外源腐胺对根际低氧胁迫下黄瓜幼苗多胺和抗氧化系统的影响

采用营养液栽培,研究了外源腐胺(Put)对根际低氧胁迫下黄瓜幼苗体内多胺含量和抗氧化系统的影响.结果显示,低氧胁迫显著刺激了黄瓜幼苗体内活性氧(ROS)和内源多胺含量的增加,提高了抗氧化酶活性;外源Put进一步提高了低氧胁迫下黄瓜幼苗体内多胺的含量和抗氧化酶活性,降低了ROS含量,从而缓解了低氧胁迫的伤害作用;Put合成抑制剂D-精氨酸(D-Arg)不仅显著抑制黄瓜幼苗体内多胺的合成,而且抑制抗氧化酶活性,同时ROS大量积累,进一步抑制黄瓜幼苗的生长;而外源Put可缓解D-Arg的抑制作用;Put转化抑制剂甲基乙二醛-双(脒基腙)(MGBG)和Put降解抑制剂氨基胍(AG)的混合施用造成游离态Put的过量积累,以及亚精胺(Spd)、精胺(Spm)含量和抗氧化酶活性的显著降低,造成ROS大量积累,进一步加重了低氧胁迫对植株的伤害.结果表明,低氧胁迫下外源Put可提高黄瓜幼苗体内游离态Put含量,促进游离态Put向Spd和Spm转化,Spd、Spm含量的增加以及(free-Spd free-Spm)/free-Put比值的升高有利于提高植株抗氧化酶活性,增强清除ROS的能力,降低膜脂过氧化的伤害,从而增强植株的低氧胁迫耐性.     

低氧胁迫下24-表油菜素内酯对黄瓜幼苗叶片光合特性及多胺含量的影响

采用1/2 Hoagland营养液培养,研究了低氧胁迫下24-表油菜素内酯(EBR)对黄瓜幼苗叶片光合特性及多胺含量的影响.结果表明：低氧胁迫下黄瓜幼苗的净光合速率(P_n)、气孔导度(g_s)、蒸腾速率(T_r)、胞间CO₂浓度(C_i)显著下降,而叶绿素含量显著提高,幼苗生长受抑;低氧胁迫显著提高了黄瓜幼苗叶片的腐胺(Put)、亚精胺(Spd)、精胺(Spm)、多胺(PAs)含量和Put/PAs,但降低了(Spd+Spm) /Put.低氧胁迫下,外源EBR不仅显著提高了黄瓜幼苗的P_n、g_s、T_r及叶绿素含量,也显著提高了黄瓜幼苗叶片的游离态Spm、结合态Spd、Spm及束缚态Put、Spd、Spm含量,促进了PAs的进一步积累,且降低了Put/PAs,提高了(Spd+Spm)/Put．可见,外源EBR调节了黄瓜幼苗内源多胺含量及形态的变化,维持了较高的光合性能,促进了叶面积和干物质量的显著增加,缓解了低氧胁迫对黄瓜幼苗的伤害.     

NaCl胁迫下大麦根系液泡膜H+-ATP酶活性受ATP和焦磷酸含量的调节

选用两个耐盐性强弱不同的大麦(Hordeumvulgare L.)品种,研究了NaCl胁迫下其幼苗根中ATP和焦磷酸(PPi)含量的变化以及PPi对液泡膜H -ATP酶活性的影响.结果表明:在含NaCl 200mmol/L的1/2 Hoagland溶液中处理2 d,耐盐品种(滩引2号)根中液泡膜H -ATP酶活性增加,然后逐渐下降,而H -PPi酶活性在NaCl处理9 d中'直下降.盐敏感品种(科品7号)在NaCl胁迫下根中H -ATP酶和H -PPi酶活性都下降(图1).与对照相比较,NaCl胁迫下耐盐品种根中ATP含量2 d时增加,4 d后下降;盐敏感品种根中ATP积累受NaCl胁迫的抑制(图2).NaCl胁迫下,两品种的PPi含量皆略有增加(图3).PPi对液泡膜H -ATP酶活性有竞争性抑制作用(图4).结果表明:ATP积累是NaCl胁迫下液泡膜H -ATP酶活性增加的原因之一,NaCl胁迫下大麦品种根中ATP含量下降和PPi对液泡膜H -ATP酶的抑制使该酶活性下降.     

