日本结缕草（Zoysia japonica）磷脂酶D基因部分 保守序列的克隆及其对机械损伤的响应

以日本结缕草Zenith为材料,通过同源克隆的方法克隆出其磷脂酶D(PLD)基因部分保守序列,序列长399 bp,并通过实时荧光定量PCR检测PLD基因在机械损伤胁迫下随时间变化的表达模式.结果表明,磷脂酶D在胁迫2h内大量表达,随后逐渐降低,在9h已接近于对照水平,说明PLD基因在损伤初期大量参与了应对机械损伤胁迫的防御与修复过程,在损伤后期其作用减弱或被代替;电导率的测定结果表明,细胞膜结构在胁迫初期遭到极大程度的破坏,胁迫9h后膜损伤程度开始减轻.结合PLD基因和电导率的表达模式,推测PLD基因在6h内会激活一系列感受信号及下游信号分子,9h后其相应防御机制发挥作用来减轻胁迫造成的损害.

Cloning of partial conservative phospholipase D gene and its response to mechanical wounding in Japan zoysia(Zoysia japonica)

Using homologous cloning method, part of conservative sequence of phospholipase D (PLD) gene was isolated from Japan zoysia Zenith, the sequence of which was 399 bp. Through the real-time fluorescent quantitative PCR, timespace expression pattern of the PLD gene was studied under the stress of mechanical wounding. Results showed that PLD gene distinctly expressed within 2 h, then gradually decreased. After 9 h, the expression was closed to the control level. PLD gene involved in the defense and repair process in the early damage and its effect reduced or be replaced in the late damage. The result of conductivity indicated that cell membrane suffered great damage in the early mechanical wounding, after 9 h, the damage began to slowly recovere. By comprehensive analysis of expression pattern of PLD and conductivity, it suggested that PLD could activate a series of signals and downstream molecules within 6 h, however, after 9 h its corresponding defense mechanism played a role in alleviating the damage caused by stress.