[关键词]
[摘要]
目的 研究介导内生真菌诱导子促进苍术悬浮细胞中苍术素生物合成的信号分子及信号转导途径,并探讨诱导子对苍术素合成途径关键酶活性的影响。方法 采用植物细胞悬浮培养法,考察内生真菌诱导子处理下苍术细胞中NO、水杨酸(SA)及苍术素量的变化。结果 内生真菌诱导子通过诱导苍术细胞中一氧化氮合酶(NOS)、苯丙氨酸解氨酶(PAL)以及乙酰辅酶A羧化酶(ACC)的活性,显著促进苍术细胞的NO迸发、SA和苍术素的合成。NOS抑制剂PBITU可以阻断诱导子对NO、SA和苍术素合成的促进作用,外源NO供体SNP及外源SA单独处理也能促进苍术素的合成,说明NO和SA是参与苍术素合成的信号分子,且NOS是参与诱导子诱发苍术细胞NO迸发的主要途径。NO猝灭剂cPITO可以有效清除诱导子诱发苍术细胞的NO迸发,显著阻断诱导子对苍术细胞中SA和苍术素合成的促进作用,外源SNP可以逆转cPITO对PAL、ACC活性以及SA、苍术素合成的抑制作用,表明NO是介导内生真菌诱导子诱发苍术细胞中苍术素和SA生物合成所必需的上游信号分子。结论 NO主要通过SA信号途径介导内生真菌诱导子激活ACC,显著促进苍术细胞中苍术素的生物合成。
[Key word]
[Abstract]
Objective To investigate the signal molecules and signal transduction involved in endophytic fungal elicitor-induced atractylodin biosynthesis and the effect of an endophytic fungal elicitor on the key enzyme activity in Atractylodes lancea. Methods Nitric oxide (NO), salicylic acid (SA), and atractylodin content changes were detected under the endophytic fungal elicitor treatment by plant cell suspension culture technology. Results The endophytic fungal elicitor remarkably promoted NO burst and the biosynthesis of SA and atractyodin by activating nitric oxide synthase (NOS), phenylalanine ammonia lyase (PAL) and acetyl coenzyme A carboxylase (ACC), respectively. NOS inhibitor PBITU could inhibit the NO and SA accumulation and the atractyodin biosynthesis induced by the elicitor. And atractyodin biosynthesis could also be triggered by exogenous NO or SA. The results indicated that NO and SA were the necessary signal molecules and NO burst was mediated by NOS induced by endophytic fungal elicitor. NO quencher cPITO could effectively remove NO burst in Atractylodes lancea cell induced by endophytic fungal elicitor and notably inhibit the biosynthesis promotion of SA and atractyodin in Atractylodes lancea cell induced by endophytic fungal elicitor. Exogenous SNP could reverse the cPITO inhibition on the activity of PAL and ACC and the synthesis of SA adn atractylodin. This suggested that NO was an upstream signal molecule mediated endophytic fungal elicitor to accelerate the biosynthesis of SA and atractyodin. Conclusion Endophytic fungal elicitor mediated through NO followed by SA could promote atractyodin biosynthesis by activating ACC in A. lancea.
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[基金项目]
国家自然科学基金资助(81102743)