外源亚精胺对盐胁迫下黄瓜幼苗游离态多胺含量和多胺合成酶基因表达的影响

以黄瓜品种‘津春2号’（Cucumis sativusL.cv.Jinchun No.2）为材料,采用营养液栽培,研究了外源亚精胺（Spd）对NaCl胁迫下黄瓜幼苗叶片游离态多胺含量和多胺合成酶基因表达的影响。结果表明,75 mmol/LNaCl胁迫下,幼苗株高、茎粗和干鲜重显著降低,外源喷施1 mmol/L Spd处理可明显缓解盐胁迫对幼苗生长的抑制。盐胁迫下叶片游离态多胺含量显著增加,外源Spd进一步促进了游离态Spd和精胺（Spm）的积累,降低了游离态腐胺（Put）的积累。多胺合成酶基因表达分析表明,盐胁迫上调了adc、odc、samdc和spds基因的表达,施用外源Spd后进一步上调了samdc基因,下调了adc、odc、spds基因的表达。表明外源Spd参与了黄瓜幼苗体内多胺代谢的调节,通过下调盐胁迫下adc、odc基因的表达,抑制游离态Put的积累,上调samdc基因的表达促进游离态Spd和Spm的积累,进而缓解盐胁迫对植物生长的抑制。     

枣品种耐盐性与叶片生物膜结合态多胺水平和H~＋-ATP酶活性的关系

120mmol·L^-1NaCl胁迫30d,耐盐性强的‘金丝小枣’叶片细胞质膜、液泡膜共价结合态腐胺（Put）、亚精胺（Spd）、精胺（Spm）含量及多胺（PAs）总水平与对照无显著性差异,但耐盐性弱的‘冬枣’叶片质膜共价结合态Put、Spd、Spm含量和PAs总水平及液泡膜Spd含量均显著降低;‘金丝小枣’叶片类囊体膜共价结合态Put含量、PAs总水平较对照显著降低,‘冬枣’则是Put、Spd、Spm含量及PAs总水平均显著降低。盐胁迫下,‘金丝小枣’叶片细胞质膜、液泡膜、类囊体膜非共价结合态Put、Spd、Spm含量及PAs总水平下降,但其中仅类囊体膜Spd含量显著低于对照,而‘冬枣’的3种膜上非共价结合态的这些多胺及其总水平均显著低于对照。与对照相比,盐胁迫下耐盐性不同的2个枣品种,叶片细胞质膜、液泡膜和类囊体膜H＋-ATP酶活性均降低,但降低幅度因枣品种和生物膜种类不同而异,且H＋-ATP酶活性与相应膜结合态多胺水平存在极紧密的正相关关系。结果表明,膜结合态多胺参与枣品种耐盐性的表达,调节盐胁迫下枣叶细胞中溶质的跨膜运输。     

硅对铵态氮胁迫下黄瓜幼苗生理特性的影响

在水培条件下,研究了外源硅对铵态氮胁迫下黄瓜幼苗生长及叶片生理特性的影响.结果表明:与硝态氮处理相比,铵态氮处理显著抑制了黄瓜幼苗地上部及根系的生长,尤其是地上部生长,在处理10 d时,铵态氮处理黄瓜单株地上部鲜质量降低6.17 g.铵态氮处理还促进了活性氧在黄瓜植株体内的积累,叶片O-·2和H2O2含量显著增加.外源硅处理可显著提高黄瓜叶片超氧化物歧化酶、过氧化物酶、过氧化氢酶、抗坏血酸过氧化物酶等抗氧化酶活性,增强清除活性氧的能力,显著降低黄瓜叶片O2-·和H2O2含量,从而减轻了活性氧对细胞膜的破坏,使黄瓜叶片电解质渗漏率及丙二醛含量降低;外源硅处理显著提高了黄瓜质膜及液泡膜H+-ATP的活性,提高细胞内外质子的运输能力;显著降低了黄瓜体内铵态氮含量,从而减轻了铵态氮的毒害.总之,外加一定浓度硅,可通过提高黄瓜抗氧化酶活性、质膜及液泡膜H+-ATP的活性以及降低植株铵态氮含量等来缓解铵态氮胁迫.     

