外源NO对低温胁迫下核桃幼苗活性氧代谢的影响

doi:10.11707/j.1001-7488.20160117

摘要/Abstract

摘要： [目的] 研究外源NO处理对低温胁迫下核桃幼苗活性氧代谢系统的影响,探讨其影响核桃抗寒性的可能作用机制,寻找通过施加外源NO提高果树抗逆性的新方法,为外源NO在未来核桃抗逆生产中的广泛应用提供理论基础。[方法] 以抗寒性不同的‘香玲’、‘鲁果12号’核桃品种为材料,采用人工气候室模拟低温处理与叶片喷施SNP相结合的方法,研究SNP(200μmol·L^-1)对低温胁迫下核桃幼苗活性氧代谢的影响。[结果] 1)正常生长条件下,喷施SNP对核桃幼苗叶片质膜透性、叶绿素含量、超氧阴离子(O₂^·-)产生速率、过氧化氢(H₂O₂)含量、膜脂过氧化产物丙二醛(MDA)含量、脯氨酸(Pro)含量和超氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)活性影响不大,显著提高了过氧化物酶(POD)、脱氢抗坏血酸还原酶(DHAR)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性和抗坏血酸(AsA)、谷胱甘肽(GSH)含量以及AsA/DHA和GSH/GSSG比值,降低了氧化型抗坏血酸(DHA)和还原型谷胱甘肽(GSH)含量,但2个品种的各项指标变化程度不同。2)低温胁迫下,SNP处理可显著提高核桃幼苗叶片POD,SOD,CAT,APX和GR等抗氧化物酶活性,同时,提高叶绿素和Pro,AsA,DHA,GSH,GSSG含量,减少H₂O₂和MDA的积累,降低O₂^·-产生速率和细胞质膜相对透性,2个品种的变化幅度不同。[结论] 低温胁迫下,外源NO处理可增强核桃幼苗叶片抗氧化物酶活性,提高抗氧化剂含量,维持抗坏血酸-谷胱甘肽(AsA-GSH)循环系统的稳定性,降低H₂O₂、MDA的积累及O₂^·-的产生速率,从而减轻活性氧对核桃叶片的伤害,保护细胞膜结构的稳定性,增强抗寒性。

关键词: 核桃, 一氧化氮, 低温胁迫, 活性氧代谢

Abstract: [Objective] Effects of exogenous nitric oxide(NO) on the active oxygen metabolism system of walnut seedlings under low temperature stress were studied, in order to investigate the possible mechanism of exogenous NO on the cold resistance of walnut.This study aimed at developing a new method for improving the resistance of fruit trees by applying exogenous NO and providing a theoretical basis for the wide application of NO on walnut in the future. [Method] The effects of sodium nitroprusside (SNP with 200 μmol·L^-1), a NO donor, on the active oxygen metabolism of walnut varieties ‘Xiangling’ and ‘Luguo No. 12’,seedlings with different cold resistance were studied in a artificial climate chamber. The seedling were subjected to low temperature treatments and the leaves were sprayed with SNP. [Result] 1) Under normal growth conditions, the effects of spraying SNP on the membrane permeability, chlorophyll content, production rate of superoxide anion radical(O₂^·-), H₂O₂, MDA and Pro content, and SOD and CAT activity in walnut leaves were not obvious. POD, DHAR, APX and GR activity, AsA and GSH content, GSH/GSSG and AsA/DHA ratio were significantly increased, while DHA and GSH content were reduced. The variation ranges of the two walnut varieties were different.2) Under low temperature stress, SNP treatment significantly increased the activity of POD, CAT, SOD, APX and GR, increased the content of chlorophyll, Pro, AsA, DHA, GSH and GSSG, and decreased the accumulation of MDA and H₂O₂, production rate of superoxide anion radical(O₂^·-) and relative permeability of plasma membrane. The variation ranges of the two walnut varieties were different. [Conclusion] Under low temperature stress, exogenous NO treatment could maintain the stability of AsA-GSH circulation system, decrease the accumulation of MDA and H₂O₂, production rate of superoxide anion radical(O₂^·-) and protect the stability of cell membrane structure by promoting antioxidant enzyme activity and antioxidant content, and there by reduce the damage of the active oxygen to the walnut leaf and enhance the cold resistance.

Key words: walnut, nitric oxide(NO), low temperature stress, active oxygen metabolism

中图分类号:

S718.43

相昆, 徐颖, 李国田, 王晓芳, 张美勇. 外源NO对低温胁迫下核桃幼苗活性氧代谢的影响[J]. 林业科学, 2016, 52(1): 143-149.

