A molecular phylogeny of the genus Alloteropsis (Panicoideae, Poaceae) suggests an evolutionary reversion from C4 to C3 photosynthesis.

BACKGROUND AND AIMS: The grass Alloteropsis semialata is the only plant species with both C(3) and C(4) subspecies. It therefore offers excellent potential as a model system for investigating the genetics, physiology and ecological significance of the C(4) photosynthetic pathway. Here, a molecular phylogeny of the genus Alloteropsis is constructed to: (a) confirm the close relationship between the C(3) and C(4) subspecies of A. semialata; and (b) infer evolutionary relationships between species within the Alloteropsis genus. METHODS: The chloroplast gene ndhF was sequenced from 12 individuals, representing both subspecies of A. semialata and all four of the other species in the genus. ndhF sequences were added to those previously sequenced from the Panicoideae, and used to construct a phylogenetic tree. KEY RESULTS: The phylogeny confirms that the two subspecies of A. semialata are among the most recently diverging lineages of C(3) and C(4) taxa currently recognized within the Panicoideae. Furthermore, the position of the C(3) subspecies of A. semialata within the Alloteropsis genus is consistent with the hypothesis that its physiology represents a reversion from C(4) photosynthesis. The data point to a similar evolutionary event in the Panicum stenodes-P. caricoides-P. mertensii clade. The Alloteropsis genus is monophyletic and occurs in a clade with remarkable diversity of photosynthetic biochemistry and leaf anatomy. CONCLUSIONS: These results confirm the utility of A. semialata as a model system for investigating C(3) and C(4) physiology, and provide molecular data that are consistent with reversions from C(4) to C(3) photosynthesis in two separate clades. It is suggested that further phylogenetic and functional investigations of the Alloteropsis genus and closely related taxa are likely to shed new light on the mechanisms and intermediate stages underlying photosynthetic pathway evolution.

Seasonal differences in photosynthesis between the C3 and C4 subspecies of Alloteropsis semialata are offset by frost and drought.

The regional abundance of C(4) grasses is strongly controlled by temperature, however, the role of precipitation is less clear. Progress in elucidating the direct effects of photosynthetic pathway on these climate relationships is hindered by the significant genetic divergence between major C(3) and C(4) grass lineages. We addressed this problem by examining seasonal climate responses of photosynthesis in Alloteropsis semialata, a unique grass species with both C(3) and C(4) subspecies. Experimental manipulation of rainfall in a common garden in South Africa tested the hypotheses that: (1) photosynthesis is greater in the C(4) than C(3) subspecies under high summer temperatures, but this pattern is reversed at low winter temperatures; and (2) the photosynthetic advantage of C(4) plants is enhanced during drought events. Measurements of leaf gas exchange over 2 years showed a significant photosynthetic advantage for the C(4) subspecies under irrigated conditions from spring through autumn. However, the C(4) leaves were killed by winter frost, while photosynthesis continued in the C(3) plants. Unexpectedly, the C(4) subspecies also lost its photosynthetic advantage during natural drought events, despite greater water-use efficiency under irrigated conditions. This study highlights previously unrecognized roles for climatic extremes in determining the ecological success of C(3) and C(4) grasses.

Low temperature effects on leaf physiology and survivorship in the C3 and C4 subspecies of Alloteropsis semialata.

The species richness of C(4) grasses is strongly correlated with temperature, with C(4) species dominating subtropical ecosystems and C(3) types predominating in cooler climates. Here, the effects of low temperatures on C(4) and C(3) grasses are compared, controlling for phylogenetic effects by using Alloteropsis semialata, a unique species with C(4) and C(3) subspecies. Controlled environment and common garden experiments tested the hypotheses that: (i) photosynthesis and growth are greater in the C(4) than the C(3) subspecies at high temperatures, but this advantage is reversed below 20 degrees C; and (ii) chilling-induced photoinhibition and light-mediated freezing injury of leaves occur at higher temperature thresholds in the C(4) than the C(3) plants. Measurements of leaf growth and photosynthesis showed the expected advantages of the C(4) pathway over the C(3) type at high temperatures. These declined with temperature, but were not completely lost until 15 degrees C, and there was no evidence of a reversal to give a C(3) advantage. Chronic chilling (5-15 degrees C) or acute freezing events induced a comparable degree of photodamage in illuminated leaves of both subspecies. Similarly, freezing caused high rates of mortality in the unhardened leaves of both subtypes. However, a 2-week chilling treatment prior to these freezing events halved injury in the C(3) but not the C(4) subspecies, suggesting that C(4) leaves lacked the capacity for cold acclimation. These results therefore suggest that C(3) members of this subtropical species may gain an advantage over their C(4) counterparts at low temperatures via protection from freezing injury rather than higher photosynthetic rates.

Drought constraints on C4 photosynthesis: stomatal and metabolic limitations in C3 and C4 subspecies of Alloteropsis semialata.

The C4 photosynthetic pathway uses water more efficiently than the C3 type, yet biogeographical analyses show a decline in C4 species relative to C3 species with decreasing rainfall. To investigate this paradox, the hypothesis that the C4 advantage over C3 photosynthesis is diminished by drought was tested, and the underlying stomatal and metabolic mechanisms of this response determined. The effects of drought and high evaporative demand on leaf gas exchange and photosynthetic electron sinks in C3 and C4 subspecies of the grass Alloteropsis semialata were examined. Plant responses to climatic variation and soil drought were investigated using a common garden experiment with well-watered and natural rainfall treatments, and underlying mechanisms analysed using controlled drying pot experiments. Photosynthetic rates were significantly higher in the C4 than the C3 subspecies in the garden experiment under well-watered conditions, but this advantage was completely lost during a rainless period when unwatered plants experienced severe drought. Controlled drying experiments showed that this loss was caused by a greater increase in metabolic, rather than stomatal, limitations in C4 than in the C3 leaves. Decreases in CO2 assimilation resulted in lower electron transport rates and decreased photochemical efficiency under drought conditions, rather than increased electron transport to alternative sinks. These findings suggest that the high metabolic sensitivity of photosynthesis to severe drought seen previously in several C4 grass species may be an inherent characteristic of the C4 pathway. The mechanism may explain the paradox of why C4 species decline in arid environments despite high water-use efficiency.

A mutant of Arabidopsis lacking the triose-phosphate/phosphate translocator reveals metabolic regulation of starch breakdown in the light.

The chloroplast envelope triose-phosphate/phosphate translocator (TPT) is responsible for carbohydrate export during photosynthesis. Using measurements of carbohydrates, partitioning of assimilated 14CO2, photosynthetic gas exchange, and chlorophyll fluorescence, we show that a mutant of Arabidopsis lacking the TPT increases synthesis of starch compared to the wild type, thereby compensating for a deficiency in its ability to export triose-phosphate from the chloroplast. However, during growth under high light, the capacity for starch synthesis becomes limiting so that the chloroplastic phosphate pool is depleted, resulting in a restriction on electron transport, a reduction in the rate of photosynthesis, and slowed plant growth. Under the same conditions but not under low light, we observe release of 14C label from starch, indicating that its synthesis and degradation occur simultaneously in the light. The induction of starch turnover in the mutant specifically under high light conditions leads us to conclude that it is regulated by one or more metabolic signals, which arise as a result of phosphate limitation of photosynthesis.