摘要
[目的]本文旨在克隆菊花中苯丙氨酸解氨酶基因(PAL),并探讨其在不同组织的表达特性以及在胁迫和激素处理下的响应模式。[方法]以菊花‘神马’为试验材料,依据其转录组数据库信息,克隆CmPAL全长,并利用生物信息学方法对其进行分析。利用荧光定量PCR检测该基因在不同组织中的相对表达量,在蚜虫和干旱胁迫以及不同激素处理下的表达变化。[结果]CmPAL基因开放阅读框(ORF)全长2 154 bp,编码氨基酸717个,理论等电点(p I)为5.75,预测蛋白相对分子质量为77.73×10~3。系统进化分析表明该基因编码的蛋白与黄花蒿和泡黄金菊的PAL亲缘关系最近,同源性分别为97.94%和91.88%,具有较高的保守性。CmPAL在菊花根中表达量最高,管状花中次之。蚜虫胁迫和茉莉酸甲酯(MeJA)可诱导CmPAL基因的表达,水杨酸(SA)、独角金内酯类似物GR24则抑制CmPAL的表达。干旱胁迫3 h处理组CmPAL基因表达量高于对照组。[结论]菊花‘神马’CmPAL基因在菊花的根中表达量最高,并受蚜虫和MeJA诱导,受SA和独角金内酯类似物GR24抑制,表明该基因可能参与多种胁迫防御反应和激素应答。
[Objectives]The purpose of this research is to clone the phenylalanine ammonia-lyase gene( PAL) in chrysanthemum and dissect it roles in response to different stresses and phytohormones. [Methods]The full-length of CmPAL was obtained from chrysanthemum'Jinba'based on its RNA-seq library. Bioinformatic technologies were used to analyze the gene. The relative expression levels of CmPAL in different tissues of plant were detected by RT-qPCR. The expression profiles of CmPAL under aphids,drought,and different plant hormones were analyzed. [Results]CmPAL gene had an open reading frame( ORF) of 2 154 bp encoding 717 amino acids. The theoretical isoelectric point( p I) was 5.75,and the predicted protein molecular weight was 77.73×10~3. Phylogenetic analysis showed that it had the closest relationship with Artemisia annua and Chrysanthemum boreale,and the amino acid sequence identity was 97.94% and 91.88%,respectively. CmPAL had the highest expression level in root,followed by that in tubular floret.CmPAL was induced by aphid and methyl jasmonate( MeJA),while it was inhibited by salicylic acid( SA) or the strigolactone analogue GR24. The expression level of CmPAL at 3 h after drought treatment was higher than that of the control group.[Conclusions]CmPAL in chrysanthemum'Jinba'was of the highest expression level in root. The expression of CmPAL positively responded to aphid and MeJA,while negatively to SA and GR24,which suggested that CmPAL might be involved in multiple stress defense and phytohormone responses.
引文
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