Xiang Kun, Xu Ying, Li Guotian, Wang Xiaofang, Zhang Meiyong. Effects of Exogenous Nitric Oxide on Reactive Oxygen Metabolism of Walnut Seedlings under Low Temperature Stress[J]. Scientia Silvae Sinicae, 2016, 52(1): 143-149.

参考文献

曹慧,王东方,王汉海,等.2015.外源NO对水分胁迫下平邑甜茶幼苗氧化损伤的缓解效应.西北植物学报,35(3):538-562.
(Cao H,Wang D F,Wang H H,2015.Alleviated effect of exogenous nitric oxide on oxidative damage in Malus hupehensis(pamp) rehd.seedlings under water stress. Acta Botanica Boreali-Occidentalia Sinica, 35(3):538-562.[in Chinese])
邓化冰,车芳璐,肖应辉.2011.开花期低温胁迫对水稻花粉性状及剑叶理化特性的影响.应用生态学报,22(1):66-72.
(Deng H B,Che F L,Xiao Y H. 2011.Effects of low temperature stress during flowering period on pollen characters and flag leaf physiological and biochemical characteristics of rice. Chinese Journal of Applied Ecology, 22(1):66-72.[in Chinese])
李凯龙,王艺潼,韩晓雪,等.2013.低钾胁迫对番茄叶片活性氧及抗氧化酶系的影响.西北植物学报,33(1):66-73.
(Li K L,Wang Y T,Han X X, et al.2013.Changes in reactive oxygen species and antioxidative defense mechanism in tomato leaves under low potassium stress. Acta Botanica Boreali-Occidentalia Sinica,33(1):66-73.[in Chinese])
李秀,巩彪,徐坤.2014.外源NO对高温胁迫下姜叶片活性氧代谢的影响.园艺学报,41(92):277-284.
(Li X,Gong B,Xu K.2014. Effects of exogenous nitric oxide on reactive oxygen metabolism in ginger leaves under heat stress. Acta Horticulturae Sinica, 41(92):277-284.[in Chinese])
李轶冰,杨顺强,任广鑫,等.2009.低温处理下不同禾本科牧草的生理变化及其抗寒性比较.生态学报, 29(3):1341-1347.
(Li Y B,Yang S Q,Ren G X,et al. 2009.Changes analysis in physiological properties of several gramineous grass species and cold-resistance comparison on under cold stress. Acta Ecologica Sinica, 29(3):1341-1347.[in Chinese])
刘慧英,朱祝军,吕国华,等.2003.低温胁迫下西瓜嫁接苗的生理变化与耐冷性关系的研究.中国农业科学,36(11):1325-1329.
(Liu H Y,Zhu Z J,Lü G H,et al. 2003.Study on relationship between physiological changes and chilling tolerance in grafted watermelon seedlings under low temperature stress.Scientia Agricultura Sinica, 36(11):1325-1329.[in Chinese])
刘柿良,潘远智,杨容孑,等.2014.外源一氧化氮对镉胁迫下长春花质膜过氧化、ATPase及矿质营养吸收的影响.植物营养与肥料学报, 20(2):445-458.
(Liu S L,Pan Y Z,Yang R J,et al. 2014.Effects of exogenous NO mineral apsorption,lipid peroxidation and ATPase of plasma membrane catharanthus rosues tissues under cadmium stress.Journal of Plant Nutrition and Fertilizer, 20(2):445-458.[in Chinese])
马金虎,邢国芳,杨小环,等.2015.外源EBR和NO信号对低温胁迫下玉米种胚抗氧化系统和低温响应基因表达的影响,应用生态学报,2015, 26(5):1411-1418.
Ma J H, Xing G F,Yang X H,et al.2015.Effects of exogenous EBR and NO signal on antioxidant system and low response gene expression under cold stress on maize embryo.Chinese Journal of Applied Ecology, 26(5):1411-1418.[in Chinese])
牟雪姣,刘理想,孟鹏鹏,等.2015.外源NO缓解蝴蝶兰低温胁迫伤害的生理机制研究.西北植物学报,35(5):978-984.
(Mu X J,Liu L X,Meng P P, et al.2015.Physiological mechanism of exogenous nitric oxide on alleviating low temperature stress of Phalaenopsis spp. Acta Botanica Boreali-Occidentalia Sinica, 35(5):978-984.[in Chinese])
倪郁,宋超,王小清. 2014.低温胁迫下拟南芥表皮蜡质的响应机制.中国农业科学,47(2):252-261.
(Ni Y, Song C,Wang X Q. 2014.Investigation on response mechanism of epicuticular wax on Arabidopsis thaliana under cold stress. Scientia Agricultura Sinica, 47(2):252-261.[in Chinese])
王爱国,罗广华.1990.植物的超氧物自由基与羟胺反应的定量关系.植物生理学通讯, (6):55-57.
(Wang A G,Luo G H.1990.Quanitaitve relaiton between the reaction of hydroxylanmine and superoxide anion radicals in plants.Plant Physiloy Communication, (6):55-57.[in Chinese])
王建,于世欣,张敏,等.2014.外源NO介导Cu胁迫下番茄GSH-PCs合成途径.应用生态学报,25(9):2629-2636.
(Wang J,Yu S X,Zhang M,et al.2014. Exogenous NO mediated GSH-PCs synthesis pathway in tomato under copper stress. Chinese Journal of Applied Ecology, 25(9):2629-2636.[in Chinese])
相昆,张美勇,徐颖,等.2011.不同核桃品种耐寒特性综合评价.应用生态学报,22(9):2325-2330.
