Endopeptidase Isoenzyme Characteristics in Cucumis sativus Leaves During Dark-induced Senescence

The changes and characteristics of endopeptidase （EP） isoenzymes in cucumber （Cucumis sativus L.） leaves during dark-induced senescence were investigated by activity staining after gradient-polyacrylamide gel electrophoresis （G-PAGE） containing co-polymerized gelatin as substrate. The results showed that both the chlorophyll and the protein contents of leaves were decreased, and the protein degradation was correlated with the increase of proteolytic activity during the course of leaf senescence. Meanwhile, nine cucumber endopeptidases isoenzymes （CEP） with 140, 120, 106, 94, 76, 55, 46, 39 and 35 kDa molecular weights were detected. Four of these, CEP2, 3, 4 and CEP9 appeared all the time, but the changes of the activity were different during incubation. Another four CEPs （CEP5, 6, 7 and CEP8） whose activities increased with dark-induced time were only detected in senescent leaves. Furthermore, the biochemical properties of these nine CEP were also characterized. All the CEPs had high activities from 35 ℃ to 45 ℃, and the optimum temperature was found to be 40 ℃. However, the activities of CEPs were not detected below 25 ℃ or over 60 ℃. The activity bands appeared at a wide range of pH from 5.0 to 9.0, but the optimum pH was found at 7.0. No CEPs were detected at pH 4 or pH 10. By inhibition analysis we concluded that CEP2, 3, 4 and CEP9 were serine endopeptidases and CEP6 was a kind of cysteine protease. It is suggested that serine endopeptidases might play a major role in cucumber leaf senescence, and for the first time, six senescencerelated endopeptidases （CEP1, 5, 6, 7, 8 and 9） were found in cucumber leaves.     

黄瓜雌花发育过程中柱头的腺特征(英文)

利用透射电镜技术研究了黄瓜（Ｃｕｃｕｍｉｓ　ｓａｔｉｖｕｓ　Ｌ．）雌花柱头发育过程中传递组织、分泌组织和乳突细胞的超微结构。在整个发育过程中,乳突细胞和分泌组织细胞的细胞质内密布很多管状及槽库膨大的内质网,产生很多分泌囊泡;在成熟柱头的传递组织和分泌组织细胞间观察到大量的胞间连丝;乳突细胞和分泌细胞高度液泡化,质膜内折;在柱头发育过程中分泌组织细胞的核周腔扩大形成裂瓣状核,到柱头成熟阶段裂瓣状核更加明显。进一步的研究显示,在成熟柱头的不同组织细胞中,　ＡＴＰａｓｅ的活性呈现在质膜和液泡膜上,随着柱头的发育,ＰＭ－Ｈ＋－ＡＴＰａｓｅ的比活性明显增强。结果表明,黄瓜雌花柱头的腺特征随发育进程而趋于显著。     

黄瓜雌花发育过程中柱头的腺特征

利用透射电镜技术研究了黄瓜(Cucumis sativus L.)雌花柱头发育过程中传递组织、分泌组织和乳突细胞的超微结构.在整个发育过程中,乳突细胞和分泌组织细胞的细胞质内密布很多管状及槽库膨大的内质网,产生很多分泌囊泡;在成熟柱头的传递组织和分泌组织细胞间观察到大量的胞间连丝;乳突细胞和分泌细胞高度液泡化,质膜内折;在柱头发育过程中分泌组织细胞的核周腔扩大形成裂瓣状核,到柱头成熟阶段裂瓣状核更加明显.进一步的研究显示,在成熟柱头的不同组织细胞中,ATPase的活性呈现在质膜和液泡膜上,随着柱头的发育,PM-H -ATPase的比活性明显增强.结果表明,黄瓜雌花柱头的腺特征随发育进程而趋于显著.     

