Crassulacean acid metabolism (CAM) is a water-use efficient adaptation of photosynthesis that has evolved independently many times in diverse lineages of flowering plants. We hypothesize that convergent evolution of protein sequence and temporal gene expression underpins the independent emergences of CAM from C3 photosynthesis. To test this hypothesis, we generate a de novo genome assembly and genome-wide transcript expression data for Kalanchoë fedtschenkoi, an obligate CAM species within the core eudicots with a relatively small genome (~260 Mb). Our comparative analyses identify signatures of convergence in protein sequence and re-scheduling of diel transcript expression of genes involved in nocturnal CO2 fixation, stomatal movement, heat tolerance, circadian clock, and carbohydrate metabolism in K. fedtschenkoi and other CAM species in comparison with non-CAM species. These findings provide new insights into molecular convergence and building blocks of CAM and will facilitate CAM-into-C3 photosynthesis engineering to enhance water-use efficiency in crops.

Summary Crassulacean acid metabolism (CAM) is a specialized mode of photosynthesis that features nocturnal CO2 uptake, facilitates increased water‐use efficiency (WUE), and enables CAM plants to inhabit water‐limited environments such as semi‐arid deserts or seasonally dry forests. Human population growth and global climate change now present challenges for agricultural production systems to increase food, feed, forage, fiber, and fuel production. One approach to meet these challenges is to increase reliance on CAM crops, such as Agave and Opuntia, for biomass production on semi‐arid, abandoned, marginal, or degraded agricultural lands. Major research efforts are now underway to assess the productivity of CAM crop species and to harness the WUE of CAM by engineering this pathway into existing food, feed, and bioenergy crops. An improved understanding of CAM has potential for high returns on research investment. To exploit the potential of CAM crops and CAM bioengineering, it will be necessary to elucidate the evolution, genomic features, and regulatory mechanisms of CAM. Field trials and predictive models will be required to assess the productivity of CAM crops, while new synthetic biology approaches need to be developed for CAM engineering. Infrastructure will be needed for CAM model systems, field trials, mutant collections, and data management.

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Increased demand for food requires us to investigate livestock forage and fodder crops that can be grown over a wide range of locations where their cultivation will not compete with that of the food supply. A large portion of the southwestern United States consists of underutilized semi‐arid land. Crops typically used for livestock fodder or forage have high‐water demands that make them uneconomical or unsustainable for semi‐arid and arid regions. The growth rate and low‐input requirements of prickly pear cactus (Opuntia ficus‐indica) make it an excellent candidate for forage or fodder supplementation or replacement in these regions. Previous reports about forage quality data on Opuntia have been scattered across multiple locations, growing conditions and cultivars. Here, we report on the forage quality and mineral content of Opuntia ficus‐indica grown under both field and greenhouse conditions. Crude protein was 71 and 264 g/kg of dry mass for field and greenhouse conditions, respectively. Field‐grown plants showed higher acid and neutral detergent fibre content than greenhouse‐grown plants reflecting higher cellulose, hemicellulose and lignin accumulation. Nutritional values were also compared to requirements of cattle to determine what deficiencies might need to be addressed through supplementation. These data suggest that Opuntia can be used in combination with other feed sources to reduce the demand of resource‐intensive forage crops for raising livestock in dryland areas.

SUMMARY Tissue succulence (ratio of tissue water/leaf area or dry mass) or the ability to store water within living tissues is among the most successful adaptations to drought in the plant kingdom. This taxonomically widespread adaptation helps plants avoid the damaging effects of drought, and is often associated with the occupancy of epiphytic, epilithic, semi‐arid and arid environments. Tissue succulence was engineered in Arabidopsis thaliana by overexpression of a codon‐optimized helix‐loop‐helix transcription factor (VvCEB1opt) from wine grape involved in the cell expansion phase of berry development. VvCEB1opt‐overexpressing lines displayed significant increases in cell size, succulence and decreased intercellular air space. VvCEB1opt‐overexpressing lines showed increased instantaneous and integrated water‐use efficiency (WUE) due to reduced stomatal conductance caused by reduced stomatal aperture and density resulting in increased attenuation of water‐deficit stress. VvCEB1opt‐overexpressing lines also showed increased salinity tolerance due to reduced salinity uptake and dilution of internal Na+ and Cl− as well as other ions. Alterations in transporter activities were further suggested by media and apoplastic acidification, hygromycin B tolerance and changes in relative transcript abundance patterns of various transporters with known functions in salinity tolerance. Engineered tissue succulence might provide an effective strategy for improving WUE, drought avoidance or attenuation, salinity tolerance, and for crassulacean acid metabolism biodesign.

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Opuntia ficus-indica (prickly pear) and Grindelia squarrosa (gumweed) are two exceptionally drought tolerant plant species capable of growing in arid and semiarid environments. Additionally, they have unique cell wall structures. Prickly pear contains pectin and high levels of ash (16.1%) that is predominantly Ca and K. Gumweed has high levels of extractives that contain grindelic acid and monoterpenoids. The objective of this paper was to evaluate how these unique cell wall components alter the pyrolysis performance of prickly pear and gumweed. Using a tandem micropyrolyzer with GC-MS/FID/TCD, a detailed account of the product slate is given for products generated between 450 and 650 °C. Pyrolysis of prickly pear showed that the high levels of ash increase the amount of organics volatilized and shifted product pools, making it possible to generate up to 7.3% carbonyls vs 3.8% for Pinus taeda (loblolly pine) and 10.5% hydrocarbons vs 1.8% for pine depending on reaction conditions. Pyrolysis of gumweed showed that the extractives were volatilized at low temperatures and led to 17.7% grindelic acid and monoterpenoids derivatives in the condensed vapor phase. At high temperatures, the extractives and other biomass components are converted to aromatics and C5–C10 hydrocarbons, giving a total yield of 16.6%, and also generate large amounts of C2–C4 hydrocarbons, 11.3%.

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