首页> 外文期刊>Plant Science: An International Journal of Experimental Plant Biology >Characterization of cDNAs differentially expressed in roots of tobacco (Nicotiana tabacum cv Burley 21) during the early stages of alkaloid biosynthesis
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Characterization of cDNAs differentially expressed in roots of tobacco (Nicotiana tabacum cv Burley 21) during the early stages of alkaloid biosynthesis

机译:生物碱生物合成早期在烟草(Nicotiana tabacum cv Burley 21)根中差异表达的cDNA的表征

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摘要

A set of 60 cDNAs were isolated by subtractive hybridization screening of a phage library using radioactively-labeled probes generated from root mRNAs isolated from tobacco (Nicotiana tabacum cv Burley 21) plants before and 3 days after topping. Among the differentially expressed gene products were full-length and partial cDNAs encoding arginine decarboxylase (ADC), ornithine decarboxylase (ODC), and S-adenosylmethionine synthetase (SAMS), enzymes involved in polyamine and alkaloid biosynthesis. The other cDNAs isolated were placed into one of several categories and encode metabolic enzymes, proteins involved in transcription and translation, components of signal transduction pathways, and homologs of genes whose expression has been shown to be regulated by phytohormones (i.e. auxin, ABA), wounding or other stress responses. RNA gel blot analysis showed that the ADC and ODC transcripts were preferentially expressed in the roots and floral tissues of mature tobacco plants, whereas SAMS transcripts were detected in all tissues examined. The steady-state levels of the ADC and ODC mRNAs increased in the roots of wild-type tobacco plants during the 24 h period after topping, whereas little change was observed in the abundance of the SAMS transcripts in these tissues. The possible factors associated with the regulation of expression of these genes are discussed. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. [References: 61]
机译:通过使用打顶前后3天从烟草(Nicotiana tabacum cv Burley 21)植物中分离的根mRNA产生的放射性标记的探针,通过放射性标记的探针对噬菌体文库进行消减杂交筛选,分离出一组60个cDNA。在差异表达的基因产物中,有编码精氨酸脱羧酶(ADC),鸟氨酸脱羧酶(ODC)和S-腺苷甲硫氨酸合成酶(SAMS)的全长和部分cDNA,这些酶参与多胺和生物碱的生物合成。分离出的其他cDNA属于以下几类之一,它们编码代谢酶,参与转录和翻译的蛋白质,信号转导途径的成分以及表达受植物激素(例如生长素,ABA)调控的基因的同源物,受伤或其他压力反应。 RNA凝胶印迹分析表明,ADC和ODC转录本优先在成熟烟草植物的根和花组织中表达,而SAMS转录本在所有检查过的组织中均被检测到。打顶后的24小时内,野生型烟草植物根中ADC和ODC mRNA的稳态水平增加,而这些组织中SAMS转录物的丰度几乎没有变化。讨论了与这些基因的表达调控相关的可能因素。 (C)2000 Elsevier Science Ireland Ltd.保留所有权利。 [参考:61]

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