Researchers from the Centre of the Region Hana for Biotechnological and Agricultural Research (CRH) in Olomouc, together with colleagues from the University of Nottingham, and the Max Planck Institute of Molecular Cell Biology in Dresden, have made another significant step in exploring plant structure and development using light sheet fluorescence microscopy. From the traditional model plant - Arabidopsis thaliana, they advanced to a much larger alfalfa, which can be observed in its natural state for up to several days. Unique experiences have been summarized in an article for the prestigious journal Nature Plants, which outlined also the perspectives of the method to the world. In the future, they intend to use it for other crops, especially barley. Obtained information is crucial, for example, for increasing plant yields or resistance to adverse environmental conditions, including drought or disease.
The stepping stone of this research were detailed protocols for long-term and gentle exploration of plants with high-resolution using commercially available laser microscopes, which CRH scientists published three years ago in Nature Protocols, attracting considerable interest in the scientific community. Subsequently, researchers have decided to develop and modify the technique for long-term and gentle exploration of plants so that they can capture objects larger than Arabidopsis, such as alfalfa embryos and plants developing from them.
"We have succeeded in further improvement of the only method that can be used to visualize living plants at different levels from intracellular to the whole plants. The fact that we have advanced to alfalfa from Arabidopsis is a great milestone. We can watch the plant development from several hours to several days. We can observe not only its development but also its interactions with external environment and microbes in defined and strictly controlled conditions. We will get much better explanations to a range of biological and biotechnological questions," said Jozef Samaj, head of Olomouc research group.
According to his colleague and a co-author Miroslav Ovecka, the transfer of the method from the model plant to alfalfa was technically quite complex. "We use a commercially available microscopic system, so we had to take into account its technical parameters. It was important to prepare conditions for long-term plant cultivation. The plant in the microscope receives fresh nutrients in a sterile environment, is supplied with the air and exposed to the normal light/dark illumination cycle. The results are not affected by secondary stress caused by microscopy itself. The significant shift in the scientific field brought about by this method lies in the fact that there has not yet been a system approach that would evaluate alfalfa at such a level. We have also described technological difficulties that other research groups often face, and suggested a way how this progressive method could be used for developmental studies for any type of plant objects. Also from this point of view, the article is unique. We are presenting new perspectives to the scientific community based on obtained results," explained Ovecka.
Olomouc scientists intend to continue to improve the methodology and adapt it for possible use in barley within project "Plants as a means of sustainable global development" under Operational Program for Research, Development and Education. Thanks to a paper in a prestigious journal, they will be able to present the progress in this technique and its possible applications in plant research at even more conferences and professional seminars. CRH brings together scientific teams from Palacky University and Olomouc worksites of the Institute of Experimental Botany of Academy of Sciences of Czech Republic, and the Crop Research Institute.
For more information:
https://www.nature.com/articles/s41477-018-0238-2