黑果枸杞LrPIP1基因的克隆鉴定及外施水杨酸对其在干旱胁迫下表达的影响

Cloning and Identification of LrPIP1 Gene of Lycium ruthenicum and Effect of Exogenous SA on Its Expression under Drought Stress

植物质膜水通道蛋白(plasma membrane intrinsic proteins,PIPs)是一类参与植物众多生理活动的多功能蛋白。为探究PIP基因在黑果枸杞(Lycium ruthenicum Murr.)响应干旱胁迫中的作用,以黑果枸杞为试验材料分离得到PIP基因,命名为LrPIP1。使用生物信息学方法对其编码蛋白的理化性质、序列进化进行分析,以及预测蛋白质的二级和三级结构并构建基因树等。将幼苗进行不同程度干旱胁迫与外施水杨酸处理,实时荧光定量PCR结果显示,无外源SA处理下,轻度干旱胁迫D1可诱导LrPIP1基因的相对表达量较CK处理显著上调,中度D2、重度胁迫D3时,LrPIP1基因的相对表达量较CK有一定程度的上调,但相较于D1表现为显著下降;外源喷施SA可影响干旱胁迫下黑果枸杞LrPIP1基因的相对表达量,轻度胁迫下,外源SA诱导LrPIP1基因表达量较CK下调;较CK处理,中、重度胁迫下SA可诱导LrPIP1基因表达量上调。上述表明,LrPIP1基因可能在黑果枸杞非生物胁迫响应过程中发挥重要调控作用。

Plasma membrane intrinsic proteins(PIPs)is an essential multifunctional protein involved in many physiological activities of plants.To investigate the role of Lycium ruthenicum Murr.PIP gene in response to drought stress.In this study,we cloned a PIP gene in Lycium ruthenicum Murr.named as LrPIP1.Bioinformatics were used to systematically analyze characteristics and phylogeny,predict secondary and three dimensional protein structure,and construct phylogenetic tree of LrPIP1 protein.Lycium ruthenicum seedlings were treated with drought stress and salicylic acid(SA)in different gradients and the expression levels of LrPIP1 gene were checked by Real-time quantitative PCR(qRT-PCR).The results showed that transcripts of LrPIP1 gene expression were significantly raised under light drought stress compared with CK treatments without SA treatments,and the expression of LrPIP1 was up-regulated compared with CK under moderate and severe stresses and down-regulated significantly compared with CK.Under drought stress,SA can regulate the expression of LrPIP1 gene,comparede with CK the transcripts of which were down-regulated under mild drought stress,but up-regulated under moderate and severe drought stresses,the expression of LrPIP1 was significantly improved.Taken together,these results demonstrated that the LrPIP1 gene might play a crucial role in regulating the abiotic stress response in Lycium ruthenicum.Thus,this study lays the foundation for further illustrating functions of LrPIP1 gene.