温水浸种对杜仲种子萌发过程中保护酶活性及内源激素含量变化的影响

doi:10.7525/j.issn.1673-5102.2020.04.006

摘要/Abstract

摘要： 探讨杜仲（Eucommia ulmoides Oliver）种子萌发过程中保护酶活性及内源激素含量的变化，为其种子破眠及萌发不同时期采取有效处理措施，从而提高出苗率提供参考。利用室温（CK）、40℃、50℃、60℃和70℃不同水温，探讨温水浸种对杜仲种子萌发的影响。种子萌发进程中，利用常规生理生化方法测定种子内超氧化物歧化酶（SOD）、过氧化物酶（POD）活性和丙二醛（MDA）含量，以及酶联免疫法测定赤霉素（GA₃）、吲哚乙酸（IAA）、脱落酸（ABA）和玉米素核苷（ZR）含量。结果表明：①温水浸种处理能提高了杜仲种子的发芽率及发芽势，其中以50℃温水处理最为显著。②与室温浸种处理相比，50℃温水浸种处理提高了种子内SOD及POD活性，减轻了质膜氧化损伤程度，致使MDA含量的持续下降，从而促使种子朝向有利于萌发的方向发展。此外，50℃温水浸种处理在提高种子内促进生长内源GA₃、IAA及ZR含量的同时，并降低了抑制生长的内源ABA含量，致使GA₃/ABA、IAA/ABA和ZR/ABA值均明显高于室内浸种处理，最终促进了杜仲种子的萌发。

关键词: 杜仲, 萌发阶段, 超氧化物歧化酶, 内源激素, 丙二醛, 脱落酸

Abstract: The experiment was conducted to investigate the regulation mechanism of enzymes and endogenous hormones of different stages of Eucommia ulmoides Oliver seed germination, aiming to provide scientific basis for taking effective treatment measures during the different stages of seed dormancy and germination to increase seedling emergence rate. The effects of warm water soaking on the germination of E.ulmoides seeds were studied with room temperature(CK), 40℃, 50℃, 60℃ and 70℃. And the activities of superoxide dismutase(SOD), peroxidase(POD), and malondialdehyde(MDA) content were determined by routine physiological and biochemical methods. Levels of 4 endogenous hormones, such as gibberellin(GA₃), indole acetic acid(IAA), zeatin riboside(ZR) and abscisic acid(ABA) were quantitatively determined by enzyme-linked immunosorbent assay. The result showed that:①The seed germination rate could be increased by soaking seeds with different temperature water, and the germination rate reached its maximum under 50℃ warm water treatment. ②Compared with room temperature water treatment, the activity of SOD and POD in the seeds were increased under 50℃ warm water, and reduced the degree of cytoplasm membrane was oxidized, and the content of MDA was decreased continuously, thus promoted the development of seeds in a direction favorable for germination. In addition, treatment with 50℃ warm water promoted the growth of endogenous GA₃, IAA and ZR in seeds, and decreased the content of endogenous ABA, and the values of GA₃/ABA, IAA/ABA and ZR/ABA were significantly higher than those of room temperature water, which ultimately promoted the germination of E.ulmoides.

Key words: Eucommia ulmoides, germinating stage, SOD, endogenous hormones, MDA, ABA

中图分类号:

Q945.7

王宁, 董莹莹, 袁美丽, 王亚楠, 曾胜蕴. 温水浸种对杜仲种子萌发过程中保护酶活性及内源激素含量变化的影响[J]. 植物研究, 2020, 40(4): 523-529.

WANG Ning, DONG Ying-Ying, YUAN Mei-Li, WANG Ya-Nan, ZENG Sheng-Yun. Effect of Warm Water Soaking on Protective Enzyme Activity and Endogenous Hormones Content of Eucommia ulmoides Seed During Different Stages of Germination[J]. Bulletin of Botanical Research, 2020, 40(4): 523-529.

