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Pereskia and the origin of the cactus life-form.

The cactus life-form is cited as an example of a tight relationship between organism form and function: a succulent, long-lived, photosynthetic stem allows cacti to survive long periods of drought while maintaining a positive tissue water status. Pereskia (Cactaceae) comprises 17 species of leafy shrubs and trees that are thought to represent the original cactus condition. Recent phylogenetic work has shown that there are two separate clades of Pereskia species, which are basal and paraphyletic with respect to the rest of the cacti. We selected seven Pereskia species, representing both clades, and characterized their water relations by measuring a suite of physiological traits in wild populations. Additionally, we estimated basic climate parameters from collection localities for all 17 Pereskia species. Extant Pereskia species exhibit ecological water use patterns that are very similar to those of the leafless, stem-succulent cacti. Ancestral trait reconstruction for the physiological and environmental data provides a preliminary assessment of the ecology and water relations of the earliest cacti and suggests that several key elements of the cactus ecological niche were established before the evolution of the cactus life-form. We interpret these ecological traits as potentially important drivers of evolutionary innovation in the cacti.

Biological Evolution↗

Ecological physiology of Pereskia guamacho, a cactus with leaves.

The specialized physiology of leafless, stem-succulent cacti is relatively well understood. This is not true, however, for Pereskia (Cactaceae), the 17 species of leafy trees and shrubs that represent the earliest diverging lineages of the cacti. Here we report on the water relations and photosynthesis of Pereskia guamacho, a small tree of the semiarid scrubland of Venezuela's Caribbean coast. Sapwood-specific xylem conductivity (Ksp) is low when compared to other vessel-bearing trees of tropical dry systems, but leaf-specific xylem conductivity is relatively high due to the high Huber value afforded by P. guamacho's short shoot architecture. P. guamacho xylem is not particularly vulnerable to drought-induced cavitation, especially considering the high leaf water potentials maintained year round. This is confirmed by the lack of significant variation exhibited in Ksp between wet and dry seasons. In the rainy season, P. guamacho exhibited C3-like patterns of stomatal conductance, but during a prolonged drought we documented nocturnal stomatal opening with a concomitant accumulation of titratable acid in leaves. This suggests that P. guamacho can perform drought-induced crassulacean acid metabolism (CAM photosynthesis), although delta 13C values imply that most carbon is assimilated via the C3 pathway. P. guamacho leaves display very low stomatal densities, and maximum stomatal conductance is low whether stomata open during the day or night. We conclude that leaf performance is not limited by stem hydraulic capacity in this species, and that water use is conservative and tightly regulated at the leaf level.

Biological Evolution↗

Antiinflammatory flavonoids from Opuntia dillenii (Ker-Gawl) Haw. flowers growing in Egypt.

Opuntia dillenii (Ker-Gawl) Haw. (Family Cactaceae), is used in folk medicine as an antidiabetic and antiinflammatory. The antiinflammatory activity of the alcohol extracts of the flowers, fruits and stems was carried out using the carrageenan-induced rat paw oedema model. The analgesic effect of the same extracts was evaluated using electric current as a noxious stimulus. The alcohol extract of the flowers revealed the most potent antiinflammatory effect and a pronounced analgesic action at a dose of 200 mg/kg. Bioassay-guided fractionation of this extract using VLC followed by Sephadex and paper chromatography, afforded three flavonoid glycosides, namely, kaempferol 3-O-alpha-arabinoside, isorhamnetin-3-O-glucoside and isorhamnetin-3-O-rutinoside. Their identification was based on physical, chemical and spectroscopic data.

Analgesics↗

Inhibition of nitric oxide synthase expression in activated microglia and peroxynitrite scavenging activity by Opuntia ficus indica var. saboten.

Activated microglia by neuronal injury or inflammatory stimulation overproduce nitric oxide (NO) by inducible nitric oxide synthase (iNOS) and reactive oxygen species (ROS) such as superoxide anion, resulting in neurodegenerative diseases. The toxic peroxynitrite (ONOO-), the reaction product of NO and superoxide anion further contributes to oxidative neurotoxicity. A butanol fraction obtained from 50% ethanol extracts of Opuntia ficus indica var. saboten (Cactaceae) stem (SK OFB901) and its hydrolysis product (SK OFB901H) inhibited the production of NO in LPS-activated microglia in a dose dependent manner (IC50 15.9, 4.2 microg/mL, respectively). They also suppressed the expression of protein and mRNA of iNOS in LPS-activated microglial cells at higher than 30 microg/mL as observed by western blot analysis and RT-PCR experiment. They also inhibited the degradation of I-kappaB-alpha in activated microglia. Moreover, they showed strong activity of peroxynitrite scavenging in a cell free bioassay system. These results imply that Opuntia ficus indica may have neuroprotective activity through the inhibition of NO production by activated microglial cells and peroxynitrite scavenging activity.

Animals↗

Genetic structure is determined by stochastic factors in a natural population of Drosophila buzzatii in Argentina.

