The effect of low temperature (LT, 10°C) on modification of plasma membrane (PM) H(+)-ATPase (EC 3.6.3.14) activity in cucumber roots was studied. Plants were grown under LT for 3 or 6 days. Some of the plants after 3 days exposure to LT were transferred to control conditions for another 3 days (post-cold, PC). The activity of PM-H(+)-ATPase was decreased in plants treated for 3 days with LT. However, the activity of PM-H(+)-ATPase was higher in plants treated with LT for a longer time and in PC plants as well. Estimation of transcript levels of cucumber PM-H(+)-ATPase in roots indicates that the action of LT involves the gene expression level. The level of PM-H(+)-ATPase mRNA was markedly decreased in roots exposed to LT for 3 days. Moreover, the increased H(+)-ATPase activity in PM isolated from plants treated for 6 days with LT and from PC plants was positively correlated with higher levels of CsHA transcripts. Western blot analysis with an anti-phosphothreonine antibody showed that modification of the activity of PM-H(+)-ATPase under LT stress did not result from phosphorylation/dephosphorylation of the enzyme protein. However, the stimulation of PM-H(+)-ATPase activity in the case of PC plants could partially have emanated from increased activity of PM NAD(P)H oxidoreductase. In addition, modification of the transcript level of proton pump genes could have resulted from the action of H(2)O(2). In PC plants, an increase in H(2)O(2) level was observed. Moreover, treatment of plants with H(2)O(2) induced expression of PM H(+)-ATPase genes.

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Plasma membrane NADPH oxidases (RBOHs, EC 1.6.3.1) are known as the main ROS generators involved in plant adaptation to stress conditions. In the present work, regulation of NADPH oxidase was analyzed in cucumber (Cucumis sativus L. var. Krak) seedlings exposed to salinity. RBOH activity and gene expression, as well as H2O2 content, were determined in the roots of plants treated with 50 or 100 mM NaCl for 1 h, and 50 mM NaCl for 1 or 6 days. It was found that enzyme activity increased in parallel with an enhancement in the H2O2 level in roots exposed to 100 mM NaCl for 1 h, and to 50 mM NaCl for 1 day. The expression of some CsRboh genes was induced by salt. Moreover, an increase in the activity of G6PDH, providing the substrate for the NADPH oxidase, was observed. In seedlings subjected to salinity for a longer time, antioxidant enzymes—including superoxide dismutase, catalase, and ascorbate peroxidase—were activated, participating in maintaining a steady-state H2O2 content in the root cells. In conclusion, NADPH oxidase and endogenous H2O2 up-regulation seem to be early events in cucumber response to salinity.

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Two genes (CsHA2, CsHA3) homolog to the plasma membrane H ? -ATPase encoding sequences have been identified in cucumber cells. The full-length cDNA of these sequences comprise an open reading frame that encodes about 950 amino acid polypeptide with several potential transmembrane domains. Both sequences share a high homology with putative plasma membrane-associated proton pumps in higher plants, ranging from 84% with that of Arabidopsis thaliana to 90% with Sesbania rostrata. Phylogenetic analysis grouped the two isoforms into one group (subfamilies II) together with Kosteletzkya virginica KVATP1, Sesbania rostrata SRHA4, tobacco PMA4 and rice OSA7. The expression of CsHA isoforms at the mRNA level showed they different organ pattern. Transcript level of CsHA2 was detected in all vegetative organs, but more abundantly in leaves than in roots, while CsHA3 expression was limited only to the roots in immature as well as in mature plants. Moreover, the RT-PCR analysis of generative organs (flowers), fruits and dry seeds revealed differential expression pattern for CsHA2 and CsHA3 suggesting that these proteins could be involved in separate physiological processes.

An antisense oligonucleotide (ASO) is a short single-stranded deoxyribonucleotide complementary to the sense strand of a selected nucleic acid. As a result, an ASO can modulate gene expression through several mechanisms. The technology based on ASO has already been applied in studies on gene function in mammalian cells and selective therapeutic strategies for many diseases. The conceptual simplicity and low cost of this method, and the developments in the field of plant genome sequencing observed in the last decades, have paved the way for the ASO method also in plant biology. It is applied in gene function analysis as well as the development of non-invasive plant production technology involving gene modifications without transgenesis. Therefore, the first part of this review provides a comprehensive overview of the structure, mechanism of action and delivery methods of ASOs in plants and shows the most important features essential for the proper design of individual experiments. We also discuss potential issues and difficulties that may arise during practical ASO implementation. The second part of this article contains an analysis of ASO applications in various studies in the field of plant biology. We presented for the first time that ASOs were also successfully applied in cucumber.

Water and nutrient deficiencies in soil are becoming a serious threat to crop production. Therefore, usable water and nutrient recovery from wastewater, such as urine and grey water, should be considered. In this work, we showed the possibility of using grey water and urine after processing in an aerobic reactor with activated sludge in which the nitrification process takes place. The resulting liquid (nitrified urine and grey water, NUG) contains three potential factors that can adversely affect plant growth in a hydroponic system: anionic surfactants, nutrient deficits, and salinity. After dilution and supplementation with small amounts of macro- and micro-elements, NUG was suitable for cucumber cultivation. Plant growth on this modified medium (enriched nitrified urine and grey water, NUGE) was similar to that of plants cultivated on Hoagland solution (HS) and reference commercial fertilizer (RCF). The modified medium (NUGE) contained a significant amount of sodium (Na) ions. Therefore, typical effects of salt stress were observed in cucumber plants, including reduced chlorophyll levels, slightly weaker photosynthesis parameters, increased H2O2 levels, lipid peroxidation, ascorbate peroxidase (APX) activity, and proline content in the leaves. In addition, reduced protein levels were observed in plants treated with recycled medium. At the same time, lower nitrate content in tissues was found, which may have resulted from their intensive use by nitrate reductase (NR), the activity of which significantly increased. Although cucumber is a glycophyte, it grew very well in this recycled medium. Interestingly, salt stress and possibly anionic surfactants promoted flower formation, which in turn could positively affect plant yield.