青海高原抗旱蚕豆对干旱胁迫的响应与干旱诱导蛋白研究

The threat of drought on agricultural production is a worldwide problem, has caused the attention of the researchers of many countries. Qinghai faba bean have its reputation as the "world's best beans", occupies a very important role in agricultural production in Qinghai province. In recent years, faba bean production area to be transfered to rainfed farming along with the irrigated agriculture area reduced. It has a social significance to cultivate drought-resistant faba bean, high-quality bean varieties to enlarge the planting area of faba bean in arid regions, meanwhile, deepening drought region planting structure adjustment, promote producing grain and oil crops in Qinghai land with a combination of social significance, needing a detailed study of Qinghai faba bean drought resistance. This study drought-tolerant faba bean "Qinghai 13" as the material, the research under different concentration gradient PEG stress faba bean response to drought stress and drought -induced proteins research. Reveal the broad bean under drought stress of morphological, physiological, biochemical, protein molecules such as comprehensive coordination ways to adapt to drought, proven bean leaf protein expression change and drought resistance related proteins of isolation, identification, clear drought induced protein play a role of function under drought stress, and through the electronic prediction of target protein gene cloning, for Qinghai faba bean art elite germplasm gene mining and germplasm innovation provides new ideas and new methods. Our research team has carried out the analysis of drought resistance of 16 genetic resource faba bean in seedling under the drought stress. Comprehensive evaluation was carried out of drought resistance for different genetic resource faba bean through the subordinate function value method、PCA（principal component analysis ）and clustering methodology. By the analysis of the comprehensive evaluation "Qinghai13" is higher drought-resistant ability, which is a earlier and professional scientific research institution to bring comprehensive evaluation into faba bean drought-resistant research in the Tibetan Plateau area. It has lay the foundation for the next further study.

干旱对农业生产的威胁是一个世界性问题，已经引起了各国研究者的高度重视。青海蚕豆有"世界上最好蚕豆"的美誉，在青海省农业生产中占非常重要的地位。近年来随着灌溉农业区面积的锐减，蚕豆的生产区域向雨养干旱农业区拓展和转移，培育抗旱、优质蚕豆品种对扩大干旱地区蚕豆的种植面积，深化干旱地区种植业结构调整，推动青海粮油作物主产区的土地用养结合具有社会意义，需要对青海蚕豆抗旱性进行详细的研究。 本研究以耐旱蚕豆"青海13号"为材料，研究不同浓度梯度PEG胁迫下蚕豆对干旱胁迫的响应与干旱诱导蛋白研究。揭示干旱胁迫下蚕豆的形态、生理生化、蛋白质分子等综合协调适应干旱的途径，探明蚕豆叶片蛋白质表达变化及抗旱相关蛋白的分离、鉴定，明确干旱诱导蛋白在干旱胁迫下发挥的功能作用，并通过电子克隆对目标蛋白基因进行预测，为青海蚕豆抗逆优异种质基因挖掘和种质资源创新提供新思路和新方法。

在干旱胁迫下植物能够产生一系列的形态、生理生化及分子生物学等方面的变化而表现出抗旱性。干旱诱导差异蛋白是干旱胁迫下新合成的一类蛋白，它们是植物抗旱发生一系列生理生化反应的基础，在植物干旱胁迫忍耐过程中起着重要的作用。“青海13号”是青海乃至西北地区干旱、半干旱地区广泛种植的耐旱蚕豆品种，深入探讨其对干旱胁迫的响应机制，揭示在干旱胁迫下蚕豆的形态、生理生化、蛋白质分子等综合协调适应干旱的途径，探明蚕豆叶片蛋白质表达变化及抗旱相关蛋白的分离、鉴定，明确干旱诱导蛋白在干旱胁迫下发挥的功能作用，有助于揭示“青海13号”抗旱性形成的原因。通过电子克隆对目标蛋白基因进行预测，为青海蚕豆抗逆优异种质基因挖掘和种质资源创新提供新思路和新方法。.应用双向电泳和质谱鉴定技术进行蚕豆叶片差异蛋白质组分析发现，32个差异表达蛋白点部分呈现上调表达，部分呈现下调表达，还有7个消失蛋白点和1个新增蛋白点。采用MALDI-TOF/TOF鉴定和生物信息学分析，成功鉴定的21个蛋白点按其所参与的代谢途径和生化功能可分为七大类，参与信号转导的2个，参与自由基清除的1个，参与防卫反应的1个，参与代谢的8个，参与蛋白加工的1个，参与光合的5个，未知功能蛋白3个。Drought inducible 22 KD protein (spot 2705)、stress-inducible protein(spot 5807)、17.5 kD一级热激蛋白(spot 3208)、超氧化物歧化酶(superoxide dismutase, spot 3106)是与抗旱性有直接关联的蛋白点，其相对表达量的上调可能是“青海13号”蚕豆具有抗旱性的重要原因。值得一提的是在上调表达中发现了一个新的干旱胁迫响应蛋白莽草酸酯脱氢酶(Shikimate dehydrogenase substrate binding, N-terminal, partial (ISS), spot 3809)，推测是青海13号蚕豆具有抗旱性机制所在。电子克隆得出的序列与蒺藜苜蓿的基因组序列进行比对，得到氨基酸序列和核酸序列。当前的研究为我们深入了解蚕豆对干旱胁迫的响应提供了新的线索，也是利用遗传等手段进一步研究这些基因功能的开始。