一氧化氮在绿豆下胚轴不定根发生过程中的作用研究
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摘要
在哺乳动物中，已证明一氧化氮参与了神经传递、血管舒张、平滑肌松弛等过程。尽管一氧化氮研究在植物方面开始较晚，但最近10年已经证明NO参与了许多重要的植物生理过程，如生长、发育、病原体防御反应、程序性细胞死亡和胁迫抗性等。前人曾证明NO参与IAA诱导的黄瓜下胚轴不定根发生，但其研究未涉及不定根发生过程中NO的时间和空间变化以及NO的来源等问题，本文以绿豆下胚轴为材料，从不同角度研究了NO在不定根发生中的作用以及IBA、NAA诱导不定根发生的机制，并对不定根发生过程中NO的来源进行了探讨。结果如下：
1．NO供体SNP可有效促进插条生根，其最适浓度为300μmol／L，浓度高时则具有伤害效应。SNP处理时间、苗龄及插条处理位置都影响生根效果。从5天龄幼苗子叶节下6cm处切取的插条，以300μmol／LSNP处理24小时生根效果最好。SNP与IBA或NAA混合处理比各自单独处理的生根效果好。NO清除剂c-PTIO和动物NOS抑制剂L-NAME单独或与IBA和NAA混合处理明显抑制插条生根，表明内源NO在不定根形成中可能起着重要作用，其来源与NOS有关。
2．利用NO特异的荧光探针对对照及IBA和NAA处理插条生根过程中内源NO的时空变化进行了检测。结果表明，无论对照还是IBA或NAA处理插条，伴随着根原基的发育，插条基部2mm区域NO荧光从无到有逐渐增强，36h主要分布于维管组织之间不定根发生区域，48h后则主要分布于根原基前端分生组织中。非生根区域无NO荧光。对照与IBA或NAA处理插条基部2mm-5mm区域NO产生及分布则完全不同，在实验期间对照插条该区域无NO产生，而IBA或NAA处理插条该区域NO产生、分布及随时间变化趋势与对照插条基部2mm区域基本一致。NO清除剂c-PTIO抑制所有处理插条的根原基形成。这些结果显示，内源NO在不定根形成中起重要作用，而且IBA和NAA诱导的不定根发生也是由NO介导的。
3．NADPH—黄递酶为NOS标志酶，本文以该酶组织化学法对不定根发生过程中的类似动物NOS活性进行了检测。结果表明，0—60h，无论对照还是IBA或NAA处理插条，伴随着根原基的发生和发育，插条基部2mm区域NADPH—黄递酶活性逐渐增强，且集中分布在维管组织之间根原基发生区域及正在形成的根原基中，其
它非生根区域基本无阳性反应。在根原基中较强阳性也集中分布于分生组织中，
这与NO的分布一致。与对照相比，相同时间IBA或NAA处理的插条NADPH一黄递
酶阳性反应略强。对照与工BA或NAA处理插条基部Zlnm一Slnln区域NADPH一黄递酶分
布也完全不同，对照插条该区域NADPH一黄递酶活性较弱，而工BA或NAA处理插条
该区域NADPH一黄递酶活性、分布及随时间变化趋势与插条基部Zllun区域基本一致。
L一NAME既抑制所有处理插条的根原基的形成，也降低NO荧光和NADPH一黄递酶活
性。以上结果表明植物中也存在类似于动物的NOS，而且该酶以及它催化产生的
NO在不定根发生过程中起着重要作用。
Several roles have been described for NO in animals, where it takes part in neurotransmission, vasorelaxation, smooth muscle relaxation, and immunoregulation of pathophysiological processes. Even though NO research in plants is not as advanced as in animals, in the last decade No was proved to participate in many key plant physiological process such as growth, development, pathogen defense reaction, programmed cell death, and stress tolerance. Though NO mediation of the adventitious rooting process induced by auxins had been demonstrated, the time and space changes of NO and the source of NO accompanied adventitious root generation and development are unknown. In this study, mung bean hypocotyls were used as materials, and the role and the source of NO of the cuttings treated by H2^O, IBA or NAA during the adventitious root formation were studied from several aspects. The results were as follows:
1. The cuttings were treated with NO-associated reagent. The results showed that NO donor-SNP significantly enhance the adventitious rooting. 300μmol/L SNP proved an optimal concentration for the initiation of organogenesis. The treatment time of SNP, the ages and the position of the cuttings also influenced the rooting. The hypocotyls of 5 days old seedlings was excised 6 cm below the cotyledonary node and then treated with 300μmol/L SNP by 24h had the optimal rooting effect. SNP obviously enhanced the effects of IBA or NAA in stimulating rooting. C-PTIO, NO specific scavenger, or NOS inhibitor L-NAME treted alone or plus IBA or NAA resulted in an inhibitory effect. It's indicated that endogenous NO appears to play a key role in the generation and development of adventitious roots, and the production of NO in this process maybe catalyzed by NOS.
