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激发子诱导番茄细胞中乙烯生物合成:特性及作为激发子活性生物测定方法的应用

Elicitor-induced ethylene biosynthesis in tomato cells: characterization and use as a bioassay for elicitor action.

作者信息

Felix G, Grosskopf D G, Regenass M, Basse C W, Boller T

机构信息

Friedrich Miescher-Institute, P.O. Box 2543, CH-4002 Basel, Switzerland.

出版信息

Plant Physiol. 1991 Sep;97(1):19-25. doi: 10.1104/pp.97.1.19.

Abstract

The induction of ethylene biosynthesis by an elicitor partially purified from yeast extract was studied in suspension-cultured tomato (Lycopersicon esculentum Mill.) cells. Unstimulated cells produced little ethylene during exponential growth and even less in stationary phase. Treatment with elicitor stimulated ethylene biosynthesis 10-fold to 20-fold in the exponentially growing cells and more than 100-fold in stationary cells. Activities of both 1-aminocyclopropane-1-carboxylate (ACC) synthase, measured in vitro, and ethylene-forming enzyme (EFE), measured in vivo, increased strongly in response to elicitor treatments. During exponential growth, cells contained large pools of ACC, and the elicitor stimulated ethylene biosynthesis primarily through induction of EFE. In the stationary phase, cells contained almost no ACC, and the elicitor stimulated ethylene biosynthesis primarily through its effect on ACC synthase activity. Cordycepin did not affect the increase in activity of ACC synthase but blocked that of EFE, indicating that the former was posttranscriptionally regulated, the latter transcriptionally regulated. Removal of elicitor by washing or inactivation of a biotinylated derivative of the elicitor by complexation with avidin caused a rapid cessation of the increase in ACC synthase activity, suggesting that continuous presence of stimulus is necessary for the response. Using induction of ethylene production to measure amounts of elicitor, it was found that the elicitor disappeared from the incubation medium during the course of the treatment.

摘要

对从酵母提取物中部分纯化得到的激发子诱导乙烯生物合成的过程,在悬浮培养的番茄(Lycopersicon esculentum Mill.)细胞中进行了研究。未受刺激的细胞在指数生长期产生少量乙烯,在稳定期产生的更少。用激发子处理后,指数生长期的细胞中乙烯生物合成被刺激了10倍至20倍,在稳定期细胞中则超过100倍。体外测定的1-氨基环丙烷-1-羧酸(ACC)合酶活性和体内测定的乙烯形成酶(EFE)活性,在激发子处理后均强烈增加。在指数生长期,细胞中含有大量的ACC库,激发子主要通过诱导EFE来刺激乙烯生物合成。在稳定期,细胞中几乎不含ACC,激发子主要通过其对ACC合酶活性的影响来刺激乙烯生物合成。虫草素不影响ACC合酶活性的增加,但阻断了EFE的活性增加,表明前者是转录后调控,后者是转录调控。通过洗涤去除激发子或通过与抗生物素蛋白络合使激发子的生物素化衍生物失活,导致ACC合酶活性的增加迅速停止,这表明持续存在刺激对于该反应是必要的。利用乙烯产生的诱导来测量激发子的量,发现激发子在处理过程中从孵育培养基中消失。

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