Antunes Mauricio S, Ha Suk-Bong, Tewari-Singh Neera, Morey Kevin J, Trofka Anna M, Kugrens Paul, Deyholos Michael, Medford June I
Department of Biology, Colorado State University, Fort Collins, CO 80523-1878, USA.
Plant Biotechnol J. 2006 Nov;4(6):605-22. doi: 10.1111/j.1467-7652.2006.00205.x.
Plants have evolved elegant mechanisms to continuously sense and respond to their environment, suggesting that these properties can be adapted to make inexpensive and widely used biological monitors, or sentinels, for human threats. For a plant to be a sentinel, a reporting system is needed for large areas and widespread monitoring. The reporter or readout mechanism must be easily detectable, allow remote monitoring and provide a re-set capacity; all current gene reporting technologies fall short of these requirements. Chlorophyll is one of the best-recognized plant pigments with an already well-developed remote imaging technology. However, chlorophyll is very abundant, with levels regulated by both genetic and environmental factors. We designed a synthetic de-greening circuit that produced rapid chlorophyll loss on perception of a specific input. With induction of the de-greening circuit, changes were remotely detected within 2 h. Analyses of multiple de-greening circuits suggested that the de-greening circuit functioned, in part, via light-dependent damage to photosystem cores and the production of reactive oxygen species. Within 24-48 h of induction, an easily recognized white phenotype resulted. Microarray analysis showed that the synthetic de-greening initiated a process largely distinct from normal chlorophyll loss in senescence. Remarkably, synthetically de-greened white plants re-greened after removal of the inducer, providing the first easily re-settable reporter system for plants and the capacity to make re-settable biosensors. Our results showed that the de-greening circuit allowed chlorophyll to be employed as a simple but powerful reporter system useful for widespread areas.
植物已经进化出精妙的机制来持续感知并响应其环境,这表明这些特性可加以利用,制造出用于监测人类威胁的廉价且广泛应用的生物监测器或哨兵。要让植物成为哨兵,就需要一个用于大面积和广泛监测的报告系统。报告或读出机制必须易于检测、能进行远程监测并具备重置能力;目前所有的基因报告技术都无法满足这些要求。叶绿素是最广为人知的植物色素之一,已有成熟的远程成像技术。然而,叶绿素含量极为丰富,其水平受遗传和环境因素共同调控。我们设计了一种合成的去绿回路,该回路在感知特定输入时会迅速导致叶绿素损失。随着去绿回路的诱导,在2小时内就能远程检测到变化。对多个去绿回路的分析表明,去绿回路部分是通过光依赖对光系统核心的损伤以及活性氧的产生来发挥作用的。在诱导后的24至48小时内,会产生易于识别的白色表型。微阵列分析表明,合成去绿引发的过程与衰老过程中正常的叶绿素损失在很大程度上不同。值得注意的是,去除诱导剂后,经合成去绿变成白色的植物会重新变绿,这为植物提供了首个易于重置的报告系统,也具备制造可重置生物传感器的能力。我们的结果表明,去绿回路使叶绿素能够用作一个简单却强大的报告系统,适用于广泛区域。