Gabrys H., Kramer D. M., Crofts A. R., Ort D. R.
Department of Plant Biology (H.G., D.R.O.), Division of Biophysics (D.M.K., A.R.C.), and Photosynthesis Research Unit (D.R.O.), United States Department of Agriculture/Agricultural Research Service, University of Illinois, Urbana, Illinois 61801-3838.
Plant Physiol. 1994 Feb;104(2):769-776. doi: 10.1104/pp.104.2.769.
We have devised a two-step screening strategy for the selection of chloroplast coupling factor reduction mutants from an M2 population of Arabidopsis thaliana. The selection strategy relies on a lowered energetic threshold for catalytic activation of the enzyme that has been shown to accompany thioredoxin-mediated reduction of a cysteine bridge on the [gamma] subunit of coupling factor. We selected first for plants that grew poorly under low irradiance but performed satisfactorily at high irradiance when the transmembrane electrochemical potential of hydrogen ions is large and competent to maintain a high level of coupling factor activation without [gamma] subunit reduction. In the second step of the screen we monitored the flash-induced electrochromic change to select putative coupling factor reduction mutants from other sorts of mutations that shared the phenotype of poor growth and vigor when transferred from high to low irradiance. Among the mutants selected, one appears incapable of reducing coupling factor, whereas another behaves as though coupling factor is at least partially reduced even in dark-adapted plants.
我们设计了一种两步筛选策略,用于从拟南芥M2群体中筛选叶绿体偶联因子还原突变体。该筛选策略依赖于酶催化激活的能量阈值降低,这已被证明与硫氧还蛋白介导的偶联因子γ亚基上半胱氨酸桥的还原有关。我们首先选择在低光照下生长不良但在高光照下表现良好的植株,此时氢离子的跨膜电化学势较大,能够在不发生γ亚基还原的情况下维持高水平的偶联因子激活。在筛选的第二步中,我们监测闪光诱导的电致变色变化,以从其他类型的突变体中筛选出假定的偶联因子还原突变体,这些突变体在从高光照转移到低光照时表现出生长不良和活力差的表型。在所选的突变体中,一个似乎无法还原偶联因子,而另一个即使在暗适应的植株中表现得好像偶联因子至少部分被还原了。