Badger Murray R, Fallahi Hossein, Kaines Sarah, Takahashi Shunichi
Molecular Plant Physiology Group and ARC Centre of Excellence in Plant Energy Biology, Research School of Biology, Australian National University, Canberra, ACT 2601, Australia.
Funct Plant Biol. 2009 Nov;36(11):867-873. doi: 10.1071/FP09199.
Exposure of Arabidopsis thaliana (L.) photorespiration mutants to air leads to a rapid decline in the F/F chlorophyll fluorescence parameter, reflecting a decline in PSII function and an onset of photoinhibition. This paper demonstrates that chlorophyll fluorescence imaging of F/F can be used as an easy and efficient means of detecting Arabidopsis mutants that are impaired in various aspects of photorespiration. This screen was developed to be sensitive and high throughput by the use of exposure to zero CO conditions and the use of array grids of 1-week-old Arabidopsis seedlings as the starting material for imaging. Using this procedure, we screened ~25 000 chemically mutagenised M2 Arabidopsis seeds and recovered photorespiration phenotypes (reduction in F/F at low CO) at a frequency of ~4 per 1000 seeds. In addition, we also recovered mutants that showed reduced F/F at high CO. Of this group, we detected a novel 'reverse photorespiration' phenotype that showed a high CO dependent reduction in F/F. This chlorophyll fluorescence screening technique promises to reveal novel mutants associated with photorespiration and photoinhibition.
将拟南芥光呼吸突变体暴露于空气中会导致叶绿素荧光参数F/F迅速下降，这反映了光系统II功能的下降和光抑制的开始。本文证明，F/F的叶绿素荧光成像可作为一种简单有效的方法，用于检测在光呼吸各个方面受损的拟南芥突变体。通过将1周龄的拟南芥幼苗阵列网格暴露于零CO₂条件下并以此作为成像的起始材料，该筛选方法被开发得既灵敏又高通量。使用此程序，我们筛选了约25000个经化学诱变的M2拟南芥种子，并以每1000粒种子约4个的频率获得了光呼吸表型（低CO₂条件下F/F降低）。此外，我们还获得了在高CO₂条件下F/F降低的突变体。在这一组中，我们检测到一种新的“反向光呼吸”表型，该表型在高CO₂条件下F/F呈现依赖性降低。这种叶绿素荧光筛选技术有望揭示与光呼吸和光抑制相关的新突变体。
