小麦TaPIP1基因克隆及其在花药开裂中潜在功能分析

Cloning of TaPIP1 gene and its potential function in anther dehiscence in wheat

二系杂交小麦育种是小麦产量提高的重要途径之一。光温敏雄性不育小麦生殖生长过程中花药的发育及开裂情况直接影响杂交小麦的制种效率和产量。植物花药的开裂与脱水活动紧密相关。水通道蛋白(aquaporins,AQPs)是高效转运水分及特异小分子的膜内在蛋白,其中质膜内在蛋白(plasma membrane intrinsic proteins,PIPs)在水分的吸收与外排中发挥着重要作用。为进一步了解水通道蛋白在小麦光温敏核雄性不育系花药开裂中的作用提供理论基础。本研究以不育系BS366花药的cDNA为模板,克隆获得了TaPIP1基因。利用生物信息学软件对TaPIP1进行分析,该基因包含一个879 bp的开放阅读框,编码292个氨基酸。TaPIP1的启动子区存在赤霉素、脱落酸、茉莉酸和光等响应元件。TaPIP1属于MIP超家族,具有典型的NPA保守结构域,亚细胞定位于细胞质膜与核膜上。miRNA互作预测发现TaPIP1受tae-miR1131与tae-miR408的剪切抑制,表明TaPIP1可能和与植物的抗氧化能力相关。通过蛋白互作预测及qPCR实验,表明TaPIP1可与热激蛋白(heat shock protein 90,TaHSP90)相互作用,在高温和干旱复合胁迫下,参与花药细胞壁膨压调控,进而调控花药的开裂。

Hybrid wheat breeding is one of the important ways to improve wheat yield.The development and dehiscence of an-thers in photoperiod-temperature sensitive genic male sterile(PTGMS)wheat affect the seed production efficiency and yield of hybrid wheat directly.Anther dehiscence is closely related to dehydration.Aquaporins(AQPs)are membrane intrinsic proteins that efficiently transport water and specific small molecules.Plasma membrane intrinsic proteins(PIPs)play an important role in water absorption and efflux in plant cells.In order to understand the roles of PIP in anther dehiscence of PTGMS.In this study,TaPIP1 was cloned from the anther of PTGMS line BS366.The gene contained an open reading frame(ORF)of 879 bp,encoding a total of 292 amino acids.There were cis-responsive elements such as gibberellin,abscisic acid,jasmonic acid,and light in the promoter region of TaPIP1.The promoter region of TaPIP1 contained elements of gibberellin,abscisic acid,jasmonic acid,and light.TaPIP1 belonged to MIP superfamily,and had a typical NPA conserved domain,subcellular localication in the plasma mem-brane and nuclear membrane.The interaction between miRNAs and TaPIP1 was predicted by bioinformatics analysis,and the results showed that TaPIP1 may be regulated by tae-miR1131 and tae-miR408 that related to the antioxidant capacity of plants.Protein-protein interaction networks(PPI)and qRT-PCR experiments revealed that TaPIP1 could interact with heat shock protein 90(TaHSP90),participating in the regulation of anther cell wall turgor under the combined stress of high temperature and drought,thus regulating anther dehiscence.