Leaf ultrastructural studies of Avicennia marina in response to salinity under natural conditions.

Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2007.

In Richards Bay Harbour, the mangrove Avicennia marina exhibits a distinct natural productivity gradient. The fringe site, which is regularly inundated twice daily by tides, supports luxuriant adult A. marina trees that are 6-10 m tall and which form a dense, well-developed canopy. The landward site which is only inundated during high spring tides, supports diminutive or dwarf A. marina that are less than 1.5 m in height. In this study we compared leaves from fringe and dwarf sites with respect to morphology, ultrastructure and ecophysiology. Alterations in leaf morphology, ultrastructure and physiology of A. marina were compared at the fringe site (35 ‰) and dwarf site (60 ‰) using morphometric measurements, light (LM), transmission (TEM) and scanning microscopy (SEM). SEM and light microscopy revealed that multicellular salt glands were located on the thick, cutinised adaxial surface from leaves of both sites. The glands appeared to be scattered and protruding from individual crypts in fringe mangrove leaves whilst they appeared sunken and occluded by cuticular material in dwarf mangrove leaves. The salt glands on the abaxial surface were not sunken but obscured by the indumentum of peltate trichomes. Ultrastructural changes observed in dwarf mangrove leaves were associated with cuticle, cell walls, chloroplasts, mitochondria of mesophyll tissue and salt glands. Fringe mangrove leaves had chloroplasts with typical well-developed grana and stroma. Ultrastructural changes of chloroplasts were evident in dwarf mangrove leaves and included swelling and separation of thylakoids, disintegration of granal stacking and integranal lamellae, as well as loss of the integrity of the chloroplast envelope. Multivesicular structures were commonly found in vacuoles and associated with chloroplasts and mitochondria in both leaf types. In fringe mangrove leaves, mitochondria appeared spherical to tubular with a relatively smooth outer membrane and a highly convoluted inner membrane. Swelling and vacuolation of mitochondrial membranes, cristae and mitochondrial clustering in the cytoplasm around the chloroplasts were evident in dwarf mangrove leaves. Extensive lipid accumulation in the form of large, dense plastoglobuli occurred in the chloroplasts of dwarf mangrove leaves. There were characteristic differences in salt gland morphology of fringe and dwarf mangrove leaves, namely in the cell walls, vacuoles, and vesicle formation. In salt glands of dwarf mangrove leaves, a distinct withdrawal of the cytoplasm from the cell wall was observed. This feature was not observed in salt glands of fringe mangrove leaves. Numerous large vacuoles were observed in the secretory cells of glands of dwarf mangrove leaves compared to those of fringe plants. Multivesicular structures, vesicles and mitochondria were common features in both leaf types. Physiological studies involved a comparison of osmotic and ionic relations as well as whole plant responses in fringe and dwarf mangrove leaves. Relative leaf water content decreased by 7.8 % and specific leaf area by 17 % in dwarf compared to those of fringe mangroves. Dwarf mangrove leaves were 27.6 % thicker and leaf cuticle thickness 37.4 % higher than those from fringe mangroves. Fringe mangrove leaves displayed higher total chlorophyll contents by 27 %, with chlorophylls a and b being 22 % and 39.6 % higher, respectively than those of dwarf mangroves. Salt gland frequencies were higher in the apex, mid-lamina and base of fringe than dwarf mangrove leaves by 36 %, 45 % and 51 %, respectively. The concentration of glycinebetaine, a compatible, N-containing osmolyte was significantly higher by 40 % in dwarf than in fringe mangrove leaves. Concentrations of proline were 27 % lower in dwarf than in fringe mangrove leaves. The predominant inorganic ion detected in mature leaves was Na+, which was 19 % higher in dwarf than fringe mangrove leaves. Phosphorus was an element that appeared deficient in dwarf mangrove leaves, being 50 % lower compared to fringe mangrove leaves. The results of this investigation indicated that there were cytomorphological alterations as well as differences in physiological responses in leaves of A. marina at fringe and dwarf sites.