Shirasawa Kenta, Takabe Tomoko, Takabe Tetsuko, Kishitani Sachie
Graduate School of Agricultural Science, Tohoku University, Aoba, Sendai 981-8555, Japan.
Ann Bot. 2006 Sep;98(3):565-71. doi: 10.1093/aob/mcl126. Epub 2006 Jun 21.
Glycinebetaine (GB), a quaternary ammonium compound, is a very effective compatible solute. In higher plants, GB is synthesized from choline (Cho) via betaine aldehyde (BA). The first and second steps in the biosynthesis of GB are catalysed by choline monooxygenase (CMO) and by betaine aldehyde dehydrogenase (BADH), respectively. Rice (Oryza sativa), which has two genes for BADH, does not accumulate GB because it lacks a functional gene for CMO. Rice plants accumulate GB in the presence of exogenously applied BA, which leads to the development of a significant tolerance to salt, cold and heat stress. The goal in this study was to evaluate and to discuss the effects of endogenously accumulated GB in rice.
Transgenic rice plants that overexpressed a gene for CMO from spinach (Spinacia oleracea) were produced by Agrobacterium-mediated transformation. After Southern and western blotting analysis, GB in rice leaves was quantified by (1)H-NMR spectroscopy and the tolerance of GB-accumulating plants to abiotic stress was investigated.
Transgenic plants that had a single copy of the transgene and expressed spinach CMO accumulated GB at the level of 0.29-0.43 micromol g(-1) d. wt and had enhanced tolerance to salt stress and temperature stress in the seedling stage.
In the CMO-expressing rice plants, the localization of spinach CMO and of endogenous BADHs might be different and/or the catalytic activity of spinach CMO in rice plants might be lower than it is in spinach. These possibilities might explain the low levels of GB in the transgenic rice plants. It was concluded that CMO-expressing rice plants were not effective for accumulation of GB and improvement of productivity.
甘氨酸甜菜碱(GB)是一种季铵化合物,是一种非常有效的相容性溶质。在高等植物中,GB由胆碱(Cho)经甜菜碱醛(BA)合成。GB生物合成的第一步和第二步分别由胆碱单加氧酶(CMO)和甜菜碱醛脱氢酶(BADH)催化。水稻(Oryza sativa)有两个BADH基因,但由于缺乏功能性CMO基因而不积累GB。水稻植株在外源施加BA时会积累GB,这导致对盐、冷和热胁迫产生显著耐受性。本研究的目的是评估和讨论水稻中内源性积累的GB的影响。
通过农杆菌介导的转化产生过表达菠菜(Spinacia oleracea)CMO基因的转基因水稻植株。经过Southern和western印迹分析后,通过(1)H-NMR光谱对水稻叶片中的GB进行定量,并研究积累GB的植株对非生物胁迫的耐受性。
具有单个转基因拷贝并表达菠菜CMO的转基因植株积累GB的水平为0.29 - 0.43微摩尔/克干重,并且在幼苗期对盐胁迫和温度胁迫的耐受性增强。
在表达CMO的水稻植株中,菠菜CMO和内源性BADHs的定位可能不同和/或菠菜CMO在水稻植株中的催化活性可能低于在菠菜中的活性。这些可能性可能解释了转基因水稻植株中GB水平较低的原因。得出的结论是,表达CMO的水稻植株对于GB的积累和生产力的提高无效。