Repeated origin of three-dimensional leaf venation releases constraints on the evolution of succulence in plants.

Succulent water storage is a prominent feature among plants adapted to arid zones, but we know little about how succulence evolves and how it is integrated into organs already tasked with multiple functions. Increased volume in succulent leaves, for example, may result in longer transport distances between veins and the cells that they supply, which in turn could negatively impact photosynthesis. We quantified water storage in a group of 83 closely related species to examine the evolutionary dynamics of succulence and leaf venation. In most leaves, vein density decreased with increasing succulence, resulting in significant increases in the path length of water from veins to evaporative surfaces. The most succulent leaves, however, had a distinct three-dimensional (3D) venation pattern, which evolved 11-12 times within this small lineage, likely via multiple developmental pathways. 3D venation "resets" internal leaf distances, maintaining moderate vein density in extremely succulent tissues and suggesting that the evolution of extreme succulence is constrained by the need to maintain an efficient leaf hydraulic system. The repeated evolution of 3D venation decouples leaf water storage from hydraulic path length, facilitating the evolutionary exploration of novel phenotypic space.

Quantifying succulence: a rapid, physiologically meaningful metric of plant water storage.

Quantification of succulence should ideally convey information about physiological function and yet also be straightforward to measure. While important aspects of succulence and its physiological consequences may be quantified using parameters derived from pressure-volume (P-V) curves, this technique applied to succulent tissues is difficult, time consuming and generally not suitable for large comparative datasets. We performed P-V curves on leaves of 25 taxa from across Caryophyllales and compared the results with direct measures of saturated water content (SWC(meas) ), the ratio of water mass at full saturation to tissue dry mass, for the same taxa. SWC(meas) was significantly related to relative capacitance, the most physiologically relevant parameter describing tissue succulence. We developed a linear model describing SWC(meas) as a function of relative capacitance and leaf volume, which is also supported when accounting for the phylogenetic relationships among taxa. These results indicate that SWC(meas) is a suitable proxy for tissue succulence, and that both cellular properties and variation in gross morphology contribute towards a plant's relative water storage capacity. Quantifying SWC(meas) across many taxa showing variation in tissue succulence will provide a new avenue for exploring the evolutionary dynamics of this important ecological adaptation.

Contemporaneous and recent radiations of the world's major succulent plant lineages.

The cacti are one of the most celebrated radiations of succulent plants. There has been much speculation about their age, but progress in dating cactus origins has been hindered by the lack of fossil data for cacti or their close relatives. Using a hybrid phylogenomic approach, we estimated that the cactus lineage diverged from its closest relatives ≈35 million years ago (Ma). However, major diversification events in cacti were more recent, with most species-rich clades originating in the late Miocene, ≈10-5 Ma. Diversification rates of several cactus lineages rival other estimates of extremely rapid speciation in plants. Major cactus radiations were contemporaneous with those of South African ice plants and North American agaves, revealing a simultaneous diversification of several of the world's major succulent plant lineages across multiple continents. This short geological time period also harbored the majority of origins of C(4) photosynthesis and the global rise of C(4) grasslands. A global expansion of arid environments during this time could have provided new ecological opportunity for both succulent and C(4) plant syndromes. Alternatively, recent work has identified a substantial decline in atmospheric CO(2) ≈15-8 Ma, which would have strongly favored C(4) evolution and expansion of C(4)-dominated grasslands. Lowered atmospheric CO(2) would also substantially exacerbate plant water stress in marginally arid environments, providing preadapted succulent plants with a sharp advantage in a broader set of ecological conditions and promoting their rapid diversification across the landscape.

Complex evolutionary transitions and the significance of c(3)-c(4) intermediate forms of photosynthesis in Molluginaceae.

C(4) photosynthesis is a series of biochemical and structural modifications to C(3) photosynthesis that has evolved numerous times in flowering plants, despite requiring modification of up to hundreds of genes. To study the origin of C(4) photosynthesis, we reconstructed and dated the phylogeny of Molluginaceae, and identified C(4) taxa in the family. Two C(4) species, and three clades with traits intermediate between C(3) and C(4) plants were observed in Molluginaceae. C(3)-C(4) intermediacy evolved at least twice, and in at least one lineage was maintained for several million years. Analyses of the genes for phosphoenolpyruvate carboxylase, a key C(4) enzyme, indicate two independent origins of fully developed C(4) photosynthesis in the past 10 million years, both within what was previously classified as a single species, Mollugo cerviana. The propensity of Molluginaceae to evolve C(3)-C(4) and C(4) photosynthesis is likely due to several traits that acted as developmental enablers. Enlarged bundle sheath cells predisposed some lineages for the evolution of C(3)-C(4) intermediacy and the C(4) biochemistry emerged via co-option of photorespiratory recycling in C(3)-C(4) intermediates. These evolutionarily stable transitional stages likely increased the evolvability of C(4) photosynthesis under selection environments brought on by climate and atmospheric change in recent geological time.

Anatomical variation in Cactaceae and relatives: Trait lability and evolutionary innovation.

The cacti have undergone extensive specialization in their evolutionary history, providing an excellent system in which to address large-scale questions of morphological and physiological adaptation. Recent molecular phylogenetic studies suggest that (1) Pereskia, the leafy genus long interpreted as the sister group of all other cacti, is likely paraphyletic, and (2) Cactaceae are nested within a paraphyletic Portulacaceae as a member of the "ACPT" clade (Anacampseroteae, Cactaceae, Portulaca, and Talinum). We collected new data on the vegetative anatomy of the ACPT clade and relatives to evaluate whether patterns in the distributions of traits may provide insight into early events in the evolutionary transition to the cactus life form. Many traits had high levels of homoplasy and were mostly equivocal with regard to infraclade relationships of ACPT, although several characters do lend further support to a paraphyletic Pereskia. These include a thick stem cuticle, prominent stem mucilage cells, and hypodermal calcium oxalate druses, all of which are likely to be important traits for stem water storage and photosynthesis. We hypothesize that high lability of many putative "precursor" traits may have been critical in generating the organismal context necessary for the evolution of an efficient and integrated photosynthetic stem.