Harada Taro, Satoh Shigeru, Yoshioka Toshihito, Ishizawa Kimiharu
Department of Developmental Biology and Neuroscience, Graduate School of Life Sciences, Tohoku University, Sendai 980-8578, Japan.
Ann Bot. 2005 Sep;96(4):683-92. doi: 10.1093/aob/mci220. Epub 2005 Jul 20.
Overwintering buds (turions) of the monocot aquatic pondweed species (Potamogeton distinctus) are highly tolerant to anoxic stress. Sucrose metabolism accompanied by enhanced activity of sucrose synthase (SuSy) operates actively during anaerobic elongation of pondweed turions. The aim of this study is to isolate SuSy genes from the turions and to investigate their transcriptional changes in response to anoxia and other stimuli.
SuSy genes were isolated from pondweed turions by PCR methods and transcript levels of SuSy genes were examined in response to anoxia, sugars and plant hormones. In addition, the effects of anoxia on SuSy activity were examined both in the soluble fraction and in the microsomal fraction.
cDNAs of two SuSy genes (PdSUS1 and PdSUS2) were cloned from pondweed turions. The levels of PdSUS1 transcripts increased under anoxia but did not with sugar treatments. Anoxia-stimulated elongation of turions was further enhanced by 2,4-dichlorophenoxyacetic acid (2,4-D) and suppressed by treatments with sorbitol, 2-deoxyglucose (2-dGlc) and abscisic acid (ABA). The levels of PdSUS1 transcripts were increased by 2,4-D and decreased by sorbitol under anoxia. The levels of PdSUS2 transcripts were not significantly affected by anoxia and any other treatments. SuSy activity of turions under anoxia was enhanced in the soluble fraction, but not in the microsomal fraction.
Up-regulation of PdSUS1 transcription under anoxia may not be attributed to sugar starvation under anoxia. A positive correlation between stem elongation and the level of PdSUS1 transcripts was observed in turions treated with anoxic conditions, 2,4-D and sorbitol. The increase in SuSy activity in the cytosol may contribute to sugar metabolism and sustain stem elongation under anoxia.
单子叶水生眼子菜属物种(菹草)的越冬芽(块茎)对缺氧胁迫具有高度耐受性。在菹草块茎的厌氧伸长过程中,伴随着蔗糖合酶(SuSy)活性增强的蔗糖代谢活跃进行。本研究的目的是从块茎中分离出SuSy基因,并研究其在缺氧及其他刺激下的转录变化。
通过PCR方法从菹草块茎中分离出SuSy基因,并检测SuSy基因在缺氧、糖类和植物激素处理下的转录水平。此外,还检测了缺氧对可溶性组分和微粒体组分中SuSy活性的影响。
从菹草块茎中克隆出两个SuSy基因(PdSUS1和PdSUS2)的cDNA。PdSUS1转录本水平在缺氧条件下升高,但在糖类处理时未升高。2,4-二氯苯氧乙酸(2,4-D)进一步增强了缺氧刺激的块茎伸长,而山梨醇、2-脱氧葡萄糖(2-dGlc)和脱落酸(ABA)处理则抑制了这种伸长。在缺氧条件下,2,4-D使PdSUS1转录本水平升高,山梨醇使其降低。PdSUS2转录本水平不受缺氧及其他任何处理的显著影响。缺氧条件下块茎的SuSy活性在可溶性组分中增强,但在微粒体组分中未增强。
缺氧条件下PdSUS1转录的上调可能并非归因于缺氧导致的糖饥饿。在用缺氧条件、2,4-D和山梨醇处理的块茎中,观察到茎伸长与PdSUS1转录本水平之间呈正相关。细胞质中SuSy活性的增加可能有助于糖代谢,并在缺氧条件下维持茎的伸长。