New method of plant hormone research is fast and sensitive

Olomouc scientists from the Institute of Experimental Botany, AS CR at Palacký University and CRH have in collaboration with Swedish colleagues developed a unique method to measure a record-high number of 101 plant hormones and their related substances in a single small sample of the root , stalk or other organ. The new procedure will accelerate hormone analyses and help scientists better understand how their levels change in different situations, for example, when crops or other plants face unfavorable conditions.

Like humans, plants also have hormones – compounds that in a small amount affect physiological processes in the body. In plants, they usually participate in managing important processes. Biologists therefore need to measure the concentrations of all involved hormones, as well as the substances from which the hormones are formed or transformed. In order to make such an extensive analysis as easy as possible, scientists are creating more and more effective detection methods. These are truly top-notch procedures for determination of 101 plant hormones and related compounds.

“Our method is fast, sensitive and works reliably with small plant samples weighing only 20 milligrams. In each sample we can simultaneously determine up to 101 substances, which is almost double than the previous methods of other authors,” said Ondřej Novák, who developed the method together with his Olomouc team and scientists from Agricultural University in Umeå, Sweden. “We use mass spectrometry because it is extremely sensitive and can accurately identify the compounds that interest us. The whole analysis takes only 32 minutes,” added another member of the Olomouc team Jan Šimura.

Practical applicability of the method was verified on the seedlings of Arabidopsis thaliana, which was exposed to salt stress. In roots samples and in above-ground parts, they found a total of 45 hormones and related substances of the 101 mentioned earlier, which they can determine thanks to the new method. Salinity affected the level of 23 of these compounds in roots and 15 in above-ground organs. Physiological responses to salinity are therefore involved in several different hormones. As found, concentrations of some hormones increased or decreased accordingly to the changes in activity of genes responsible for their synthesis or degradation. Linking data on the response of genes and hormones to salinity or other stimuli will provide a more comprehensive picture of the processes that take place in the plant.

“Drought, frost, or soil salinity cause great losses in agriculture each year. We are therefore trying to understand the biological processes that lead to activation of defensive mechanisms that allow the plant to survive. We have been working with scientists from the Czech Republic and the whole world to analyze hormones in various plants including crops. We will certainly use our “101” method in these projects. But we have other plans in the future. We would like to measure plant hormone levels in different cell types as well as in individual parts of the cell – organelles,” Novák said.

An article describing the newly developed method has been published by Plant Physiology. The text is freely available on the magazine website.

Mr. Novák leads the Department of Metabolomics in the Laboratory of Growth Regulators, which is a joint workplace of Palacký University and the Institute of Experimental Botany of the Academy of Sciences of the Czech Republic. At the same time, it is part of Center of teh Region Haná for Biotechnological and Agricultural Research.