(Xiang K, Zhang M Y,Xu Y,et al. 2011.Cold-tolerance of walnut cultivars:a comprehensive evaluation.Chinese Journal of Applied Ecology, 22(9):2325-2330.[in Chinese])
许楠,孙广玉.2009.低温锻炼后桑树幼苗光合作用和抗氧化酶对冷胁迫的响应.应用生态学报, 20(4):761-766.
(Xu N, Sun G Y.2009.Responses of mulberry seedlings photosynthesis and antioxidant enzymes to chilling stress after low-temperature acclimation.Chinese Journal of Applied Ecology, 20(4):761-766.[in Chinese])
岳海,李国华,李国伟,等.2010.澳洲坚果不同品种耐寒特性的研究.园艺学报,37(1):31-38.
(Yue H,Li G H,Li G W,et al.2010.Studieson cold resistance of different macadamia cultivars.Acta Horticulturae Sinica, 37(1):31-38.[in Chinese])
杨美森,王雅芳,干秀霞,等.2012.外源一氧化氮对冷害胁迫下棉花幼苗生长、抗氧化系统和光合特性的影响.中国农业科学,45(15):3058-3067.
(Yang M S,Wang Y F,Gan X X,et al.2012.Effects of exogenous nitric oxide on growth, antioxidant system and photosynthetic characteristics in seedling of cotton cultivar under chilling injury stress.Scientia Agricultura Sinica, 45(15):3058-3067.[in Chinese])
张宪政,陈凤玉,王荣富.1994.植物生理学实验技术.沈阳:辽宁科学技术出版社.
(Zhang X Z,Chen F Y,Wang R F. 1994.Guide of experimental technology of physiology.Shenyang:Liaoning Science and Technology Press.[in Chinese])
张志良,瞿伟菁,李小方.2009.植物生理学实验指导.北京:高等教育出版社.
(Zhang Z L, Qu W J,Li X F. 2009.Laboratory guide of plant physiology.Beijing:Higher Education Press.[in Chinese]
Anderson J V,Chevone B I,Hess J L.1992.Seasonal variation in the antioxidant system of eastern white pine needles.Plant Physiology,98(2):501-508.
Bates L,Waldren R,Teare I.1973.Rapid determination of free proline for water-stress studies.Plant and Soil,39(1):205-207.
Beauchamp C, Fridovich I.1971.Superoxide dismutase:improved assays and an assay applicable to acrylamide gels.Analytical Biochemistry,44(1):276-287.
Bradford M.1976.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry,72(1/2):248-254.
Cakmak I,Marschner H.1992.Magnesium deficiency and high light intensity enhance activities of superoxide dismutase,ascorbate peroxidase,and glutathione reductase in bean leaves.Plant Physiology,98(4):1222-1227.
Hayat S,Hayat Q,Alyemeni M N,et al.2012.Role of proline under changing environments:a review.Plant Signal Behav,7 (11):1-11.
Kramer G, Norman H, Krizek D, et al.1991.Influence of UV-B radiation on polyamines lipid peroxidation and membranelipids in cucumber. Phytochemistry, 30(7):2101-2108.
Laspina N V,Groppa M D,Tomaro M L,et al.2005.Nitric oxide protects sunflower leaves against Cd-induced oxidative stress. Plant Sciences,169(2):323-330.
Law M Y,Charles S A,Halliwell B.1983.Glutathione and ascorbic acid in spinach(Spinacia oleracea)chloroplasts. The effect of hydrogen peronide and of paraguat.Biochemical Journal,210(2):899-903.
Liu F,Guo F Q. 2013.Nitric oxide deficiency accelerates chlorophyll breakdown and stability loss of thylakoid membranes during dark-induced leaf senescence in Arabidopsis. PLoS One, 8(2):1-12.
Nakano Y,Asada K.1981.Hydragen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. J Plant Cell Physiol,22(5):867-880.
Pinheiro H A, DaMatta F M, Chaves A R M, et al. 2004.Drought tolerance in relation to protection against oxidative stress in clones of Coffea canephora subjected to long-term drought. Plant Science,167(6):1307-1314.
Singh H P,Batish D R,Kaur G,et al.2008.Nitric oxide(as sodium nitroprusside) supplementation ameliorates Cd toxicity in hydroponically grown wheat roots.Environmental and Experimental Botany,63(1/3):158-167.
Szabados L, Savoure A.2010.Proline:a multifunctional amino acid.Trends Plant Sci,15(2):89-97.
Uchida A,Jagendorf A T,Hibino T.2002.Effects of hydrogen peroxide and nitric oxide on both salt and heat stress tolerance in rice. Plant Science,163(3):515-523.

