Basu Chhandak, Kausch Albert P, Luo Hong, Chandlee Joel M
Department of Plant Sciences, University of Rhode Island, 9 East Alumni Avenue, Suite 7, Kingston, Rhode Island 02881, USA.
J Plant Physiol. 2003 Oct;160(10):1233-9. doi: 10.1078/0176-1617-01104.
Transient expression profiles for several chimeric beta-glucuronidase (GUS) gene constructs were determined in tissues (young leaves, mature leaves and roots) of creeping bentgrass (Agrostis palustris, cv. Penn A4) following microprojectile bombardment. The constructs analyzed consisted of the uidA (GUS) reporter gene driven by four different promoters (ubiquitin 3-potato, ubiquitin corn, ubiquitin rice and CaMV 35S). The total number of GUS hits (or transient expression units; TEUs) were determined manually under a dissecting scope after histochemical staining for GUS. Results suggest that the ubiquitin rice promoter is most active in cells of turfgrass, regardless of the developmental stage or tissue-type. The ubiquitin corn promoter was the next best. Of the four promoter used, except for ubiquitin 3-potato, reporter gene activity was dramatically higher in mature leaves compared to young leaves. The relative efficiency of each promoter was about the same in roots and leaves. We have also analyzed uidA (GUS) reporter gene activity following microprojectile bombardment in transient expression assays with callus from two cultivars (Providence or Penn A4) of creeping bentgrass. Differences in the frequency of GUS positive hits were observed between cultivars up to 72 hours post-bombardment. However, this difference between cultivars disappeared after 72 hours post-bombardment. This information describing promoter functionality in bentgrass will be important when designing gene constructs for trait modification and when choosing appropriate cultivars for improvement through gene transfer experiments. This is the first in depth report on organ-specific and developmental gene expression profiles for transgenes in a turfgrass species.
在对匍匐翦股颖（Agrostis palustris，品种Penn A4）进行微粒轰击后，测定了几种嵌合β-葡萄糖醛酸酶（GUS）基因构建体在其组织（幼叶、成熟叶和根）中的瞬时表达谱。所分析的构建体由受四种不同启动子（泛素3-马铃薯、泛素玉米、泛素水稻和花椰菜花叶病毒35S）驱动的uidA（GUS）报告基因组成。在对GUS进行组织化学染色后，在解剖显微镜下手动确定GUS命中数（或瞬时表达单位；TEUs）的总数。结果表明，无论发育阶段或组织类型如何，泛素水稻启动子在草坪草细胞中最活跃。泛素玉米启动子次之。在所使用的四种启动子中，除泛素3-马铃薯外，成熟叶中的报告基因活性明显高于幼叶。每个启动子在根和叶中的相对效率大致相同。我们还在对匍匐翦股颖两个品种（Providence或Penn A4）的愈伤组织进行的瞬时表达分析中，测定了微粒轰击后uidA（GUS）报告基因的活性。在轰击后长达72小时的时间内，观察到不同品种之间GUS阳性命中频率存在差异。然而，在轰击后72小时后，不同品种之间的这种差异消失了。在设计用于性状改良的基因构建体以及通过基因转移实验选择合适的品种进行改良时，这些描述翦股颖启动子功能的信息将非常重要。这是关于草坪草物种中转基因的器官特异性和发育基因表达谱的第一份深入报告。