不同倍性黄瓜的形态和一些生理生化指标比较

由同一基因型产生的单倍体、二倍体和四倍体黄瓜的叶面积和花大小等形态性状、单位面积叶绿素含量和苹果酸脱氢酶(MDH)表达量等生理生化指标随染色体倍性的增加而增加,过氧化物酶(POD)活性则随倍性的增加而降低,剂量效应明显;其他形态指标和可溶性蛋白含量等与染色体倍性无明显相关性.     

离体培养黄瓜子叶诱导雌花的研究

采用添加Spd和IAA的MS培养基培养离体黄瓜子叶,研究了Spd和IAA对雌花诱导的协同作用,及昼夜温差、培养基中N素和pH值对雌花诱导的影响。结果表明,分别添加Spd、IAA时的雌花诱导率和单株雌花数偏低或为0,12 mg·L^-1 Spd与0.01mg·L^-1 IAA 配合时的诱导效果明显高于单独处理的,而对照组未见雌花,说明Spd和IAA对雌花诱导的协同作用显著。在0、2、6、10℃昼夜温差,60、70、80、90 mmol·L^-1的N素含量和pH 5.4、5.8、6.2、6.6的培养条件下,70 mmol·L^-1 N、6℃温差和pH 6.2时的雌花诱导效果较好,表明适当提高昼夜温差、培养基中N素和pH值有利于黄瓜子叶的雌花诱导。     

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.     

Development of EST‐SSRs in Cucumis sativus from sequence database

Simple sequence repeat markers derived from expressed sequence tags (EST‐SSR) are potentially valuable tools for plant breeding and germplasm collection conservation, and increasingly, efforts have been made for developing this type of marker. We have identified 20 polymorphic SSR markers from cucumber ESTs deposited in public sequence database. The average allele number was 3.3 per locus, ranging from two to six alleles during screening 20 cucumber genotypes with the mean expected heterozygosity of 0.477. Amplification products were also detected by 13 pairs of primer in Cucumis melo. These informative EST‐SSR markers can be used in cucumber genetic improvement projects.     

多裂骆驼蓬的生物碱对黄瓜种子萌发的影响

经多裂骆驼蓬中生物碱处理过的黄瓜种子吸水量、呼吸速率、α-淀粉酶活性、GA1_( 3)和ATP含量均下降,种子膜透性增大,丙二醛含量提高;子叶中异柠檬酸裂解酶和羟基丙酮酸还原酶活性以及超弱发光(UWL)值也受到明显抑制。其抑制或促进作用的程度均随着多裂骆驼蓬中生物碱浓度的提高而增强。     

NaCl胁迫对黄瓜根系边缘细胞发生的影响

以黄瓜为试材,研究了NaCl处理对植株生物量、根长、根系活力、根边缘细胞的数目和活性及黏胶层厚度的影响。结果表明,NaCl处理降低了植株生物量与根系长度,增加了黄瓜幼苗的根系活力。黄瓜边缘细胞的出现几乎与根同时发生,当根长达到25mm时,边缘细胞的数目与活性均达到最大值。NaCl处理对边缘细胞的数目与活性表现出一定的抑制作用。离体根尖的边缘细胞活性也随NaCl处理浓度与处理时间的增加而逐渐减小,但根边缘细胞黏胶层厚度却随NaCl处理浓度的增加而增加。总之,NaCl对黄瓜幼苗造成一定伤害,但根系边缘细胞可通过降解死亡与增加黏胶的分泌量在一定程度上减轻这种伤害程度。     

Immunolocalization of Arabinogalactan Proteins and Pectins in Floral Buds of Cucumber (Cucumis sativus L.) During Sex Determination

