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Effective Strategies for Handling Troublesome Plant Material in Botanical Studies

This guide outlines practical steps to improve the handling of troublesome plant materials in botanical research. Key strategies include selecting alternative isolation and staining buffers, incorporating antioxidants and preservatives, choosing different plant tissues, and utilizing advanced techniques like image cytometry. Specific buffer recipes and chemical preservatives such as antioxidants and organic acids are provided to enhance tissue preservation, ensuring better results in histological analysis. The work focuses on common challenges faced in plant studies, highlighting effective solutions for researchers.

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Effective Strategies for Handling Troublesome Plant Material in Botanical Studies

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  1. Troublesome material: how to deal with? Jan Suda Department of Botany, Faculty of Science, Charles University Institute of Botany, Academy of Sciences of the Czech Republic

  2. Steps towards the improvement 1) choosing of another isolation / staining buffer 2) inclusion of antioxidants or other preservatives 3) selection of different plant tissue 4) employment of different technique (image cytometry)

  3. B u f f e r s e l e c t i o n O t t o b u f f e r s Otto I Citric acid . H2O 4.2 g Tween 20 1 ml Otto II Na2HPO4 . 12H2O 28.65 g Water to 200 ml L B 0 1 b u f f e r Tris 363.4 mg Na2EDTA 148.9 mg spermine . 4HCl 34.8 mg KCl 1.193 g NaCl 233.8 mg Triton X-100 200 μl Water to 200 ml, pH 7.5 Tris - MgCl2 b u f f e r Tris 4.84 g MgCl2 . 6H20 162.64 mg Triton X-100 1 ml Water to 200 ml, pH 7.5 M a t z k b u f f e r Tris 1.211 g MgCl2 . 6H20 0.107 g NaCl 0.5 g Triton X-100 0.1 ml Water to 200 ml, pH 7.5

  4. C h e m i c a l p r e s e r v a t i v e s Antioxidants 2-mercaptoethanol (2μl / ml) Different fluorochrome DAPI generally less problematic Preservatives PVP-10, PVP-40 (min. 10 mg / ml) Rosaceae (Rosa, Rubus)

  5. Plant group tannins (Marcetella) mucilaginous compounds (Viola, Lythrum, Lavatera, Ocotea) organic acids (Aeonium, Monanthes) ??? (Boraginaceae - Echium) Reynoutria (tannins, organic acids?, only with PI) Teline, Spartocytisus, Chamaecytisus Helianthemum (mucilaginous compounds) Geranium (tannins) Improvement cotyledons petioles, young stems, inflorescence axes, cotyledons stems, very young leaves cotyledons youngest (in-rolled) leaves young stems ??? (seeds) T i s s u e s e l e c t i o n

  6. leaves cotyledons Echium leaves young stems Lythrum H i s t o g r a m s d e m o n s t r a t i o n

  7. T r o u b l e s o m e h i s t o g r a m Dactylorhiza

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