2. The temporal and spatial change of NO of the cuttings treated by H2O, IBA or NAA during the generation and the development of adventitious roots of mung bean hypocotyl cuttings were detected by NO-sensitive fluorescence probe DAF-2DA. The
results indicated that, with the development of root primordium, the fluorescence of NO appeared and increased gradually in 2mm region of hypocotyl basal part in all treatments. The green fluorescence of NO was detected in the region between the vascular tissues of the hypocotyls at 36h after cutting. The cytosol of DAF-2DA-positive cells stained uniformly for NO. With the development and elongation of root primordium, the number of DAF-2DA-positive cells increased gradually, and the root apical meristem cells show intense fluorescence. No NO was observed in non-rooting areas. In 2mm-5mm region of hypocotyl basal part, the generation and distribution of NO of the cuttings treated by IB A or NAA were different with that of treated by H2O. No NO was detected in control. The generation, distribution and change of NO of cuttings treated by IBA or NAA in this region were in accordance with that of in 2mm region of hypocotyl basal part. C-PTIO, the specific NO scavenger, suppressed the fluorescence and inhibited the formation of root primordial clearly. The results strongly argue that that endogenous NO appears to play a key role in the adventitious rooting process. It also suggested that NO could also mediate the IBA and NAA response during the rooting process in mung bean.
3. NOS-like enzyme activity was detected during adventitious root formation by NADPH-diaphorase histochemical method. With the origination and development of adventitious roots, not in control but in the cuttings treated by IBA or NAA, the activity of NADPH-diaphorase and the number of cell expressing NADPH-diaphorase-positive increased rapidly in the forming root in 2mm region of hypocotyl basal part. The non-rooting areas haven't positive cells. In accordance with the distribution of NO, intense staining mainly distributed in root meristem. The NADPH-diaphorase activity of the cuttings treated by IBA or NAA were more intense than the control in the same time. In 2mm-5mm region of hypocotyl basal part, the distribution of NADPH-diaphorase of the cuttings treated by IBA or
1．NO供体SNP可有效促进插条生根，其最适浓度为300μmol／L，浓度高时则具有伤害效应。SNP处理时间、苗龄及插条处理位置都影响生根效果。从5天龄幼苗子叶节下6cm处切取的插条，以300μmol／LSNP处理24小时生根效果最好。SNP与IBA或NAA混合处理比各自单独处理的生根效果好。NO清除剂c-PTIO和动物NOS抑制剂L-NAME单独或与IBA和NAA混合处理明显抑制插条生根，表明内源NO在不定根形成中可能起着重要作用，其来源与NOS有关。
2．利用NO特异的荧光探针对对照及IBA和NAA处理插条生根过程中内源NO的时空变化进行了检测。结果表明，无论对照还是IBA或NAA处理插条，伴随着根原基的发育，插条基部2mm区域NO荧光从无到有逐渐增强，36h主要分布于维管组织之间不定根发生区域，48h后则主要分布于根原基前端分生组织中。非生根区域无NO荧光。对照与IBA或NAA处理插条基部2mm-5mm区域NO产生及分布则完全不同，在实验期间对照插条该区域无NO产生，而IBA或NAA处理插条该区域NO产生、分布及随时间变化趋势与对照插条基部2mm区域基本一致。NO清除剂c-PTIO抑制所有处理插条的根原基形成。这些结果显示，内源NO在不定根形成中起重要作用，而且IBA和NAA诱导的不定根发生也是由NO介导的。
3．NADPH—黄递酶为NOS标志酶，本文以该酶组织化学法对不定根发生过程中的类似动物NOS活性进行了检测。结果表明，0—60h，无论对照还是IBA或NAA处理插条，伴随着根原基的发生和发育，插条基部2mm区域NADPH—黄递酶活性逐渐增强，且集中分布在维管组织之间根原基发生区域及正在形成的根原基中，其
它非生根区域基本无阳性反应。在根原基中较强阳性也集中分布于分生组织中，
这与NO的分布一致。与对照相比，相同时间IBA或NAA处理的插条NADPH一黄递
酶阳性反应略强。对照与工BA或NAA处理插条基部Zlnm一Slnln区域NADPH一黄递酶分
布也完全不同，对照插条该区域NADPH一黄递酶活性较弱，而工BA或NAA处理插条
该区域NADPH一黄递酶活性、分布及随时间变化趋势与插条基部Zllun区域基本一致。
L一NAME既抑制所有处理插条的根原基的形成，也降低NO荧光和NADPH一黄递酶活
性。以上结果表明植物中也存在类似于动物的NOS，而且该酶以及它催化产生的
NO在不定根发生过程中起着重要作用。
Several roles have been described for NO in animals, where it takes part in neurotransmission, vasorelaxation, smooth muscle relaxation, and immunoregulation of pathophysiological processes. Even though NO research in plants is not as advanced as in animals, in the last decade No was proved to participate in many key plant physiological process such as growth, development, pathogen defense reaction, programmed cell death, and stress tolerance. Though NO mediation of the adventitious rooting process induced by auxins had been demonstrated, the time and space changes of NO and the source of NO accompanied adventitious root generation and development are unknown. In this study, mung bean hypocotyls were used as materials, and the role and the source of NO of the cuttings treated by H2^O, IBA or NAA during the adventitious root formation were studied from several aspects. The results were as follows:
1. The cuttings were treated with NO-associated reagent. The results showed that NO donor-SNP significantly enhance the adventitious rooting. 300μmol/L SNP proved an optimal concentration for the initiation of organogenesis. The treatment time of SNP, the ages and the position of the cuttings also influenced the rooting. The hypocotyls of 5 days old seedlings was excised 6 cm below the cotyledonary node and then treated with 300μmol/L SNP by 24h had the optimal rooting effect. SNP obviously enhanced the effects of IBA or NAA in stimulating rooting. C-PTIO, NO specific scavenger, or NOS inhibitor L-NAME treted alone or plus IBA or NAA resulted in an inhibitory effect. It's indicated that endogenous NO appears to play a key role in the generation and development of adventitious roots, and the production of NO in this process maybe catalyzed by NOS.