Arabinogalactan proteins (AGPs) and pectins were detected in the floral buds of cucumber (Cucumis sativus L.) during its sex determination using the following monoclonal antibodies: MAC 207 (recognizes AGP epitopes); JIM 8 (recognizes a subset ofAGP epitopes); and JIM 5 and JIM 7 (epitopes of pectins esterified to various degrees). In the stem apex meristem (SAM) of the cucumber, epitopes of MAC 207, JIM 7, and JIM 5 were localized in the cells from second to third peripheral layers when the sex organ primodium began to differentiate; epitopes of MAC 207 and JIM 5 were also detected in the ragged edge cells. A very dense labeling signal with MAC 207 was observed in the carpel and pistil primodium. The AGP epitopes recognized by JIM 8 were localized in the anther of the male flower and the anther-like portion of the stagnant stamen of the female flower. This suggests that the AGPs and pectins in the SAM of the cucumber are closely associated with the differentiation of the SAM, from meristematic cells to floral primodium. The subset of AGPs recognized by JIM 8 may play an important role in stamen formation.     

Immunolocalization of Arabinogalactan Proteins and Pectins in Floral Buds of Cucumber （Cucumis sativus L.） During Sex Determination

Arabinogalactan proteins (AGPs) and pectins were detected in the floral buds of cucumber(Cucumis sativus L.) during its sex determination using the following monoclonal antibodies: MAC 207(recognizes AGP epitopes); JIM 8 (recognizes a subset ofAGP epitopes); and JIM 5 and JIM 7 (epitopes of pectins esterified to various degrees). In the stem apex meristem (SAM) of the cucumber, epitopes of MAC 207, JIM 7, and JIM 5 were localized in the cells from second to third peripheral layers when the sex organ primodium began to differentiate; epitopes of MAC 207 and JIM 5 were also detected in the ragged edge cells. A very dense labeling signal with MAC 207 was observed in the carpel and pistil primodium. The AGP epitopes recognized by JIM 8 were localized in the anther of the male flower and the anther-like portion of the stagnant stamen of the female flower. This suggests that the AGPs and pectins in the SAM of the cucumber are closely associated with the differentiation of the SAM, from meristematic cells to floral primodium. The subset of AGPs recognized by JIM 8 may play an important role in stamen formation.     

Effect of benzoic acid hydroxyl- and methoxy-ring substituents on cucumber (Cucumis sativus L.) root membrane potential

Abstract

The allelopathic effect of some benzoic acid (BA) OH- and OCH₃-ring substituents was studied on cucumber root transmembrane potential difference (Vm). Most of the methoxy-BAs induced a rapid Vm depolarization, followed by a Vm hyperpolarization, with the only exception for p-anisic acid (pA). On the other hand, salicylic acid (SA) and 3,4-dimethoxybenzoic acid (DHB) strongly depolarized Vm. A positive correlation was found between Vm hyperpolarization and lipophilicity of methoxylated BAs, whereas a positive correlation was found between lipophilicity and Vm depolarization of hydroxylated BAs. The influence of BAs on K⁺ was studied by means of specific blocking with Cs⁺ indicating a possible direct interaction of SA, gallic acid (GA), vanillic acid (VA) and 3,4-dimethoxybenzoic acid (DMB). Interference of BAs with the Vm hyperpolarizing effect of root perfusion with the fungal toxin fusicoccin were also observed.     

黄瓜的冷害及耐冷性

随着蔬菜反季节栽培面积的不断扩大,如何提高黄瓜(Cucumis sativus L.)耐冷性已成为选育新品种的研究重点.系统地综述近几年黄瓜耐冷性的鉴定、获得途径、冷害机理以及遗传和分子遗传学等方面的研究,以促进对黄瓜冷害机制的研究,加速耐冷品种的培育.耐冷性鉴定时要从耐冷指数、低温发芽能力、MDA(丙二醛)含量和电解质渗漏率等几个方面综合鉴定.耐冷性的获得途径主要有冷驯化、激素处理、热激处理和培育耐低温品种,最重要的途径是耐冷品种选育.黄瓜冷害机理包括细胞膜的流动性降低及透性增加,光合作用被抑制,根系吸收减弱,可溶性糖含量减少,淀粉粒积累增加,微管的稳定性受到破坏等.黄瓜低温发芽能力由非加性基因决定,而幼苗时期主要由加性基因控制.黄瓜耐冷的分子遗传学研究进展缓慢,目前已克隆出在低温锻炼中特异表达的功能未知的基因CCRl8.今后还应研究黄瓜低温胁迫时的信号转导系统,以进一步揭示黄瓜的冷害机理;利用野生资源的抗逆性状,拓宽栽培黄瓜的遗传基础,选育适于保护地栽培的耐低温品种.     

