High-Nitrate-Supply-Induced Transcriptional Upregulation of Ascorbic Acid Biosynthetic and Recycling Pathways in Cucumber.

Nowadays open field and protected vegetable cultivation practices require and use genotypes which are precisely tailored to their intended growth environments. Variability of this kind provides a rich source of material to uncover molecular mechanisms supporting the necessarily divergent physiological traits. In this study, typical field-optimized and glasshouse-cultivated cucumber F1 hybrids were investigated, and displayed slower growth ('Joker') and faster growth ('Oitol') in seedlings. Antioxidant capacity was lower in 'Joker' and higher in 'Oitol', pointing to a potential redox regulation of growth. The growth response of seedlings to paraquat treatment indicated stronger oxidative stress tolerance in the fast-growing 'Oitol'. To test whether protection against nitrate-induced oxidative stress was also different, fertigation with increasing potassium nitrate content was applied. This treatment did not change growth but decreased the antioxidant capacities of both hybrids. Bioluminescence emission revealed stronger lipid peroxidation triggered by high nitrate fertigation in the leaves of 'Joker' seedlings. To explore the background of the more effective antioxidant protection of 'Oitol', levels of ascorbic acid (AsA), as well as transcriptional regulation of relevant genes of the Smirnoff-Wheeler biosynthetic pathway and ascorbate recycling, were investigated. Genes related to AsA biosynthesis were strongly upregulated at an elevated nitrate supply in 'Oitol' leaves only, but this was only reflected in a small increase in total AsA content. High nitrate provision also triggered expression of ascorbate-glutathion cycle genes with stronger or exclusive induction in 'Oitol'. AsA/dehydro-ascorbate ratios were higher in 'Oitol' for all treatments, with a more pronounced difference at high nitrate levels. Despite strong transcriptional upregulation of ascorbate peroxidase genes (APX) in 'Oitol', APX activity only increased significantly in 'Joker'. This suggests potential inhibition of APX enzyme activity specifically in 'Oitol' at a high nitrate supply. Our results uncover an unexpected variability in redox stress management in cucumbers, including nitrate inducibility of AsA biosynthetic and recycling pathways in certain genotypes. Possible connections between AsA biosynthesis, recycling and nitro-oxidative stress protection are discussed. Cucumber hybrids emerge as an excellent model system for studying the regulation of AsA metabolism and the roles of AsA in growth and stress tolerance.

Downregulation of Polyamine and Diamine Oxidases in Silicon-Treated Cucumber.

Silicon (Si) is a ubiquitous element in soil with well-known beneficial effects under certain conditions, in several plant species, if supplied in available form for uptake. It may alleviate damage in various stress situations and may also promote growth when no obvious stressors are applied. Effects of Si are often linked to mitigation of oxidative stress, in particular to the induction of antioxidant defense mechanisms. In the work presented, the impact of silicon provision on pro-oxidant systems was investigated in cucumber. Plants of the F1 cultivar hybrid 'Joker' were grown under in vitro conditions in the absence of any applied external stressor. Silicon provision decreased H2O2 content and lowered lipid peroxidation in the leaves of the treated plants. This was paralleled by declining polyamine oxidase (PAO) and diamine oxidase (DAO) activities. Several PAO as well as lipoxygenase (LOX) genes were coordinately downregulated in Si-treated plants. Unlike in similar systems studied earlier, the Si effect was not associated with an increased transcript level of gene coding for antioxidant enzymes. These results suggest an inhibitory effect of Si provision on pro-oxidant amine oxidases, which may decrease the level of reactive oxygen species by retarding their production. This extends the molecular mechanisms linked to silicon effects onto redox balance in plants.

Identification of a novel UDP-glycosyltransferase gene from Rhodiola rosea and its expression during biotransformation of upstream precursors in callus culture.

Roseroot (Rhodiola rosea L.) is a medicinal plant with adaptogenic properties and several pharmaceutically important metabolites. In this study, a full length cDNA encoding a UDPG gene of roseroot was identified, cloned and characterized. Its ORF (1425 bp) was transferred into E. coli, where the expression of the recombinant enzyme was confirmed. To monitor the enzyme activity, 3 precursors (tyramine, 4-hydroxyphenylpyruvate & tyrosol) of salidroside biosynthesis pathway were added to roseroot callus cultures and samples were harvested after 1, 6, 12, 24, 48 & 96 h. Along with the controls (without precursor feeding), each sample was subjected to HPLC and qRT-PCR for phytochemical and relative UDP-glycosyltransferase gene expression analysis, respectively. The HPLC analysis showed that the salidroside content significantly increased; reaching 0.5% of the callus dry weight (26-fold higher than the control) after 96 h when 2 mM tyrosol was given to the media. The expression of the UDP-glycosyltransferase increased significantly being the highest at 12 h after the feeding. The effect of tyramine and 4-hydroxyphenylpyruvate was not as pronounced as of tyrosol. Here, we introduce a R. rosea specific UDPG gene and its expression pattern after biotransformation of intermediate precursors in in vitro roseroot callus cultures.

Influence of genotype and explant source on the in vitro regeneration ability of different melon varieties.

Nine genotypes of melon (Cucumis melo L.) were selected for the investigation of regeneration. Most of the tested varieties showed regeneration ability on medium containing 0.5 mg l⁻¹ or 1 mg l⁻¹ BA, but following the appearance of shoot buds, only six varieties produced leafy shoots. The effect of combinations of BA with different auxins (IAA, NA, 2,4-D) and ABA in the culture medium on shoot regeneration was tested on cotyledon explants of 'Hógolyó' and 'Hale's Best'. To establish optimal conditions for the adventitious shoot induction six types of seedling-derived explants were prepared from seedlings of four different ages. The best results for shoot forming capacity were achieved with cotyledons followed by decapitated seedlings and hypocotyls derived from 4-day-old seedlings. Cotyledon segments of 'Hógolyó' and 'Hale's Best' were also cultivated on media with different concentrations of IAA and BA supplemented with 0.26 mg l⁻¹ ABA. The highest number of well-formed plantlets was counted for 'Hógolyó' on the medium supplemented with 0.9 mg l⁻¹ BA+ 0.6 mg l⁻¹ IAA+ 0.26 mg l⁻¹ ABA. This is the first report on the in vitro regeneration of 'Hógolyó' from decapitated seedling and hypocotyl explants and of 'JavÍtott Zentai', 'Muskotály', 'Hógolyó', 'Tétényi csereshéjú' and 'Magyar Kincs' from cotyledon explants.