2. The temporal and spatial change of NO of the cuttings treated by H2O, IBA or NAA during the generation and the development of adventitious roots of mung bean hypocotyl cuttings were detected by NO-sensitive fluorescence probe DAF-2DA. The
results indicated that, with the development of root primordium, the fluorescence of NO appeared and increased gradually in 2mm region of hypocotyl basal part in all treatments. The green fluorescence of NO was detected in the region between the vascular tissues of the hypocotyls at 36h after cutting. The cytosol of DAF-2DA-positive cells stained uniformly for NO. With the development and elongation of root primordium, the number of DAF-2DA-positive cells increased gradually, and the root apical meristem cells show intense fluorescence. No NO was observed in non-rooting areas. In 2mm-5mm region of hypocotyl basal part, the generation and distribution of NO of the cuttings treated by IB A or NAA were different with that of treated by H2O. No NO was detected in control. The generation, distribution and change of NO of cuttings treated by IBA or NAA in this region were in accordance with that of in 2mm region of hypocotyl basal part. C-PTIO, the specific NO scavenger, suppressed the fluorescence and inhibited the formation of root primordial clearly. The results strongly argue that that endogenous NO appears to play a key role in the adventitious rooting process. It also suggested that NO could also mediate the IBA and NAA response during the rooting process in mung bean.
3. NOS-like enzyme activity was detected during adventitious root formation by NADPH-diaphorase histochemical method. With the origination and development of adventitious roots, not in control but in the cuttings treated by IBA or NAA, the activity of NADPH-diaphorase and the number of cell expressing NADPH-diaphorase-positive increased rapidly in the forming root in 2mm region of hypocotyl basal part. The non-rooting areas haven't positive cells. In accordance with the distribution of NO, intense staining mainly distributed in root meristem. The NADPH-diaphorase activity of the cuttings treated by IBA or NAA were more intense than the control in the same time. In 2mm-5mm region of hypocotyl basal part, the distribution of NADPH-diaphorase of the cuttings treated by IBA or
引文
[1] Koshkand DE, The molecule of the year, Science, 1992, 258:1861-1861
[2] Cultta E, Koshland DE, NO news is good news, Science, 1992, 258: 1862-1863
[3] Butler AR, Williams DLH, The physiological role of nitric oxide, Chem Soc Rev, 1993:233-235
[4] Dawson TM, Dawson VL, Nitric oxide: action and pathological roles, Neuroscientist, 1995, 1(1): 7-9
[5] Dawson VL, Dawson TM, Nitric oxide action in neurochemistry, Neurochem Int, 1996,29(2):97-100
[6] 叶建山，朱传征 一氧化氮分子——打开神秘生命科学大门的一把钥匙，化学教育，1997，11：3-4
[7] 吴国庆，一氧化氮神奇生物化学作用正在揭示中，化学教育，1997，11：4-5
[8] Klaus-DK, Karin F, Victoria KB, Nitric oxide: cytotoxicity versus cytoprotection-how, why, when, and where? Nitric oxide: Biology and Chemistry, 1997, 1(2): 107-120
[9] Beligni MY, Lamattina L, 2000. Nitric oxide stimulates seed germination and de-etiolation, and inhibits hypocotyl elongation, three light-inducible responses in plants. Planta, 210:215-221
[10] Giba Z, Grubiic D, Todorobic S, Sajc L, Stojakovic D, Konjecic R, Effect of nitric oxide-releasing compounds on phytochrome-controlled germination of Empress tree seeds, Plant Growth Reg, 1998, 26: 175-181
[11] Pagnussat GC, Simontacchi M, Puntarulo S, Lamattina L, Nitric oxide is required for root organogenesis, Plant Physio., 2002, 129:954-956
[12] Leshem YY, Nitric oxide in biological systems, Plant Growth Regul, 1996, 18:155-159
[13] Ribeiro EA Jr, Cunha FQ, Tamashiro WMSC et al., Growth phase-depedent subcellular localization of nitric oxide synthase in maize cells, FEBS Lett, 1999, 445:283-286
[14] Delledonne M, Xia YJ, Dixon RA et al., Nitric oxide functions as a signal in plant disease resistance, 1998, Nature, 394:585-588
[15] Pedroso MC, Magalhaes JR, Durzan A, A nitric oxide burst precedes apoptosis in angiosperm and gymnosperm callus cells and foliar tissues, J Exp Bot., 2000, 51: 1027-1036
[16] Clark A, Desikan R, Hurst RD et al, NO way back: nitric oxide and programmed cell death in Arabidopsis thaliana suspension cultures, The Plant J, 2000, 24: 667-677
