大豆中硒的生理生化及其含硒蛋白的初步分离
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摘要
硒是重要的生命微量营养元素之一，在地球上分布广泛而又极不均匀，全世界有2／3的地区缺硒，我国缺硒地区也在2／3之上，其中1／3为世界公认的严重缺硒区。已知与硒营养缺乏有关的疾病达40余种，缺硒严重威胁人类健康和畜牧业的发展。人畜硒营养水平取决于摄入食物的含硒量，植物是自然界硒循环生态链和无机硒转化为有机硒的关键环节，又是人畜摄入硒最重要的直接硒源。
本文以大豆为实验材料，采用盆栽土培，通过多种研究方法，多角度、多方位地研究了硒的应用。试验以恩施本地大豆品种中豆32为样本，采用土培，运用原子吸收分光光度法作为硒含量的检测手段，研究了大豆对硒的吸收、积累、分布的动态；对环境硒的耐受能力；不同供硒水平对大豆生长发育、干物质和硒累积量及动态；产量、含硒量的影响。通过对根系氧化还原能力的测定；谷胱甘肽过氧化物酶、超氧化物歧化酶活性以及还原性谷胱甘肽、脂质过氧化产物丙二醛含量的测定；叶绿素、可溶性蛋白质、维生素C含量的分析，研究了硒的生理功能及机体内部生理生化功能和抗逆作用。运用分级盐析沉淀结合电泳、柱层析方法进行了从大豆中提取分离含硒蛋白的初步研究，结果显示：
大豆对坏境中的硒有一定的生物富集作用，能主动吸收环境即使是高硒环境中的硒。富集作用随着植物生长发育成熟而增强；根系吸收的硒经同化被运送到各个器官积累，各器官的含硒量亦随该器官的成熟而提高，收获时各器官的含硒量依次为：根＞种子＞叶＞茎；植株从环境中吸收的无机硒大部分被转化为有机态，有总硒的3／5存在于蛋白质中。硒对植物的生物效应符合微量元素生物效应剂量规律，在适宜的浓度硒范围内随着硒浓度的增加，根系氧化还原能力、叶绿素含量、可溶性蛋白质含量、维生素含量提高；谷胱甘肽过氧化物酶、超氧化物歧化酶等抗氧化酶类的活性增强，同时脂质过氧化产物丙二醛的含量下降，能显著促进大豆生长发育，维持旺盛的生命活力，延缓衰老，增强抗逆性。高浓度的硒使上述诸项指标反向变化，对大豆生长产生毒害作用。经过对分离措施的筛选，建立起分级盐析沉淀结合聚丙烯酰胺凝胶电泳、分子筛柱层析从大豆蛋白质中分离含硒蛋白组分的初步分离分组方法，为进一步纯化含硒蛋白提供了依据
和基础。
根据本研究结果，结合他人在植物硒研究方面己经做过的工作，作者认为:硒是植
物必需微量营养元素;建议把硒作为农牧业生产中提高产量、改善品质的重要措施，在
广大的缺硒和低硒地区推广，以此来改善农牧业生产的硒生态环境，提高人畜硒营养水
平。同时充分利用高硒地区丰富的生物硒资源，开发含硒量准确的硒治疗剂和各类补硒
保健产品，为治疗和预防缺硒相关疾病，提供硒含量科学合理、高利用度无毒副作用的
硒生物制品，也使高硒地区的宝贵资源得以开发增值，促进地方经济发展。
Selenium is one of the most important nutrient elements to life and it is distributed wildly but extremely unevenly on the earth. 2/3 of area in the world lacks selenium and so does in china, of which 1/3 is considered to be critically lack of selenium. Above 40 kinds of diseases and cancers are related to the lack of selenium and this seriously threat the health of human and the development of animal husbandry. The selenium content of intaken food determinate the nutritive level of selenium in human and poultry. Plant is the key link through which the ecological selenium and the inorganic selenium are converted into the organic selenium in the natural selenium cycle and it is also the most important direct selenium resource to boot the human and the poultry.
