沙丘芦苇所含独特化合物对细胞膜流动性的影响
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摘要
质膜在细胞内外物质运输、维持细胞内外的离子平衡、信息传递、细胞间的识别及细胞运动等重要的生命过程中起着关键性的作用。
本实验以绿豆(Vigna radiata L.)黄化下胚轴和水生芦苇(Phragmites communistrin.)为材料,用荧光染料8-苯胺基-1-萘磺酸盐(8-Anilino-1-naphthalene sulfonic,ANS)和1,6-二苯基-1,3,5-己三烯(1,6-Diphenyl-1,3,5-hexatriene,DPH)为荧光探针研究外源PAAC处理后线粒体膜的流动性的变化。荧光探剂ANS与膜结合后主要有两种因素可引起ANS荧光的变化,一是它所处环境的极性改变,二是与ANS合的膜脂流动性变化。如果前者不变,则与膜结合的ANS荧光强度即能反映膜脂的流动性,荧光强度越大,膜脂流动性越小。此荧光强度反映的是整个脂区运动状态的平均值。DPH是研究膜脂流动性比较敏感的一种常用探针。DPH掺入到膜脂的膜脂酸烃链区后,介质粘度变大,顺反异构化受到抑制,成为全反构型,即唯一能发荧光的构型。由于DPH分子长轴接近于与膜脂酸链分子长轴平行,所以,荧光偏振度(P)能很好地反映膜脂区域的微粘度,定量地说明膜脂分子的运动情况。
结果表明:PAAC处理对线粒体膜的流动性有降低的作用,并且和PAAC的浓度呈一定的量效关系。在实验范围内,高温胁迫下同样降低膜的流动性,最重要的是能够稳定膜的流动性,但对相变温度的改变似乎不起作用。因此推测,在夏季自然高温环境下,PAAC可以通过降低膜的流动性而对植物膜的功能起到稳定和保护作用。
Membrane is playing a cruical role in substance transportation between outside and inside of cell, maintain ion balance, information transmission, cell identification and cell movement and also other impotant life process.
Using etiolated hypocotyledonary axis of mung bean and swamp reed as materials, 8-anilinonaphthalene-l-sulfonic and 1,6-diphenyl-1,3,5-hexatriene as probes, a fluorescence spectrophotometry method was performed to research the change of membrane fluidity of mitochondia treated by extraneous PAAC. Two factors will change the fluorescence of ANS when ANS combine with membrane. One is the polarity of ANS's environment changes, the other is the fluidity of membrane lipoid combined with ANS changes. If the first is invariable, that the intensity of ANS's fluorescence can reflect the fluidity of membrane lipoid. Fluorescence more intensity, slower of fluidity. And this fluorescence reflects the average value of all the lipoid region's movement. DPH is a sensitivity and commom probe when learn about the fluidity of membrane lipoid. When DPH mixes into the hydrocarbon chain region of membrane fat acid, medium will be more sticky and the transform of cis isomer will be suppressed, so that all are resver, that is can launch fluoresence. Owing to the long axis of DPH molecule is nearly paralleling with the long axis of membrane lipiod acid molecule. Therefore the degree of fluoresence polarization(P) can welly show the min-viscosity of membrane lipoid region, and quantificationally illuminate the movement of membrane lipoid molecule.
The results of the study suggest that membrane fluidity of mitochondia have been reduced after treated by PAAC, furthermore, there is a dose-effect relationship. Confined within our experiment, PAAC reduces the membrane fluidity under the high temperature stress, more importantly, it can stabilize the fluidity of membrance, however, it has no effect on the change of the transition temperature. Therefore, it can be suppose that PAAC can stabilize and protect the function of the biomembrane in plants by reduceing the fludity under the high temprature in summer.
本实验以绿豆(Vigna radiata L.)黄化下胚轴和水生芦苇(Phragmites communistrin.)为材料,用荧光染料8-苯胺基-1-萘磺酸盐(8-Anilino-1-naphthalene sulfonic,ANS)和1,6-二苯基-1,3,5-己三烯(1,6-Diphenyl-1,3,5-hexatriene,DPH)为荧光探针研究外源PAAC处理后线粒体膜的流动性的变化。荧光探剂ANS与膜结合后主要有两种因素可引起ANS荧光的变化,一是它所处环境的极性改变,二是与ANS合的膜脂流动性变化。如果前者不变,则与膜结合的ANS荧光强度即能反映膜脂的流动性,荧光强度越大,膜脂流动性越小。此荧光强度反映的是整个脂区运动状态的平均值。DPH是研究膜脂流动性比较敏感的一种常用探针。DPH掺入到膜脂的膜脂酸烃链区后,介质粘度变大,顺反异构化受到抑制,成为全反构型,即唯一能发荧光的构型。由于DPH分子长轴接近于与膜脂酸链分子长轴平行,所以,荧光偏振度(P)能很好地反映膜脂区域的微粘度,定量地说明膜脂分子的运动情况。
结果表明:PAAC处理对线粒体膜的流动性有降低的作用,并且和PAAC的浓度呈一定的量效关系。在实验范围内,高温胁迫下同样降低膜的流动性,最重要的是能够稳定膜的流动性,但对相变温度的改变似乎不起作用。因此推测,在夏季自然高温环境下,PAAC可以通过降低膜的流动性而对植物膜的功能起到稳定和保护作用。
Membrane is playing a cruical role in substance transportation between outside and inside of cell, maintain ion balance, information transmission, cell identification and cell movement and also other impotant life process.
