低氧胁迫下钙对网纹甜瓜幼苗多胺含量及多胺氧化酶活性的影响

以低氧性敏感的网纹甜瓜(Cucumis melo var. raticulalus)品种`西域一号'(Xiyu 1)为材料,研究了钙对低氧胁迫下幼苗生长、游离态、结合态和束缚态的腐胺(Put)、亚精胺(Spd)、精胺(Spm)含量、二胺氧化酶(DAO)和多胺氧化酶(PAO)活性、H₂O₂含量的影响。结果表明:与通气对照相比,低氧胁迫处理幼苗鲜重和干重显著降低,根、叶中3种形态的Put、Spd、Spm含量显著增加,DAO和PAO活性、H₂O₂含量显著提高;营养液加钙处理不但缓解了低氧胁迫对幼苗生长的抑制作用,而且幼苗游离态、结合态和束缚态Put、Spd、Spm含量显著高于单纯低氧胁迫处理,DAO和PAO活性、H₂O₂含量显著低于低氧胁迫处理;低氧胁迫下缺钙处理加重了低氧胁迫对网纹甜瓜幼苗的伤害。表明在低氧胁迫下,钙参与了网纹甜瓜幼苗多胺的代谢过程,对增强幼苗耐低氧性起着重要的作用。     

多胺氧化酶检测方法的改进及其在低氧水培黄瓜根系中的应用

对多胺氧化酶活性检测方法进行了改进,多胺氧化产物H2O2浓度的最低检测量可从1.25μmol·L-1降低到0.65μmol·L-1,其显色产物具有良好的稳定性与重现性.用改进的方法检测的低氧水培下黄瓜根系多胺氧化酶活性比通气对照提高2.45倍,酶氧化底物以精胺(Spm)为最佳,最适pH值为6.5.     

大豆初生幼苗多胺氧化酶活性的细胞化学定位

对大豆(Glycinemax(L.)Merril“l)垦农4号”萌发种子和初生幼苗中的多胺氧化酶(polyamineoxidase,PAO,EC1.4.3.6)的活性和分布进行了研究。结果表明,PAO活性仅在种子萌发起始后(吸胀后24h)才检测到,然后随着种子萌发进程,PAO活性快速升高。但是,在萌发种子(吸胀后72h)和初生幼苗(吸胀后120h)中,PAO活性在各器官中的分布有明显差异。在萌发种子中,PAO活性在胚根最高(5.17±0.91Ug-1FW),胚轴次之,胚芽再次之,子叶活性最低(0.12±0.03Ug-1FW);在初生幼苗中,PAO活性在下胚轴中最高(5.47±0.66Ug-1FW),幼根次之,顶芽再次之,子叶最低(0.10±0.03Ug-1FW)。这种差异对种子萌发和幼苗形态建成有积极意义。运用细胞化学定位在透射电镜下观察初生幼苗PAO在各部位的分布,发现PAO主要定位在顶芽细胞的液泡膜上、子叶细胞的细胞壁及其外侧表面、下胚轴细胞的细胞壁及其表面,且PAO与细胞壁表面结合较紧;根细胞的细胞壁、细胞间隙、细胞膜、液膜上均有分布,但以液泡膜分布居多。本研究结果进一步证实了PAO在细胞壁和细胞间隙有着较广泛的分布。首次报道PAO在细胞膜和液泡膜上有分布。     

干旱胁迫下小麦幼苗根、叶多胺含量和多胺氧化酶活性的变化

 以抗旱性不同的两个小麦品种（‘晋麦33’和‘温麦8’）（Triticum aestivum cv. Jinmai 33 and Wenmai 8）为材料,研究了干旱胁迫下多胺含量和多胺氧化酶活性的变化。结果表明：旱过程中,幼苗根、叶中腐胺（Put）、亚精胺（Spd）、精胺（Spm）3种多胺含量和多胺氧化酶（PAO）活性先迅速升高,而后下降。与抗旱性弱的‘晋麦33’相比,抗旱性强的品种‘温麦8’幼苗根、叶中Spd、Spm 含量初期升高幅度大,之后下降速率减慢;PAO活性的变化与之相反,‘晋麦33’的PAO活性提高的幅度大于‘温麦8号’。多胺含量和PAO活性在小麦幼苗的根与叶之间呈极显著正相关。干旱初期,小麦根、叶中多胺迅速积累可能是干旱胁迫反应的一个信号,随后较高的Spd、Spm 水平有利于增强小麦幼苗的抗旱性。     

