Grass species within South Africa show a photosynthetic subtype and phylogenetic response to rainfall gradients, with Panicoideae species (NADP-Me and NAD-Me) inhabiting mesic environments, while Aristidoideae species (NADP-Me) inhabit more arid environments. It is predicted that climate change will alter rainfall patterns within southern Africa, which could have implications for grassland distributions and functional composition. Globally, and in South Africa, species distributions indicates that NAD-Me species have a preference for more arid environments, but this may be complicated by phylogeny as most NAD-Me species belong to the Chloridoideae subfamily. Additionally, differences in the metabolism and energetic requirements of different carboxylation types are expected to confer different ecological advantages, such as drought tolerance, but the role of these different pathways is not well understood. Based on natural distribution and photosynthetic subtype differences, it was hypothesised that Panicoideae NADP-Me species would be less drought tolerant than Panicoideae NAD-Me and Aristidoideae NADP-Me species and that subtypes and lineages would show different drought recovery rates. Furthermore, drought sensitivity would be of a metabolic and not a stomatal origin and plants that maintained favourable leaf water status would be more drought tolerant and recover faster. This was tested experimentally by comparing Panicoideae species (NADP-Me and NAD-Me) and NADP-Me species (Panicoideae and Aristidoideae). Plants were subjected to a progressive 58 day drought period and a recovery phase where gas exchange, chlorophyll fluorescence and leaf water relations were measured at select intervals. In conjunction with this, a rapid drought experiment was performed on Zea mays (NADP-Me: Panicoideae) plants where similar parameters were measured. Photosynthetic drought and recovery responses showed both a subtype and phylogenetic response. Panicoideae species were less drought tolerant than Aristidoideae species, although Panicoideae NAD-Me showed better recovery rates than Panicoideae NADP-Me species, while Aristidoideae species recovered the quickest. Panicoideae NAD-Me and Aristidoideae species maintained higher leaf water status during drought which contributed to the maintenance of PSII integrity and thus facilitated rapid photosynthetic recovery. During drought Panicoideae species showed greater metabolic limitations over Aristidoideae species and for the first time, lower metabolic limitations were associated with osmotic adjustment. This is a novel finding whereby osmotic adjustment and the subsequent maintenance of leaf water are key to preventing metabolic limitations of photosynthesis in C₄ grasses. Results from the Z. mays rapid drought study showed the limitations to photosynthesis were exclusively metabolic and unlikely to be a direct consequence of turgor loss. It was apparent that the response to drought was stronger amongst lineages, as NADP-Me species from different subfamilies showed a significant difference in drought tolerances. Aristidoideae species’ exceptional drought tolerance and predicted increased aridification could favour these species over Panicoideae species under future climates.

The success of C₄ plants lies in their ability to concentrate CO₂ at the site of Rubisco thereby conferring greater efficiencies of light, water and nitrogen. Such characteristics should advantage C₄ plants in arid, hot environments. However, not all C₄ subtypes are drought tolerant. The relative abundance of NADP-ME species declines with increasing aridity. Furthermore, selected species have been demonstrated as being susceptible to severe drought showing metabolic limitations of photosynthesis. However there is a lack of phylogenetic control with many of these studies. The aims of this study were to determine whether the NADP-ME subtype was inherently susceptible to drought by comparing six closely related C₃ and C₄ (NADP-ME) Panicoid grasses. Gas exchange measurements were made during a natural rainless period and a controlled drought / rewatering event. Prior to water stress, the C₄ species had higher assimilation rates (A), and water use efficiencies (WUE[subscript leaf]) than the C₃ species, while transpiration rates (E) and stomatal conductances (g[subscript s]) were similar. At low soil water content, the C₃ species reduced gs by a greater extent than the C₄ species, which maintained higher E during the driest periods. The C₄ species showed proportionally greater reductions in A than the C₃ species and hence lost their WUE[subscript leaf] and photosynthetic advantage. CO₂ response curves showed that metabolic limitation was responsible for a greater decrease in A in the C₄ type than the C₃ type during progressive drought. Upon re-watering, photosynthetic recovery was quicker in the C species than the C₄ species. Results from whole plant measurements showed that the C₄ type had a significant whole plant water use efficiency advantage over the C₃ type under well-watered conditions that was lost during severe drought due to a greater loss of leaf area through leaf mortality rather than reductions in plant level transpiration rates. The C₃ type had xylem characteristics that enhanced water-conducting efficiency, but made them vulnerable to drought. This is in contrast to the safer xylem qualities of the C₄ type, which permitted the endurance of more negative leaf water potentials than the C₃ type during low soil water content. Thus, the vulnerability of photosynthesis to severe drought in NADP-ME species potentially explains why NADP-ME species abundance around the world decreases with decreasing rainfall.