盐胁迫对大麦幼苗质膜,液泡膜上共价和非共价结合多胺含量的影响

用不同浓度ＮａＣｌ处理７ｄ龄大麦（Ｈｏｒｄｅｕｍ ｖｕｌｇａｒｅＬ．）幼苗３ｄ。以非共价键和共价键形式分别与质膜和液泡膜微囊及膜蛋白结合的多胺含量受低 度盐的促进而被高浓度盐所抑制。以非共价键形式与膜微囊结合的各种多胺中亚精胺（Ｓｐｄ）含量最高,占膜上多胺总量的４０％－７０％,与膜蛋白共价结合的各种多胺中腐胺（Ｐｕｔ）含量占主导地位,占膜蛋白上多胺总量的３５％－６０％。在根系液泡膜上发现一种含量丰     

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。     

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。     

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.     

盐胁迫对大麦叶片类囊体膜上两种形态多胺含量和膜蛋白合成的影响

 用不同浓度NaCl处理6天龄大麦（Hordeum vulgare）幼苗3 d,以非共价键形式与叶片类囊体膜结合的多胺含量在低浓度盐处理时含量不变,随NaCl处理浓度的上升含量明显下降。在检测到的3种多胺中Spd（亚精胺）含量最高,约占膜上非共价键结合多胺总量的40%。以共价键形式与类囊体膜蛋白结合的多胺含量随NaCl处理浓度的上升一直呈下降趋势。在检测到的3种多胺中Put（腐胺）含量最高,约占膜上共价键结合多胺总量的60%以上。低浓度盐处理对14C-Glu向类囊体膜蛋白的掺入没有影响,随NaCl处理浓度的上升14C-Glu向类囊体膜蛋白的掺入明显下降。     

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.     

Relationship between seed quality and changes in conjugated polyamine in plasma membrane purified from wheat embryos during grain ripening

In order to explore the relationship between seed quality and changes in conjugated polyamine in plasma membrane purified from wheat embryos during grain ripening. Plasma membrane (PM) vesicles were isolated from the embryos of ripening wheat (Triticum aestivum L.) grains by the gradient centrifugation method. The contents of polyamines conjugated (covalently and non-covalently) to the PM vesicles were investigated. Results showed that after pollination, from the 22nd to the 32nd day, the embryos of wheat grains underwent dehydration, as judged by the decrease of embryo relative water content (ERWC). During embryo ripening, non-covalently conjugated (NCC)-Spd and NCC-Spm, covalently conjugated (CC)-Put and CC-Spd contents increased markedly, while relative embryo cell vigor (RECV) decreased slightly. The treatment with methylglyoxyl-bis (guanylhydrazone) (MGBG), an inhibitor of S-adenosylmethionine decarboxylase (SAMDC), inhibited the increases of the NCC-Spd and NCC-Spm contents, enhanced the decrease of RECV simultaneously, and decreased mature seed relative germination potential (RGP). The effects of MGBG were reversed by exogenous Spd and Spm. Phenanthrolin (o–Phen), an inhibitor of transglutaminase (TGase), inhibited the increases of CC-Put and CC-Spd contents, enhanced the decrease of RECV simultaneously, and decreased mature seed RGP. These results suggest that during embryo ripening, the levels of NCC-Spd, NCC-Spm, CC-Put, and CC-Spd increase, and these increases might affect embryo cell vigor and seed germination potential.     

盐适应过程中香根草内源游离态、结合态、束缚态多胺含量的变化

研究了不同浓度NaCl胁迫下,香根草（Vetiteria zizanioides）根、叶中的游离态、结合态、束缚态多胺（PAs）[包括腐胺（Put）,尸胺（Cad）,亚精胺（Sod）和精胺（Spm）]含量的变化。在中度盐胁迫（100,200mmol L^-1NaCl）9天时,香根草基本能够正常生长,但在重度盐胁迫（300mmol L^-1NaCl）下,其生长受到严重抑制。在上述3个不同浓度的NaCl胁迫下,香根草根、叶中游离态Put,Cad,spd,Stma和总的游离态PAs含量明显下降,在高盐浓度下下降幅更大;结合态Put,Cad,Sod,Spm和总的结合态PAs含量显著上升,但在重度盐胁迫下升幅较小或与对照相当;束缚态Put,Cad和总的束缚态PAs含量均减少,而束缚态Spd和Spm含量在叶中是下降的,在根中则增加,且在中度盐胁迫下更明显。对根和叶片而言,除游离态（Spd＋Spm）,Put比值在重度盐胁迫下较对照显著下降外,其它游离态、结合态、束缚态和总的（Spd＋Spm）／Put比值在不同盐胁迫下均上升,在中度盐胁迫下更明显。这表明,维持多胺总量的稳态和较高的（Spd＋Spm）／Put比值是香根草适应中度盐胁迫的一个重要机制。     