外源亚精胺对根际低氧胁迫下黄瓜幼苗根系多胺含量和抗氧化酶活性的影响

采用营养液水培,研究了根际低氧胁迫下外源亚精胺对两个抗低氧能力不同的黄瓜(Cucumis sativus)品种(‘中农8号’和‘绿霸春4号’)根系中多胺含量和超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性的影响。结果表明,外源亚精胺能显著提高低氧胁迫下黄瓜幼苗根系中亚精胺和精胺含量,降低腐胺含量,同时,根系中SOD、POD、CAT活性也相应提高,使得幼苗鲜重和干重明显增加;单纯低氧处理及外源亚精胺的加入,抗低氧能力较强的品种‘绿霸春4号’较抗低氧能力较弱的品种‘中农8号’根系中抗氧化酶活性高。黄瓜幼苗根系中较高的亚精胺、精胺含量和较低的腐胺含量可能有利于提高抗氧化酶活性,增强幼苗的低氧逆境适应能力。     

钙对低氧胁迫下黄瓜幼苗体内多胺及多胺氧化酶的影响

以‘中农8号’黄瓜品种为实验材料,采用营养液栽培法研究了钙对根际低氧胁迫下黄瓜幼苗体内多胺(PAs)含量及多胺氧化酶(PAO)活性的影响。结果表明:(1)各处理黄瓜幼苗根系和叶片中的PAs含量以及3种形态的腐胺(Put)、亚精胺(Spd)、精胺(Spm)含量均表现为低氧高钙(8 mmol.L-1Ca2 )>低氧常钙(2 mmol.L-1Ca2 )>低氧缺钙(0 mmol.L-1Ca2 )>通气常钙(2 mmol.L-1Ca2 )处理,而PAO活性却表现出相反的趋势(通气常钙>低氧缺钙>低氧常钙>低氧高钙),且处理间大多存在显著差异(P<0.05);根系中的PAs含量明显高于叶片,而PAO活性明显低于叶片。(2)黄瓜幼苗体内3种形态的PAs以游离态含量最高,其次是结合态,最低为束缚态;游离态和结合态PAs在叶片中均以Spd为主,在根系中均以Put为主,束缚态PAs含量在根系和叶片中均为Spd>Put>Spm。研究表明,在低氧胁迫下,营养液加钙引起黄瓜幼苗体内多胺含量的上升和PAO活性下降,钙参与了黄瓜幼苗体内多胺的代谢过程,对缓解低氧胁迫有重要作用。     

一氧化氮调节盐胁迫下小麦幼苗根部质膜H+-ATPase和焦磷酸酶活性提高耐盐性

采用外源一氧化氮(NO)供体硝普钠(SNP)研究了NO对盐胁迫下小麦(Triticum aestivum L.)幼苗耐盐性的影响.结果表明,0.1 mmol/L SNP处理显著缓解了1 50 mmol/L NaCl胁迫对小麦幼苗生长的抑制效应,包括水分丧失以及叶绿素降解,从而提高了小麦幼苗的耐盐性.进一步结合1 mg/mL血红蛋白处理则显著逆转了SNP诱导的上述效应;利用亚硝酸钠和铁氰化钾作为对照也证实了NO对小麦幼苗耐盐性的专一性调节作用,并可能与NO对小麦幼苗根部质膜H -ATPase和焦磷酸酶活性诱导有关.此外,尽管NO显著提高了盐胁迫下小麦幼苗根部细胞质膜H -ATPase和焦磷酸酶的ATP水解活性,但是对跨膜H 转运则没有明显影响.应用外源CaSO4和EGTA处理也证实,Ca2 可能在NO诱导的质膜H -ATPase和焦磷酸酶活性的提高过程中起信号作用.另外,分析盐胁迫下小麦幼苗根部Na 和K 含量的变化也发现,NO对Na 含量没有明显影响,但是却显著提高了K 水平和K /Na 比,这可能也是NO提高小麦幼苗耐盐性的原因之一.     

