Abstract
BackgroundSmall 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.ResultsWe 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 %.ConclusionsIn 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-016-0875-4) contains supplementary material, which is available to authorized users.
Highlights
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
In order to verify the applicability of this correlation, we analyzed 4′,6-diamidino-2-phenylindole (DAPI)-stained pollen taken from flower buds of different sizes
We have shown that a small chemical screen using pollen harvested from few flowering plants is one of the fastest systems for high-throughput screening (HTS), and versatile as pollen from virtually any plant can be used
Summary
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. In some cases, associated gene targets have been identified [14,15,16,17,18,19,20] Despite of these achievements, further progress in plant chemical biology largely depends on to what extent image-based screening pipelines can be improved and applied to increase the spatio-temporal phenotypic resolution of fast growing plant systems, and enable the rapid and sensitive screening of large small molecule libraries [14, 21, 22]. We hypothesize that any compound, found to inhibit or stimulate pollen germination and tube growth is likely to affect other plant processes, such as seed germination, growth or differentiation of roots or shoots
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