中文摘要
水通道蛋白(AQP)能在植物体内实现水分快速跨膜运输,但其在梨自交不亲和性(SI)中的作用尚不清楚,国内外尚无报道。前期我们已证实,AQP与梨SI之间有直接联系。本项目拟在前期工作基础上,利用已知S基因型梨品种:‘丰水’、‘奥嗄’、‘今村秋’、‘奥嗄二十世纪’为研究对象,用AQP抑制剂、激活剂、培养基渗透压调节剂等处理,结合分子生物学与生物信息学手段阐明梨花粉细胞AQP的性质及作用,研究AQP的表达与调控,确定AQP的种类与分布情况,进一步探明AQP在梨花粉萌发和花粉管伸长过程中的作用机制。为进一步阐明植物SI提供新的理论支持,为研究梨SI反应的生理机制提供新的思路。
英文摘要
Aquaporin(AQP) can transport water in a high speed transmembrane type. But the role it playing in the pear self-incompatibility(SI) has not been found clearly so far. There is not any reports all over the word. At the very beginning, we have confirmed that AQP has direct relations with pear SI. This program is based on what we have done before. We use 'Imamuraaki', 'Osa', 'Housui' and 'Osa-nijisseiki' as research objects. Through the way disposing them by AQP inhibitor, activitor and culture medium osmolarity modulator, combining with molecular biological and bioinformatics technologies, we will make the function and properties of AQP clear, study on the expression and regulation of AQP, and make sure the distribution of AQP. Find out the effect mechanism of AQP taking part in pear pollen tube elongation and pollen germination. Pear pollen will be dealed with S-RNase combinating with field pollination validation to ascertain the mechanism of AQP affecting in the SI reaction. This study will provide a new theory surpport for SI in plant.We can also give new ideas to the physiological mechanism of SI reaction.
结题摘要
水通道蛋白(AQPs)能在植物体内实现水分快速跨膜运输。本项目通过利用水通道蛋白抑制剂,激活剂处理‘丰水’、‘今村秋’、‘奥嘎’‘奥嘎20世纪’花粉,结果表明其对花粉萌发影响显著。同时,利用Blast、Swiss-Model, MEME, Network Protein Sequence Analysis等生物信息学软件,筛选出梨水通道蛋白PbrPIP,PbrTIP基因家族,并对其系统树,基因结构和功能域以及其表达蛋白质的空间结构等进行了分析。结果表明,共有16个PIP类梨水通道蛋白基因、15个TIP类水通道蛋白基因,PbrPIP基因含有3-4个内含子,PbrTIP基因含有2-3个内含子,其表达的蛋白质氨基酸序列长度在198-301之间,理论等电点在5.16-7.69之间,均为稳定性蛋白。亚细胞定位分析表明,PbrPIP均定位在细胞膜上,而PbrTIP中除PbrTIP2、PbrTIP13 PbrTIP14以及PbrTIP15定位在液泡膜和细胞膜上以外,其它蛋白质亚细胞定位均定位在液泡膜上。二级结构由α螺旋,无规则卷曲和延伸链等结构元件组成,空间结构高度相似。qRT-PCR分析表明,梨PbrPIP基因在根、茎、叶、花粉及花柱中表达情况的差异说明虽然它们在结构上具有很高的同源性,但是这些基因的功能发生了分化,从而广泛参与植物各种重要生命活动。
