A tryptophan decarboxylase from cucumber seedlings

The formation of tryptamine from tryptophan by extracts of cucumberhypocotyls is mediated by a tryptophan decarboxylase. The enzymerequires pyridoxal-5'-phosphate, and the pH optimum of thisenzyme is 7.0. The activity of the enzyme is inhibited by potassiumcyanide, but not by sodium azide or sodium fluoride; indicatingthat this enzyme is a decarboxylase rather than a peroxidase.The removal of contaminating epiphytic bacteria does not significantlyaffect the enzyme activity, and preincubation of enzyme extractsin streptomycin is also without effect. Neither aerobic noranaerobic cultures of internal bacteria which contaminate cucumberhypocotyb exhibit enzymic activity at pH 7.0. ¹Present address: P.O. Box 59 Prineville, Oregon 97754, U.S.A. (Received August 1, 1970; )     

Morphogenesis in cucumber seedlings is negatively controlled by gravity

 Seedlings of most cucurbitaceous plants develop a peg (protuberance caused by cell outgrowth) on the transition zone between the hypocotyl and root. The peg is necessary for removing the seed coat after germination. In our spaceflight experiments on the STS-95 space shuttle, Discovery, we found that cucumber (Cucumis sativus L.) seedlings grown under microgravity conditions developed two pegs symmetrically at the transition zone. Thus, cucumber seedlings potentially develop two pegs and do not require gravity for peg formation itself, but on the ground the development of one peg is suppressed in response to gravity. This may be considered as negative control of morphogenesis by gravity. Received: 17 August 1999 / Accepted: 4 October 1999     

Enhancement of stress tolerance in cucumber seedlings by proanthocyanidins

Proanthocyanidins (PAs) are the main products of the flavonoid biosynthetic pathway in many plants. However, their biological function during environmental stresses in plants is rarely reported. In the present study, the effects of pretreatment with PAs on the response of cucumber (Cucumis sativus L.) seedlings to high irradiance (HI), polyethylene glycol (PEG), and cold stress were investigated. The PAs pretreament alleviated stress-induced oxidative damage in plant cells and increased the activity of alternative oxidase (AOX) and content of abscisic acid (ABA). Furthermore, PAs-pretreated seedlings suffered less damage by the stress conditions, maintained higher content of chlorophyll a+b and AOX proteins in comparison with the control. Therefore, our findings suggest that PAs might contribute to plant tolerance to environmental stresses.     

Gravity-regulated formation of the peg in developing cucumber seedlings

It has been proposed that peg formation in the vascular transition region (TR zone) between the hypocotyl and the root in Cucurbitaceae seedlings is a gravimorphogenetic phenomenon. Initiation of the peg became visible 36 h after imbibition when cucumber (Cucumis sativus L. cv. Burpee Hybrid II) seeds were germinated in a horizontal position at 24°C in the dark. Simultaneously, sedimented amyloplasts (putative statoliths) were apparent in the sheath cells surrounding the vascular strands, and in the cortical cells immediately adjacent to them, in the TR zone. In contrast, the other cortical cells, some of which were destined to develop into the peg, contained amyloplasts which were not sedimented. These results suggest that the graviperception mechanism for peg formation may be like that of statoliths in shoot gravitropism. By 48 h following imbibition, the cells of the TR zone still had sedimented amyloplasts but had lost their sensitivity to gravity, possibly because of their maturation.     

Amelioration of chilling stress by triadimefon in cucumber seedlings

Cucumber (Cucumis satvus L.) seeds were imbibed in distilled water (control) and 10 mg l^–1 triadimefon (TDM) for 10 h and then grown in a plant growth chamber with a light/dark temperature of 28/20 °C and a photoperiod of 14 h with a light intensity of 60 µmol m^–2 s^–1. 14-day-old seedlings were exposed to chilling stress with a light/dark temperature of 6/3 °C for 4 d. TDM improved the growth rate of cucumber seedling subjected to chilling stress and increased photosynthetic pigments contents and relative water content compared with the control at the end of chilling stress. Chilling stress decreased protein content and the activities of SOD, CAT and POD, but it increased proline, H₂O₂ and MDA accumulation, and relative electrical conductivity. TDM ameliorated the injury caused by chilling stress by preventing decreases in protein content and the activities of SOD, CAT and POD and by inhibiting increases in proline, H₂O₂ and MDA contents, and relative electrical conductivity, which suggested that TDM ameliorated the negative effect of chilling stress.     