[17] Mata CG, Lamattina L, Nitric oxide induces stomatal closure and enhances the adaptive plant response against drought stress, Plant Physiol., 2001, 126: 1196-1204
[18] Barroso JB, Corpas FJ, Carteras A et al, Localization of nitric oxide synthase in plant peroxisomes, J Biol Chem, 1999, 274(51): 36729-36733
[19] Pedroso MC, Magalhaes JR, Durzan D, Nitric oxide induces cell death in Taxus cells, Plant Sci, 2000, 157(2): 173-180
[20] Sen S, Cheema IR, Nitric oxide synthase and calmodulin immunoreactivity in plant embryonic tissue, Biochemical Archives, 1995, 11: 221-227
[21] Modlol LV, Cunha FQ, Braga MR et al, Nitric oxide synthase-mediated phytoalexin accumulation in soybean cotyledons in response to the Diaporthe phaseolorum f. sp. Meridionalis Elicitor, Plant Physiol, 2002, 130: 1288-1297
[22] Dean JV, Harper JE, The conversion of nitrite to nitrogen oxide(s) by the constitutive NAD(P)H-nitrate reductase enzyme from soybean, Plant Physiol, 1988, 88: 389-395
[23] Yamasak H, Sakiham Y, Simultaneous production of nitric oxide and peroxynitrite by plant nitrate reductase: in vitro evidence for the NR-dependent formation of active nitrogen species, FEBS Lett, 2000, 468: 89-92
[24] Yamasak H, Sakiham Y, Takahash S, An alternative pathway for nitric
oxide production in plants: New features of an old enzyme, Trends Plant Sci, 1999, 4(4):128-129
[25] Rochel P, Strube F, Rockel A et al, Regulation of nitric oxide(NO) production by plant nitrate reductase in vivo and in vitro, J Exp Bot, 2002,53:103-110
[26] 方芳，刘景生，《细胞信息与调控》，北京医科大学中国协和医科大学出版社，1998：207—227
[27] 孙存普，张建中，段绍瑾，《自由基生物学导论》，中国科学技术大学出版社，1999，94-111
[28] 归莉琼，魏东芝，NO：一种重要的生物信使分子，生命科学，1998，10(4)：188-190
[29] Schmidt HHWH, Walter U, NO at work, Cell, 1994, 78:919-925
[30] Wojtaszek P, Nitric oxide in plants-to NO or not to NO, Phytochemistry, 2000, 54(1): 1-4
[31] Caro A, Puntarulo S, Nitric oxide generation by soybean embryonic axes, Possible effect on mitochondrial function, Free Radic Res, 1999, 31(Sup): S205-212
[32] Leshem YY, Pinchasov Y, Non-invasive photoacoustic speectroscopic determination of relative endogenous nitric oxide and ethylene content stoichiometry during the ripening of strawberries Fragaria anannasa(Duch.) and avocados Persea americana(Mill.), J Exp Bot, 2000,51:1471-1473
[33] Cueto M, Hernandez-Perera O, Martin R et al, Presence of nitric oxide synthase activity in roots and nodules of Lupinus albus, FEBS Lett, 1996, 398: 159-164
[34] Marletta MA, Nitric oxide synthase structure and mechanism, J Biol Chem, 1993, 268: 12231-12234
[35] Nathan C, Xie QW, Nitric oxide synthase: roles tolls, and controls, Cell, 1994, 78: 915-918
[36] Durner J, Wendehenne D, Klessig DF, Defense gene induction in tobacco
by nitric oxide, cyclic GMP, and cyclic ADP-ribose, Proc Natl Acad Sci USA, 1998, 95(17): 10328-10333
[37] Foissner I, Wendehenne D, hangebartels C et al, In vivo imaging of an elicitor or induced nitric oxide burst in tobacco, Plant J, 2000, 23: 817-824
[38] Cho MJ, Vaghy PL, Kondo R et al, Reciprocal regulation of mammalian nitric oxide synthase and calcineurin by plant calmodulin isoforms, Biochemistry, 1998, 37(45): 15593-15597
[39] Ninnemann H, Maier J, Indications for occurrence of nitric oxide synthase in fungi and plants and involvement in photoconidiation of Neurospora crassa, Phorochem Photobiol, 1996, 64: 393-398
[40] Zhao Z, Chen G, Zhang C, Interaction between reactive oxygen species and nitric oxide in drought-induced abscisic acid synthesis in root tips of wheat seedlings, Aust J Plant Physiol, 2001, 28: 1055-1061
[41] Pedroso MC, Durzan DJ, Effect of different gravity environments on DNA fragmentation and cell death in Kalanchoe leaves, Ann Bot, 2000, 86: 983-994
[42] Dean JV, Harper JE, Nitric oxide and nitrous oxide production by soybean and winged bean during the in vivo nitrate reductase assay. Plant Physiol, 1986, 82: 718-723
[43] Yamasaki H, Sakihama Y and Takahashi S. An alterative pathway for nitric oxide production in plants: new features of an old enzyme. Trends Plant Sci, 1999, 4(4): 128—129
[44] Yamasaki H, Sakihama Y. Simultaneous production of nitric oxide and peroxynitrite by plant nitrate reductase: in vitro evidence for the NR-dependent formation of active nitrogen species. FEBS Lett, 2000, 468(1): 89—92
[45] Rochel P, Strube F, Rockel A, Wildt J, Kaiser WM. Regulation of nitric oxide(NO)production by plant nitrate reductase in vivo and in vitro.J Exp Bot, 2002, 53: 103-110