多胺浸种改善盐胁迫大麦根系液泡膜功能的机理

研究了 0 .1mmol/L腐胺 (Put)和 0 .5mmol/L亚精胺 (Spd)浸种对 2 0 0mmol/LNaCl胁迫下大麦 (HordeumvulgareL .)幼苗生长速率、干物质积累、离子分布、液泡膜蛋白结合多胺含量以及液泡膜膜脂组分与功能的影响。结果表明 ,Put和Spd浸种均可缓解盐胁迫对大麦幼苗的盐害 ,促进生长和干物质积累 ,降低大麦幼苗体内 [Na ]/[K ]。与盐处理的对照植株相比 ,Put和Spd浸种均可提高大麦幼苗根系液泡膜磷脂含量 ,降低糖脂结合半乳糖含量 ,而膜上非共价结合多胺含量Spd PAx (一种未知多胺 )与Put Dap (二氨基丙烷 )之比 ( (Spd PAx) / (Put Dap) )、共价和非共价结合多胺总量均上升。统计分析结果表明 ,液泡膜非共价结合多胺 (Spd PAx) / (Put Dap)与H _ATPase和H _PPase活性呈显著正相关关系。     

Effects of Calcium on Absorption and Distribution of Ions and H+-ATPase Activity in Barley and Wheat Under Salt Stress

Calcium decreased Na+ absorption and transportation to the shoots,increased K+ and Ca2+ absorption and transportation ,decreased the leakage of electrolyties,and increased the accumulation of dry matter in barley and wheat seedlings under NaC1 stress. Calcium ion promoted the H+-ATPase activities in plasma membrane and tonoplast vesicles isolated from the young roots of the two plants, and increased respiration of the roots. This is in consistent with the results that calcium regulates ion absorption and distribution via its enhancement of H+-ATPase activities in plasma membrane and tonoplast.     

等渗盐胁迫对番茄抗氧化酶和ATP酶及焦磷酸酶活性的影响

用Ca(NO3)2 80 mmol/L和NaCl 120 mmol/L等渗溶液处理番茄幼苗后,细胞质和叶绿体中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)的活性升高,并且NaCl胁迫的作用明显高于Ca(NO3)2胁迫.Ca(NO3)2处理提高了线粒体中SOD、CAT、APX的活性,而NaCl处理降低了它们的活性.根系质膜H -ATPase、液泡膜H -ATPase、焦磷酸酶(H -PPase)的活性和叶片丙二醛(MDA)及脯氨酸含量在两种盐胁迫后明显增加.NaCl处理对植株生长的抑制程度明显高于Ca(NO3)2处理.     

Increased Polyamines Conjugated to Tonoplast Vesicles Correlate with Maintenance of the H<Superscript>+</Superscript>-ATPase and H<Superscript>+</Superscript>-PPase Activities and Enhanced Osmotic Stress Tolerance in Wheat

