Interactions of drought and low temperature stress on lipid and fatty acid composition of cucumber genotypes differing in growth response at suboptimal temperature

The effects of drought stress and/or low temperature stress on total lipid and phospholipid content and fatty acid composition of leaves of cucumber ( Cucumis sativus L.) genotypes differing in growth response at suboptimal temperature were studied. Both drought and low temperature resulted in reduced growth, especially in cv. Farbio, the genotype least tolerant to low temperature. Drought resulted in an increase in total lipid and phospholipid per g fresh weight. On a lipid basis no change in phospholipids or fatty acid content was observed. The fatty acid composition was changed by drought and low temperature, resulting in an increase in the degree of unsaturation. The genotype-specific reaction to treatment for total lipid content and the degree of unsaturation point to the possibility of a genetic origin for drought-induced lipid changes, which may be used in a breeding program for improved growth at suboptimal temperature.     

Re-examination of the localization of Mg-chelatase within the chloroplast

In a plastid-free assay, Mg-chelatase from pea ( Pisum sativum L. cv. Spring) and cucumber ( Cucumis sativus L. cv. Sumter) chloroplasts is inhibited to equal extents by the mercurial reagents. p -chloromercuribenzoate (PCMB) and p -chloromercuribenzene sulfonate (PCMBS). However, in intact chloroplasts PCMB inhibits Mg-chelatase fourfold more strongly than does PCMBS. Since PCMBS cannot penetrate membranes as readily as PCMB, Mg-chelatase may be localized interior to the inner chloroplast envelope. When Mg-chelatase is assayed with photosynthetically generated ATP, the presence of an external ATP trap does not inhibit activity, suggesting that the enzyme is not located in the interenvelope space. None of the components of Mg-chelatase are integral membrane proteins: Mg-chelatase activity is readily solubilized by washing the total chloroplast membranes in buffers of low MgCl₂ content. This precludes localization by purifying individual thylakoid and envelope membranes which requires low MgCl₂ concentrations.     

Determination of 4-chloroindoleacetic acid methyl ester in Vicieae species by gas chromatography-mass spectrometry

The natural chlorinated auxin, 4-chloroindoleacetic acid methyl ester, was identified in immature seeds of Lathyrus sativus L., Lathyrus maritimus (L.) Bigel and Lathyrus odoratus L. by thin layer chromatography and gas chromatography-mass spectrometry. In immature seeds of Vicia sativa L. and Lens culinaris Medik. the hormone was identified by selected ion monitoring. The hormone was determined quantitatively using pentadeuterated 4-chloroindoleacetic acid methyl ester as internal standard. Contents varied from 1 mg/kg fresh weight in Lathyrus sativus to 0.02 mg/kg in Lens culinaris. Lathyrus maritimus also contained indoleacetic acid methyl ester (0.3 mg/kg) besides the chlorinated analogue.     

Role of thermal dissipation in the photoprotection in cucumber plants after exposure to a chill stress

Experiments were carried out to investigate the changes in CO₂ assimilation, photon allocation, and photosynthetic electron flux in leaves of cucumber (Cucumis sativus L.) plants after chilling stress. Chilling significantly decreased CO₂ assimilation, the energy flux via linear electron transport (J _PS2) and non-constitutive thermal dissipation (J _NPQ) but increased fluorescence and constitutive thermal dissipation (J _f,D) in chilling-sensitive genotype Jinyan No. 4. In contrast, chilling had little effects on J _NPQ and J _f,D although CO₂ assimilation and J _PS2 were inhibited in chilling-tolerant genotype Jinchun No. 3. In parallel with the reduction in J _PS2, electron flux to oxygenation and carboxylation by ribulose-1,5-bisphosphate carboxylase/oxygenase all significantly decreased while electron flux to O₂ significantly increased, especially in chilling-sensitive genotype. Thermal and fluorescence dissipation were the main energy dissipation pathways whilst water-water cycle was an important electron sink when photosynthetic carbon reduction was suppressed after chilling. Chilling sensitivity of the photosynthetic apparatus was related to the operation of different photoprotection mechanisms.     