[46] Wildt J, Kley D, Rockel A, Rockel P, Segschneider HJ, Emission of NO from higher plant species, J Geo Res, 1997, 102: 5919-5927
[47] Crawford NM, Nitrate: nutrient and signal for plant growth. Plant Cell, 1995, 7: 859-868
[48] Garcia-Mata C, Lamattina L, Nitric oxide and abscisic acid cross talk in guard cells, Plant Physiol, 2002,128:790-792
[49] Wildt J, Kley D, Rockel A et al., Emission of NO from higher plant species, J Geo Res, 1997, 102: 5919-5927
[50] Durner J, Klessig DF, Nitric oxide as a signal in plants. Curr Opin Plant Biol, 1999, 2: 369-374
[51] Cooney RV, Harwood PJ, Custer IJ et al., Light mediated conversion of nitrogen dioxide to nitric oxide by carotenoids. Environ Health Persp, 1994, 102: 460-462
[52] Wojtaszek P, Nitric oxide in plants—To NO or not to NO, Phyto Chemistry, 2000, 54(1): 1-4
[53] Leshem YY, Haramaty E, Plant aging: the emission of NO and ethylene and the effect of NO-releasing compounds on growth of pea(Pisum sativum) foliage, L Plant Physiol, 1996, 148: 258-263
[54] Gouva CMCP, Souza JF, Magalhes ACN, Martins IS, NO-releasing substances that induce growth elongation in maize root segments, Plant Growth Reg, 1997,21:183-187
[55] Pagnussat GC, Lanteri ML, Lamattina L, Nitric oxide and cyclic GMP are messengers in the indole acetic-induced adventitious rooting process. Plant Physio, 2003,132:1241-1248
[56] Klessig DF, Durner J, Noad R, Navarre DA, Wendehenne D, Kumar D, Zhou JM, Shah J, Zhang SQ, Kachroo P, Trifa Y, Pontier D, Lam E, Sliva H, Nitric oxide and salicylic acid signaling in plant defense, Proc Natl Acad Aci USA, 2000, 97: 8849-8855
[57] Magalhaes JR, Monte DC, Durzan D, Nitric oxide and ethylene emission in Arabidopsis thaliana, Physiol Mol Biol Plant, 2000, 6: 117-127
[58] Zhao Z, Chen G, Zhang C, Interaction between reactive oxygen species and nitric oxide in drought-induced abscisic acid synthesis in root tips of wheat seedlings, Aust J Plant Physiol, 2001, 28: 1055-1061
[59] Neill SJ, Desikan R, Clarke A, Hancock JT, Nitric oxide is a novel component of abscisic acid signaling in stomatal guard cells, Plant Physiol, 2002, 128: 13-16
[60] Leshem YY, Wills RBH, Ku UVV. Evidence for the function of the free radical gas-nitric oxide(NO)-as an endogenous maturation and senescence regulating factor in higher plants. Plant Physio Biochem, 1998, 36: 825-833
[61] Wills RBH, Leshem YY. Method: Use of nitric oxide to extend the postharvest life of fruit, vegetables and flowers, po9370, Australian Provisional Patent, 1997.
[62] Mata CG, Lamattina L, Nitric oxide and abscisic acid cross talk in guard cells. Plant Physiol, 2002, 128: 790-792
[63] Tun NN, Holk A, Soberer GFE, Rapid increase of NO release in plant cell cultures induced by cytokinin, FEBS Lett, 2001, 509: 174-176
[64] Beligni MY, Fath A, Bethke PC, Lamattina L and Jones RL, Nitric oxide acts as an antioxidant and delays programmed cell death in barley aleurone layers, Plant Physiol, 2002, 129: 1642-1650
[65] Beligni MV, Lamattina L, Nitric oxide: a non-traditional regulator of plant growth, Trends in Plant Science, 2001,6:508-509
[66] Clarke A, Desikan R, Hurst RD, Hancock JT, Neill SJ, NO way back: nitric oxide and programmed cell death in Arabidopsis thaliana suspension cultures, Plant J, 2000, 24: 667-677
[67] Navarre DA, Wendehenne D, Durner J, Noad R, Klessig DF, Nitric oxide modulates the activity of tobacco aconitase, Plant Physiol, 2000, 122: 573-582
[68] 潘瑞炽，李铃，植物生长发育的化学调控，广东高等教育出版社，1995，52-54
[69] Javis BC., Endogenous control of adventitious rooting in non-woody cuttings. In:
Jackson MB. ed. New Root Formation in Plants and Cuttings. The Netherlands: Martinus Nijhoff Publishers, 1986,191-222
[70] Brune B, Lapetina EG, Activation of a cytosolic ADP-ribosyltransferase by nitric oxide-generating agents, The J. of Biol. Chem., 1989, 264, 8455-8458.
[71] Beligni MV, Lamattina L, Is nitric oxide toxic or protective? Trends Plant Sci., 1999, 4, 299-300.
[72] Pan RC, Zhao ZJ, Synergistic effects of plant growth retardants and IBA on the formation of adventitious roots in hypocotyl cuttings of mung bean, Plant Growth Regul, 1994, 14, 15-19.
[73] 黄爱霞，佘小平，硝普钠(SNP)对绿豆下胚轴插条生根的影响，西北植物学报，2003，23(12)，2196-2199．
[74] Nag S, Saha K, Choudhuri MA, Role of auxin and polyamines in adventitious root formation in relation to changes in compounds involved in rooting, J. Plant Growth Regul., 2001, 20, 182-194.
