Genotype assessment of grape regenerants from floral explants

A molecular typing of regenerant vines based on co-dominant simple sequence repeat (SSR) markers was applied for checking recombination events during somatic embryogenesis from floral explants. Twenty-one samples of somatic embryos and plantlets from embryogenic callus of both anthers and ovaries of the V. vinifera cv. Chardonnay, the rootstock Kober 125 AA, and the accession V. rupestris du Lot were randomly chosen from a number of regenerant lines. The genotype at polymorphic VVS2, VVMD5, VVMD7, VVMD27, VrZAG62 and VrZAG79 loci was produced and compared with reference patterns. No recombination events were detected in the cells involved in the somatic embryogenesis induction of all the checked samples, since all of them generated the same SSR profile of the grape variety from which explants were isolated.

Leaf monoterpene emission limits photosynthetic downregulation under heat stress in field-grown grapevine

Rising temperature is among the most remarkably stressful phenomena induced by global climate changes with negative impacts on crop productivity and quality. It has been previously shown that volatiles belonging to the isoprenoid family can confer protection against abiotic stresses. In this work, two Vitis vinifera cv. ‘Chardonnay’ clones (SMA130 and INRA809) differing due to a mutation (S272P) of the DXS gene encoding for 1-deoxy-D-xylulose-5-phosphate (the first dedicated enzyme of the 2C-methyl-D-erythritol-4-phosphate (MEP) pathway) and involved in the regulation of isoprenoids biosynthesis were investigated in field trials and laboratory experiments. Leaf monoterpene emission, chlorophyll fluorescence and gas-exchange measurements were assessed over three seasons at different phenological stages and either carried out in in vivo or controlled conditions under contrasting temperatures. A significant (p < 0.001) increase in leaf monoterpene emission was observed in INRA809 when plants were experiencing high temperatures and over two experiments, while no differences were recorded for SMA130. Significant variation was observed for the rate of leaf CO2 assimilation under heat stress, with INRA809 maintaining higher photosynthetic rates and stomatal conductance values than SMA130 (p = 0.003) when leaf temperature increased above 30 °C. At the same time, the maximum photochemical quantum yield of PSII (Fv/Fm) was affected by heat stress in the non-emitting clone (SMA130), while the INRA809 showed a significant resilience of PSII under elevated temperature conditions. Consistent data were recorded between field seasons and temperature treatments in controlled environment conditions, suggesting a strong influence of monoterpene emission on heat tolerance under high temperatures. This work provides further insights on the photoprotective role of isoprenoids in heat-stressed Vitis vinifera, and additional studies should focus on unraveling the mechanisms underlying heat tolerance on the monoterpene-emitter grapevine clone

RAPD markers in wild and cultivated Vitis vinifera

Some Vitis vinifera cultivars and V. vinifera ssp. silvestris individuals have been subjected to the RAPD analysis in order to estimate the genetic diversity existing within this germplasm. 44 decamer primers of arbitrary sequence have been used for PCR and reproducible band profiles have been obtained. The distribution of the individualized polymorphic DNA markers has not turned out to be different in a remarkable way between cultivated and wild grapevines but this RAPD approach provides for some characteristics useful to analyze genetic relationships even within the Vitis vinifera species

Genetic characterization and relationships of traditional grape cultivars from Serbia

Reference genetic profiles were generated for 12 traditional grapevine cultivars of Serbia through a genotyping approach that included the "core set" of 9 SSR markers for genetic identification and further 13 common microsatellites for strengthening genetic relationship analysis. Consistent matching with SSR markers of grapevines cultivated in neighbouring countries or maintained in European germplasm collections was found for most of the genotypes, suggesting possible synonyms and revealing that 'Muskat Krokan' corresponds to 'Muscat fleur d’Oranger' and two 'Tamjanika' cultivars are identical to 'Moscato Giallo' and 'Moscato Rosa'. When compared with germplasm representing the classical eco-geographic grouping of grapevine cultivars, Serbian non-Muscat genotypes clustered within the Convar pontica subconvar balcanica taxon thus supporting their indigenous origin.

In vitro screening of interspecific hybrids (Malus spp.) for resistance to apple proliferation

A breeding programme was set up six years ago in Trentino as part of the Project “Scopazzi del Melo - Apple Proliferation” (SMAP) in order to obtain AP resistant apple rootstocks.Twenty-six hybrids generated from the crossings (Malus sieboldii, second generation, x Malus domestica) were micropropagated and studied in standardised conditions. An in vitro screening system for AP resistance previously set for the parents of the crosses was adopted and modified. Specific symptoms of the disease, as well as height and basal proliferation of the shoot and size of the leaves, were recorded in vitro at 3 months post-inoculation. At the same time, phytoplasma concentration was determined in the whole shoot by quantitative RT-PCR. An in vitro disease index taking into account all the above-mentioned parameters was developed.Each healthy genotype was graft-inoculated in triplicate with two different phytoplasma strains after plant rooting and acclimatisation. Phenotype and phytoplasma titre were evaluated in the roots the year after infection.Preliminary results indicated that the resistance trait segregates in the progenies. The resistant genotypes had lower phytoplasma concentrations than the susceptible controls, did not show AP-specific symptoms and their growth was not affected by infection. By comparing the resistant behaviour of the same genotypes, the in vitro screening allows for a quick selection of genotypes that are worth evaluating in the field for agronomic traits.Keywords: Apple Proliferation, Malus sieboldii, resistance screening, quantitative real-time PCR, disease inde

