Nodulation studies in the model legume Medicago truncatula:: Advantages of using the constitutive EF1α promoter and limitations in detecting fluorescent reporter proteins in nodule tissues

被引:40
作者
Auriac, Marie-Christine [1 ]
Timmers, Antonius C. J. [1 ]
机构
[1] INRA, CNRS, Lab Plant Microorganism Interact, UMR 2594, F-31326 Castanet Tolosan, France
关键词
Sinorhizobium meliloti; RHIZOBIUM-MELILOTI; GENE-EXPRESSION; 35S PROMOTER; ALFALFA; TRANSGENES; DIFFERENTIATION; ELONGATION; INITIATION; VISUALIZE; VARIANTS;
D O I
10.1094/MPMI-20-9-1040
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The Cauliflower mosaic virus 35S promoter currently is being used in RNAi-based approaches for attenuating host gene expression during legume root nodule development and also for the expression of fluorescent reporters in nodule tissues. In this study, we have evaluated the expression of this promoter in the indeterminate nodules of the model plant Medicago truncatula. Our results clearly show that the 35S promoter is inactive in both the nodule meristem and in bacteroid-containing cells of the nodules. On the other hand, the Arabidopsis thaliana EF1 alpha promoter was found to be strongly expressed both in the nodule meristem and in all nodule-invaded cells. Therefore, we conclude that the constitutive EF1 alpha promoter is far superior for mRNAi or overexpression studies in nodule tissues compared with the commonly used 35S promoter. In addition, our experiments have revealed that the intensity of fluorescent markers such as green fluorescent protein is severely attenuated within invaded cells in the nitrogen-fixation zone of the nodule, most likely by fluorescence quenching. This phenomenon may hinder the use of these tools for live-cell imaging in nodule tissue.
引用
收藏
页码:1040 / 1047
页数:8
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