NaCl胁迫对菜用大豆种子多胺代谢的影响

采用蛭石栽培,在100 mmol·L-1NaCl胁迫下,对耐盐性不同的两个品种菜用大豆种子的丙二醛(MDA)含量和多胺(PAs)代谢进行了研究.结果表明:NaCl胁迫显著增加了菜用大豆种子的MDA含量,但耐盐品种‘绿领特早’(LL)的增幅低于盐敏感品种‘理想高产95-1’(LX).与LX相比,LL种子在整个NaCl胁迫期间均维持了相对较高的游离态精胺(Spm)、结合态Spm、结合态亚精胺(Spd)、束缚态Spd和束缚态腐胺(Put)含量,较高的(Spd +Spm )/Put 和(cPAs+bPAs)/fPAs值及较低的Put/PAs值,在胁迫中、后期(9～15 d)维持了相对较高的游离态Spd含量;胁迫期间,LL的精胺酸脱羧酶(ADC)长时期(6～15 d)保持相对较高的活性,而多胺氧化酶(PAO)则长时期(6～15 d)维持相对较低的活性.综上,LL具有较强的多胺合成能力及较强的Put向Spd和Spm以及游离态多胺向结合态和束缚态多胺转化的能力,进而有效抑制了细胞的膜脂过氧化,这可能是其耐盐性较强的重要原因之一.     

Changes in content of free,conjugated and bound polyamines and osmotic adjustment in adaptation of vetiver grass to water deficit

Osmotic adjustment and alteration of polyamines (PAs) have been suggested to play roles in plant adaptation to water deficit/drought stress. In this study, the changes in cell intactness, photosynthesis, compatible solutes and PAs [including putrescine (Put), spermidine (Spd) and spermine (Spm) each in free, conjugated and bound forms] were investigated in leaves of vetiver grass exposed to different intensity of water deficit stress and subsequent rewatering. The results showed that, when vetiver grass was exposed to the moderate (20% and 40% PEG-6000 solutions) and severe (60% PEG solution) water deficit for 6 days, the plant injury degree (expressed as the parameters of plant growth, cell membrane integrity, water relations and photosynthesis) increased and contents of free and conjugated Put decreased with the rise of PEG concentration. Under the moderate water deficit, the plants could survive by the reduced osmotic potential (ψ_s), increased free and conjugated Spd and Spm in leaves. After subsequent rewatering, the osmotic balance was re-established, most of the above investigated physiological parameters were fully or partly recovered to the control levels. However, it was not the case for the severely-stressed and rewatering plants. It indicates that, vetiver grass can cope well with the moderate water deficit/drought stress by using the strategies of osmotic adjustment and maintenance of total contents of free, conjugated and bound PAs in leaves.     

外源多胺对低氧胁迫下黄瓜幼苗根系生长及H+-ATP酶和H+-焦磷酸酶活性的影响

采用营养液水培方式,研究了根际低氧胁迫下外源多胺对黄瓜幼苗植株根系生长,内源多胺含量与质膜H -ATP酶、液泡膜H -ATP酶和焦磷酸酶活性的影响.结果表明,根际低氧胁迫显著抑制黄瓜幼苗根系的生长,外源Put(腐胺)和Spd(亚精胺)可缓解低氧胁迫对根系的生长抑制,多胺主要以Spd的形式发挥促进性的生理作用,Put通过转化为Spd发挥作用;低氧胁迫下黄瓜根系内源多胺含量略有提高,外源多胺处理可增加内源多胺的含量;低氧胁迫下外源Put和Spd处理后质膜H -ATP酶活性显著提高,外源多胺对黄瓜根系液胞膜H -ATP酶和H -焦磷酸酶活性没有明显影响,说明低氧胁迫下外源多胺主要通过提高质膜H -ATP酶活性而发挥生理作用.     

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.     

