欧美杨水分利用效率相关基因PdEPF1的克隆及表达  

The cloning and expression of WUE-related gene (PdEPF1) in Populus deltoides×Populus nigra

作　　者：郭鹏[1] 金华[1] 尹伟伦[2] 夏新莉[2] 姜国斌[1] 

机构地区：[1]大连民族学院环境与资源学院,大连116600 [2]北京林业大学生物学院,北京100083

出　　处：《生态学报》2012年第14期4481-4487,共7页Acta Ecologica Sinica

基　　金：国家自然科学基金(31100489);辽宁省重点农业攻关计划项目"耐旱;耐盐碱速生杨树品种吴屯杨(WTI)的选育"(2008207001);中央高校基本科研业务费资助项目(DC12010204)

摘　　要：提高植物水分利用效率(WUE)是未来解决我国甚至世界干旱缺水的最重要手段之一。在对植物WUE的众多研究方法中,大多集中在生理手段,但通过分子生物学手段研究其表达调控机制的较少。欧美杨(Populus deltoides×Populus nigra)是中纬度地区最适合种植的短轮伐期工业用材集约经营树种之一。近年来我国引进了许多优良的欧美杨无性系用于营造大面积的速生丰产林并取得很好的经济和社会效益。但高耗水量的缺点限制了其进一步的推广。通过基因芯片技术从欧美杨中找到一个可能调控WUE的基因-PdEPF1.荧光定量表达进一步验证了这一结果。荧光定量表达分析表明该基因受ABA、盐、冷、干旱等胁迫诱导表达。组织特异表达分析说明PdEPF1基因在顶端叶和根中有表达,成熟叶衰老叶中则无表达。克隆到启动子分析表明该启动子含有多种干旱响应元件(drought response elememt),赤霉素响应元件(GA response elememt),低温响应元件(coldresponse elememt),ABA响应元件(ABA response elememt)等逆境相关的作用元件。Water-use efficiency（WUE） is crucial to plant survival,yield and vegetation dynamics.Many researches trended to reduce water use by more crop per drop.Althoughmuch progresses have been achieved in determining the physiological mechanisms of WUE,there is few report on the genetic controls of WUE.The first gene ERECTA,which t promotes WUE,was shown to decrease stomatal density and be involved in the positive regulation of photosynthetic capacity. The EPIDERMAL PATTERNING FACTOR 1（EPF1） gene identified in Arabidopsis is expressed specifically in stomatal cells and precursors.EPF1 controls stomatal patterning through the regulation of asymmetric cell division,its activity depends on the TOO MANY MOUTHS receptor-like protein and ERECTA family receptor kinases,suggesting that EPF1 provides a positional cue interpreted by these receptors.A mutation of EPF1 results in increased stomatal density and violation of the one-cell-spacing rule（clustering of stomata）.However,there is no report on the relationship between AtEPF1 and WUE. The cottonwood poplar,Populus deltoides×Populus nigra,is one of the most important species in large-scale forestation in China.However,its high water consumption is a key factor limiting its growth in arid and semiarid regions.Identifcation of genes increasing WUE in this species is important for understanding stress responses and improvement of this species. In this study,microarray analysis between poplar genotypes NE-19（P.nigra×（P.deltoides×P.nigra）） with the higher WUE and DN-2（P.deltoides×P.nigra） with a lower WUE,was performed.PdEPF1 from NE-19,one of the WUE-related genes,was identified,strongly expressed in immature leaves and roots,but not in senescent leaves,functional leaves and the stem.This gene is strongly induced by drought,abscisic acid and high salt levels,and enhanced by gibberellic acid.To unvarel the mechanism responsible for the inducible expression of PdEPF1 by different stresses,a 1090-bp PdEPF1 promoter fragment was isolated by the adaptor PCR w

关 键 词：水分利用效率 欧美杨 PdEPF1基因 逆境 

分 类 号：S792.11[农业科学—林木遗传育种]