桑树磷脂酶MaPLA1-2D亚型4个基因的克隆与胁迫应答分析

磷脂酶(phospholipase)是一类在植物生长发育和胁迫应答中起重要调控作用的磷脂水解酶,也是一类重要的信号转导酶。而磷脂酶A1(PLA1)在植物应答生物胁迫和非生物胁迫中的功能研究鲜见报道。研究从桑树(Morus alba L.)中克隆了磷脂酶PLA1的1个亚型MaPLA1-2D基因,对其进行了序列分析、组织表达、胁迫诱导表达和蛋白亚细胞定位分析。结果表明,桑树PLA1-2D亚型基因包括4个成员,命名为MaPLA1-2D.1~MaPLA1-2D.4。4个基因在桑树根和叶中高水平表达,蛋白亚细胞定位在叶绿体。序列和进化分析表明MaPLA1-2D基因4个成员与拟南芥AtDAD1基因的保守结构域序列具有较高相似度且进化关系紧密。MaPLA1-2D基因4个成员的启动子含有多种胁迫应答顺式元件和激素响应元件;胁迫诱导表达模式分析表明MaPLA1-2D基因表达受干旱和脱落酸处理显著诱导。以上结果说明,MaPLA1-2D基因与拟南芥DAD同源,可能在桑树非生物胁迫应答中发挥重要功能。

Gene Cloning and Stress Responsive Analysis of Phospholipase A1-2D in Mulberry (Morus alba L.)

Phospholipase is a class of phospholipase hydrolases and signal transducers that play an important role in regulating plant growth and development and stress response. However, the function of phospholipase A1 (PLA1) in plant response to biological and abiotic stresses is rarely reported. In this study, a subtype of phospholipase PLA1, MaPLA1-2D was cloned in mulberry, and the sequence feature, tissue expression, stress-induced expression of MaPLA1-2D, and subcellular localization of the protein were analyzed. The results showed that MaPLA1-2D contains 4 isoforms, named MaPLA1-2D.1- MaPLA1-2D.4. The four genes were highly expressed in the roots and leaves of mulberry, and the proteins were mainly localized in chloroplasts. The MaPLA1-2D members had high sequence similarity and close evolutionary relationship with Arabidopsis AtDAD1 in the conserved domain. It was showed that the promoters of the four MaPLA1-2D genes contained a variety of stress-responsive cis-elements and hormone responsive elements, and the gene expressions of MaPLA1-2D were significantly induced by drought stress and abscisic acid (ABA) treatments. These results indicate that MaPLA1-2D genes are homologous to Arabidopsis DAD, which may play an important role in abiotic stress response of mulberry.