The effects of osmotic stress on H⁺-ATPase and H⁺-PPase activities and the levels of covalently conjugated polyamines (CC-PAs) and noncovalently conjugated polyamines (NCC-PAs) were investigated using tonoplast vesicles isolated from the roots of wheat (Triticum aestivum L.) seedlings differing in drought-tolerance. The results showed that after polyethylene glycol (PEG) 6,000 (–0.55MPa) treatment for 7 days, seedling leaf relative water content (LRWC), relative dry weight increase rate (RDWIR) and root H⁺-ATPase and H⁺-PPase activities from the drought-sensitive cultivar Yangmai No. 9 decreased more markedly than those from the drought-tolerant cultivar Yumai No. 18. At the same time, the increase of the NCC-spermidine (NCC-Spd) and CC-putrescine (CC-Put) levels in root tonoplast vesicles from Yumai No. 18 was more obvious than that from Yangmai No. 9. Exogenous Spd treatment alleviated osmotic stress injury to Yangmai No. 9 seedlings, coupled with marked increases of tonoplast NCC-Spd levels and H⁺-ATPase and H⁺-PPase activities. Treatments with methylglyoxyl bis (guanyl hydrazone) (MGBG), an inhibitor of S-adenosylmethionine decarboxylase (SAMDC), and phenanthrolin, an inhibitor of transglutaminase (TGase), significantly inhibited the osmotically induced increases of NCC-Spd and CC-Put levels, respectively, in root tonoplast vesicles from Yumai No. 18 seedlings. Both MGBG and phenanthrolin treatments markedly promoted osmotically induced decreases of tonoplast H⁺-ATPase and H⁺-PPase activities and osmotic stress tolerance of seedlings of this cultivar. These results suggest that the NCC-Spd and CC-Put present in tonoplast vesicles isolated from wheat seedling roots might enhance the adaptation of seedlings to osmotic stress via maintenance of tonoplast H⁺-ATPase and H⁺-PPase activities.     

Visualization by Freeze-Fracture Electron Microscopy of Intramembraneous Particles corresponding to the Tonoplast H+-Pyrophosphatase and H+-ATPase of Kalanchoë daigremontiana Hamet et Perrier de la Bâthie*

The H⁺-PPase and the H⁺-ATPase of the vacuolar membrane were separated during purification of tonoplast proteins of Kalanchoë daigremontiana Hamet et Perrier de la Bǎthie. Three membrane protein fractions prepared contained firstly, the H⁺-PPase protein without any subunits of the H⁺-ATPase, secondly, the H⁺-PPase protein with only minute traces of the intramembraneous 16 kDa c-subunit of the H⁺-ATPase, and thirdly, the H⁺-ATPase subunits without H⁺-PPase peptides as verified by SDS-PAGE. These three preparations were reconstituted into soybean (Glycine max L.)-phospholipid vesicles, and compared with proteoliposomes obtained by reconstitution of total solubilized tonoplast proteins as well as with native tonoplast vesicles. Analysis of freeze-fracture replicas prepared from these five different types of vesicles showed that there are two populations of intramembraneous particles, one with a diameter of 6.7-7.2 nm corresponding to the H⁺-PPase, and one with an average diameter of 9.1 nm belonging to the H⁺-ATPase. Thus, freeze-fracture electron microscopy allows one to visualize H⁺-PPase particles in addition to H⁺-ATPase particles in the tonoplast of Kalanchoë daigremontiana.     

Effect of root-applied spermidine on growth and respiratory metabolism in roots of cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>) seedlings under hypoxia