[75] Noritake T, Kawakita K, Doke N, Nitric oxide induces phytoalexin accumulation in potato tuber tissues, Plant Cell Physiol., 1996,37:113-116
[76] Davis TD, Haissig BE, Sankhla N, Adventitious root formation in cutting, In:Dudley TR, editor , Advances in plant science series, Vol. Ⅱ. Portland, Oregon: Dioscorides Press. P, 1988
[77] Gasper T, Kevers C, Hausman JF, Berthon JY, Ripetti V, Practical uses of peroxidase activity as a predictive marker of rooting performance of micropropagated shoots, Agronmie, 1992,12:757-765
[78] Gasper T, Kevers C, Hausman JF, Ripetti V, Peroxdase activity and endogenous free auxin during adventitious root formation, In: Lumsden PJ, Nicholas JR, Davis WJ, editor, Physiology, growth and development of plants in culture, Dordrecht: Kluwer Academic Publisher. p, 1994,
[79] Gasper T, Kevers C, Hausman JF, Indissociable chief factors in the inductive phase of adventitious rooting, In:Altmanm Waisel, editors,
[2] Cultta E, Koshland DE, NO news is good news, Science, 1992, 258: 1862-1863
[3] Butler AR, Williams DLH, The physiological role of nitric oxide, Chem Soc Rev, 1993:233-235
[4] Dawson TM, Dawson VL, Nitric oxide: action and pathological roles, Neuroscientist, 1995, 1(1): 7-9
[5] Dawson VL, Dawson TM, Nitric oxide action in neurochemistry, Neurochem Int, 1996,29(2):97-100
[6] 叶建山，朱传征 一氧化氮分子——打开神秘生命科学大门的一把钥匙，化学教育，1997，11：3-4
[7] 吴国庆，一氧化氮神奇生物化学作用正在揭示中，化学教育，1997，11：4-5
[8] Klaus-DK, Karin F, Victoria KB, Nitric oxide: cytotoxicity versus cytoprotection-how, why, when, and where? Nitric oxide: Biology and Chemistry, 1997, 1(2): 107-120
[9] Beligni MY, Lamattina L, 2000. Nitric oxide stimulates seed germination and de-etiolation, and inhibits hypocotyl elongation, three light-inducible responses in plants. Planta, 210:215-221
[10] Giba Z, Grubiic D, Todorobic S, Sajc L, Stojakovic D, Konjecic R, Effect of nitric oxide-releasing compounds on phytochrome-controlled germination of Empress tree seeds, Plant Growth Reg, 1998, 26: 175-181
[11] Pagnussat GC, Simontacchi M, Puntarulo S, Lamattina L, Nitric oxide is required for root organogenesis, Plant Physio., 2002, 129:954-956
[12] Leshem YY, Nitric oxide in biological systems, Plant Growth Regul, 1996, 18:155-159
[13] Ribeiro EA Jr, Cunha FQ, Tamashiro WMSC et al., Growth phase-depedent subcellular localization of nitric oxide synthase in maize cells, FEBS Lett, 1999, 445:283-286
[14] Delledonne M, Xia YJ, Dixon RA et al., Nitric oxide functions as a signal in plant disease resistance, 1998, Nature, 394:585-588
[15] Pedroso MC, Magalhaes JR, Durzan A, A nitric oxide burst precedes apoptosis in angiosperm and gymnosperm callus cells and foliar tissues, J Exp Bot., 2000, 51: 1027-1036
[16] Clark A, Desikan R, Hurst RD et al, NO way back: nitric oxide and programmed cell death in Arabidopsis thaliana suspension cultures, The Plant J, 2000, 24: 667-677
[17] Mata CG, Lamattina L, Nitric oxide induces stomatal closure and enhances the adaptive plant response against drought stress, Plant Physiol., 2001, 126: 1196-1204
[18] Barroso JB, Corpas FJ, Carteras A et al, Localization of nitric oxide synthase in plant peroxisomes, J Biol Chem, 1999, 274(51): 36729-36733
[19] Pedroso MC, Magalhaes JR, Durzan D, Nitric oxide induces cell death in Taxus cells, Plant Sci, 2000, 157(2): 173-180
[20] Sen S, Cheema IR, Nitric oxide synthase and calmodulin immunoreactivity in plant embryonic tissue, Biochemical Archives, 1995, 11: 221-227
[21] Modlol LV, Cunha FQ, Braga MR et al, Nitric oxide synthase-mediated phytoalexin accumulation in soybean cotyledons in response to the Diaporthe phaseolorum f. sp. Meridionalis Elicitor, Plant Physiol, 2002, 130: 1288-1297
[22] Dean JV, Harper JE, The conversion of nitrite to nitrogen oxide(s) by the constitutive NAD(P)H-nitrate reductase enzyme from soybean, Plant Physiol, 1988, 88: 389-395
[23] Yamasak H, Sakiham Y, Simultaneous production of nitric oxide and peroxynitrite by plant nitrate reductase: in vitro evidence for the NR-dependent formation of active nitrogen species, FEBS Lett, 2000, 468: 89-92
[24] Yamasak H, Sakiham Y, Takahash S, An alternative pathway for nitric
oxide production in plants: New features of an old enzyme, Trends Plant Sci, 1999, 4(4):128-129