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.     

氯化钠胁迫对嫁接黄瓜叶片多胺含量的影响

以日本耐盐品种‘帝王新土佐’南瓜为砧木,以’新泰密刺’黄瓜为接穗,在100 mmol·L^-1 NaCl胁迫下,对黄瓜嫁接和自根植株不同时期叶片中不同形态多胺含量的变化进行了研究.结果表明：NaCl胁迫下黄瓜嫁接植株游离态腐胺（Put）含量在胁迫2 d时与自根植株无显著差异,其余时间均显著高于自根植株;游离态亚精胺（Spd）和游离态精胺（Spm）含量在整个胁迫期间均显著高于自根植株;游离态多胺总量（PAs）在胁迫第4天出现峰值;嫁接植株游离态Put/PAs值在胁迫4 d时与自根植株无显著差异,其余胁迫时间均显著低于自根植株,而(Spd+Spm)/Put值在整个胁迫期间均显著高于自根植株;嫁接植株结合态和束缚态Put、Spd和Spm含量在整个胁迫期间均显著高于自根植株,结合态和束缚态PAs在胁迫第6天出现峰值;结合态多胺的Put/PAs值和(Spd+Spm)/Put值变化趋势与游离态多胺一致;嫁接植株束缚态Put/PAs值在胁迫6 d时与自根植株无显著差异,其余时间均显著低于自根植株,而(Spd+Spm)/Put值在整个胁迫期间均显著高于自根植株.表明黄瓜嫁接植株表现出较强的耐盐特征.     

Excretion of polyamines in alfalfa and tobacco suspension-cultured cells and its possible role in maintenance of intracellular polyamine contents

Changes in polyamines (PAs) in cells and cultivation media of alfalfa (Medicago sativa L.) and tobacco bright yellow 2 (BY-2) (Nicotiana tabacum L.) cell suspension cultures were studied over their growth cycles. The total content of PAs (both free and conjugated forms) was nearly 10 times higher in alfalfa, with high level of free putrescine (Put) (in exponential growth phase it represented about 65-73% of the intracellular Put pool). In contrast, the high content of soluble Put conjugates was found in tobacco cells (in exponential phase about 70% of the intracellular Put). Marked differences occurred in the amount of PAs excreted into the cultivation medium: alfalfa cells excreted at the first day after inoculation 2117.0, 230.5, 29.0 and 88.0 nmol g(-1) of cell fresh weight (FW) of Put, spermidine (Spd), spermine (Spm) and cadaverine (Cad), respectively, while at the same time tobacco cells excreted only small amount of Put and Spd (12.7 and 2.4 nmol g(-1) FW, respectively). On day 1 the amounts of Put, Spd, Spm and Cad excreted by alfalfa cells represented 21, 38, 12 and 15% of the total pool (intra- plus extra-cellular contents) of Put, Spd, Spm and Cad, respectively. In the course of lag-phase and the beginning of exponential phase the relative contents of extracellular PAs continually decreased (with the exception of Cad). On day 10, the extracellular Put, Spd, Spm and Cad still represented 11.3, 10.9, 2.1 and 27% of their total pools. The extracellular PAs in tobacco cells represented from day 3 only 0.1% from their total pools. The possible role of PA excretion into the cultivation medium in maintenance of intracellular PA contents in the cells of the two cell culture systems, differing markedly in growth rate and PA metabolism is discussed.     

NaCl处理下大麦根系质膜微囊结合多胺与Na+/H+逆向运输的关系

NaCl10 0mmol/L处理结合外施Spd和Put以及多胺代谢抑制剂邻二氮杂菲和MGBG ,以改变大麦根系质膜结合多胺种类和数量 ,研究了大麦根系质膜上两种形态多胺与质子泵和Na /H 逆向运输活性的关系。结果发现 ,NaCl处理后大麦根系质膜微囊上存在Na /H 逆向运输活性。质膜H ATPase活性与膜上非共价键结合多胺数量间呈显著正相关 ,其中 ,Spd对H ATPase的激活程度大于Put。膜蛋白上共价键结合多胺数量与Na /H 逆向运输活性间呈极显著正相关关系 ,说明大麦根系质膜Na /H 逆向运输的盐诱导似乎与Na /H 逆向运输蛋白的从头合成有关。此外 ,质膜Na /H 逆向运输活性仅与膜蛋白上共价键结合多胺数量有关 ,而与多胺种类关系不大。     

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.     