参考文献

[1] 吴可心,吴斌,彭晓曦,等.不同产地杜仲叶片活性成分及杜仲胶质量分数差异[J].东北林业大学学报,2019,47(10):40-43,74.Wu K X,Wu B,Peng X X,et al.Active constituents and eucommia gum content of Eucommia ulmoides Oliver from multiple origins[J].Journal of Northeast Forestry University,2019,47(10):40-43,74.
[2] 冯风,梁志荣.我国历史上对杜仲的认识和利用[J].西北林学院学报,1996,11(2):84-89.Feng F,Liang Z R.Understanding and utilization of Eucommia ulmoides in Chinese history[J].Journal of Northwest Forestry College,1996,11(2):84-89.
[3] 阴健,郭力弓.中药现代研究与临床应用[M].上海:学苑出版社,1993.Yin J,Guo L G.Modern research and clinical application of traditional Chinese medicine[M].Shanghai:Shanghai Science and Technology Press,1993.
[4] 李杨.杜仲籽中桃叶珊瑚苷的生物转化及药代动力学研究[D].西安:西北大学,2008.Li Y.The research on bioconversion and pharmacokinetics of aucubin from the seeds of Eucommia ulmoides Oliv.[D].Xi'an:Northwest University,2008.
[5] Kimura Y,Okuda H,Arichi S.Effects of geniposide isolated from Gardenia jasminoides on metabolic alterations in high sugar diet-fed rats[J].Chemical and Pharmaceutical Bulletin,1982,30(12):4444-4447.
[6] Kim D H,Kim B R,Kim J Y,et al.Mechanism of covalent adduct formation of aucubin to proteins[J].Toxicology Letters,2000,114(1-3):181-188.
[7] 李家实,阎玉凝.杜仲皮与叶化学成分的初步研究[J].中药通报,1986,11(8):41-42.Li J S,Yan Y N.Preliminary study on the chemical constituents of Eucommia ulmoides Oliv.[J].Chinese Medicine Journal,1986,11(8):41-42.
[8] 唐建军,张禄源,何鸣筱.杜仲的研究与应用进展[J].植物学通报,1998,15(6):47-51.Tang J J,Zhang L Y,He M X.Advances in research of Eucommia ulmoides Oliver and its utilization[J].Chinese Bulletin of Botany,1998,15(6):47-51.
[9] Hamann C,Rodgers P A,Alenius H,et al.Cross-reactivity between gutta-percha and natural rubber latex:assumptions vs.reality[J].The Journal of the American Dental Association,2002,133(10):1357-1367.
[10] Chen C H,Shen Y K,Hsieh S C.The investigation of gutta-percha temperature and compaction force change when using the vertical compaction of warm gutta-percha technique[J].Journal of Polymer Engineering,2014,34(3):219-223.
[11] 杜红岩,谢碧霞,邵松梅.杜仲胶的研究进展与发展前景[J].中南林学院学报,2003,23(4):95-99.Du H Y,Xie B X,Shao S M.Prospects and research progress of gutta-percha[J].Journal of Central South Forestry University,2003,23(4):95-99.
[12] 林坚,郑光华,张庆昌.杜仲种子休眠原因及发芽特性的研究[J].种子,1989(2):8-10.Lin J,Zheng G H,Zhang Q C.Study on dormancy and germination characteristics of Eucommia ulmoides seeds[J].Seed,1989(2):8-10.
[13] 申延,何泼.杜仲种子发芽试验分析[J].林业科技开发,2005,19(4):32-33.Shen Y,He P.Test on seeds germination of Eucommia ulmoides[J].China Forestry Science and Technology,2005,19(4):32-33.
[14] 吴楚材,黄炎.杜仲种子贮藏方法及发芽试验研究[J].中南林学院学报,1989,9(1):62-65.Wu C C,Huang Y.Study on storage method and germination test of Eucommia ulmoides seeds[J].Journal of Central-South Forestry College,1989,9(1):62-65.
[15] 邹仁双.温水及生长调节剂处理对杜仲种子发芽率的影响[J].防护林科技,2016(7):38-39,45.Zou R S.Effects of warm water and growth regulator on seed germination of Eucommia ulmoides[J].Protection Forest Science and Technology,2016(7):38-39,45.
[16] 李合生,孙群,赵世杰,等.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000.Li H S,Sun Q,Zhao S J,et al.Principle and technology of plant physiological biochemical experiment[M].Beijing:Higher Education Press,2000.
[17] 邹琦.植物生理生化实验指导[M].北京:中国农业出版社,2003.Zou Q.The guidance of plant physiological and biochemical[M].Beijing:China Agriculture Press,2003.
[18] 段承俐,段银妹,萧凤回.三七种子后熟过程中内源激素的动态变化[J].中草药,2011,42(4):779-782.Duan C L,Duan Y M,Xiao F H.Dynamic changes of endogenous phytohormones during after-ripening process of Panax notoginseng seeds[J].Chinese Traditional and Herbal Drugs,2011,42(4):779-782.
[19] 杨荣超,张海军,王倩,等.植物激素对种子休眠和萌发调控机理的研究进展[J].草地学报,2012,20(1):1-9.Yang R C,Zhang H J,Wang Q,et al.Regulatory mechanism of plant hormones on seed dormancy and germination(Review)[J].Acta Agrestia Sinica,2012,20(1):1-9.
