NO缓解玉米幼苗盐胁迫伤害的生理机制

以玉米幼苗为材料,通过NO的供体硝普纳(SNP)的合成抑制剂L-NAME和NaN3、清除剂cPTIO组合处理,分析外源NO和IAA对盐胁迫下玉米幼苗生长,以及NO对盐胁迫下玉米幼苗叶片和根尖IAA含量、IOD和POD活性的影响,以探讨NO与IAA在提高植物抗盐性中的关系.结果表明,盐胁迫下,SNP和IAA均能显著促进玉米幼苗株高、主根长和侧根数的增加;SNP能显著提高玉米幼苗叶片和根尖IAA含量,降低IOD和POD活性;L-NAME和NaN3及cPTIO均能有效减弱SNP诱导的IAA含量的增加.由此可见,在盐胁迫条件下,NO信号可能位于IAA信号的上游,它通过促进玉米幼苗內源IAA的积累缓解盐胁迫对其生长的抑制.     

NaCl胁迫下玉米幼苗中一氧化氮与茉莉酸积累的关系

以三叶一心期的玉米幼苗为材料,研究了NaCl胁迫下玉米幼苗根尖和叶片中一氧化氮(NO)和茉莉酸(JA)积累之间的关系.结果表明:NaCl胁迫下玉米幼苗根尖和叶片中NO和JA的含量均增加,且NO积累的时间早于JA;根尖中脂氧合酶(LOX)活性逐渐降低,而叶片中LOX活性显著升高.硝普钠(SNP,NO供体)处理使幼苗的JA含量和LOX活性亦增加;用NO清除剂cPTIO及NO合成的抑制剂L-NAME、NaN3处理幼苗时,可抑制NaCl胁迫诱导的JA积累以及叶片中LOX活性的增加.可见,玉米幼苗在盐胁迫下爆发的NO可能通过调控LOX活性来调节其JA的积累.     

NO对盐胁迫下苜蓿根系生长抑制及氧化损伤的缓解效应

以"甘农4号"苜蓿品种为材料,采用水培法,用NO供体硝普钠(SNP)、硝普钠类似物亚铁氰化钠(不产生NO)、NO特异清除剂c-PTIO、一氧化氮合酶(NOS)活性抑制剂N-硝基-L-精氨酸甲脂(L-NAME)、硝酸还原酶(NR)活性抑制剂钨酸盐处理苜蓿植株,研究NO对盐胁迫下苜蓿幼苗根系生长、根系活力、根系中渗透调节物质、膜脂过氧化、活性氧含量及抗氧化酶活性等的影响,探讨NO调控苜蓿幼苗根系耐盐性的生理机制。结果表明:盐胁迫下SNP处理提高了根系活力,促进了苜蓿幼苗根系生长,降低游离脯氨酸含量,促进可溶性蛋白含量增加;增强超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、愈创木酚过氧化物酶(GPX)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性,提高还原型抗坏血酸(As A)和还原型谷胱甘肽(GSH)含量,降低过氧化氢(H2O2)、羟自由基(OH·)含量、超氧阴离子(O·-2)产生速率和膜脂过氧化产物丙二醛(MDA)含量;同时,SNP处理显著促进了苜蓿幼苗根系内源NO的积累。NO供体SNP的类似物亚铁氰化钠对盐胁迫下苜蓿根系各项生理生化指标无明显影响;盐胁迫下添加c-PTIO、L-NAME和钨酸盐进一步降低了苜蓿幼苗根系活力和根系生长,抑制了根系抗氧化系统活性,加剧了根系膜脂过氧化作用,降低了内源NO积累,添加SNP则能缓解该抑制效应;表明外源SNP处理能明显缓解盐胁迫对苜蓿幼苗根系生长的抑制和氧化损伤,且通过NOS和NR途径产生的内源NO也可能在苜蓿根系适应盐胁迫的调节中起关键作用;该研究结果为苜蓿耐盐机制及NO在苜蓿耐盐育种、化学调控和盐碱地栽培利用等提供了理论依据。     

一氧化氮参与调节盐胁迫诱导的玉米幼苗脱落酸积累

以三叶一心期的玉米幼苗为实验材料,研究了盐胁迫下玉米幼苗根尖和叶片中一氧化氮（NO）和脱落酸（ABA）积累之间的关系。结果表明,盐胁迫下玉米幼苗NO和ABA的含量均增加,用NO供体硝普钠（Sodium nitroprusside,SNP）处理时,ABA含量亦增加,且累积的时间较盐胁迫下早。用NO合成的抑制剂L-NAME （Nω-nitro-L-arginine methyl ester hydrochloride）和NaN,处理时,可减弱盐胁迫诱导的ABA含量的增加,用NO清除剂cPTIO处理时,这种盐胁迫诱导的ABA增加减少。推测盐胁迫下产生的NO参与调节ABA的积累及逆境下植物的防御反应。     