Changes of free, soluble conjugated and bound polyamine titers of jojoba explants under sodium chloride salinity in vitro

Jojoba (Simmondsia chinensis L.) single node explants were cultured in a basal medium supplemented with 17.8 microM 6-benzyladenine and four levels of sodium chloride concentration (0, 56.41, 112.82 and 169.23 mM). The free, the soluble conjugated and the insoluble bound forms of polyamines (PAs) (putrescine (Put), spermidine (Spd) and spermine (Spm)) were determined monthly during a 3-month proliferation stage. Free Put and Spd were found in higher levels in the control treatment, while Spm content was higher in the salt treatments. All soluble conjugated PAs were found to be in lower concentrations in explants growing on medium supplemented with salt, while the opposite was true for the insoluble bound PAs. It appeared that certain PAs and PAs forms could play a significant role in the adaptation mechanism of jojoba under saline conditions.     

Isolation of plasma membrane and tonoplast fractions from spinach leaves by preparative free-flow electrophoresis and effect of photoinduction

A membrane fraction enriched in plasma membrane and tonoplast vesicles was isolated from green leaves of Spinacia oleracea L. and subjected to subfractionation by free-flow electrophoresis. The most electronegative membrane vesicle fraction collected after the free-flow electrophoretic separation was identified as derived from tonoplast, while the least electronegative fraction was identified as derived from plasma membrane. The identification of the fractions was based on membrane morphology, and on the presence or absence of biochemical markers. The plasma membrane fraction was enriched in thick (9–11 nm) membranes which bound N-1-naphthylphthalamic acid (NPA), and reacted with phosphotungstic acid at low pH on thin sections for electron microscopy. The tonoplast fraction was enriched in vesicles with 7–9 nm thick membranes that neither bound NPA nor reacted with phosphotungstic acid at low pH. Both the plasma membrane and the tonoplast fraction were about 90% pure, with a cross-contamination of not more than 2%. Membrane vesicles originating from dictyosomes, endoplasmic reticulum, mitochondria, plastids, or peroxisomes contaminated the plasma membrane and the tonoplast fractions by a few % only. In leaves of photoinduced plants (24 h light period), the plasma membranes were thicker than in control leaves (8 h light, 16 h dark). The plasma membrane fraction obtained from photo-induced leaves by free-flow electrophoresis retained this increase in thickness, showing not only that photoinduction alters plasma membrane structure, but also that this change is stable to isolation.     

Cytological changes and alterations in polyamine contents induced by cadmium in tobacco BY-2 cells.

Changes in cell viability, proliferation, cell and nuclear morphology including nuclear and DNA fragmentation induced by 0.05 and 1 mM CdSO4 (Cd2+) in tobacco BY-2 cell line (Nicotiana tabacum L.) were studied in the course of 7 days. Simultaneously changes in endogenous contents of both free and conjugated forms of polyamines (PAs) were investigated for 3 days. The application of 0.05 mM Cd2+ evoked decline of cell viability to approximately 60% during the first 24 h of treatment. Later on degradation of cytoplasmic strands, formation of the stress granules and vesicles, modifications in size and shape of the nuclei, including their fragmentation, were observed in the surviving cells. Their proliferation was blocked and cells elongated. Beginning the first day of treatment TUNEL-positive nuclei were detected in cells cultivated in medium containing 0.05 mM Cd2+. Treatment with highly toxic 1 mM Cd2+ induced fast decrease of cell viability (no viable cells remained after 6-h treatment) and cell death occurred before DNA cleavage might be initiated. The exposure of tobacco BY-2 cells to 0.05 mM Cd2+ resulted in a marked accumulation of total PAs (represented by the sum of free PAs and their perchloric acid (PCA)-soluble and PCA-insoluble conjugates) during 3-day treatment. The increase in total PA contents was primarily caused by the increase in putrescine (Put) concentration. The accumulation of free spermidine (Spd) and spermine (Spm) at 12 and 24 h in 0.05 mM Cd2+ treated BY-2 cells and high contents of Spd and especially Spm determined in dead cells after I mM Cd2+ application was observed. The participation of PA conjugation with hydroxycinnamic acids and PA oxidative deamination in maintaining of free PA levels in BY-2 cells under Cd2+-induced oxidative stress is discussed.