Induction of chilling tolerance in cucumber (Cucumis sativus) seedlings by endogenous and applied ethanol

Five-day-old etiolated cucumber ( Cucumis sativus L.) seedlings cv. Marketmore held at 2°C for 72 h developed chilling injury, resulting in desiccation and collapse of the hypocotyl tissue and eventual plant death. Hypoxia-induced accumulation of ethanol and acetaldehyde led to tolerance of subsequent chilling, as evidenced by continued hypocotyl growth and freedom from injury. Attenuated accumulation of volatiles by applied bisulfite reduced the development of hypoxia-induced chilling tolerance in seedlings. In seedlings held in normoxia cold tolerance was induced by applied ethanol vapors, whereas acetaldehyde had a marginal effect, suggesting that hypoxia-induced cold tolerance may arise from the accumulation and activity of ethanol. Cold tolerance was also induced by exposure of seedlings to volatile anesthetics including n -propanol, n -butanol, chloroform and halothane, suggesting that ethanol activity may result from fluidization of membrane lipids. This view is consistent with results which showed that ethanol activity was not associated with lipid metabolism. However, development of cold tolerance in ethanol-enriched tissues was time dependent, indicating that ethanol activity probably also entails biosynthetic event(s).     

光合菌对黄瓜光合及抗氧化同工酶的影响

以2种基因型的黄瓜（Cucumis sativus）为材料,研究光合菌（PSB）的喷施对植物生物量、净光合速率（Pn）、叶片PSII的最大光化学效率（Fv／Fm）及抗氧化同工酶代谢的影响。结果表明,喷施PSB均能诱导2种基因型黄瓜的生物量显著增加,并伴随Pn的显著提高。但是,2种基因型黄瓜的Fv/Fm并不受PSB喷施的影响：PSB能使总过氧化物歧化酶（soo）和抗坏血酸过氧化酶（APX）活性提高,并使它们的多数同工酶的活性上调,这些同工酶活性的增加在叶绿体中表现更为明显（如Cu／Zn-SOD、Fe-SOD和sAPX）。研究结果表明,PSB能通过增强黄瓜抗氧化酶体系的活性改善植株的抗氧化能力,从而在植株的生长和光合作用方面起到促进作用。     

Correlation of iron content, spectral forms of chlorophyll and chlorophyll-proteins in iron deficient cucumber (Cucumis sativus)

Leaves and chloroplast suspensions of severely and slightly iron deficient cucumber ( Cucumis sativus L.) plants were characterized by low-temperature fluorescence emission spectroscopy and Deriphat polyacrylamide gel electrophoresis. The emission spectra of the chloroplast suspensions were resolved into Gaussian components and those changes induced by iron deficiency were related to the variations in the chlorophyll-protein pattern. The symptoms described with these methods were also correlated with the iron content of the leaves. It was concluded that the lack of physiologically active iron caused a relative decrease of photosystem I (PSI) and light harvesting complex I (LHCI), together with the long wavelength fluorescence, especially the 740 nm Gaussian component, and. to a much lesser extent, of the photosystem II (PSII) core complexes (relative increase of 685, 695 nm components). However, the relative decrease in the amount of light harvesting complex II (LHCII) was followed by a relative increase in its fluorescence band at 680 nm, showing that energy transfer from LHCII to core complex II (CCII) was partly disturbed. Thus iron deficiency affected the photosynthetic apparatus in a complex way: it decreased the synthesis of chlorophylls (Chls) and influenced the expression and assembly of Chl-binding proteins.     