Cd胁迫下SA和NO互作对小麦幼苗根生长和叶绿素含量的影响

以"陇春27"号水培小麦幼苗为研究材料，外源添加水杨酸（SA）、一氧化氮（NO）清除剂（carboxy-PTIO，c-PTIO）、NO供体硝普钠（SNP）、硝酸还原酶（NR）抑制剂钨酸盐（Tungstate）及NO合成酶（NOS）抑制剂（L-NAME）进行不同预处理，分析其在镉（Cd）胁迫下根的生长和叶片叶绿素含量的变化，探讨SA和NO互作对小麦幼苗Cd胁迫的缓解机制。结果表明：随着Cd处理时间的延长，小麦幼苗根中SA含量显著降低，NO含量则呈现先增加（6 h和12 h）后减少（24 h和48 h）的趋势；Cd胁迫抑制了小麦幼苗根的生长，减少了叶片叶绿素的含量，而一定浓度的SA或SNP预处理可以缓解Cd胁迫对小麦幼苗根长的抑制作用，增加叶绿素的含量。c-PTIO、L-NAME和Tungstate单独预处理显著抑制了小麦幼苗根的生长，减少了NO的含量，但不影响叶绿素含量。SA400+L-NAME预处理可以缓解Cd胁迫对小麦幼苗根长的抑制作用以及叶绿素和NO含量的减少作用；SA400+c-PTIO或SA400+Tungstate预处理可增加Cd胁迫下叶绿素的含量，但对根的伸长无影响。进一步研究发现，Cd胁迫抑制了NR的活性，而SA400预处理可以使Cd胁迫下NR的活性增强，不同处理对NOS的活性均无影响。综上所述，Cd胁迫导致小麦幼苗根内源SA含量降低和NO含量先升高再降低；外源添加SA或SNP预处理缓解了Cd胁迫对根生长的抑制和叶绿素含量降低的作用；外源SA通过影响NO的产生从而提高小麦幼苗对Cd胁迫的耐受性，最终缓解了Cd对小麦幼苗的毒害作用。     

NO参与γ-氨基丁酸促进白三叶种子的萌发过程

白三叶种子用不同浓度γ-氨基丁酸（GABA）处理后,低浓度GABA处理的白三叶种子萌发速率和一氧化氮（NO）含量显著提高;NO清除剂cPTIO和硝酸还原酶（NR）抑制剂叠氮钢（NaN3）显著抑制GABA促进的白三叶种子萌发过程,而一氧化氮合酶（NOS）抑制剂L-NAME对GABA促进白三叶种子萌发的影响不显著。据此推测,GABA促进白三叶种子萌发过程中可能有NO参与,并且NO的产生主要依赖于硝酸还原酶途径。     

NO对He-Ne激光和增强UV-B辐射小麦幼苗气孔运动的作用机理研究

为探讨NO对He-Ne激光和增强UV-B辐射小麦（Triticum aestivuml）气孔运动的作用机理,采用低剂量（5 mW.mm-2）He-Ne激光和增强（10.08 kJ.m-2.d-1）UV-B辐射并结合药理学实验和激光共聚焦显微技术,对ML7113小麦的叶片及表皮条进行不同的处理,结果显示：（1）UV-B辐射既可诱导小麦叶片气孔关闭,又能够明显增加气孔保卫细胞和叶片的NO水平,且NO清除剂明显抑制了UV-B辐射诱导的小麦叶片气孔关闭,同时气孔保卫细胞和叶片内的NO含量明显减少。（2）一氧化氮合酶（NOS）抑制剂L-NAME对经UV-B辐射诱导的小麦幼苗气孔开度及保卫细胞和叶片内NO含量的抑制程度明显大于硝酸还原酶（NR）抑制剂NaN3对其的抑制程度,说明一氧化氮合酶（NOS）合成途径是小麦叶片经UV-B辐射后NO的主要产生途径。（3）就气孔开度而言,L〉CK〉BL〉B。就小麦叶片及保卫细胞内NO含量而言,B〉BL〉CK〉L。就硝酸还原酶（NR）和一氧化氮合酶（NOS）的活性而言,B组NR活性最低,NOS活性最高,L组NR活性最高,NOS活性最低。表明经He-Ne激光和增强UV-B辐射诱导的小麦气孔开度的变化确实与保卫细胞及叶片中NO含量的多少有关,气孔开度的减小及增大对应于NO含量的增多或减少,同时进一步证实了小麦叶片经He-Ne激光单独辐照后,NO的主要合成途径也来源于NOS途径。     

