Acta Botanica Sinica Volume 44 Issue 3, Pages 314-317.
Expression of Recombinant Tryptophan Decarboxylase in Different Subcellular Compartments in Tobacco Plant

Wamg Miao Li Qiu Rong Stefano Di FIORE Rainer FISCHER

Abstract:

The gene encoded for tryptophan decarboxylase (TDC), which is the key enzyme in terpenoil indole alkaloids pathway, was targeted to different subcellular compartments and stably expressed in transgenic tobacco (Nicotiana tabacum L.) plants at the levels detected by Western blot and tryptamine accumulation analysis. It was shown that the TDC was located in subcellular compartments, the chloroplasts and cytosol. The recombinant TDC targeted to chloroplasts and cytosol in tobacco plants was effectively expressed as soluble protein by Western blot analysis and enzymatic assay. The level of tryptamine accumulation in chloroplast was higher than that in cytosol and very low in vacuole and endoplasmic reticulum (ER) to be hardly detected by Western blot analysis. It was indicated that the highest amount of tryptamine was in chloroplasts, lower in endoplasmic reticula and the lowest in vacuoles as compared to those in wild type plants. The TDC targeted to different subcellular compartments of tobacco plants and its expression level were studied by different nucleotide sequences coding signal peptides at 5′end of tdc gene in order to know the effects of the TDC in compartmentation on its functionality.

重组色氨酸脱羧酶在烟草不同亚细胞区室的表达

王淼1*  李秋荣2  Stefano Di DIORE3  Rainer FISCHER3

(1.中国科学院沈阳应用生态研究所,沈阳,1100162.沈阳军区总医院,沈阳,110016; 3.Botanical Institute of Rjeinisch -Westfaliusch Techniische Hochschule Aachen,D-52074 Aachen ,Germany)

 

摘要:将萜烯类吲哚生物碱代谢关键酶--色氨酸脱羧酶(TDC)的编码基因转到烟草(Nicotiana tabacum L.)植物体内,标定在不同的亚细胞区室表达。通过蛋白免疫印迹法和色胺在植物体内的累积量测定分析,对转基因植物进行筛选。结果表明,TDC在叶绿体和胞液中高效表达,TDC在叶绿体中的表达水平最高,高于在胞液中的表达,在内质网和液泡中表达水平很低,用蛋白免疫印迹法未检出。

关键词 色氨酸脱羧酶 生物碱代谢途径 烟草 亚细胞区室

*通讯作者 Email: wangmiao@iae.ac.cn

 


Keywords:
tryptophan decarboxylase; alkaloids biosynthetic pathway;Nicotiana tabacum; subcellular compartment

PROMOTIONS

    Photo Gallery
Scan with iPhone or iPad to view JIPB online
Scan using WeChat with your smartphone to view JIPB online
Editorial Office, Journal of Integrative Plant Biology, Institute of Botany, CAS
No. 20 Nanxincun, Xiangshan, Beijing 100093, China
Tel: +86 10 6283 6133 Fax: +86 10 8259 2636 E-mail: jipb@ibcas.ac.cn

Copyright © 2017 by the Institute of Botany, the Chinese Academy of Sciences
Online ISSN: 1744-7909 Print ISSN: 1672-9072 CN: 11-5067/Q