黄瓜花原基的形成及与Ca~(2+)的关系

以黄瓜为试验材料研究了肉眼可见的花原基突起之前 ,花原基早期的分化过程。结果显示花原基分化和开花的起始节位是第一真叶节 ;肉眼可见花原基突起前早期的分化是在叶腋亚表皮部位形成一个球形的花原基起始细胞团 ,此细胞团进一步分裂、扩大形成肉眼可见的花原基突起 ;第一真叶节的花原基起始细胞团分化集中发生于 6～ 7d苗龄时期 ;Ca2 + 在花原基起始细胞团细胞中主要分布在细胞壁和细胞间隙 ,而在非起始细胞团的叶腋亚表皮细胞则主要分布在液泡中 ,并对Ca2 + 在花原基起始细胞团分化中的作用进行了讨论。     

Leaf Growth,Photosynthetic Rate,and Phytohormone Contents in Cucumis sativus Plants under Progressive Soil Drought

Effects of progressive soil drought on leaf growth, the rate of photosynthesis, and phytohormone contents were followed in the experiments with cucumber (Cucumis sativus L.) plants. Suppression of photosynthesis by drought did not immediately cause growth retardation, because the latter was observed one day earlier than the inhibition of photosynthesis. In the meantime, growth retardation could be caused by a decline in IAA and cytokinin contents, rather than ABA accumulation, because ABA accumulated when the growth has been already suppressed.     

Cucumis sativus and Ipomoea purpurea: New hosts for Alternaria hydrangeae in China

Alternaria leaf spot is a common disease on various plants worldwide. In this study, an Alternaria species, A. hydrangeae causing leaf spot on Cucumis sativus (cucumber) and Ipomoea purpurea in China was identified based on morphology and multi-locus analysis of the partial ITS, GAPDH, Alt a 1, TEF1, and RPB2 gene regions. The pathogenicity of the present isolates and a representative isolate of A. hydrangeae were assessed on living leaves of C. sativus, I. purpurea and Hydrangea paniculata. Similar symptoms were observed on every plant inoculated with the isolates. The same fungus was re-isolated from inoculated leaves fulfilling Koch's postulates. Morphology of the original and re-isolated isolates showed characteristics similar to A. hydrangeae with some variations. Multi-locus analysis indicated that the present isolate fell into A. hydrangeae clade. This study is the first report of A. hydrangeae as an agent of Alternaria leaf spot in C. sativus and I. purpurea in China, which extends the host range of the fungus.     

Influence of bicarbonate and metal ions on the development of root Fe(III) reducing capacity by Fe-deficient cucumber (Cucumis sativus) plants

The effect of bicarbonate and selected metal ions on the development of enhanced root Fe(III) reducing capacity (a response to Fe deficiency of dicotyledons) was studied in young plants of cucumber ( Cucumis sativus L. cv. Ashley) grown in nutrient solution. Pretreatment of 11-day-old Fe-deficient cucumber plants with 20 m M NaHCO₃, for at least 23 h prior to determination of root Fe(III) reducing capacity, markedly inhibited this response. The inhibitory effect of bicarbonate could be partly reversed by a 4- to 8-h treatment with either 10 μ M MnSO₄, 10 μ M FeEDDHA, 2 μ M ZnSO₄, 0.5 μ M NiCl₂, or 0.25 μ M , or CoSO₄ (final concentrations), added to the nutrient solution. By contrast, the addition of other salts of metal ions, like CuSO₄ and Cd(NO₃)₂, at 0.25, 0.5 or 1 μ M , or MgSO₄, at 0.5, 1 or 2 m M (final concentrations), had no beneficial effect. The results suggest that bicarbonate may inhibit the development of root Fe(III) reducing capacity by diminishing the availability of certain metal ions required for this response.