NR和NOS在CTK延缓离体小麦叶片衰老中的作用

用一氧化氮(NO)清除剂血红蛋白(Hb)、硝酸还原酶(NR)抑制剂钨酸钠(Na2WO4)、一氧化氮合酶(NOS)抑制剂L-硝基精氨酸甲酯(L-NAME)并结合激动素(KT)和玉米素(ZT)两种细胞分裂素(CTK)处理离体小麦叶片,测定分析各处理的相关生理生化指标,以明确NR和NOS在CTK延缓离体小麦叶片衰老过程中的作用.结果显示:KT和ZT单独处理均能显著延缓离体小麦叶片衰老过程中叶绿素、可溶性蛋白含量的降低,抑制丙二醛(MDA)的积累,促进NR和NOS活性升高;在Hb、Na2WO4或L-NAME存在时,上述KT和ZT延缓衰老的效应均显著减弱,同时NR和NOS活性的升高也分别被Na2WO4和L-NAME显著抑制.该结果暗示CTK延缓离体小麦叶片衰老可能与其诱导了NR和NOS活性的提高,进而促进NO的生成有关.     

EDDS对Cd胁迫下三叶鬼针草幼苗NO产生的影响

【摘要】通过室内盆栽试验, 研究了40 mg·kg-1 Cd(CdCl2·2.5 H2O)胁迫下, 不同浓度乙二胺二琥珀酸(EDDS)(0、0.5、1.5、2.5、5.0 mmol·L-1)单施及EDDS与一氧化氮(NO)供体硝普钠(SNP)(0、0.25、0.5、1.0 mmol·L-1)联合施加对三叶鬼针草(Bidens pilosa L.)幼苗应激信号分子NO产生量和一氧化氮合酶(NOS)活性的影响。结果表明: 单施EDDS, 植株不同部位NO生成量随EDDS浓度的升高呈增加趋势, 5.0 mmol·L-1时达到最大; 0.5 mmol·L-1的EDDS可增强根、叶中NOS活性。在探究NO产生较多和NOS活性增强显著的EDDS处理浓度与SNP联合施加的研究中发现, 随SNP浓度的升高, 根中NO生成量先升高后降低, 茎和叶中持续升高; 适宜浓度的SNP可进一步增强植株体内NOS活性。EDDS诱导NO的生成会被硝酸还原酶(NR)抑制剂(NaN3)和NOS抑制剂(L-NAME)抑制, 对EDDS处理下NOS活性影响较小。NO清除剂(c-PTIO)能有效清除部分NO, 增强根和叶中NOS活性。因此, 在Cd胁迫下, 适宜浓度的EDDS单施及与SNP联合施加都会增加三叶鬼针草幼苗体内NO产生量。     

镉胁迫下紫花苜蓿幼苗内源一氧化氮和活性氧的生成

以"甘农三号"紫花苜蓿幼苗为材料,在水培条件下,研究了不同浓度镉(Cd)胁迫下紫花苜蓿根、茎和叶内源一氧化氮(NO)和活性氧(ROS)的生成机制以及根系活力的变化.结果表明:在0~2.0 mmol·L-1范围内,随着Cd浓度的增加,幼苗内NO含量呈现先升高后降低的趋势,最后可维持在略高或持平于对照的水平.幼苗内一氧化氮合成酶(NOS)活性、硝酸还原酶(NR)活性、亚硝酸根离子(NO2-)含量和类胡萝卜素(Car)含量的变化与NO含量变化规律相似却又不全相同.NOS和NR是影响幼苗茎中NO含量的主要因素,NOS、NO2-和NR则是影响叶中NO含量的主要因素,而根中NO含量主要与NOS活性和NO2-含量有较大相关性.随着Cd浓度的增加,幼苗内过氧化氢(H2 O2)含量、丙二醛(MDA)含量、超氧阴离子(O-2·)含量和相对电导率(REC)呈现显著升高趋势,说明高浓度的Cd处理会使ROS大量积累,细胞膜遭破坏,细胞质外流,进而引发膜脂过氧化.随着Cd浓度的增加,紫花苜蓿根系活力的变化为先升高后降低,指示了低浓度Cd处理会促进植物代谢,增强其生命力;而高浓度Cd会致使植株代谢受抑制,细胞受损害.NO和ROS的相关性不大,说明二者虽同为自由基,但它们产生和变化方式大有差别.     

