Disentangling the genetic bases of Saccharomyces cerevisiae nitrogen consumption and adaptation to low nitrogen environments in wine fermentation

The budding yeast Saccharomyces cerevisiae has been considered for more than 20 years as a premier model organ- ism for biological sciences, also being the main microorganism used in wide industrial applications, like alcoholic fermentation in the winemaking process. Grape juice is a challenging environment for S. cerevisiae , with nitrogen deficiencies impairing fermentation rate and yeast biomass production, causing stuck or sluggish fermentations, thus generating sizeable economic losses for wine industry. In the present review, we summarize some recent efforts in the search of causative genes that account for yeast adaptation to low nitrogen environments, specially focused in wine fermentation conditions. We start presenting a brief perspective of yeast nitrogen utilization under wine fermentative conditions, highlighting yeast preference for some nitrogen sources above others. Then, we give an outlook of S. cerevisiae genetic diversity studies, paying special attention to efforts in genome sequencing for population structure determination and presenting QTL mapping as a powerful tool for phenotype-genotype correlations. Finally, we do a recapitulation of S. cerevisiae natural diversity related to low nitrogen adaptation, specially showing how different studies have left in evidence the central role of the TORC1 signalling pathway in nitrogen utilization and positioned wild S. cerevisiae strains as a reservoir of beneficial alleles with potential industrial applications (e.g. improvement of industrial yeasts for wine production). More studies focused in disentangling the genetic bases of S. cerevisiae adaptation in wine fermentation will be key to determine the domestication effects over low nitrogen adaptation, as well as to definitely proof that wild S. cerevisiae strains have potential genetic determinants for better adaptation to low nitrogen conditions.

Effect of arbuscular mycorrhizal fungi on young vines in copper-contaminated soil

Abstract High copper (Cu) levels in uprooted old vineyard soils may cause toxicity in transplanted young vines, although such toxicity may be reduced by inoculating plants with arbuscular mycorrhizal fungi (AMF). The objective of this study was to evaluate the effects of AMF on the plant growth, chlorophyll contents, mycorrhizal colonization, and Cu and phosphorus (P) absorption in young vines cultivated in a vineyard soil contaminated by Cu. Commercial vineyard soil with high Cu levels was placed in plastic tubes and transplanted with young vines, which were inoculated with six AMF species (Dentiscutata heterogama, Gigaspora gigantea, Acaulospora morrowiae, A. colombiana, Rhizophagus clarus, R. irregularis) and a control treatment on randomized blocks with 12 replicates. After 130 days, the mycorrhizal colonization, root and shoot dry matter (DM), height increment, P and Cu absorption, and chlorophyll contents were evaluated. The height increment, shoot DM and chlorophyll contents were not promoted by AMF, although the root DM was increased by R. clarus and R. irregularis, which had the greatest mycorrhizal colonization and P uptake. AMF increased Cu absorption but decreased its transport to shoots. Thus, AMF species, particularly R. clarus and R. irregularis, contribute to the establishment of young vines exposed to high Cu levels.

Impact of pectolytic enzyme on the fermentation time and free-run juice rate of Sharad red wine.

We evaluated the effect of addition of commercially available pectolytic enzyme preparations in the must from Sharad variety, cultivated in India and its influence on some oenological parameters of red wine. The small scale fermentations demonstrated marked difference of organoleptic (colour) and rheologic characteristics (filterability, settling rates) and aroma between enzyme treated samples and control samples. We observed 29% enhancement in free-run juice yield and a remarkable 43% reduction in the fermentation time compared to the control. This biotechnological approach has demonstrated the economic feasibility and the benefits of adding 0.05 g. Kg-1 grapes pectolytic enzyme in terms of yield, aroma, colour, clarity and fermentation period.

The effects of cadmium chloride on secondary metabolite production in Vitis vinifera cv. cell suspension cultures

BACKGROUND: Plant secondary metabolites are possess several biological activities such as anti-mutagenic, anti-carcinogenic, anti-aging, etc. Cell suspension culture is one of the most effective systems to produce secondary metabolites. It is possible to increase the phenolic compounds and tocopherols by using cell suspensions. Studies on tocopherols production by cell suspension cultures are seldom and generally focused on seed oil plants. Although fresh grape, grape seed, pomace and grape seed oil had tocopherols, with our best knowledge, there is no research on tocopherol accumulation in the grape cell suspension cultures. In this study, it was aimed to determine the effects of cadmium chloride treatments on secondary metabolite production in cell suspension cultures of grapevine. Cell suspensions initiated from callus belonging to petiole tissue was used as a plant material. Cadmium chloride was applied to cell suspension cultures in different concentration (1.0 mM and 1.5 mM) to enhance secondary metabolite (total phenolics, total flavanols, total flavonols, trans-resveratrol, and α-, ß-, γ- δ-tocopherols) production. Cells were harvested at two days intervals until the 6th day of cultures. Amounts of total phenolics, total flavanols and total flavonols; trans-resveratrol and tocopherols (α-, ß-, γ- and δ-tocopherols) and dry cell weights were determined in the harvested cells. RESULTS: Phenolic contents were significantly affected by the sampling time and cadmium concentrations. The highest values of total phenolic (168.82 mg/100 g), total flavanol (15.94 mg/100 g), total flavonol (14.73 mg/100 g) and trans-resveratrol (490.76 µg/100 g) were found in cells treated with 1.0 mM CdCl2 and harvested at day 2. Contents of tocopherols in the cells cultured in the presence of 1.0 mM CdCl2 gradually increased during the culture period and the highest values of α, ß and γ tocopherols (145.61, 25.52 and 18.56 µg/100 g) were detected in the cell cultures collected at day 6. CONCLUSIONS: As a conclusion, secondary metabolite contents were increased by cadmium chloride application and sampling time, while dry cell weights was reduced by cadmium chloride treatments.

