RESUMO
Humic acids have been widely used for centuries to enhance plant growth and productivity. The beneficial effects of humic acids have been attributed to different functional groups and phytohormone-like compounds enclosed in macrostructure. However, the mechanisms underlying the plant growth-promoting effects of humic acids are only partially understood. We hypothesize that the bio-stimulatory effect of humic acids is mainly due to the modulation of innate pathways of auxin and cytokinin biosynthesis in treated plants. A physiological investigation along with molecular characterization was carried out to understand the mechanism of bio-stimulatory effects of humic acid. A gene expression analysis was performed for the genes involved in auxin and cytokinin biosynthesis pathways in wheat seedlings. Furthermore, Arabidopsis thaliana transgenic lines generated by fusing the auxin-responsive DR5 and cytokinin-responsive ARR5 promoter to ß-glucuronidase (GUS) reporter were used to study the GUS expression analysis in humic acid treated seedlings. This study demonstrates that humic acid treatment improved the shoot and root growth of wheat seedlings. The expression of several genes involved in auxin (Tryptophan Aminotransferase of Arabidopsis and Gretchen Hagen 3.2) and cytokinin (Lonely Guy3) biosynthesis pathways were up-regulated in humic acid-treated seedlings compared to the control. Furthermore, GUS expression analysis showed that bioactive compounds of humic acid stimulate endogenous auxin and cytokinin-like activities. This study is the first report in which using ARR5:GUS lines we demonstrate the biostimulants activity of humic acid.
RESUMO
To develop crops capable of withstanding challenges posed by climate change, breeding strategies must focus on addressing multiple stresses occurring concurrently in plants. Leaf epidermal structures such as trichomes, stomata, and epidermal cells play an important role in mediating plant defense and could be essential traits that impart wide-ranging tolerance to biotic and abiotic stresses. Consequently, it is important to inform on the underlying diversity in these traits in lentil germplasm (Lens spp.). In this study, we characterized foliar microstructures of 12 genotypes belonging to seven wild and cultivated Lens species. We performed scanning electron microscopy on leaflet and pod surfaces for their qualitative characterization. For quantitative characterization, we observed surface imprints via light microscopy and quantified trichome density (TD), trichome length (TL), stomatal density (SD), epidermal cell density (ECD), and stomatal index (SI) on adaxial and abaxial leaflet surfaces for each genotype. We also assessed the heritability of trichome traits by evaluating interspecific recombinant inbred lines (RILs) derived from the cross Lens culinaris CDC Redberry × Lens tomentosus IG 72805. Comparing foliar microstructures, we found that TD and TL varied widely among cultivated and wild lentil genotypes. However, in most lentil genotypes, the adaxial leaflet surface had lower TD and longer trichomes compared to the abaxial surface. Pubescence on pods comprised five major phenotypes: no trichomes or glabrous pods, very short trichomes at low density, short trichomes at high density, medium-length trichomes at high density, and long trichomes at high density. Leaves of all species were amphistomatous, and SI, SD, and ECD were all higher on the adaxial compared to the abaxial surface. Adaxial surfaces had slightly sunken stomata, which might be an adaptive trait to conserve water. Quantifying TD and TL on the leaflets of interspecific RILs revealed transgressive segregation of these traits, suggesting that TD and TL are quantitative in nature. While taxonomic implications of this study are limited, a detailed description of agronomically relevant morphophysiological traits presented in this paper along with the mode of inheritance of trichomes may serve as a resource for scientists developing lentil adapted to concurrent biotic and abiotic stresses of the future.