Role of nitric oxide dependence on nitric oxide synthase-like activity in the water stress signaling of maize seedling

Nitric oxide (NO) has been known as an important signal in plant antioxidative defense but its production and roles in water stress are less known. The present study investigated whether NO dependence on a NO synthase-lika (NOS) activity is involved in the signaling of drought-induced protective responses in maize seedlings. NOS activity, rate of NO release and drought responses were analyzed when NO donor sodium nitroprusside (SNP), NO scavenger c-PTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramathylimidazoline-1-oxyl-3-oxide) and NOS inhibitor L-NAME (NG-nitro-L-arginine methyl ester) were applied to both detached maize leaves and whole plants. Both NOS activity and the rate of NO release increased substantially under dehydration stress. The high NOS activity induced by c-PTIO as NO scavenger and NO accumulation Inhibited by NOS inhibitor L-NAME In dehydration-treated maize seedlings Indicated that most NO production under water deficit stress may be generated from NOS-like activity. After dehydration stress for 3 h, detached maize leaves pretreated with NO donor SNP maintained more water content than that of control leaves pretreated with water. This result was consistent with the decrease in the transpiration rate of SNP-treated leaves subjected to drought treatment for 3 h. Membrane permeability, a cell injury index, was lower in SNP-trested maize leaves under dehydration stress for 4 h when compared with the control leaves. Also, superoxide dismutsse (SOD) activity of SNP combined drought treatment maize leaves was higher than that of drought treatment alone, indicating that exogenous NO treatment alleviated the water loss and oxidative damage of maize leaves under water deficit stress. When c-PTIO as a specific NO scavenger was applied, the effects of applied SNP were overridden. Treatment with L-NAME on leaves also led to higher membrane permeability, higher transpiration rate and lower SOD activities than those of control leaves, indicating that NOS-like activity was involved in the antioxidative defense under water stress. These results suggested that NO dependence on NOS-like activity serves as a signaling component in the induction of protective responses and is associated with drought tolerance in maize seedlings.     

外源NO对缺镁胁迫下玉米幼苗生长和离子平衡的影响

研究了在缺镁胁迫下,外源NO对缺镁玉米幼苗生长、根系活力和离子含量的影响。结果表明,缺镁胁迫使玉米幼苗株高、根长和干鲜重下降,根系活力降低,N元素在地上部和根部分配失调,新叶和老叶中Mg2＋、Cu2＋、Fe3＋、Mn2＋等离子含量下降,Ca2＋、K＋、Zn2＋等离子含量上升。根中Mg2＋离子含量下降,Ca2＋、K＋、Zn2＋、Cu2＋、Fe3＋、Mn2＋等离子含量上升。用100μmol·L-1一氧化氮供体硝普钠（SNP）处理后,玉米幼苗株高、根长、干重和鲜重均提高,根系活力增强,改善了N代谢,新叶中Ca2＋、K＋和Zn2＋等离子含量下降,Mg2＋、Cu2＋、Fe3＋和Mn2＋等离子含量提高,老叶中Mg2＋、Ca2＋、K＋和Zn2＋等离子含量下降,Cu2＋、Fe3＋和Mn2＋等离子含量提高,根中Mg2＋、Ca2＋、K＋、Cu2＋、Zn2＋、Fe3＋和Mn2＋离子含量均下降。实验结果表明,NO保护玉米幼苗免受缺镁胁迫的影响。     

Cadmium decreases crown root number by decreasing endogenous nitric oxide,which is indispensable for crown root primordia initiation in rice seedlings

Cadmium (Cd) is toxic to crown roots (CR), which are essential for maintaining normal growth and development in rice seedlings. Nitric oxide (NO) is an important signaling molecule that plays a pivotal role in plant root organogenesis. Here, the effects of Cd on endogenous NO content and root growth conditions were studied in rice seedlings. Results showed that similar to the NO scavenger, cPTIO, Cd significantly decreased endogenous NO content and CR number in rice seedlings, and these decreases were recoverable with the application of sodium nitroprusside (SNP, a NO donor). Microscopic analysis of root collars revealed that treatment with Cd and cPTIO inhibited CR primordia initiation. In contrast, although SNP partially recovered Cd-caused inhibition of CR elongation, treatment with cPTIO had no effect on CR elongation. l-NMMA, a widely used nitric oxide synthase (NOS) inhibitor, decreased endogenous NO content and CR number significantly, while tungstate, a nitrate reductase (NR) inhibitor, had no effect on endogenous NO content and CR number. Moreover, enzyme activity assays indicated that treatment with SNP inhibited NOS activity significantly, but had no effect on NR activity. All these results support the conclusions that a critical endogenous NO concentration is indispensable for rice CR primordia initiation rather than elongation, NOS is the main source for endogenous NO generation, and Cd decreases CR number by inhibiting NOS activity and thus decreasing endogenous NO content in rice seedlings.     

