干旱胁迫下白芥差异表达蛋白的研究
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摘要
环境胁迫常引起植物基因表达和细胞代谢的改变,从而影响作物的生长发育和产量。光照、干旱、盐、温度是限制作物产量最主要的胁迫因素,而干旱造成的损失最大,其损失量超过其他逆境造成损失的总和。因而改良作物遗传特性、提高作物品种的抗旱性成为作物品种高产、稳产的关键。白芥具有抗逆性强等很多优良的农艺性状,尤其是所表现出来的抗旱性能已得到重视和应用,相关研究逐步由一般性的抗旱性鉴定、评价逐步走向基因和分子水平,白芥逐渐成为十字花科作物育种的优良种质资源类型。
本研究以白芥为研究对象,对其干旱胁迫下叶片中多胺含量的变化、叶片差异蛋白表达情况进行研究,旨在探讨和揭示白芥对干旱胁迫的抗性机理。试验通过PEG6000处理白芥幼苗来模拟干旱对白芥幼苗的影响,发现PEG6000完全可以成功的模拟干旱胁迫,使幼苗表现出干旱条件下的生长状态。通过对不同干旱程度下白芥幼苗叶片中多胺含量变化的测定,发现多胺含量随干旱程度的增加呈现上升趋势,但随干旱程度加剧,叶片中的多胺含量开始逐步下降,细胞受伤害程度加剧。由此可以推断,干旱胁迫会调动植物体内多胺积累以提高植株的抗旱性。试验通过蛋白质双向电泳和质谱技术研究了白芥干旱胁迫与对照幼苗叶片的蛋白差异表达情况,发现干旱胁迫较对照相比产生58个差异蛋白质。其中,对照组中有16个特有蛋白质,处理组中有42个特有蛋白质,共有有43个蛋白出现表达量的变化。可以推测这些差异蛋白与白芥的抗旱机理密切相关。试验选取10个差异蛋白点进行质谱分析,分析结果显示有5个蛋白点在NCBI和SP数据库中获得相关匹配,对这些蛋白进行初步鉴定和相关功能推测。有5个蛋白点在NCBI和SP数据库中没有获得相关匹配,预测可能为新的抗旱性相关蛋白。结果表明,本研究方法成熟可靠,结果较为理想,可以为以后进一步深入研究白芥抗旱机理、抗旱基因的克隆以及作物的抗旱育种提供重要的理论依据。
Sinapis alba L. is an important Chinese herbal medicines, with a lot of good agronomic traits, such as yellow seed and crack pod. Especially demonstrated by the drought resistance has been widespread attention and conducted in-depth study, cruciferous breeding excellent germplasm resources.
Environmental stress often caused by changes in plant gene expression and cell metabolism, thus affecting the growth and yield of crops. Biotic and abiotic stress are often accompanied by the growth of the plants, in which light, drought, salt, temperature is the most important stress factors limiting crop production, while the losses caused by drought, and its loss more than the sum of the losses of other adversities. Thus increasing the drought resistance of crop varieties has become the key to high-yield crop varieties and stable yield.
The polyamines are a presence in plants, the regulation of plant growth and development of low molecular weight aliphatic nitrogen-containing alkali. Putrescine, spermidine, spermidine is the common polyamines. Recent studies show that polyamines and plant resistance are closely linked, when plants experience drought and other environmental stresses, the cells increase in the sky a protective effect through the polyamine content. In addition, the plants subjected to drought stress, total body protein synthesis rate dropped, some of the original protein synthesis is inhibited, while at the same time new synthesis of some proteins (stress-inducible protein). Stress-inducible protein to adapt to the adversity of plants play an important role, can significantly improve plant resistance to stress ability.
For current research on the arid sinapis alba L. proteome level at home and abroad are still rarely reported that this experiment by PEG6000treatment of seedlings to simulate the impact of the drought seedlings, found that PEG6000can simulate drought stress, so that seedling performanceout of the state of growth under drought conditions. Before the relevant reports, drought can affect the accumulation of polyamines in the leaves. we measured polyamine content in leaves of sinapis alba L. seedlings under different drought levels, polyamine content with the increase of the degree of aridity on the rise, but withdrought intensified, the polyamine content in leaves decreased cell damage intensified. Thus, we can initially infer that drought stress to mobilize the accumulation of polyamines, polyamine accumulation helps to improve the drought resistance of sinapis alba L.. Study the relationship of the degree of aridity and polyamine accumulation, suggesting that polyamines play an important role in the mechanism of sinapis alba L. drought, but the exact mechanism still needs further exploration; Drought mechanism in order to more in-depth study of2D-PAGE., we identified58differentially expressed proteins under drought stress. Among them, the control group,16unique proteins,42unique proteins in the treatment group. Speculate that these proteins are closely related to the drought resistance of the sinapis alba L.. We selected10protein spots to mass spectrometry analysis showed that five protein spots match in the database of NCBI and SP. These proteins have been identified, and speculated that its function.For further research to study the mechanism of drought, drought Cloning and crop drought breeding provides an important theoretical basis.