[25] Rochel P, Strube F, Rockel A et al, Regulation of nitric oxide(NO) production by plant nitrate reductase in vivo and in vitro, J Exp Bot, 2002,53:103-110
[26] 方芳，刘景生，《细胞信息与调控》，北京医科大学中国协和医科大学出版社，1998：207—227
[27] 孙存普，张建中，段绍瑾，《自由基生物学导论》，中国科学技术大学出版社，1999，94-111
[28] 归莉琼，魏东芝，NO：一种重要的生物信使分子，生命科学，1998，10(4)：188-190
[29] Schmidt HHWH, Walter U, NO at work, Cell, 1994, 78:919-925
[30] Wojtaszek P, Nitric oxide in plants-to NO or not to NO, Phytochemistry, 2000, 54(1): 1-4
[31] Caro A, Puntarulo S, Nitric oxide generation by soybean embryonic axes, Possible effect on mitochondrial function, Free Radic Res, 1999, 31(Sup): S205-212
[32] Leshem YY, Pinchasov Y, Non-invasive photoacoustic speectroscopic determination of relative endogenous nitric oxide and ethylene content stoichiometry during the ripening of strawberries Fragaria anannasa(Duch.) and avocados Persea americana(Mill.), J Exp Bot, 2000,51:1471-1473
[33] Cueto M, Hernandez-Perera O, Martin R et al, Presence of nitric oxide synthase activity in roots and nodules of Lupinus albus, FEBS Lett, 1996, 398: 159-164
[34] Marletta MA, Nitric oxide synthase structure and mechanism, J Biol Chem, 1993, 268: 12231-12234
[35] Nathan C, Xie QW, Nitric oxide synthase: roles tolls, and controls, Cell, 1994, 78: 915-918
[36] Durner J, Wendehenne D, Klessig DF, Defense gene induction in tobacco
by nitric oxide, cyclic GMP, and cyclic ADP-ribose, Proc Natl Acad Sci USA, 1998, 95(17): 10328-10333
[37] Foissner I, Wendehenne D, hangebartels C et al, In vivo imaging of an elicitor or induced nitric oxide burst in tobacco, Plant J, 2000, 23: 817-824
[38] Cho MJ, Vaghy PL, Kondo R et al, Reciprocal regulation of mammalian nitric oxide synthase and calcineurin by plant calmodulin isoforms, Biochemistry, 1998, 37(45): 15593-15597
[39] Ninnemann H, Maier J, Indications for occurrence of nitric oxide synthase in fungi and plants and involvement in photoconidiation of Neurospora crassa, Phorochem Photobiol, 1996, 64: 393-398
[40] Zhao Z, Chen G, Zhang C, Interaction between reactive oxygen species and nitric oxide in drought-induced abscisic acid synthesis in root tips of wheat seedlings, Aust J Plant Physiol, 2001, 28: 1055-1061
[41] Pedroso MC, Durzan DJ, Effect of different gravity environments on DNA fragmentation and cell death in Kalanchoe leaves, Ann Bot, 2000, 86: 983-994
[42] Dean JV, Harper JE, Nitric oxide and nitrous oxide production by soybean and winged bean during the in vivo nitrate reductase assay. Plant Physiol, 1986, 82: 718-723
[43] Yamasaki H, Sakihama Y and Takahashi S. An alterative pathway for nitric oxide production in plants: new features of an old enzyme. Trends Plant Sci, 1999, 4(4): 128—129
[44] Yamasaki H, Sakihama Y. Simultaneous production of nitric oxide and peroxynitrite by plant nitrate reductase: in vitro evidence for the NR-dependent formation of active nitrogen species. FEBS Lett, 2000, 468(1): 89—92
[45] Rochel P, Strube F, Rockel A, Wildt J, Kaiser WM. Regulation of nitric oxide(NO)production by plant nitrate reductase in vivo and in vitro.J Exp Bot, 2002, 53: 103-110
[46] Wildt J, Kley D, Rockel A, Rockel P, Segschneider HJ, Emission of NO from higher plant species, J Geo Res, 1997, 102: 5919-5927
[47] Crawford NM, Nitrate: nutrient and signal for plant growth. Plant Cell, 1995, 7: 859-868
[48] Garcia-Mata C, Lamattina L, Nitric oxide and abscisic acid cross talk in guard cells, Plant Physiol, 2002,128:790-792
[49] Wildt J, Kley D, Rockel A et al., Emission of NO from higher plant species, J Geo Res, 1997, 102: 5919-5927
[50] Durner J, Klessig DF, Nitric oxide as a signal in plants. Curr Opin Plant Biol, 1999, 2: 369-374
[51] Cooney RV, Harwood PJ, Custer IJ et al., Light mediated conversion of nitrogen dioxide to nitric oxide by carotenoids. Environ Health Persp, 1994, 102: 460-462
[52] Wojtaszek P, Nitric oxide in plants—To NO or not to NO, Phyto Chemistry, 2000, 54(1): 1-4
[53] Leshem YY, Haramaty E, Plant aging: the emission of NO and ethylene and the effect of NO-releasing compounds on growth of pea(Pisum sativum) foliage, L Plant Physiol, 1996, 148: 258-263
[54] Gouva CMCP, Souza JF, Magalhes ACN, Martins IS, NO-releasing substances that induce growth elongation in maize root segments, Plant Growth Reg, 1997,21:183-187