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。     

γ-氨基丁酸对低氧胁迫下甜瓜幼苗多胺代谢的影响

以‘西域一号’甜瓜为试验材料,采用营养液水培法,研究了低氧胁迫下外源添加γ-氨基丁酸(GABA)对甜瓜幼苗多胺代谢的影响.结果表明：与通气对照相比,低氧胁迫处理的甜瓜幼苗谷氨酸脱羧酶(GAD)活性和GABA含量显著提高,同时多胺合成酶活性提高诱导多胺含量显著增加,但二胺氧化酶(DAO)和多胺氧化酶(PAO)活性也显著提高;根系精氨酸脱羧酶(ADC)活性提高幅度较大,导致根系游离态腐胺含量较高,而叶片鸟氨酸脱羧酶(ODC)和S-腺苷甲硫氨酸脱羧酶(SAMDC)活性提高幅度较大,导致叶片游离态亚精胺(Spd)含量较高;根系游离态DAO和PAO活性显著低于叶片,其细胞壁结合态PAO活性显著高于叶片.与低氧胁迫处理相比,低氧胁迫下外源添加GABA处理的甜瓜幼苗叶片和根系中GABA和谷氨酸含量均显著提高,而GAD活性显著降低;精氨酸、鸟氨酸、甲硫氨酸含量的提高促使多胺合成酶活性显著提高,从而诱导多胺含量显著增加,DAO和PAO活性显著降低.     

γ-氨基丁酸对低氧胁迫下甜瓜幼苗多胺代谢的影响

以‘西域一号’甜瓜为试验材料,采用营养液水培法,研究了低氧胁迫下外源添加γ-氨基丁酸(GABA)对甜瓜幼苗多胺代谢的影响.结果表明:与通气对照相比,低氧胁迫处理的甜瓜幼苗谷氨酸脱羧酶(GAD)活性和GABA含量显著提高,同时多胺合成酶活性提高诱导多胺含量显著增加,但二胺氧化酶(DAO)和多胺氧化酶(PAO)活性也显著提高;根系精氨酸脱羧酶(ADC)活性提高幅度较大,导致根系游离态腐胺含量较高,而叶片乌氨酸脱羧酶(ODC)和S-腺苷甲硫氨酸脱羧酶(SAMDC)活性提高幅度较大,导致叶片游离态亚精胺(Spd)含量较高;根系游离态DAO和PAO活性显著低于叶片,其细胞壁结合态PAO活性显著高于叶片.与低氧胁迫处理相比,低氧胁迫下外源添加GABA处理的甜瓜幼苗叶片和根系中GABA和谷氨酸含量均显著提高,而GAD活性显著降低;精氨酸、鸟氨酸、甲硫氨酸含量的提高促使多胺合成酶活性显著提高,从而诱导多胺含量显著增加,DAO和PAO活性显著降低.     

氯化钠胁迫下野生和栽培大豆幼苗体内的多胺水平变化

以通用的较耐盐的栽培大豆Lee68品种和对盐敏感的野生大豆N23232种群为参照,研究了盐胁迫下耐盐野生大豆BB52种群幼苗体内多胺(PAs)组分、含量及多胺氧化酶(PAO)活性的变化。结果表明,盐胁迫下BB52幼苗根PAs中Put和Spm含量下降较Lee68和N23232显著,但Spd含量下降较少．BB52叶片PAs中Put含量下降,Spd上升,(Spd+Spin)／Put值增加和Put／PAs值降低幅度与耐盐性呈正相关趋势．盐胁迫下,各材料根和叶中PAO活性上升,N23232上升最明显．探讨了多胺水平与BB52耐盐性的关系。     

盐胁迫下大麦幼苗多胺的种类和状态与多胺氧化酶活性的关系

200 mmol/L的NaCl胁迫8 d大麦幼苗叶片和根系中的三种形态多胺都有不同程度地下降,其中游离态多胺含量的下降幅度最大;高氯酸不溶性结合态多胺含量变化较小.根系中PAO的活性先上升后下降,而叶片中PAO的活性先下降后上升.游离态多胺中,亚精胺和精胺(Spd Spm)的含量变化与相应部位PAO的活性变化趋势相反,表明PAO在盐胁迫下可能调节了游离态多胺的含量从而影响高氯酸可溶结合态与高氯酸不溶结合态多胺的含量.     