本研究以白芥为研究对象,对其干旱胁迫下叶片中多胺含量的变化、叶片差异蛋白表达情况进行研究,旨在探讨和揭示白芥对干旱胁迫的抗性机理。试验通过PEG6000处理白芥幼苗来模拟干旱对白芥幼苗的影响,发现PEG6000完全可以成功的模拟干旱胁迫,使幼苗表现出干旱条件下的生长状态。通过对不同干旱程度下白芥幼苗叶片中多胺含量变化的测定,发现多胺含量随干旱程度的增加呈现上升趋势,但随干旱程度加剧,叶片中的多胺含量开始逐步下降,细胞受伤害程度加剧。由此可以推断,干旱胁迫会调动植物体内多胺积累以提高植株的抗旱性。试验通过蛋白质双向电泳和质谱技术研究了白芥干旱胁迫与对照幼苗叶片的蛋白差异表达情况,发现干旱胁迫较对照相比产生58个差异蛋白质。其中,对照组中有16个特有蛋白质,处理组中有42个特有蛋白质,共有有43个蛋白出现表达量的变化。可以推测这些差异蛋白与白芥的抗旱机理密切相关。试验选取10个差异蛋白点进行质谱分析,分析结果显示有5个蛋白点在NCBI和SP数据库中获得相关匹配,对这些蛋白进行初步鉴定和相关功能推测。有5个蛋白点在NCBI和SP数据库中没有获得相关匹配,预测可能为新的抗旱性相关蛋白。结果表明,本研究方法成熟可靠,结果较为理想,可以为以后进一步深入研究白芥抗旱机理、抗旱基因的克隆以及作物的抗旱育种提供重要的理论依据。
Sinapis alba L. is an important Chinese herbal medicines, with a lot of good agronomic traits, such as yellow seed and crack pod. Especially demonstrated by the drought resistance has been widespread attention and conducted in-depth study, cruciferous breeding excellent germplasm resources.
Environmental stress often caused by changes in plant gene expression and cell metabolism, thus affecting the growth and yield of crops. Biotic and abiotic stress are often accompanied by the growth of the plants, in which light, drought, salt, temperature is the most important stress factors limiting crop production, while the losses caused by drought, and its loss more than the sum of the losses of other adversities. Thus increasing the drought resistance of crop varieties has become the key to high-yield crop varieties and stable yield.
The polyamines are a presence in plants, the regulation of plant growth and development of low molecular weight aliphatic nitrogen-containing alkali. Putrescine, spermidine, spermidine is the common polyamines. Recent studies show that polyamines and plant resistance are closely linked, when plants experience drought and other environmental stresses, the cells increase in the sky a protective effect through the polyamine content. In addition, the plants subjected to drought stress, total body protein synthesis rate dropped, some of the original protein synthesis is inhibited, while at the same time new synthesis of some proteins (stress-inducible protein). Stress-inducible protein to adapt to the adversity of plants play an important role, can significantly improve plant resistance to stress ability.
For current research on the arid sinapis alba L. proteome level at home and abroad are still rarely reported that this experiment by PEG6000treatment of seedlings to simulate the impact of the drought seedlings, found that PEG6000can simulate drought stress, so that seedling performanceout of the state of growth under drought conditions. Before the relevant reports, drought can affect the accumulation of polyamines in the leaves. we measured polyamine content in leaves of sinapis alba L. seedlings under different drought levels, polyamine content with the increase of the degree of aridity on the rise, but withdrought intensified, the polyamine content in leaves decreased cell damage intensified. Thus, we can initially infer that drought stress to mobilize the accumulation of polyamines, polyamine accumulation helps to improve the drought resistance of sinapis alba L.. Study the relationship of the degree of aridity and polyamine accumulation, suggesting that polyamines play an important role in the mechanism of sinapis alba L. drought, but the exact mechanism still needs further exploration; Drought mechanism in order to more in-depth study of2D-PAGE., we identified58differentially expressed proteins under drought stress. Among them, the control group,16unique proteins,42unique proteins in the treatment group. Speculate that these proteins are closely related to the drought resistance of the sinapis alba L.. We selected10protein spots to mass spectrometry analysis showed that five protein spots match in the database of NCBI and SP. These proteins have been identified, and speculated that its function.For further research to study the mechanism of drought, drought Cloning and crop drought breeding provides an important theoretical basis.
引文
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