Suzuki M, Miyamoto R, Hattori T, Nakamura K, Asahi T
Laboratory of Biochemistry, School of Agriculture, Nagoya University, Japan.
Plant Cell Physiol. 1995 Mar;36(2):273-9. doi: 10.1093/oxfordjournals.pcp.a078759.
The regulatory functions of the 5'-flanking regions of two genes for catalase (cat1 and cat2) from castor bean were analyzed in transgenic tobacco plants that carried fusion constructs that included the gene for beta-glucuronidase (GUS) for Escherichia coli. Dry mature seeds from transgenic plants carrying the CAT1-GUS or CAT2-GUS constructs, in which the GUS gene was fused to the 5'-flanking region of cat1 or cat2, respectively, contained significant GUS activity, indicating that the promoters of cat1 and cat2 were active during seed development. GUS activity increased in response to germination in the seeds of transgenic tobacco that carried CAT1-GUS, as well as in those that carried CAT2-GUS. During the post-germinative stage the GUS activity directed by CAT2-GUS increased still further, whereas that directed by CAT1-GUS decreased. The changes in GUS activity in the transgenic tobacco plants that carried CAT1-GUS and CAT2-GUS were similar to the changes in the levels of transcripts of cat1 and cat2, respectively, in castor bean. The results suggest that the expression of cat1 and cat2 in the germinating seeds and post-germinative seedlings is regulated mainly at the level of transcription. However, the distribution of GUS activity among the organs of the transgenic tobacco seedlings and plantlets, which was examined by histochemical staining and by enzymatic assays of tissue extracts, was not identical to that of transcripts of cat1 and cat2 in castor bean. Histochemical analysis also revealed the interesting spatial regulation of the expression of the promoter of cat2 in the transgenic tobacco seedlings.
对蓖麻中过氧化氢酶的两个基因(cat1和cat2)5'侧翼区域的调控功能进行了分析,分析对象是携带融合构建体的转基因烟草植株,这些构建体包含来自大肠杆菌的β-葡萄糖醛酸酶(GUS)基因。携带CAT1-GUS或CAT2-GUS构建体的转基因植物的干燥成熟种子中含有显著的GUS活性,在这些构建体中,GUS基因分别与cat1或cat2的5'侧翼区域融合,这表明cat1和cat2的启动子在种子发育过程中具有活性。携带CAT1-GUS的转基因烟草种子以及携带CAT2-GUS的转基因烟草种子在萌发过程中GUS活性均增加。在萌发后阶段,由CAT2-GUS指导的GUS活性进一步增加,而由CAT1-GUS指导的GUS活性则下降。携带CAT1-GUS和CAT2-GUS的转基因烟草植株中GUS活性的变化分别与蓖麻中cat1和cat2转录本水平的变化相似。结果表明,cat1和cat2在萌发种子和萌发后幼苗中的表达主要在转录水平上受到调控。然而,通过组织化学染色和组织提取物的酶活性测定对转基因烟草幼苗和植株各器官中GUS活性分布的检测结果与蓖麻中cat1和cat2转录本的分布不同。组织化学分析还揭示了转基因烟草幼苗中cat2启动子表达的有趣空间调控。