盐碱混合胁迫对裸燕麦幼苗叶片脯氨酸和多胺代谢的影响

为探讨裸燕麦对盐碱混合胁迫的脯氨酸(Pro)和多胺(PAs)响应机制,采用温室砂培试验,研究了低(25 mmol·L-1)、高(75 mmol·L~(-1))浓度盐碱混合胁迫0、1、3、5和7 d幼苗叶片Pro和PAs含量及相关代谢酶活性的变化。结果表明:与对照相比,盐碱混合胁迫在整个胁迫期间提高了裸燕麦叶片Pro含量,高浓度胁迫提高的幅度大于低浓度胁迫;低浓度胁迫下吡咯啉-5-羧酸合成酶(P5CS)活性提高,而鸟氨酸-δ-氨基转移酶(δ-OAT)和脯氨酸脱氢酶(Pro DH)活性受抑;高浓度胁迫下δ-OAT活性提高,而P5CS和Pro DH活性下降,说明低浓度胁迫下Pro合成以谷氨酸途径为主,高浓度胁迫下以鸟氨酸途径为主;低、高浓度盐碱混合胁迫下,游离态腐胺(Put)、亚精胺(Spd)和精胺(Spm)及结合态Spd和束缚态Spd、Spm含量提高,结合态Put、Spm和束缚态Put含量在胁迫1~5 d时提高,胁迫7 d时下降,且高浓度胁迫的变幅大于低浓度胁迫;Put/PAs和(c PAs+b PAs)/f PAs在胁迫1~3 d时提高,胁迫5~7 d时下降或变化不明显,(Spd+Spm)/Put的变化却相反;整个胁迫期间低浓度胁迫保持了较低的Put/PAs和较高的(Spd+Spm)/Put,(c PAs+b PAs)/f PAs除胁迫1 d时低浓度胁迫低于高浓度胁迫外,其余胁迫时间两浓度处理间差异不显著;精氨酸脱羧酶(ADC)和鸟氨酸脱羧酶(ODC)活性在低、高浓度盐碱混合胁迫期间提高,二胺氧化酶(DAO)活性在胁迫1~3 d下降或变化不大,胁迫5~7 d时升高,多胺氧化酶(PAO)活性的变化与DAO相反,且高浓度胁迫下ADC、ODC、DAO和PAO的变幅大于低浓度胁迫。说明盐碱混合胁迫初期(1~3 d)游离态PAs(f PAs)向结合态和束缚态PAs(cPAs+bPAs)转化增强,低浓度胁迫后期(5~7 d)Put向Spd和Spm转化的能力强于高浓度胁迫,但高浓度胁迫在整个胁迫期间PAs合成和分解的速率更高。     

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.     

Nitric oxide enhances salt tolerance in cucumber seedlings by regulating free polyamine content

Nitric oxide (NO), an endogenous signaling molecule in plants and animals, mediates responses to abiotic and biotic stresses. This study was conducted in nutrient solution to investigate the effects of exogenous sodium nitroprusside (SNP), an NO donor, on plant growth and free polyamine content in cucumber leaves and roots under NaCl stress. The results showed that 100 μM SNP in solution significantly improved the growth of cucumber seedlings under NaCl stress for 8 days, as indicated by increased, plant height, stem thickness, fresh weight and increased dry matter accumulation. Further analysis demonstrated that the content of free polyamines and the activity of polyamine oxidase (PAO) in cucumber seedling leaves and roots initially increased dramatically under NaCl stress, although they decreased over a longer period of stress. Throughout the treatment period, the value of (spermine + spermidine)/putrescine [(Spd + Spm)/Put] also decreased under NaCl stress compared to the control. In contrast, the application of 100 μM SNP in the nutrient solution decreased the content of free Put, Spd, total free polyamines and PAO activity under NaCl stress. It also caused an increase in the content of Spm and the value of (Spd + Spm)/Put, adjusted the ratio of three kinds of free polyamines (Put, Spd, Spm) in cucumber seedlings. The high (Spd + Spm)/Put value and the accumulation of Spm were beneficial to improving the salt tolerance of plants. Therefore, NO alleviated the damage to cucumber seedlings caused by salt stress. NO enhanced the tolerance of cucumber seedlings to NaCl stress by regulating the content and proportions of the different types of free polyamines.     

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。     

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.     

D-精氨酸对低氧胁迫下黄瓜幼苗根系多胺含量和无氧呼吸代谢的影响

采用营养液水培法,研究了根际低氧胁迫下D-精氨酸（D-Arg）对两个抗低氧能力不同的黄瓜品种根系中多胺含量和无氧呼吸代谢的影响.结果表明,低氧处理下,黄瓜幼苗根系中多胺含量显著增加,无氧呼吸代谢能力提高;与抗低氧能力弱的‘中农八号’相比,抗低氧能力强的‘绿霸春四号’根系中乙醇发酵活性较高,乳酸发酵活性较低;低氧胁迫下,D-精氨酸能显著降低黄瓜幼苗根系中腐胺、亚精胺和精胺含量,根系中乙醇脱氢酶（ADH）和乳酸脱氢酶（LDH）活性增加,乙醇和乳酸含量升高,植株生长受到抑制,而外源腐胺能缓解D-精氨酸的这种作用.说明黄瓜幼苗根系中较高的多胺含量可能有利于缓解低氧胁迫对植株造成的伤害.     

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.