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比较了不同低温(14℃和4℃)胁迫对烟草(Nicotiana rustica L.)愈伤组织抗氰交替途径诱导和交替氧化酶表达的影响。 结果显示,不同低温胁迫处理能显著诱导烟草愈伤组织交替途径容量和实际运行的增加,且都呈现出基本相同的变化模式:在胁迫的初期(1～3 d)持续增加,在3 d时达到最高,而后下降到一个相对恒定的水平。但交替途径容量增加的幅度与温度下降的程度密切相关,而交替途径实际运行量的诱导程度在不同低温胁迫下的差异却很小。 表明交替途径容量和实际运行对低温胁迫的响应是不同的。免疫印迹分析结果表明:低温胁迫明显诱导了交替氧化酶总蛋白的增加,且其随低温胁迫进程的变化与交替途径容量的变化基本一致;而对交替氧化酶单体与二聚体在低温胁迫下的含量变化检测结果则显示,烟草愈伤组织中交替氧化酶主要以二聚体形式存在,且这一存在形式并不随低温胁迫程度的加深而发生改变。两种形式的交替氧化酶蛋白含量都能被低温胁迫诱导增加,但其单体水平在两种不同的低温胁迫下并无明显差别,而4℃低温胁迫诱导的二聚体交替氧化酶蛋白含量明显高于14℃。表明不同程度低温对抗氰交替途径发生的不同影响主要是由于对交替氧化酶蛋白二聚体形式的不同诱导程度所致;而高活性的交替氧化酶单体形式则不因低温胁迫程度的
The effects of different low temperature (14 ℃ and 4 ℃) stress on the induction of resistance to cyanidation and the alternation of oxidase in Nicotiana rustica L. callus were compared. The results showed that different treatments of low temperature stress could significantly induce the increase of alternate pathway capacity and actual operation of tobacco callus, and all showed the same pattern of change: increasing continuously in the early stage of stress (1-3 days) Reached the highest, then dropped to a relatively constant level. However, the magnitude of increase in capacity of alternative routes is closely related to the degree of temperature decrease, while the degree of induction of actual operation quantities of alternative routes under different low temperature stresses is very small. It shows that the capacity of alternating path and the actual operation are different from the response to low temperature stress. The result of immunoblot analysis showed that cold stress significantly induced the increase of total protein of alternating oxidase, and its change with the course of low temperature stress was basically the same as that of alternative pathway. However, the changes of alternan oxidase monomer and dimer under low temperature stress The results showed that the alternation of oxidase mainly existed in the dimer form in the callus of tobacco and this existed form did not change with the deepening of the degree of cold stress. Both forms of alternative oxidase protein content can be induced by low temperature stress, but the monomer level in two different low temperature stress did not differ significantly, and 4 ℃ low temperature induced dimer alternating oxidase protein content Significantly higher than 14 ° C. The results showed that the different effects of different degrees of hypothermia on the alternate ways of cyanogen exchange were mainly attributed to the different degrees of induction of alternative oxidase protein dimer forms, while the high activity of alternative oxidase monomeric forms did not result from low temperature stress