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Elicitor-Induced Ethylene Biosynthesis in Tomato Cells: Characterization and Use as a Bioassay for Elicitor Action

机译:番茄细胞中诱导子诱导的乙烯生物合成:表征和用作诱导子作用的生物测定

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摘要

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.
机译:在悬浮培养的番茄(Lycopersicon esculentum Mill。)细胞中研究了从酵母提取物中部分纯化的引发剂对乙烯生物合成的诱导作用。未经刺激的细胞在指数生长过程中几乎不产生乙烯,而在固定相中则更少。用激发子处理后,在成指数增长的细胞中刺激了10到20倍的乙烯生物合成,在静止细胞中刺激了100倍以上的乙烯生物合成。响应诱导剂处理,在体外测量的1-氨基环丙烷-1-羧酸(ACC)合酶和在体内测量的乙烯形成酶(EFE)的活性均大大增强。在指数生长期间,细胞包含大量的ACC,并且引发剂主要通过诱导EFE刺激乙烯的生物合成。在固定期,细胞几乎不包含ACC,并且引发剂主要通过其对ACC合酶活性的影响来刺激乙烯的生物合成。虫草素不影响ACC合酶活性的增加,但阻断了EFE的活性,表明前者是转录后调控的,后者是转录调控的。通过洗涤的去除引发剂或通过与抗生物素蛋白的复合作用使引发剂的生物素化衍生物失活而使引发剂的ACC合酶活性的增加迅速停止,这表明刺激的连续存在是必需的。使用乙烯产生的诱导来测量引发剂的量,发现该引发剂在处理过程中从孵育培养基中消失。

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