Genetic relationships among local Vitis vinifera cultivars from Campania (Italy)

A total of 114 accessions putatively corresponding to 69 local grape cultivars from Campania (Southern Italy) were analysed with 8 microsatellite markers (VVS2, VVMD5, VVMD7, VVMD25, VVMD27, VVMD31, VrZAG62 and VrZAG79) in order to evaluate their genetic diversity and relationships. According to their unique genotype at SSR loci finally 56 varieties were found. Interesting cases of synonymy, i.e. Greco di Tufo and Asprinio, Palummina and Piedirosso, and homonymy were disclosed. Pairwise genetic distances were calculated between all cultivars. Clustering of cultivars did not reflect their current distribution and this suggests that grape cultivars of Campania might have been introduced from various and distinct geographic areas

Ampelometric evaluation of wild grape (Vitis vinifera L. ssp. sylvestris (C.C. Gmel.) Hegi) accessions in the germplasm collection of FEM-IASMA, Italy

In this paper, 45 wild grapevine accessions collected during two consecutive years were compared for 36 ampelometric traits using digital image analysis. The sample set contained male and female individuals from different geographic regions: Germany, North Italy, Central Italy, South Italy, Sardinia and Turkey. The leaf morphological data from the collected samples suggest that geographic origin, gender and vintage could have an effect on ampelometric traits in this species

The parentage of 'Sangiovese', the most important Italian wine grape

A previous microsatellite study pointed out a possible parent-offspring relationship between 'Sangiovese', the most widespread red grape cultivar in Italy, and 'Ciliegiolo', an ancient Tuscan variety. Testing 'Sangiovese' as a parent of 'Ciliegiolo', we searched for the putative other parent in our extensive, private and standardized database, but we did not find any candidate. Testing 'Ciliegiolo' as a parent of 'Sangiovese', we found four candidate cultivars. After the analysis of 50 microsatellites, only one stood the paternity test and we established with a strong statistical support that 'Sangiovese' is a progeny of 'Ciliegiolo' and 'Calabrese di Montenuovo', an obscure grapevine from Campania, Italy. This cultivar does not have a registered name and is supposed to have been introduced from Calabria. Among 180 additional local grape cultivars from Calabria, Campania or Tuscany, we did not find any matching variety. As a consequence, we propose to adopt the name 'Calabrese di Montenuovo' for this grape cultivar. In addition, we found relatives of 'Sangiovese' and 'Calabrese di Montenuovo' in Calabria, thus strongly suggesting a Calabrian origin for 'Calabrese di Montenuovo' and indicating that 'Sangiovese' has ancestors and/or progenies in Tuscany and in Southern Italy.

Differentially expressed genes between drought-tolerant and drought-sensitive barley genotypes in response to drought stress during the reproductive stage

Drought tolerance is a key trait for increasing and stabilizing barley productivity in dry areas worldwide. Identification of the genes responsible for drought tolerance in barley (Hordeum vulgare L.) will facilitate understanding of the molecular mechanisms of drought tolerance, and also facilitate the genetic improvement of barley through marker-assisted selection or gene transformation. To monitor the changes in gene expression at the transcriptional level in barley leaves during the reproductive stage under drought conditions, the 22K Affymetrix Barley 1 microarray was used to screen two drought-tolerant barley genotypes, Martin and Hordeum spontaneum 41-1 (HS41-1), and one drought-sensitive genotype Moroc9-75. Seventeen genes were expressed exclusively in the two drought-tolerant genotypes under drought stress, and their encoded proteins may play significant roles in enhancing drought tolerance through controlling stomatal closure via carbon metabolism (NADP malic enzyme, NADP-ME, and pyruvate dehydrogenase, PDH), synthesizing the osmoprotectant glycine-betaine (C-4 sterol methyl oxidase, CSMO), generating protectants against reactive-oxygen-species scavenging (aldehyde dehydrogenase,ALDH, ascorbate-dependent oxidoreductase, ADOR), and stabilizing membranes and proteins (heat-shock protein 17.8, HSP17.8, and dehydrin 3, DHN3). Moreover, 17 genes were abundantly expressed in Martin and HS41-1 compared with Moroc9-75 under both drought and control conditions. These genes were possibly constitutively expressed in drought-tolerant genotypes. Among them, seven known annotated genes might enhance drought tolerance through signalling [such as calcium-dependent protein kinase (CDPK) and membrane steroid binding protein (MSBP)], anti-senescence (G2 pea dark accumulated protein, GDA2), and detoxification (glutathione S-transferase, GST) pathways. In addition, 18 genes, including those encoding Δl-pyrroline-5-carboxylate synthetase (P5CS), protein phosphatase 2C-like protein (PP2C), and several chaperones, were differentially expressed in all genotypes under drought; thus they were more likely to be general drought-responsive genes in barley. These results could provide new insights into further understanding of drought-tolerance mechanisms in barley