Leaf glandular trichomes in Empetrum nigrum: morphology, histochemistry, ultrastructure and secondary metabolites.

The morphology, histochemistry and ultrastructure of the glandular trichomes on Empetrum nigrum leaves have been studied and more than a third of the metabolites were identified. Samples of the leaves were fixed and processed for light and electron microscopy. Glandular trichomes are situated on the inner surface of the rolled leaves. They have a clavate head and a short stalk. Histochemical tests and fluorescent microscopy demonstrate differentiated staining of the various cell types in the glandular trichome for proteins, pectins, lipids, tannins and phenylpropanoids. During secretion, the secretory cells contain rough and smooth endoplasmic reticulum, Golgi stacks with large vesicles, diversiform leucoplasts in contact with a reticular sheath and opaque deposits in the vacuoles. There are ultrastructural and functional differences between the secretory cells in the trichome head: synthesis of hydrophilic substances predominately occurs in the upper and middle secretory cells, whereas synthesis of lipophilic compounds takes place in the middle and lower secretory cells. Gas chromatography-mass spectrometry was used to determine the content of metabolites in the methanol extracts from the leaves. Many phenolic compounds (phenolic acids, bibenzyls, catechins, flavanones and flavan-3-ols) as well as several terpenoids were found. Two chalcones (2′,4′-dimethoxydihydrochalcone and 2′,4′,6′-trihydroxydihydrochalcone), one bibenzyl (batatasin III), one flavanone (7-hydroxyflavanone) and 8 terpenoids (including phytol, α-tocopherol, ß-sitosterol, α-amyrin, uvaol, oleanolic acid, ursolic acid and dehydroursolic acid) were identified in E. nigrum leaf extracts. The total yield of phenolic compounds is five to six times higher than the yield of terpenoids. It has been established that chalcones have no hydroxyl groups in ring B whereas bibenzyls have a hydroxyl group in the 3-position in ring B. On the basis of the histochemistry, fluorescent microscopy, ultrastructure and chemical analysis, it may be concluded that synthesis and accumulation of phenolic substances and terpenoids takes place in the clavate glandular trichomes. Secondary metabolites synthesized in the trichomes protect leaf tissues from viruses, bacteria and pathogenic fungi. [ABSTRACT FROM AUTHOR]