Using many chemical, physiological and biochemical method with soil culture in pot, this article studied the selenium application on soybean from various aspects. Taking nature soybean of Enshi "zhongdou32" as sample, using atom absorption spectrometry to detect selenium, the author studied the following items in soybean: the trends of absorption, accumulation, distribution of selenium in soybean and the endurance ability of soybean with the selenium in the environment; the effect of different concentration selenium treatment on the growth and development, the accumulation of dry matter and selenium, producing and selenium content. According to the measurement on the activity of root system, GSH-Px, SOD, the content of GSH, lipid peroxide production, and the analyzing on the content of chlorophyll, water-soluble protein, Vc, this article studied the physiological function and exterior mechanism of selenium in organic body and its effect of resistance. By using fractional salting-out precipitation and column
chromatography combining PAGE electrophoresis, the author managed to extract and separate the protein containing selenium from soybean seeds. The result indicated:
The soybean could accumulate and positively absorb selenium in the environment even with high selenium content and thus made the selenium concentration higher than that of the environment. The accumulation became stronger with the plant development. The selenium
was absorbed by the root system and transferred to different organ and accumulated there. The selenium contents in different organ increased with the maturity of the organ. In the harvest time, selenium content in different organ was root system > seed >leaves>stem. The most part of selenium absorbed by the plant was converted into organic selenium, about 3/5 of total selenium in soybean seeds was stored in protein. The biological effect of selenium on the soybean conforms to the general dose rule of microelement. Within proper selenium concentration, with the selenium concentration increasing, there were the following changes: the oxidation-reduction ability of root system and so did the content of chlorophyll, water-soluble, Vc; the activity of anti-oxidase such as GSH-Px and SOD etc increased. At the same time, GSH content also increased while lipid peroxide production MDA content decreased. In all, proper selenium concentration could improve the growth and development of soybean, made soybean keeping vig
orous life activity, delayed decrepit, and enhanced the ability of anti-adversity. While high selenium concentration, it can change the above-mentioned items conversely. According to sieving on several separation measures, developed the method to extract the selenium-containing protein. This method provided the basis to the further purification the selenium-containing protein.
According to the result and the work on the selenium in plant by the other researcher. The author concludes: Selenium is an essential microelement in plant and it should be used as micro manure to increase the production and improve quality in agriculture and husbandry. To ameliorate the ecological environment of selenium and increase the nutrient level of human and poultry, this method should be spread. Meanwhile the biological selenium resource in the are
本文以大豆为实验材料，采用盆栽土培，通过多种研究方法，多角度、多方位地研究了硒的应用。试验以恩施本地大豆品种中豆32为样本，采用土培，运用原子吸收分光光度法作为硒含量的检测手段，研究了大豆对硒的吸收、积累、分布的动态；对环境硒的耐受能力；不同供硒水平对大豆生长发育、干物质和硒累积量及动态；产量、含硒量的影响。通过对根系氧化还原能力的测定；谷胱甘肽过氧化物酶、超氧化物歧化酶活性以及还原性谷胱甘肽、脂质过氧化产物丙二醛含量的测定；叶绿素、可溶性蛋白质、维生素C含量的分析，研究了硒的生理功能及机体内部生理生化功能和抗逆作用。运用分级盐析沉淀结合电泳、柱层析方法进行了从大豆中提取分离含硒蛋白的初步研究，结果显示：
大豆对坏境中的硒有一定的生物富集作用，能主动吸收环境即使是高硒环境中的硒。富集作用随着植物生长发育成熟而增强；根系吸收的硒经同化被运送到各个器官积累，各器官的含硒量亦随该器官的成熟而提高，收获时各器官的含硒量依次为：根＞种子＞叶＞茎；植株从环境中吸收的无机硒大部分被转化为有机态，有总硒的3／5存在于蛋白质中。硒对植物的生物效应符合微量元素生物效应剂量规律，在适宜的浓度硒范围内随着硒浓度的增加，根系氧化还原能力、叶绿素含量、可溶性蛋白质含量、维生素含量提高；谷胱甘肽过氧化物酶、超氧化物歧化酶等抗氧化酶类的活性增强，同时脂质过氧化产物丙二醛的含量下降，能显著促进大豆生长发育，维持旺盛的生命活力，延缓衰老，增强抗逆性。高浓度的硒使上述诸项指标反向变化，对大豆生长产生毒害作用。经过对分离措施的筛选，建立起分级盐析沉淀结合聚丙烯酰胺凝胶电泳、分子筛柱层析从大豆蛋白质中分离含硒蛋白组分的初步分离分组方法，为进一步纯化含硒蛋白提供了依据
和基础。
根据本研究结果，结合他人在植物硒研究方面己经做过的工作，作者认为:硒是植
物必需微量营养元素;建议把硒作为农牧业生产中提高产量、改善品质的重要措施，在
广大的缺硒和低硒地区推广，以此来改善农牧业生产的硒生态环境，提高人畜硒营养水
平。同时充分利用高硒地区丰富的生物硒资源，开发含硒量准确的硒治疗剂和各类补硒