ATP involvement in plant tissue responses to low temperature

The effects of chilling and freezing temperature on membrane permeability and ATP content were studied in the leaves of cucumber ( Cucumis sativus L.) and winter rape ( Brassica napus L. var. oleifera L.) leaves, grown at different temperatures. In the winter rape leaves, the endogenous ATP content was modified by application of dinitrophenol (DNP) solutions of different concentrations. The low temperature-induced changes in membrane permeability (as monitored by the conductivity method) were found to be associated with ATP decrease, both in the chilling-sensitive and chilling-resistant (subjected to freezing) plants. In tissues showing reversible injuries, changes in ATP content preceded those in membrane permeability and the adenylate energy charge was affected slightly. In tissues showing irreversible membrane damage, the ATP content was always below 0.4 μmol (g dry weight)⁻¹ and the adenylate energy charge was near 0.5. DNP treatment increased freezing sensitivity of winter rape leaves. In the cold-hardened winter rape leaves, however, freezing and thawing did not significantly affect ATP content or the energy charge, although the specimen showed a rather large increase in membrane permeability. In these leaves ATP content recovered about 20 h after a freezing and thawing treatment. It is proposed that a decrease in ATP supply might be the primary reason for the membrane leakiness at low temperature, both in chilling-sensitive and chilling-resistant (subjected to freezing) plants. The conclusion is, however, not true for the cold-acclimated, frostadapted cells.     

Casein kinase activity in etiolated Cucumis sativus cotyledons

Two calcium- and light-dependent protein kinases have been reported in etiolated Cucumis sativus cotyledons (Vidal et al. 2007). In the present work, we studied casein kinase (CK) activity in etiolated cucumber cotyledons of in-gel and in vitro kinase assays, using specific CK inhibitors, and ATP and GTP as phosphate donors. Two proteins with CK activity were detected in both casein gels and autophosphorylation assays. One of them, with a molecular mass of approximately 36 kDa, showed biochemical CK1 characteristics: it was inhibited by specific CK1 inhibitors and only used ATP as phosphate donor. The second, with a molecular mass of approximately 38 kDa, had CK2 characteristics; it used both ATP and GTP as phosphate donors, was inhibited by all specific CK2 inhibitors, and was recognized by a polyclonal antibody directed against the α catalytic subunit of a CK2 from tobacco. The kinase activity of the CK2 detected in etiolated cucumber cotyledons showed circadian rhythmicity in both in vitro and in-gel casein phosphorylation and in autophosphorylation assays. Thus, our results suggest that the CK2 of approximately 38 kDa could be related to the circadian oscillator of C. sativus cotyledons.     

青海芨芨草属一新变种

与原变种的区别：花序紧缩；颖宽披针形，顶端较钝；基盘钝圆，长0.5mm；外稃顶端没有较长的毛。     

Accumulation of a 22-kDa protein and its mRNA in the leaves of Raphanus sativus in response to salt stress or water deficit

The response of 10-day-old seedlings of Raphanus sativus L. cv. Fakir to salt stress (100 m M to 200 m M NaCl) was investigated. Three weeks after initiation of salt treatment, the fresh weight of the shoots of salt-treated plants was half that of untreated plants. The salt stress resulted in the accumulation of Na⁺, preferably in the old leaves. The K⁺ level was reduced by as much as 50% in the old leaves of NaCl-treated plants, whereas this reduction was only 20–25% in the young leaves. Free proline accumulated in all aerial organs, and the highest levels were found in the young leaves. Patterns of total proteins extracted from the leaves of control or salt-treated plants were compared. The most obvious change concerned a 22-kDa, pl 7.5 polypeptide, which accumulated after exposure of the plants to NaCl. The appearance of this polypeptide was also mediated by a rapid drought stress, and sequencing indicated that it is related to the Künitz protease inhibitor family. A cDNA clone corresponding to the radish 22-kDa polypeptide was obtained and sequenced. Northern blot analysis showed that salt stress induces a large accumulation of this mRNA in the leaves of radish.     

TINY BRANCHED HAIR functions in multicellular trichome development through an ethylene pathway in Cucumis sativus L.

The fruit trichomes of Cucurbitaceae are widely desired in many Asian countries and have been a key determinant of cucumber (Cucumis sativus L.) cultivar selection for commercial production and breeding. However, our understanding of the initiation and development of cucumber trichomes is still limited. Here, we found that the cucumber TINY BRANCHED HAIR (TBH) gene is preferentially expressed in multicellular trichomes. Overexpression of CsTBH in tbh mutants restored the trichome phenotype and increased the percentage of female flowers, whereas silencing of CsTBH in wild-type plants resulted in stunted trichomes with a lower rate of female flowers. Furthermore, we provide evidence that CsTBH can directly bind to the promoters of cucumber 1-Aminocyclopropane-1-Carboxylate Synthase (CsACS) genes and regulate their expression, which affects multicellular trichome development, ethylene accumulation, and sex expression. Two cucumber acs mutants with different trichome morphology and sex morphs compared with their near-isogenic line further support our findings. Collectively, our study provides new information on the molecular mechanism of CsTBH in regulating multicellular trichome development and sex expression through an ethylene pathway.