Wu L L, Song I, Kim D, Kaufman P B
Department of Biology, University of Michigan, Ann Arbor 48109-1048, USA.
J Plant Physiol. 1993 Aug;142(2):179-83. doi: 10.1016/s0176-1617(11)80960-0.
An asymmetric (top vs. bottom) increase in invertase activity is elicited by gravistimulation in oat-shoot pulvini starting within 3 h after treatment. In order to analyze the regulation of invertase gene expression in this system, we examined the effect of gravistimulation on invertase mRNA induction. Total RNA and poly (A)+RNA, isolated from oat pulvini, and two oligonucleotide primers, corresponding to two conserved amino-acid sequences (NDPNG and WECPD) found in invertase from other species, were used for the polymerase chain reaction (PCR). A partial-length cDNA (550 base pairs) was obtained and characterized. There was a 52% deduced amino-acid sequence homology to that of carrot beta-fructosidase and a 48% homology to that of tomato invertase. Northern blot analysis showed that there was an obvious transient accumulation of invertase mRNA elicited by gravistimulation of oat pulvini. The mRNA was rapidly induced to a maximum level at 1 h following gravistimulation treatment and gradually decreased afterwards. The mRNA level in the bottom half of the oat pulvinus was significantly higher (five-fold) than that in the top half of the pulvinus tissue. The induction of invertase mRNA was consistent with the transient enhancement of invertase activity during the graviresponse of the pulvinus. These data indicate that the expression of the invertase gene(s) could be regulated by gravistimulation at the transcriptional and/or translational levels. Southern blot analysis showed that there were four genomic DNA fragments hybridized to the invertase cDNA. This suggests that an invertase gene family may exist in oat plants.
重力刺激可在处理后3小时内引发燕麦茎节叶枕中转化酶活性的不对称(顶部与底部)增加。为了分析该系统中转化酶基因表达的调控,我们研究了重力刺激对转化酶mRNA诱导的影响。从燕麦叶枕中分离出的总RNA和聚腺苷酸RNA,以及与其他物种转化酶中发现的两个保守氨基酸序列(NDPNG和WECPD)相对应的两个寡核苷酸引物,用于聚合酶链反应(PCR)。获得并鉴定了一个部分长度的cDNA(550个碱基对)。其推导的氨基酸序列与胡萝卜β-果糖苷酶的序列有52%的同源性,与番茄转化酶有48%的同源性。Northern印迹分析表明,燕麦叶枕的重力刺激可引发转化酶mRNA明显的瞬时积累。重力刺激处理后1小时,mRNA迅速诱导至最高水平,随后逐渐下降。燕麦叶枕下半部分的mRNA水平明显高于(五倍)叶枕组织上半部分。转化酶mRNA的诱导与叶枕重力反应过程中转化酶活性的瞬时增强一致。这些数据表明,转化酶基因的表达可能在转录和/或翻译水平上受到重力刺激的调控。Southern印迹分析表明,有四个基因组DNA片段与转化酶cDNA杂交。这表明燕麦植物中可能存在一个转化酶基因家族。