[55] Pagnussat GC, Lanteri ML, Lamattina L, Nitric oxide and cyclic GMP are messengers in the indole acetic-induced adventitious rooting process. Plant Physio, 2003,132:1241-1248
[56] Klessig DF, Durner J, Noad R, Navarre DA, Wendehenne D, Kumar D, Zhou JM, Shah J, Zhang SQ, Kachroo P, Trifa Y, Pontier D, Lam E, Sliva H, Nitric oxide and salicylic acid signaling in plant defense, Proc Natl Acad Aci USA, 2000, 97: 8849-8855
[57] Magalhaes JR, Monte DC, Durzan D, Nitric oxide and ethylene emission in Arabidopsis thaliana, Physiol Mol Biol Plant, 2000, 6: 117-127
[58] Zhao Z, Chen G, Zhang C, Interaction between reactive oxygen species and nitric oxide in drought-induced abscisic acid synthesis in root tips of wheat seedlings, Aust J Plant Physiol, 2001, 28: 1055-1061
[59] Neill SJ, Desikan R, Clarke A, Hancock JT, Nitric oxide is a novel component of abscisic acid signaling in stomatal guard cells, Plant Physiol, 2002, 128: 13-16
[60] Leshem YY, Wills RBH, Ku UVV. Evidence for the function of the free radical gas-nitric oxide(NO)-as an endogenous maturation and senescence regulating factor in higher plants. Plant Physio Biochem, 1998, 36: 825-833
[61] Wills RBH, Leshem YY. Method: Use of nitric oxide to extend the postharvest life of fruit, vegetables and flowers, po9370, Australian Provisional Patent, 1997.
[62] Mata CG, Lamattina L, Nitric oxide and abscisic acid cross talk in guard cells. Plant Physiol, 2002, 128: 790-792
[63] Tun NN, Holk A, Soberer GFE, Rapid increase of NO release in plant cell cultures induced by cytokinin, FEBS Lett, 2001, 509: 174-176
[64] Beligni MY, Fath A, Bethke PC, Lamattina L and Jones RL, Nitric oxide acts as an antioxidant and delays programmed cell death in barley aleurone layers, Plant Physiol, 2002, 129: 1642-1650
[65] Beligni MV, Lamattina L, Nitric oxide: a non-traditional regulator of plant growth, Trends in Plant Science, 2001,6:508-509
[66] Clarke A, Desikan R, Hurst RD, Hancock JT, Neill SJ, NO way back: nitric oxide and programmed cell death in Arabidopsis thaliana suspension cultures, Plant J, 2000, 24: 667-677
[67] Navarre DA, Wendehenne D, Durner J, Noad R, Klessig DF, Nitric oxide modulates the activity of tobacco aconitase, Plant Physiol, 2000, 122: 573-582
[68] 潘瑞炽，李铃，植物生长发育的化学调控，广东高等教育出版社，1995，52-54
[69] Javis BC., Endogenous control of adventitious rooting in non-woody cuttings. In:
Jackson MB. ed. New Root Formation in Plants and Cuttings. The Netherlands: Martinus Nijhoff Publishers, 1986,191-222
[70] Brune B, Lapetina EG, Activation of a cytosolic ADP-ribosyltransferase by nitric oxide-generating agents, The J. of Biol. Chem., 1989, 264, 8455-8458.
[71] Beligni MV, Lamattina L, Is nitric oxide toxic or protective? Trends Plant Sci., 1999, 4, 299-300.
[72] Pan RC, Zhao ZJ, Synergistic effects of plant growth retardants and IBA on the formation of adventitious roots in hypocotyl cuttings of mung bean, Plant Growth Regul, 1994, 14, 15-19.
[73] 黄爱霞，佘小平，硝普钠(SNP)对绿豆下胚轴插条生根的影响，西北植物学报，2003，23(12)，2196-2199．
[74] Nag S, Saha K, Choudhuri MA, Role of auxin and polyamines in adventitious root formation in relation to changes in compounds involved in rooting, J. Plant Growth Regul., 2001, 20, 182-194.
[75] Noritake T, Kawakita K, Doke N, Nitric oxide induces phytoalexin accumulation in potato tuber tissues, Plant Cell Physiol., 1996,37:113-116
[76] Davis TD, Haissig BE, Sankhla N, Adventitious root formation in cutting, In:Dudley TR, editor , Advances in plant science series, Vol. Ⅱ. Portland, Oregon: Dioscorides Press. P, 1988
[77] Gasper T, Kevers C, Hausman JF, Berthon JY, Ripetti V, Practical uses of peroxidase activity as a predictive marker of rooting performance of micropropagated shoots, Agronmie, 1992,12:757-765
[78] Gasper T, Kevers C, Hausman JF, Ripetti V, Peroxdase activity and endogenous free auxin during adventitious root formation, In: Lumsden PJ, Nicholas JR, Davis WJ, editor, Physiology, growth and development of plants in culture, Dordrecht: Kluwer Academic Publisher. p, 1994,
[79] Gasper T, Kevers C, Hausman JF, Indissociable chief factors in the inductive phase of adventitious rooting, In:Altmanm Waisel, editors,