Brisson L F, Zelitch I, Havir E A
Department of Biochemistry and Genetics, Connecticut Agricultural Experiment Station, New Haven 06504, USA.
Plant Physiol. 1998 Jan;116(1):259-69. doi: 10.1104/pp.116.1.259.
Constructs containing the cDNAs encoding the primary leaf catalase in Nicotiana or subunit 1 of cottonseed (Gossypium hirsutum) catalase were introduced in the sense and antisense orientation into the Nicotiana tabacum genome. The N. tabacum leaf cDNA specifically overexpressed CAT-1, the high catalatic [corrected] form, activity. Antisense constructs reduced leaf catalase specific activities from 0.20 to 0.75 times those of wild type (WT), and overexpression constructs increased catalase specific activities from 1.25 to more than 2.0 times those of WT. The NADH-hydroxypyruvate reductase specific activity in transgenic plants was similar to that in WT. The effect of antisense constructs on photorespiration was studied in transgenic plants by measuring the CO2 compensation point (gamma) at a leaf temperature of 38 degrees C. A significant linear increase was observed in gamma with decreasing catalase (at 50% lower catalase activity gamma increased 39%). There was a significant temperature-dependent linear decrease in gamma in transgenic leaves with elevated catalase compared with WT leaves (at 50% higher catalase gamma decreased 17%). At 29 degrees C, gamma also decreased with increasing catalase in transgenic leaves compared with WT leaves, but the trend was not statistically significant. Rates of dark respiration were the same in WT and transgenic leaves. Thus, photorespiratory losses of CO2 were significantly reduced with increasing catalase activities at 38 degrees C, indicating that the stoichiometry of photorespiratory CO2 formation per glycolate oxidized normally increases at higher temperatures because of enhanced peroxidation.
将含有编码烟草初生叶过氧化氢酶或棉籽(陆地棉)过氧化氢酶亚基1的cDNA的构建体以正义和反义方向导入烟草基因组。烟草叶片cDNA特异性过表达CAT-1,即高催化活性形式。反义构建体使叶片过氧化氢酶比活性降低至野生型(WT)的0.20至0.75倍,而过表达构建体使过氧化氢酶比活性增加至WT的1.25至2.0倍以上。转基因植物中NADH-羟基丙酮酸还原酶的比活性与WT相似。通过在38℃叶片温度下测量CO2补偿点(γ),研究了反义构建体对转基因植物光呼吸的影响。随着过氧化氢酶活性降低,γ值显著线性增加(过氧化氢酶活性降低50%时,γ值增加39%)。与WT叶片相比,过氧化氢酶活性升高的转基因叶片中γ值随温度显著线性降低(过氧化氢酶活性升高50%时,γ值降低17%)。在29℃时,与WT叶片相比,转基因叶片中γ值也随过氧化氢酶活性增加而降低,但该趋势无统计学意义。WT和转基因叶片的暗呼吸速率相同。因此,在38℃时,随着过氧化氢酶活性增加,光呼吸CO2损失显著降低,这表明由于过氧化作用增强,在较高温度下,每氧化一分子乙醇酸产生的光呼吸CO2的化学计量通常会增加。