Hosy Eric, Vavasseur Alain, Mouline Karine, Dreyer Ingo, Gaymard Frédéric, Porée Fabien, Boucherez Jossia, Lebaudy Anne, Bouchez David, Very Anne-Aliénor, Simonneau Thierry, Thibaud Jean-Baptiste, Sentenac Hervé
Biochimie et Physiologie Moléculaires des Plantes, Unité Mixte de Recherche 5004, Agro-M/Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/UM2, F-34060 Montpellier Cedex 1, France.
Proc Natl Acad Sci U S A. 2003 Apr 29;100(9):5549-54. doi: 10.1073/pnas.0733970100. Epub 2003 Apr 1.
Microscopic pores present in the epidermis of plant aerial organs, called stomata, allow gas exchanges between the inner photosynthetic tissue and the atmosphere. Regulation of stomatal aperture, preventing excess transpirational vapor loss, relies on turgor changes of two highly differentiated epidermal cells surrounding the pore, the guard cells. Increased guard cell turgor due to increased solute accumulation results in stomatal opening, whereas decreased guard cell turgor due to decreased solute accumulation results in stomatal closing. Here we provide direct evidence, based on reverse genetics approaches, that the Arabidopsis GORK Shaker gene encodes the major voltage-gated outwardly rectifying K(+) channel of the guard cell membrane. Expression of GORK dominant negative mutant polypeptides in transgenic Arabidopsis was found to strongly reduce outwardly rectifying K(+) channel activity in the guard cell membrane, and disruption of the GORK gene (T-DNA insertion knockout mutant) fully suppressed this activity. Bioassays on epidermal peels revealed that disruption of GORK activity resulted in impaired stomatal closure in response to darkness or the stress hormone abscisic acid [corrected]. Transpiration measurements on excised rosettes and intact plants (grown in hydroponic conditions or submitted to water stress) revealed that absence of GORK activity resulted in increased water consumption. The whole set of data indicates that GORK is likely to play a crucial role in adaptation to drought in fluctuating environments.
植物地上器官表皮中存在的微小孔隙称为气孔,它允许内部光合组织与大气之间进行气体交换。气孔孔径的调节可防止过多的蒸腾水分流失,这依赖于围绕气孔的两个高度分化的表皮细胞(保卫细胞)的膨压变化。溶质积累增加导致保卫细胞膨压升高,从而引起气孔开放;而溶质积累减少导致保卫细胞膨压降低,进而引起气孔关闭。在此,我们基于反向遗传学方法提供直接证据,证明拟南芥GORK震荡基因编码保卫细胞膜上主要的电压门控外向整流钾离子通道。在转基因拟南芥中表达GORK显性负突变多肽,发现其可强烈降低保卫细胞膜上外向整流钾离子通道的活性,而GORK基因的破坏(T-DNA插入敲除突变体)则完全抑制了该活性。对表皮条的生物测定表明,GORK活性的破坏导致气孔在黑暗或应激激素脱落酸作用下关闭受损[已修正]。对离体莲座叶和完整植株(水培条件下生长或遭受水分胁迫)的蒸腾测量表明,缺乏GORK活性会导致水分消耗增加。所有数据表明,GORK可能在波动环境中的干旱适应中起关键作用。