Induction of secondary metabolite production by UV-C radiation in Vitis vinifera L. Öküzgözü callus cultures

BACKGROUND: The aim of the present work was to examine the role of UV-C irradiation on the production of secondary metabolites (total phenolic, total flavanols, total flavonols, catechin, ferulic acid and trans-resveratrol in phenolic compounds and α-, ß-, γ- δ-tocopherols) in callus cultures. Studies on the effects of UV-C treatment on callus culture are seldom and generally focused on UV-B. However UV-C radiation play an important role in accumule secondary metabolites. RESULTS: In this study, callus cultures from Öküzgözü grape cultivar were initiated from leaf petiole explants. Calli formed after 6 weeks on the medium supplemented with 0.5 mg L-1 benzylaminopurine (BA), 0.5 mg L-1 indole acetic acid (IAA) on B5 media. Callus tissues were exposed to UV-C irradiation at 10, 20 and 30 cm distances from the UV source for 5 and 10 minutes and samples were collected at hours 0, 24 and 48. CONCLUSIONS: The greatest total phenolic content (155.14 mg 100 g-1) was detected in calli exposed to UV-C for 5 min from 30 cm distance and sampled after 24 h. 24 h and 48 h incubation times, 30 cm and 5 min were the most appropriate combination of UV-C application in total flavanol content. Maximum total flavonol content (7.12 mg 100 g-1) was obtained on 0 h, 5 min and 20 cm combination. The highest (+)- catechin accumulation (8.89 mg g-1) was found in calli with 10 min UV-C application from 30 cm distance and sampled after 48 h. Ferulic acid content increased 6 fold in Öküzgözü callus cultures (31.37 µg g-1) compared to the control group. The greatest trans-resveratrol content (8.43 µg g-1) was detected in calli exposed to UV-C for 5 min from 30 cm distance and sampled after 24 h. The highest α-tocopherol concentration was found in calli exposed to UV-C for 10 min from 30 cm distance and sampled after 24 h. As a conclusion, it was showed that UV-C radiation had remarkable promoting effects on the accumulation of secondary metabolites in the calli of Öküzgözü grape cultivar.

Stimulation of trans-resveratrol biosynthesis in Vitis vinifera cv. Kyoho cell suspension cultures by 2, 3-dihydroxypropyl jasmonate elicitation

Background: Plant cell suspension culture of Vitis vinifera is a promising technology for investigating different factors that are able to induce and/or modify stilbenes biosynthesis. Jasmonates have been reported to play an important role in a signal transduction pathway that regulates defence responses as well as the production of secondary metabolites. In this study, 2, 3-dihydroxypropyl jasmonate (DHPJA) was used to investigate its effect on stimulating trans-resveratrol (t-R) accumulation and the plant defence responses in Vitis vinifera cv. Kyoho cell suspension cultures for the first time. Results: It demonstrated that DHPJA had superior effects on stilbenoids accumulation over methyl jasmonate (MeJA). The optimal condition was 150 uM DHPJA added on day 15 of cultivation period, with the highest level of t-R accumulation which was increased 1.8-fold and 1.3-fold compared with the control and 150 uM MeJA respectively. DHPJA induced stronger plant defence responses, including oxidative burst and activation of L-phenylalanine ammonia lyase (PAL) than MeJA. H2O2 generation induced by DHPJA played a significant role in enhancing t-R accumulation. Adding a specific inhibitor of H2O2 signalling pathway inhibited DHPJA-induced t-R accumulation, but had no effects on DHPJA-induced other metabolites accumulation, which resulted in regulations of product diversity. Conclusions: This study demonstrated that DHPJA was an efficient elicitor to enhance t-R accumulation by activating stronger oxidative burst, and H2O2 signalling pathway could regulate product diversity in DHPJA-induced V. vinifera cv. Kyoho cell suspension cultures.

Phytoplasma and virus detection in commercial plantings of Vitis vinifera cv. Merlot exhibiting premature berry dehydration

A new and devastating physiological disorder of Vitis vinifera cv. Merlot was recently reported, known as premature berry dehydration (PBD), which is characterized by plant growth reduction, induction of general senescence and pedicel necrosis in the fruit, causing significant reductions in vineyard production. The causes of this disease remain unclear and previous reports suggest that it may be associated with phloem disruption and water provision. For this reason, any factor causing phloem disturbances could cause an important change in the berry water status. As some micro-organisms have been reported to disrupt phloem flow, we analyzed the occurrence of phytoplasma and viruses in commercial vineyards presenting PBD. In this study, a phytoplasma was detected by electron microscopy and nested PCR while virus infections were diagnosed by RT-PCR in samples collected during two growing seasons. The presence of phytoplasma only in samples from grape plants with PBD suggests that this pathogen may be one of the causal agents of this disorder. We suggest that the influence of other factors, such as virus infections, agronomic handling and environmental conditions also modulate berry dehydration. This is the first study at the microscopic and molecular levels that correlates phytoplasma presence with PBD.