[20] 陈翔,孙云波,陈垣,等.浸种温度对红芪(Hedysarum polybotrys)种子发芽的影响[J].中国沙漠,2015,35(3):632-638.Chen X,Sun Y B,Chen Y,et al.Effects of soaking temperature on seed germination characteristics of Hedysarum polybotrys[J].Journal of Desert Research,2015,35(3):632-638.
[21] 程瑶,方向文,蒋志荣,等.温水浸种对蒙古黄芪种子萌发特性的影响[J].植物科学学报,2017,35(3):413-420.Cheng Y,Fang X W,Jiang Z R,et al.Effect of warm water treatment on seed germination characteristics of Astragalus membranaceus var.mongholicus[J].Plant Science Journal,2017,35(3):413-420.
[22] 王进,罗光宏,颜霞,等.荒漠植物苦豆子和苦马豆种子硬实特性与活力关系研究[J].中国沙漠,2011,31(4):974-980.Wang J,Luo G H,Yan X,et al.Relationship between seed hardness and seed vigor of desert plant Sophora alopecuroides and Swainsonia salsula[J].Journal of Desert Research,2011,31(4):974-980.
[23] 杨艳,黎云祥,胥晓.珙桐种子休眠解除和萌发过程中主要抗氧化酶活性和代谢产物含量的变化[J].植物分类与资源学报,2015,37(6):779-787.Yang Y,Li Y X,Xu X.The activity of principal antioxidant enzymes and the content of metabolites in dormancy breaking and germination of Davidia involucrate seeds[J].Plant Diversity and Resources,2015,37(6):779-787.
[24] 李江,吴黄铭,陈惠萍.外源CO和NO对水稻种子萌发过程中干旱胁迫损伤的缓解效应[J].西北植物学报,2011,31(4):731-738.Li J,Wu H M,Chen H P.Exogenous carbon monoxide and nitric oxide alleviate the oxidative damage in rice seed germination under drought stress[J].Acta Botanica Boreali-Occidentalia Sinica,2011,31(4):731-738.
[25] 张怀山,赵桂琴,栗孟飞,等.中型狼尾草幼苗对PEG、低温和盐胁迫的生理应答[J].草业学报,2014,23(2):180-188.Zhang H S,Zhao G Q,Li M F,et al.Physiological responses of Pennisetum longissimum var.intermedium seedlings to PEG,low temperature and salt stress treatments[J].Acta Prataculturae Sinica,2014,23(2):180-188.
[26] 赖华燕,王友凤,吴凯,等.外源激素对福建青冈种子易萌性和抗氧化酶活性的影响[J].生态学杂志,2017,36(2):382-388.Lai H Y,Wang Y F,Wu K,et al.Effect of exogenous hormones on germination and antioxidant enzyme activity of seeds of Cyclobalanopsis chungii[J].Chinese Journal of Ecology,2017,36(2):382-388.
[37] Priestly D A.Seed aging[M].London:Cornell University Press,1986.
[28] Ogawa M,Hanada A,Yamauchi Y,et al.Gibberellin biosynthesis and response during Arabidopsis seed germination[J].The Plant Cell,2003,15(7):1591-1604.
[29] Khan A A.Primary,preventive and permissive roles of hormones in plant systems[J].The Botanical Review,1975,41(4):391-420.
[30] Brady S M,Mccourt P.Hormone cross-talk in seed dormancy[J].Journal of Plant Growth Regulation,2003,22(1):25-31.
[31] Kucera B,Cohn M A,Leubner-Metzger G.Plant hormone interactions during seed dormancy release and germination[J].Seed Science Research,2005,15(4):281-307.
[32] Rohde A,Bhalerao R P.Plant dormancy in the perennial context[J].Trends in Plant Science,2007,12(5):217-223.
[33] 孙敬爽,贾桂霞.朝鲜冷杉种子层积过程中萌发特性和内源激素ABA、IAA含量的变化[J].林业科学研究,2006,19(1):117-120.Sun J S,Jia G X.Germination characteristic and endogenous ABA,IAA content change in the seed of Abies koreana during stratifying[J].Forest Research,2006,19(1):117-120.
[34] 陆秀君,梅梅,刘月洋,等.GA₃和变温层积对天女木兰种子萌发及内源激素的影响[J].西北植物学报,2014,34(9):1828-1835.Lu X J,Mei M,Liu Y Y,et al.Effect of treatment with GA₃ and variable temperature stratification on germination and endogenous hormones of Magnolia sieboldii seeds[J].Acta Botanica Boreali-Occidentalia Sinica,2014,34(9):1828-1835.
[35] 吴元奇,冷亦峰,夏超,等.不同休眠玉米种子内源激素的含量测定和分析[J].草业学报,2015,24(12):213-219.Wu Y Q,Leng Y F,Xia C,et al.Comparative analysis of endogenous hormones in maize seeds with different dormancy characteristics[J].Acta Prataculturae Sinica,2015,24(12):213-219.
[36] 苏海兰,周先治,李希,等.云南重楼种子萌发过程内源激素含量及酶活性变化研究[J].核农学报,2018,32(1):141-149.Su H L,Zhou X Z,Li X,et al.Dynamic changes of enzyme and endogenous of Paris polyphylla smith var.yunnanensis seed during different stages of germination[J].Journal of Nuclear Agricultural Sciences,2018,32(1):141-149.