保健产品，为治疗和预防缺硒相关疾病，提供硒含量科学合理、高利用度无毒副作用的
硒生物制品，也使高硒地区的宝贵资源得以开发增值，促进地方经济发展。
Selenium is one of the most important nutrient elements to life and it is distributed wildly but extremely unevenly on the earth. 2/3 of area in the world lacks selenium and so does in china, of which 1/3 is considered to be critically lack of selenium. Above 40 kinds of diseases and cancers are related to the lack of selenium and this seriously threat the health of human and the development of animal husbandry. The selenium content of intaken food determinate the nutritive level of selenium in human and poultry. Plant is the key link through which the ecological selenium and the inorganic selenium are converted into the organic selenium in the natural selenium cycle and it is also the most important direct selenium resource to boot the human and the poultry.
Using many chemical, physiological and biochemical method with soil culture in pot, this article studied the selenium application on soybean from various aspects. Taking nature soybean of Enshi "zhongdou32" as sample, using atom absorption spectrometry to detect selenium, the author studied the following items in soybean: the trends of absorption, accumulation, distribution of selenium in soybean and the endurance ability of soybean with the selenium in the environment; the effect of different concentration selenium treatment on the growth and development, the accumulation of dry matter and selenium, producing and selenium content. According to the measurement on the activity of root system, GSH-Px, SOD, the content of GSH, lipid peroxide production, and the analyzing on the content of chlorophyll, water-soluble protein, Vc, this article studied the physiological function and exterior mechanism of selenium in organic body and its effect of resistance. By using fractional salting-out precipitation and column
chromatography combining PAGE electrophoresis, the author managed to extract and separate the protein containing selenium from soybean seeds. The result indicated:
The soybean could accumulate and positively absorb selenium in the environment even with high selenium content and thus made the selenium concentration higher than that of the environment. The accumulation became stronger with the plant development. The selenium
was absorbed by the root system and transferred to different organ and accumulated there. The selenium contents in different organ increased with the maturity of the organ. In the harvest time, selenium content in different organ was root system > seed >leaves>stem. The most part of selenium absorbed by the plant was converted into organic selenium, about 3/5 of total selenium in soybean seeds was stored in protein. The biological effect of selenium on the soybean conforms to the general dose rule of microelement. Within proper selenium concentration, with the selenium concentration increasing, there were the following changes: the oxidation-reduction ability of root system and so did the content of chlorophyll, water-soluble, Vc; the activity of anti-oxidase such as GSH-Px and SOD etc increased. At the same time, GSH content also increased while lipid peroxide production MDA content decreased. In all, proper selenium concentration could improve the growth and development of soybean, made soybean keeping vig
orous life activity, delayed decrepit, and enhanced the ability of anti-adversity. While high selenium concentration, it can change the above-mentioned items conversely. According to sieving on several separation measures, developed the method to extract the selenium-containing protein. This method provided the basis to the further purification the selenium-containing protein.
According to the result and the work on the selenium in plant by the other researcher. The author concludes: Selenium is an essential microelement in plant and it should be used as micro manure to increase the production and improve quality in agriculture and husbandry. To ameliorate the ecological environment of selenium and increase the nutrient level of human and poultry, this method should be spread. Meanwhile the biological selenium resource in the are
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