de Veau E J, Burris J E
Department of Biology, The Pennsylvania State University, University Park, Pennsylvania 16802.
Plant Physiol. 1989 Jun;90(2):500-11. doi: 10.1104/pp.90.2.500.
A method was devised to quantify short-term photorespiratory rates in terrestrial plants using (18)O-intermediates of the glycolate pathway, specifically glycolate, glycine, and serine. The pathway intermediates were isolated and analyzed on a GC/MS to determine molecular percent (18)O-enrichment. Rates of glycolate synthesis were determined from (18)O-labeling kinetics of the intermediates, derived rate equations, and nonlinear regression techniques. Glycolate synthesis in wheat (Triticum aestivum L.), a C(3) plant, and maize (Zea mays L.), a C(4) plant, was stimulated by high O(2) concentrations and inhibited by high CO(2) concentrations. The synthesis rates were 7.3, 2.1, and 0.7 micromoles per square decimeter per minute under a 21% O(2) and 0.035% CO(2) atmosphere for leaf tissue of wheat, maize seedlings, and 3-month-old maize, respectively. Photorespiratory CO(2) evolution rates were estimated to be 27, 6, and 2%, respectively, of net photosynthesis for the three groups of plants under the above atmosphere. The results from maize tissue support the hypothesis that C(4) plants photorespire, albeit at a reduced rate in comparison to C(3) plants, and that the CO(2)/O(2) ratio in the bundle sheath of maize is higher in mature tissue than in seedling tissue. The pool size of the three photorespiratory intermediates remained constant and were unaffected by changes in either CO(2) or O(2) concentrations throughout the 10-minute labeling period. This suggests that photorespiratory metabolism is regulated by other mechanism besides phosphoglycolate synthesis by ribulose-1,5-bisphosphate carboxylase/oxygenase, at least under short-term conditions. Other mechanisms could be alternate modes of synthesis of the intermediates, regulation of some of the enzymes of the photorespiratory pathway, or regulation of carbon flow between organelles involved in photorespiration. The glycolate pool became nearly 100% (18)O-labeled under an atmosphere of 40% O(2). This pool failed to become 100% (18)O-enriched under lower O(2) concentrations.
设计了一种方法,利用乙醇酸途径的(18)O中间体,特别是乙醇酸、甘氨酸和丝氨酸,来量化陆生植物的短期光呼吸速率。分离出途径中间体并通过气相色谱/质谱进行分析,以确定分子(18)O富集百分比。根据中间体的(18)O标记动力学、推导的速率方程和非线性回归技术来确定乙醇酸合成速率。在C3植物小麦(Triticum aestivum L.)和C4植物玉米(Zea mays L.)中,乙醇酸合成受到高O2浓度的刺激,并受到高CO2浓度的抑制。在21% O2和0.035% CO2气氛下,小麦叶片组织、玉米幼苗和3个月大玉米的乙醇酸合成速率分别为每分钟每平方分米7.3、2.1和0.7微摩尔。在上述气氛下,三组植物的光呼吸CO2释放速率估计分别为净光合作用的27%、6%和2%。玉米组织的结果支持了以下假设:C4植物进行光呼吸,尽管与C3植物相比速率较低,并且玉米成熟组织中维管束鞘中的CO2/O2比率高于幼苗组织。在整个10分钟的标记期内,三种光呼吸中间体的库大小保持恒定,不受CO2或O2浓度变化的影响。这表明,至少在短期条件下,光呼吸代谢是由除1,5-二磷酸核酮糖羧化酶/加氧酶合成磷酸乙醇酸之外的其他机制调节的。其他机制可能是中间体的替代合成模式、光呼吸途径中某些酶的调节,或参与光呼吸的细胞器之间碳流的调节。在40% O2气氛下,乙醇酸库几乎100%被(18)O标记。在较低的O2浓度下,该库未能达到100%的(18)O富集。