Carpenter J L, Ploense S E, Snustad D P, Silflow C D
Department of Genetics and Cell Biology, University of Minnesota, St. Paul 55108-1095.
Plant Cell. 1992 May;4(5):557-71. doi: 10.1105/tpc.4.5.557.
The pool of tubulin protein in tissues of Arabidopsis is provided by the expression of multiple alpha-tubulin (TUA) and beta-tubulin genes. Whereas most tubulin genes are expressed in many tissues, previous evidence suggested that the TUA1 gene might be expressed primarily in pollen. We now report a detailed analysis of TUA1 expression during Arabidopsis development. In RNA from tissues of dissected flowers, TUA1 transcripts were detected only in stamens and mature pollen. Chimeric genes containing TUA1 5' flanking DNA fused to the beta-glucuronidase (GUS) coding region were used to create transgenic Arabidopsis plants. Plants containing a chimeric gene with 533 bp of 5' flanking sequence were analyzed by histochemical assay to localize GUS expression within the plant. The blue product of GUS enzyme activity accumulated very rapidly in postmitotic pollen grains. Much lower levels of GUS activity were detected in anthers with uninucleate pollen grains, in flower receptacles, and in a few vegetative tissues. Analysis of 5' deletions of the TUA1 promoter suggested that 97 bp of 5' flanking DNA is sufficient to drive GUS expression in pollen and young anthers, whereas at least 380 bp is required to detect GUS expression in the receptacle. Examination of the TUA1 promoter sequence revealed several motifs that are repeated within the TUA1 promoter and are similar to sequences in other pollen-specific promoters.
拟南芥组织中的微管蛋白池由多个α-微管蛋白(TUA)基因和β-微管蛋白基因的表达提供。虽然大多数微管蛋白基因在许多组织中都有表达,但先前的证据表明TUA1基因可能主要在花粉中表达。我们现在报告了拟南芥发育过程中TUA1表达的详细分析。在解剖花组织的RNA中,仅在雄蕊和成熟花粉中检测到TUA1转录本。包含与β-葡萄糖醛酸酶(GUS)编码区融合的TUA1 5'侧翼DNA的嵌合基因被用于创建转基因拟南芥植株。通过组织化学分析对含有533 bp 5'侧翼序列的嵌合基因的植株进行分析,以定位植物体内的GUS表达。GUS酶活性的蓝色产物在有丝分裂后的花粉粒中迅速积累。在具有单核花粉粒的花药、花托和一些营养组织中检测到的GUS活性水平要低得多。对TUA1启动子5'缺失的分析表明,97 bp的5'侧翼DNA足以驱动花粉和幼嫩花药中的GUS表达,而在花托中检测到GUS表达至少需要380 bp。对TUA1启动子序列的检查揭示了几个在TUA1启动子内重复且与其他花粉特异性启动子中的序列相似的基序。