The effects of exogenous spermidine (Spd) application to hypoxic nutrient solution on the contents of endogenous polyamines (PAs) and respiratory metabolism in the roots of cucumber (Cucumis sativus L.) seedlings were investigated. Cucumber seedlings were grown hydroponically in control and hypoxic nutrient solutions with and without addition of Spd at a concentration of 0.05 mM. The activities of key enzymes involved in the tricarboxylic acid cycle (TCAC), such as succinate dehydrogenase (SDH) and isocitrate dehydrogenase (IDH), were significantly inhibited under root-zone hypoxia with dissolved oxygen (DO) at 1 mg/l. In contrast, the activities of enzymes involved in the process of fermentation, such as pyruvate decarboxylase (PDC), alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), and alanine aminotransferase (AlaAT), were significantly increased. Thus, aerobic respiration was inhibited and fermentation was enhanced in the roots of cucumber seedlings as a result of decreasing ATP content to inhibit the dry weight of seedlings under hypoxic stress. Moreover, the contents of free, soluble conjugated, and insoluble bound putrescine (Put), Spd, and spermine (Spm) in the roots of cucumber seedlings were significantly increased under hypoxia stress. Interestingly, application of Spd to hypoxic roots markedly suppressed the accumulation of free Put and, in contrast, promoted an increase in free Spd and Spm, as well as soluble conjugated and insoluble bound Put, Spd, and Spm contents. From these data, we deduced that exogenous Spd promotes the conversion of free Put into free Spd and Spm, and soluble conjugated and insoluble bound PAs under hypoxia stress. Furthermore, the activities of LDH, PDC, and ADH were suppressed and, in contrast, the activities of SDH and IDH were enhanced by application of exogenous Spd to hypoxic roots. As a result, aerobic respiration was enhanced but fermentation metabolism was inhibited in the roots of cucumber seedlings, leading to an increase in ATP content to alleviate the inhibited dry weight of seedlings due to hypoxia stress. These results suggest that application of Spd to hypoxic nutrient solution promoted conversion of free Put into free Spd and Spm as well as soluble conjugated and insoluble bound PAs, further enhanced IDH and SDH activities, and inhibited ethanol fermentation and lactate fermentation, resulting in increased ATP content and eventually enhanced tolerance of cucumber plants to root-zone hypoxia.     

Effects of spermidine and spermine levels on salt tolerance associated with tonoplast H+-ATPase and H+-PPase activities in barley roots

The effects of polyamines (Putrescine— Put; Spermidine—Spd; and Spermine—Spm) on␣salt tolerance of seedlings of two barley (Hordeum vulgare L.) cultivars (J4, salt-tolerant; KP7, salt-sensitive) were investigated. The results showed that, the salt-tolerant cultivar J4 seedlings accumulated much higher levels of Spd and Spm and lower Put than the salt-sensitive cultivar KP7␣under salt stress. At the same time, the dry weight of KP7 decreased significantly than that of␣J4. After methylglyoxal bis(guanylhydrazone) [MGBG, an inhibitor of S-adenosylmethionine decarboxylase (SAMDC)] treatment, Spd and Spm levels together with the dry weight of both cultivars were reduced, but the salt-caused dry weight reduction in two cultivars could be reversed by the concomitant treatment with Spd. MGBG decreased the activities of tonoplast H⁺-ATPase and H⁺-PPase too, but the experiments in vitro indicated that MGBG was not able to affect the above two enzyme activities. However, the polyamines, especially Spd, promoted their activities obviously. These results suggested that the conversion of Put to Spd and Spm and maintenance of higher levels of Spd and Spm were necessary for plant salt tolerance.     

多胺浸种改善盐胁迫大麦根系液泡膜功能的机理

研究了0.1 mmol/L 腐胺 (Put) 和0.5 mmol/L 亚精胺 (Spd) 浸种对200 mmol/L NaCl胁迫下大麦(Hordeum vulgare L.)幼苗生长速率、干物质积累、离子分布、液泡膜蛋白结合多胺含量以及液泡膜膜脂组分与功能的影响.结果表明,Put和Spd浸种均可缓解盐胁迫对大麦幼苗的盐害,促进生长和干物质积累,降低大麦幼苗体内[Na+]/[K+].与盐处理的对照植株相比,Put和Spd浸种均可提高大麦幼苗根系液泡膜磷脂含量,降低糖脂结合半乳糖含量,而膜上非共价结合多胺含量Spd+PAx (一种未知多胺) 与 Put+Dap (二氨基丙烷)之比((Spd+PAx)/(Put+Dap))、共价和非共价结合多胺总量均上升.统计分析结果表明,液泡膜非共价结合多胺(Spd+PAx)/(Put+Dap)与H+-ATPase和H+-PPase活性呈显著正相关关系.