Using etiolated hypocotyledonary axis of mung bean and swamp reed as materials, 8-anilinonaphthalene-l-sulfonic and 1,6-diphenyl-1,3,5-hexatriene as probes, a fluorescence spectrophotometry method was performed to research the change of membrane fluidity of mitochondia treated by extraneous PAAC. Two factors will change the fluorescence of ANS when ANS combine with membrane. One is the polarity of ANS's environment changes, the other is the fluidity of membrane lipoid combined with ANS changes. If the first is invariable, that the intensity of ANS's fluorescence can reflect the fluidity of membrane lipoid. Fluorescence more intensity, slower of fluidity. And this fluorescence reflects the average value of all the lipoid region's movement. DPH is a sensitivity and commom probe when learn about the fluidity of membrane lipoid. When DPH mixes into the hydrocarbon chain region of membrane fat acid, medium will be more sticky and the transform of cis isomer will be suppressed, so that all are resver, that is can launch fluoresence. Owing to the long axis of DPH molecule is nearly paralleling with the long axis of membrane lipiod acid molecule. Therefore the degree of fluoresence polarization(P) can welly show the min-viscosity of membrane lipoid region, and quantificationally illuminate the movement of membrane lipoid molecule.
The results of the study suggest that membrane fluidity of mitochondia have been reduced after treated by PAAC, furthermore, there is a dose-effect relationship. Confined within our experiment, PAAC reduces the membrane fluidity under the high temperature stress, more importantly, it can stabilize the fluidity of membrance, however, it has no effect on the change of the transition temperature. Therefore, it can be suppose that PAAC can stabilize and protect the function of the biomembrane in plants by reduceing the fludity under the high temprature in summer.
引文
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[5]季宇彬,万梅绪,等.龙葵碱对荷瘤小鼠红细胞免疫功能的影响【J】.中草药,2007年3月第38卷第3期:412-414.
[6]LIU SM,JI YL,CHEN KS,et al.Effect of choline chloride on membrane fluidity of mitocondria isolated from etiolated seedlings of wheat 【J】.Acat.Biophysica Sinica,2002,18(1):19-22.
[7]Haslam SM.Variation of population types in Phragmites communis Trin.Ann Bot,1970.34:147-158.
[8]王洪亮,张承烈,陈国仓.河西走廊芦苇不同生态型生境适应的渗透调节物质.生态学报,1994,14:56-60.
[9]Zheng W J,Zheng XP,Zhang CL.A survey of photosynthetic carbon metabolism in 4 ecotypes of Phragmites communis in northwest Chain:leaf anatomy,ultrastructure,and activities of ribulose 1,5-bisphosphate carboxylase,phosphoenolpyruvate carboxylase and glycollate oxidase.Physiol Plant 2000,110:201-208.
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[12]Vance N,Copes DO,Zaerr JB.Differences in proteins synthesized in needles of unshaded andshaded Pi nus ponderosa var.scopulorum seedlings during prolonged drought.Plant Physiol 1990,92:1244-1248.
[13]王洪乔.植物逆境生理.植物生理学通讯,1981,(6):72-81.
[14]Bohnert HJ,Jensen RG.Strategies for engineering water-stress tolerance in plants.Trends Biotechnol 1996,14:89-97.
[15]Cheng YF,Pu TL,Zhang CL et al.PcTGD,a highly expressed gene in stem,is related to waters4tress in reed(Phragmites communis Trin.).Chin Sci Bull,2001,46(10):850-854.
[16]Smirnoff N.Antioxidant systems and plant response to the environment.In:Smirnoff N(ed).Environment and Plant Metabolism.Oxiford:Bios Sci Pub,1995.217-243.
[17]张承烈,周瑞莲,陈国仓．芦苇耐脱水能力的生理生态学分析．植物生态学及地植物学学报,1992,16(4):311-316．
[18] Wang J G, Zhang CL. DNA damage and repair of two ecotypes of Phragmites communis subjectedto water stress. Acta Bot Sin., 2001, 43(5):490-494.
[19] Bohnert H J, Nelson D E, Jensen R G. Adaptation to environmental stress. Plant Cell 1995,7:1099-1111.
[20] Csonkal L. Physiological and genetic responses of bacteria to osmotic stress. Microbiol. Rev 1989,53:121-147.
[21] Bartels D, Nelson D. Approaches to improve stress tolerance using molecular genetics. Plant Cell Environ 1994, 17:659-667.
[22] Anderson JM. Jasmonic acid-dependent increases in the level of specific polypeptides in soybean suspension cultures and seedlings. J. Plant Growth Regul 1988,7:203-11.
[23] Creelman RA, Mul et JE. Biosynthesis and action of jasmonates in plants. Ann. Rev. Plant Physiol. Plant Mol. Biol. 1997,48:355-381.
[24] Shena B, Jensen RG, Bohnert HJ. Increased resistance to oxidative stress in transgenic plants by targeting mannitol biosynthesis to chloroplasts. Plant Physiology 1997, 113:1177-1183.
[25] Shenb B, Jensen RG, Bohnert HJ. Mannitol protects against oxidation by hydroxyl radicals.Plant Physiol 1997, 115:527-532.
[26] Smirnoff N, Cumbes Qj. Hydroxyl radical scavenging activity of compatible solutes.Phytochem, 1989,28:1057-1060.
[27] Paleg LG, Steward GR, Bradbeer JW. Antioxidant systems and plant response to the environment. Plant Physiol 1984,75:974.
[28] Yancy PH, Somero GN J In Environment and Plant Metabolism. Exp Zool 1980, 212:205.
[29] Hare P D, Cress W A, Staden J V. Dissecting the role of osmolyte accumulation during stress.Plant Cell and Environ., 1998, 21:535-553.
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