工厂化黄瓜穴盘育苗昼温适应性

在人工气候室内以黄瓜穴盘苗为材料,测定不同昼温处理下(昼温分别为30℃、27℃、24℃、21℃、18℃、15℃,夜温均为15℃)黄瓜幼苗下胚轴长、下胚轴粗、第一叶片和第二叶片的长和宽、地上部和地下部干物质积累量、叶片含水率及叶片的叶绿素荧光特性,并用主成分分析法和聚类分析法对不同昼温处理下的黄瓜穴盘苗质量进行分析.结果表明:不同昼温处理下黄瓜穴盘苗各生长指标存在显著性差异,幼苗质量的昼温反应表现为24℃＞21℃＞27℃＞30℃＞18℃＞15℃;通过主成分分析和系统聚类可以把各温度处理分为:最适温度处理(24℃/15℃)、适宜温度处理(21℃/15℃)和不适宜温度处理3类;不适宜温度处理又可分为高温抑制类(27C/15℃,30℃/15℃)和低温抑制类(15℃/15℃,18℃/15℃)2类.     

Auxin-mediated response of cucumber seedlings to gravity]

Gravity regulates peg formation because cucumber seedlings grown in a horizontal position develop a peg on the lower side of the transition zone (TR zone) but not on the upper side. Studies on peg formation have suggested the regulation of peg formation by gravity as follows. Cucumber seedlings potentially develop a peg on both the lower and upper sides of the TR zone. The development of the peg on upper side of the TR zone is suppressed in response to gravity. A phytohormone, auxin, induces peg formation. Upon gravistimulation the auxin concentration on the upper side of the TR zone is reduced to a level below the threshold value necessary for peg formation. The unequally distributed auxin across TR zone is caused by a change in accumulation of auxin influx carrier (CsAUX1) protein and auxin efflux carrier (CsPIN1) protein in response to gravity. In addition, TR zone before peg initiation expresses both CsARF2 (putative activator of auxin response factor) and CsIAA1 (putative repressor of auxin-inducible gene expression), by which TR zone could respond the auxin gradient regulated by gravity.     

Chilling sensitivity of cucumber cotyledon protoplasts and seedlings

Cucumber (Cucumis sativus L.) seedlings are more sensitive to chilling stress when transferred to low temperature from the night cycle than from the day cycle. However, greater damage occurs when chilling is carried out in light than in dark. Freshly isolated protoplasts are extremely sensitive to damage when chilled at 4°C in light, but suffer significantly less injury when chilled in dark. If freshly isolated protoplasts are pre-chill conditioned at 27°C in either light or dark for a few hours prior to exposure to various chilling stresses, subsequent chilling damage is markedly reduced. Damage to chilled protoplasts also is reduced if cultures are placed in dark instead of light immediately following removal from low temperature. Experiments utilizing the cell wall synthesis inhibitor, dichlorobenzonitrile, showed that cell wall regeneration during the pre-chill conditioning period at 27°C does not appear to be associated with the enhanced chilling tolerance observed in these cultures. The results obtained in this investigation suggest that the physiological properties of cucumber cotyledon protoplasts accurately reflect those of intact seedlings, and hence provide a good system for studies into the mechanism of chilling damage in plants.     

Auxin biogenesis: subcellular compartmentation of indoleacetaldehyde reductases in cucumber seedlings

Subcellular fractionation of cucumber (Cucumis sativus L.) seedlings was achieved, and two of the enzymes in the auxin biosynthetic pathway were localized. NADH-specific indoleacetaldehyde reductase activity was observed only in the cytosol fractions obtained from separated hypocotyl and cotyledon tissue. In contrast, a portion of the NADPH-specific indoleacetaldehyde reductase activity was associated with a microsomal fraction derived from these tissues. The NADPH-specific indoleacetaldehyde reductase was consistently found to be more firmly associated with the microsomal fraction derived from hypocotyls than with that from the cotyledons. These results indicate a division of the terminal steps of auxin biogenesis into at least two subcellular compartments.     

