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通过染色体步移法分离桃[Prunus persica(L.)Batch]PpPGIP1 上游启动子序列,利用生物信息学方法对其功能进行初步预测;构建该启动子植物表达载体并通过农杆菌介导法转化烟草,研究不同激素诱导条件下GUS蛋白瞬时表达情况;并利用实时荧光定量 RT-PCR 技术分析了PpPGIP1在水杨酸(SA)、茉莉酸甲酯(MeJA)、脱落酸(ABA)和 1-氨基环丙烷-1-羧酸(ACC)诱导下的表达特性,获得了长度为1018bp的桃PpPGIP1启动子序列。该序列与梅和中国李PGIP启动子序列的同源性分别为90%和89%;该启动子序列含有SA、MeJA、ABA和ETH诱导调控相关的抗病与胁迫顺式作用元件;ABA和ACC可以诱导PpPGIP1启动子调控GUS基因的表达;实时荧光定量RT-PCR分析结果表明:SA、MeJA、ABA和ACC都可以诱导桃叶片中PpPGIP1的表达。PpPGIP1可能参与SA、MeJA、ABA和ETH的信号转导,在桃抵抗病原菌和逆境胁迫方面起着非常重要的作用。
The upstream promoter sequence of Prunus persica (L.) Batch] PpPGIP1 was isolated by chromosome walking and its function was preliminarily predicted by bioinformatics method. The promoter was constructed and transformed into tobacco by Agrobacterium tumefaciens , To study the transient expression of GUS under different hormone-induced conditions. The effects of PpPGIP1 on the expression of GUS protein in saliva, MeJA, ABA and 1-amino groups were analyzed by real-time fluorescence quantitative RT- Cyclopropane-1-carboxylic acid (ACC), the peach PpPGIP1 promoter sequence was obtained with a length of 1018 bp. This sequence has 90% and 89% homology with Mei and Li PGIP promoter sequences, respectively. The promoter sequence contains the cis-acting elements for resistance and stress induced by SA, MeJA, ABA and ETH. ABA and ACC could induce PpPGIP1 promoter to regulate the expression of GUS gene. Real-time fluorescent quantitative RT-PCR analysis showed that SA, MeJA, ABA and ACC could induce the expression of PpPGIP1 in peach leaf. PpPGIP1 may be involved in the signal transduction of SA, MeJA, ABA and ETH and plays an important role in the resistance of peach to pathogenic bacteria and stress.