Chuprov-Netochin Roman, Neskorodov Yaroslav, Marusich Elena, Mishutkina Yana, Volynchuk Polina, Leonov Sergey, Skryabin Konstantin, Ivashenko Andrey, Palme Klaus, Touraev Alisher
Moscow Institute of Physics and Technology, Dolgoprudny, 141700, Moscow region, Russian Federation.
Research Centerof Biotechnology of the Russian Academy of Science, 117312, Moscow, Russian Federation.
BMC Plant Biol. 2016 Sep 6;16(1):192. doi: 10.1186/s12870-016-0875-4.
Small synthetic molecules provide valuable tools to agricultural biotechnology to circumvent the need for genetic engineering and provide unique benefits to modulate plant growth and development.
We developed a method to explore molecular mechanisms of plant growth by high-throughput phenotypic screening of haploid populations of pollen cells. These cells rapidly germinate to develop pollen tubes. Compounds acting as growth inhibitors or stimulators of pollen tube growth are identified in a screen lasting not longer than 8 h high-lighting the potential broad applicability of this assay to prioritize chemicals for future mechanism focused investigations in plants. We identified 65 chemical compounds that influenced pollen development. We demonstrated the usefulness of the identified compounds as promotors or inhibitors of tobacco and Arabidopsis thaliana seed growth. When 7 days old seedlings were grown in the presence of these chemicals twenty two of these compounds caused a reduction in Arabidopsis root length in the range from 4.76 to 49.20 % when compared to controls grown in the absence of the chemicals. Two of the chemicals sharing structural homology with thiazolidines stimulated root growth and increased root length by 129.23 and 119.09 %, respectively. The pollen tube growth stimulating compound (S-02) belongs to benzazepin-type chemicals and increased Arabidopsis root length by 126.24 %.
In this study we demonstrate the usefulness of plant pollen tube based assay for screening small chemical compound libraries for new biologically active compounds. The pollen tubes represent an ultra-rapid screening tool with which even large compound libraries can be analyzed in very short time intervals. The broadly applicable high-throughput protocol is suitable for automated phenotypic screening of germinating pollen resulting in combination with seed germination assays in identification of plant growth inhibitors and stimulators.
小型合成分子为农业生物技术提供了有价值的工具，可避免基因工程的需求，并为调节植物生长发育带来独特益处。
我们开发了一种方法，通过对花粉细胞单倍体群体进行高通量表型筛选来探索植物生长的分子机制。这些细胞迅速萌发以形成花粉管。在不超过8小时的筛选中，鉴定出了作为花粉管生长抑制剂或刺激剂的化合物，这凸显了该测定法在为未来植物机制重点研究确定化学物质优先级方面的潜在广泛适用性。我们鉴定出65种影响花粉发育的化合物。我们证明了所鉴定的化合物作为烟草和拟南芥种子生长促进剂或抑制剂的有用性。当7天大的幼苗在这些化学物质存在的情况下生长时，与在无化学物质条件下生长的对照相比，其中22种化合物导致拟南芥根长度减少4.76%至49.20%。两种与噻唑烷结构同源的化学物质分别刺激根生长并使根长度增加129.23%和119.09%。刺激花粉管生长的化合物（S-02）属于苯并氮杂䓬类化学物质，使拟南芥根长度增加了126.24%。
在本研究中，我们证明了基于植物花粉管的测定法在筛选小型化合物库以寻找新的生物活性化合物方面的有用性。花粉管代表了一种超快速筛选工具，利用它甚至可以在非常短的时间间隔内分析大型化合物库。这种广泛适用的高通量方案适用于对萌发花粉进行自动化表型筛选，与种子萌发测定相结合可用于鉴定植物生长抑制剂和刺激剂。