一氧化氮在乙烯诱导蚕豆气孔关闭中的作用

以蚕豆为材料研究了一氧化氮(nitric oxide,NO)和乙烯对气孔运动的影响。结果表明,10μmol/L的NO供体硝普钠(sodium nitroprusside,SNP)以及0.04%的乙烯能明显诱导蚕豆气孔关闭,并且二者共同处理后,能够增强其促进气孔关闭的作用。乙烯合成抑制剂AVG可以减弱NO诱导气孔关闭的程度,NO清除剂c-PTIO和NR抑制剂NaN3也可减弱乙烯诱导气孔关闭的程度,而一氧化氮合酶(nitric oxide synthase,NOS)抑制剂L-NAME对乙烯诱导气孔关闭的作用不明显。推测,在调控蚕豆气孔关闭过程中,NO可能主要通过NR途径参与乙烯调控气孔关闭过程。     

Glucose-6-phosphate dehydrogenase plays a pivotal role in nitric oxide-involved defense against oxidative stress under salt stress in red kidney bean roots

The pivotal role of glucose-6-phosphate dehydrogenase (G-6-PDH)-mediated nitric oxide (NO) production in the tolerance to oxidative stress induced by 100 mM NaCl in red kidney bean (Phaseolus vulgaris) roots was investigated. The results show that the G-6-PDH activity was enhanced rapidly in the presence of NaCl and reached a maximum at 100 mM. Western blot analysis indicated that the increase of G-6-PDH activity in the red kidney bean roots under 100 mM NaCl was mainly due to the increased content of the G-6-PDH protein. NO production and nitrate reductase (NR) activity were also induced by 100 mM NaCl. The NO production was reduced by NaN(3) (an NR inhibitor), but not affected by N(omega)-nitro-L-arginine (L-NNA) (an NOS inhibitor). Application of 2.5 mM Na(3)PO(4), an inhibitor of G-6-PDH, blocked the increase of G-6-PDH and NR activity, as well as NO production in red kidney bean roots under 100 mM NaCl. The activities of antioxidant enzymes in red kidney bean roots increased in the presence of 100 mM NaCl or sodium nitroprusside (SNP), an NO donor. The increased activities of all antioxidant enzymes tested at 100 mM NaCl were completely inhibited by 2.5 mM Na(3)PO(4). Based on these results, we conclude that G-6-PDH plays a pivotal role in NR-dependent NO production, and in establishing tolerance of red kidney bean roots to salt stress.     

外源NO对NaCl胁迫下黄瓜幼苗生长和根系谷胱甘肽抗氧化酶系统的影响

以津春2号黄瓜为材料,采用营养液水培的方法,研究了外源一氧化氮(NO)对黄瓜幼苗生长和根系谷胱甘肽抗氧化酶系统的影响.结果表明,(1)正常生长条件下添加NO能促进黄瓜幼苗生长,而添加亚甲基蓝(MB-1)显著抑制黄瓜幼苗的生长;(2)添加NO显著缓解了NaCl胁迫对黄瓜幼苗生长的抑制,提高根系还原型谷胱甘肽(GSH)含量、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性,而氧化型谷胱甘肽(GSSG)含量略有下降,同时缓解了NaCl胁迫下抗坏血酸(ASA)含量的下降幅度;(3)NaCl胁迫下添加NO的同时添加MB-1可部分解除NO的作用,与NaCl胁迫下单独添加NO处理比较,GR活性、GSH和ASA含量均降低,GSSG含量提高,APX先升高后下降.研究发现,外源NO可能通过鸟苷酸环化酶(cGC)介导来调节NaCl胁迫下黄瓜幼苗根系GR活性和GSH、GSSG、ASA含量,提高抗氧化酶活性和非酶抗氧化物质含量,增强植株对活性氧的清除能力,减少膜脂过氧化,缓解NaCl胁迫对黄瓜幼苗造成的伤害.