Effect of paclobutrazol and chilling on leaf membrane lipids in cucumber seedlings

The effects of paclobutrazol on the leaf membrane lipid composition of seedlings of cucumber ( Cucumis sativus L. cv. Victory) subjected to chilling temperatures were assessed. At a non-injurious temperature (12.5°C), there was no difference in the polar lipid fatty acid composition or in the glycolipid, phospholipid or free sterol content of leaves from treated vs untreated seedlings, regardless of whether paclobutrazol was administered 1 or 7 days prior to analysis. In the latter case (7 days pretreatment), there were clear effects of the bioregulator on plant growth and morphology as well as on leaf chlorophyll content. At an injurious chilling temperature (5°C), desaturation of leaf polar lipid fatty acids was markedly reduced in both treated and untreated seedlings. Chilling at 5°C resulted in losses of fresh weight and membrane lipids in leaves of both groups of plants. These losses were either reversible or irreversible, depending upon the duration of chilling and of pretreatment with paclobutrazol. Seedlings pretreated with 10 μg ml⁻¹ paclobutrazol generally sustained less chilling injury than untreated controls, as judged by the extent of wilting, necrosis and desiccation. This correlated with reduced losses of leaf fresh Weight and membrane lipids.     

Changes of polyamines and ethylene in cucumber seedlings in response to chilling stress

Cucumber ( Cucumis sativus L. cv. Victory) seedlings were exposed to chilling at 5°C and endogenous levels of polyamines and 1-aminocyclopropane-1-carboxylic acid (ACC) were measured after chilling and after warming at 20°C. The level of spermidine was higher in the chilled seedlings than in the non-chilled seedlings. Treatment with a plant bioregulator, (2RS,3RS)-1-(4-cholorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)pentan-3-ol (paclobutrazol), reduced the chilling injury and the levels of spermidine in the chilled seedlings. The levels of ACC and production of ethylene showed sharp increases after warming following exposure to chilling. These increases were suppressed by the application of aminooxyacetic acid (AOA). However, AOA treatment did not reduce chilling injury or affect the levels of polyamines in the tissue. These data indicate that the increase in ACC and ethylene is a response of the tissue to the chilling exposure and is not a cause of the injury. The data also suggest that the syntheses of polyamines and ethylene are not competitive with each other even under chilling stress conditions.     

Photophysiology and phytochrome content of long-hypocotyl mutant and wild-type cucumber seedlings

Photomorphogenetic responses have been studied in a cucumber (Cucumis sativus L.) mutant (lh), which has long hypocotyls in white light (WL). While etiolated seedlings of this mutant have a similar phytochrome content and control of hypocotyl elongation as wild type, deetiolation is retarded and WL-grown seedlings show reduced phytochrome control. Spectrophotometric measurements exhibit that WL-grown tissues of the lh mutant (flower petals and Norflurazon-bleached leaves) contain 35 to 50% of the phytochrome level in the wild type. We propose that this is a consequence of a lack of light-stable phytochrome, in agreement with our hypothesis proposed on the basis of physiological experiments. The lh mutant lacks an end-of-day far-red light response of hypocotyl elongation. This enables the end-of-day far-red light response, clearly shown by the wild type, to be ascribed to the phytochrome, deficient in the lh mutant. Growth experiments in continuous blue light (BL) and continuous BL + red light (RL) show that when RL is added to BL, hypocotyl growth remains inhibited in the wild type, whereas the lh mutant exhibits significant growth promotion compared to BL alone. It is proposed that the hypocotyls fail to grow long in low fluence rate BL because photosynthesis is insufficient to sustain growth.     

Transport and metabolism of h-gibberellin a(1) in dioecious cucumber seedlings

The transport of ³H-GA₁ through hypocotyl segments of cucumber (Cucumis sativus L.) was found to be nonpolar. The transport of ³H-GA₁ was increased by pretreatment with relatively high concentrations of either IAA or Ethephon (2-chloroethylphosphonic acid). Hypocotyl segments from plants of a gynoecious genotype transported more ³H-GA₁ than those of an androecious. The metabolism of ³H-GA₁ in hypocotyl segments was neither related to the sex genotype of the cucumber plant nor influenced by pretreatment with Ethephon. The primary metabolite of GA₁ was suggested to be GA₈. Two other suspected metabolites were not identified. Differences in the endogenous GA of gynoecious and androecious plants could not be accounted for by transport differences.