Nitrogen and carbon costs of growth and antioxidant production during acclimation to environmental stress in two species of Gethyllis
Gethyllis multifolia L. Bolus and G. villosa Thunb. are winter-growing, summer blooming, deciduous and bulbous geophytes that grow naturally in the semi-arid succulent Karoo biome of South Africa. Both species grow under full sun conditions and have four distinctive growth phases: a winter (cold and wet) growing phase, leaf senescence phase towards spring, flowering phase during the hot and dry summer months, and fruit and leaf formation phase in autumn. The medicinal uses of this genus (including G. multifolia "Kukumakranka" and G. villosa "hairy kukumakranka") range from cures for colic, digestive disturbances, teething problems, fatigue, boils, bruises and insect bites, to being used as an aphrodisiac. Gethyllis multifolia is threatened in its natural habitat and is listed in the 'Vulnerable' category of the 'Red Data List of Southern African Plants' and the 'IUCN-World Conservation Union List of Plants'. The literature indicate that the habitats of both species are being exposed to drier conditions and is further threatened by the encroachment of invasive indigenous plant species. It is not known to which extent these factors may pose a threat to the existence of both species. The first objective of this investigation was to determine the costs of vegetative and reproductive growth during the seasonal life cycle of the plant, using carbon (C) and nitrogen (N) as a physiological currency. The second objective was to elucidate a functional basis to explain the difference in the conservation status of both species in their natural habitat. Both species were subjected to drought and shading as environmental stresses and the plant physiological performance was investigated via photosynthetic gas exchange. The third objective of the study was to evaluate the antioxidant content (total polyphenol, flavonol/flavone and flavanone content) and antioxidant capacity [ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC) and 2,2'-azino-di-3- ethylbenzthiazoline sulphonate (ABTS) radical cation scavenging ability] of natural populations and plant samples that were exposed to photo- and -drought environmental stresses. This study was done to elucidate the antioxidant profile of plant parts of natural populations as well as providing farmers, traditional healers and pharmaceutical companies with cultivation environmental conditions to enhance the antioxidant properties of the species. This investigation also attempted to isolate and characterize, by means of thin-layer chromatography (TLC) and column chromatography (CC), natural compounds from both species to lend support to the purported antioxidant benefit of both species and to further lend support to claims made by traditional healers of the medicinal potential of the genus. This study, however, did not engage in any in viva studies or human trials to support published literature of the medicinal benefits of the genus. The photosynthetic adaptation studies indicated that G. villosa had a better photosynthetic performance than G. multifolia during both drought and low light conditions because of the inability of G. multifolia to adapt to a wider range of environmental extremes. The C and N cost of growth and reproduction studies revealed that G. villosa had a more efficient resource utilisation strategy for both growth and reproduction. These physiological responses suggest that G. villosa, in general, has a more efficient survival strategy and that G. multifolia will struggle to adapt to drier environmental conditions, as well as growing in the shade of encroaching invasive plant species. To conclude, this could be a contributing factor as to why G. multifolia is threatened in its natural habitat and G. villosa not. The antioxidant content-and -capacity study on natural populations of both species revealed the highest total polyphenol content, FRAP and ORAC values for the flowers and fruits of G. multifolia and G. villosa compared to other plant parts. These values were found to be in line with and in some cases higher than most commercial fruits and vegetables. The antioxidant activity during drought and photo-stress of the leaves, bulbs and roots was found to be highest in the roots of both species during drought stress. Gethyllis multifolia, in general, exhibited higher total polyphenol content than G. villosa, with the highest content measured during drought stress in the roots of G. multifolia. Phytochemical investigation of the leaves, bulbs and roots of G. multifolia and G. villosa revealed the presence of tannins, flavonoids, phenolics, saponins, glycosides as well as essential oils, while alkaloids were absent. The chromatographic profiles of the leaves, bulbs and roots of both species further indicated that the roots of G. multifolia contained the highest concentration of natural products, compared to G. villosa and other plant parts. Further in-depth studies on the roots of G. multifolia led to the isolation and characterization of three known flavonoids, of which one was also isolated as its endogenously acetylated derivative. In contrast to the fact that both species had a high polyphenol content and exhibited high antioxidant activity, the isolated compounds in this study revealed very low antioxidant activity. However, the literature revealed that some of these isolated compounds exhibit antifungal, antibacterial, anti angiogenic and anti carcinogenic properties in vitro, which could be ascribed to the medicinal applications of plant parts of certain species belonging to this genus. Furthermore, this study suggests that further chemistry and pharmaceutical research on the genus, Gethyllis, in specific the flowers and fruit of these two species, be pursued.