcripts associated with fatty acid, flavonoid, and phenylpropanoid biosynthesis were over-represented in the early stages, while those of lipid storage were over-represented in the late stages. Expression analyses of different maturation stages of J. curcas seed showed that most changes in transcript abundance occurred between the two last stages, suggesting that the timing of metabolic pathways during seed maturation in J. curcas occurs in late stages. The co-expression results showed that the hubs (CB5-D, CDR1, TT8, DFR, HVA22) with the highest number of edges, associated with fatty acid and flavonoid biosynthesis, are showing a decrease in their expression during seed maturation. Furthermore, seed development and hormone pathways are significantly well connected. CONCLUSION: The obtained results revealed differentially expressed sequences (DESs) regulating important pathways related to seed maturation, which could contribute to the understanding of the complex regulatory network during seed maturation with the focus on lipid, flavonoid and phenylpropanoid biosynthesis. This study provides detailed information on transcriptional changes during J. curcas seed maturation and provides a starting point for a genomic survey of seed quality traits. The results highlighted specific genes and processes relevant to the molecular mechanisms involved in Jatropha seed maturation. These data can also be utilized regarding other Euphorbiaceae species.

The Effect of Crystallinity on the Toughness of Cast Polyamide 6 Rods with Different Diameters.

The present paper concentrates on the toughness and the degree of crystallinity of the magnesium-catalyzed polyamide 6 rods cast in different diametres, which are commonly used for gear manufacturing. Its toughness cannot be regarded as a constant feature due to the casting technology. The mechanical properties of the semi-finished products are sensitive to the manufactured dimension, e.g., cast diameter, which are investigated by the Charpy impact test and tensile impact test. It is generally accepted that the impact strength and tensile-impact strength correlate with the degree of crystallinity beside many other material's feature. Crystallinity is evaluated by Differential Scanning Calorimetry. The aim of this study is to determine the relationship between toughness and crystallinity of the magnesium-catalyzed cast PA6 rods with different diameters. For the research cast rods between 40 and 300 mm diameter were selected in seven-dimensional steps. Based on the results, it was found that the toughness depends strongly on the diameter size. Furthermore, it is proved that the crystallinity explains 62.3% of the variation of the Charpy's impact strengths, while the tensile impact method was not suitable to detect the difference between the test samples.

Differentiation of grapevine (Vitis vinifera L.) conculta members based on molecular tools.

Twenty-seven grapevine (Vitis vinifera L.) varieties within 12 putative berry colour variation groups (conculta) were genotyped with 14 highly polymorphic microsatellite (simple sequence repeats (SSR)) markers. Three additional oligonucleotide primers were applied for the detection of the Gret1 retroelement insertion in the promoter region of VvMybA1 transcription factor gene regulating the UFGT (UDP-glucose: flavonoid 3-O-glucosyltransferase) activity. UFGT is the key enzyme of the anthocyanin biosynthetic pathway. SSR results proved that the analysed cultivars can be grouped only into nine concultas, the other three putative berry colour variant groups consist of homonyms as a consequence of misnaming. In the case of Sárfehér-Sárpiros, Delaware red-Delaware white and Járdovány fekete-Járdovány fehér, it was attested that they are not bud sports, but homonyms. Some conculta members could be differentiated according to the presence or the absence of the Gret1 retroelement (Chasselas, Furmint and Lisztes), while others, Bajor, Bakator, Gohér and Traminer conculta members, remained indistinguishable either by the microsatellites or the Gret1-based method.