[1]	周乃富, 宋晓波, 张俊佩, 常英英, 裴东. 核桃芽接愈合的组织学机制[J]. 林业科学, 2019, 55(6): 37-43.
[2]	张佳琦, 胡恒康, 徐川梅, 胡渊渊, 黄有军, 夏国华, 黄坚钦, 常英英, 叶磊, 娄和强, 张启香. 核桃JrGA2ox基因的克隆、亚细胞定位及功能验证[J]. 林业科学, 2019, 55(2): 50-60.
[3]	黄仁, 张韵, 张启香, 王正加, 夏国华, 黄坚钦, 胡渊渊. 异源授粉山核桃果皮光合能力差异的转录组分析[J]. 林业科学, 2019, 55(1): 128-137.
[4]	曾楚楚, 娄钧翼, 郭明, 王冰璇, 卢闻君. 防治核桃干腐病的新型纳米生物基农药缓释胶囊制备及缓释性能[J]. 林业科学, 2018, 54(5): 87-100.
[5]	包文泉, 乌云塔娜, 杜红岩, 李铁柱, 刘慧敏, 王淋, 白玉娥. 基于SSR标记的西藏光核桃群体遗传多样性和遗传结构分析[J]. 林业科学, 2018, 54(2): 30-41.
[6]	梁永富, 王康才, 薛启, 隋利, 叶军, 陈兴忠. 生长调节剂对低温胁迫下酸橙幼苗抗逆生理指标的影响[J]. 林业科学, 2017, 53(3): 68-75.
[7]	李世亮, 倪张林, 莫润宏, 屈明华, 汤富彬, 刘毅华. 云贵川主产区核桃中重金属污染水平及其风险评估[J]. 林业科学, 2017, 53(11): 52-59.
[8]	徐川梅, 黄坚钦, 王正加, 夏国华, 张启香, 黄有军, 张守攻. 山核桃花粉母细胞减数分裂及核型[J]. 林业科学, 2017, 53(11): 77-84.
[9]	陈丹丹, 李天凤, 王琦琦, 唐光辉. 核桃举肢蛾成虫触角的超微结构观察[J]. 林业科学, 2017, 53(10): 118-132.
[10]	徐沁怡, 王标, 赵建文, 吴建峰, 曹亦润, 杨先有, 夏国华, 王正加, 黄坚钦, 胡渊渊. 2种花粉授粉山核桃果皮光合特性的差异比较[J]. 林业科学, 2017, 53(1): 38-46.
[11]	沈一凡, 钱进芳, 郑小平, 袁紫倩, 黄坚钦, 温国胜, 吴家森. 山核桃中心产区林地土壤肥力的时空变化特征[J]. 林业科学, 2016, 52(7): 1-12.
[12]	王娇莉, 南小宁, 任争争, 明洁, 唐光辉. 核桃举肢蛾和桃蛀螟幼虫肠道细菌多样性的PCR-DGGE和T-RFLP分析[J]. 林业科学, 2016, 52(6): 76-85.
[13]	张俊佩, 王滋, 周贤武, 赵荣军, 徐慧鸽, 贾志明, 裴东. 不同品系美国黑核桃木材物理力学性质的差异[J]. 林业科学, 2016, 52(6): 108-114.
[14]	徐永杰, 韩华柏, 王滑, 陈凌娜, 马庆国, 裴东. 大巴山区核桃实生居群的坚果表型和遗传多样性[J]. 林业科学, 2016, 52(5): 111-119.
[15]	徐凤华, 程龙军, 魏晓玲, 窦锦青. 巨桉低温胁迫响应基因EgrCR的表达与功能[J]. 林业科学, 2016, 52(3): 59-67.