D. buzzatii is a cactophilic species associated with several cactaceae in Argentina. This particular ecological niche implies that this species is faced with a non-uniform environment constituted by discrete and ephemeral breeding sites, which are colonized by a finite number of inseminated females. The genetic consequences of this population structure upon the second chromosome polymorphism were investigated by means of F-statistics in a natural endemic population of Argentina. The present study suggests that differentiation of inversion frequencies in third instar larvae among breeding sites has taken place mainly at random and selection is not operating to determine the structure of this population. The average number of parents breeding on a single pad seems to be similar to the number colonizing Opuntia ficus indica rotting cladodes in Carboneras, a derived population from Spain. There is no significant excess of heterokaryotypes within pads or in the population as a whole. The results obtained in the present study suggest that the potential role of selective versus stochastic factors relative to the among pad heterogeneity in the population here studied is different from that of the Spanish population previously reported. Potential mechanisms responsible for these differences are discussed.

Animals↗

Extensive ribosomal DNA genic variation in the columnar cactus Lophocereus.

Sequence analysis of the hypervariable internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA) is commonly used to gain insights into plant and animal population structure and phylogeny. We characterized ITS1, ITS2, and the 5.8S coding region of 18 senita (Lophocereus) individuals from 12 different populations in Baja as well as from closely related cactus species. Analyses of multiple clones demonstrated extensive paralogy in the senita rDNA gene family. We identified at least two putatively non-recombining rDNA operons in senita as well as multiple paralogous sequences within each operon. Usage of PCR, reverse transcriptase (RT)-PCR, Southern blot, primary sequence analyses of the 18S rDNA gene, and secondary structure analyses of the 5.8S rRNA showed that one of the operons encodes rDNA pseudogenes in a low copy-number (Truncated), whereas the second operon encodes an expressed rRNA (Functional). Surprisingly, we found extensive paralogy not only in the ITS regions but also in the 5.8S coding regions in senita both within and between operons. Phylogenetic analyses suggest that the second rDNA operon originated prior to the divergence of Lophocereus. A significant (p < 0.05) divergence-rate acceleration was found in the Lophocereus 5.8S rDNA coding region in the Functional operon in comparison to Pereskiopsis porteri (Cactaceae) and Portulaca molokiniensis (Portulacaceae) with Silene dioica and Spinacia oleracea as the outgroups.

Base Sequence↗

Species variation in the intracellular localization of pyruvate, Pi dikinase in leaves of crassulacean-acid-metabolism plants: an immunogold electron-microscope study.

In malic enzyme-dependent crassulacean-acid-metabolism (ME-CAM) plants, malic acid is decarboxylated by NADP-ME and NAD-ME and generates pyruvate with CO2. Pyruvate is phosphorylated to phosphoenolpyruvate by pyruvate, Pi dikinase (PPDK) and is then conserved in gluconeogenesis. Although PPDK was considered to be located in chloroplasts (e.g., Mesembryanthemum crystallinum), it has recently been found to accumulate in both the chloroplasts and the cytosol in two Kalanchoë species. In this study, the intracellular localization of PPDK was investigated in 22 ME-CAM species in 13 genera of 5 families by immunogold labeling and electron microscopy. This revealed that the pattern of intracellular localization of PPDK varies among the ME-CAM plants and is divided into three types: Chlt, in which PPDK accumulates only in the chloroplasts; Cyt-Chlt, in which PPDK accumulates in both chloroplasts and cytosol; and Cyt, in which PPDK accumulates predominantly in the cytosol. Members of a particular genus tend to have a common PPDK-localization type. In the Cactaceae, all species from seven genera were classified as Cyt. The photosynthetic tissues of all ME-CAM species, including the Cyt type, had substantial PPDK activity, suggesting that PPDK in the cytosol is active and probably plays a functional role. In the Chlt species, NADP-ME activity was relatively greater than NAD-ME activity. In the Cyt-Chlt and Cyt species, however, either the activity of NAD-ME was higher than that of NADP-ME or they were approximately the same. The species variation in the intracellular localization of PPDK is discussed in relation to CAM function and to molecular and phylogenetic aspects.

Blotting, Western↗

How prevalent is crassulacean acid metabolism among vascular epiphytes?

The occurrence of crassulacean acid metabolism (CAM) in the epiphyte community of a lowland forest of the Atlantic slope of Panama was investigated. I hypothesized that CAM is mostly found in orchids, of which many species are relatively small and/or rare. Thus, the relative proportion of species with CAM should not be a good indicator for the prevalence of this photosynthetic pathway in a community when expressed on an individual or a biomass basis. In 0.4 ha of forest, 103 species of vascular epiphytes with 13,099 individuals were found. As judged from the C isotope ratios and the absence of Kranz anatomy, CAM was detected in 20 species (19.4% of the total), which were members of the families Orchidaceae, Bromeliaceae, and Cactaceae. As predicted, the contribution of CAM epiphytes to the total number of individuals and to total biomass (69.6 kg ha(-1)) was considerably lower (3.6% or 466 individuals and, respectively, 3.0% or 2.1 kg ha(-1)).

Biomass↗

Presence of arbuscular mycorrhizal fungi in South Florida native plants.

The roots of 27 species of South Florida plants in 15 families (including one cycad, six palms, one Smilax, and 19 dicotyledons) native to pine rockland and tropical hardwood hammock communities were examined for arbuscular mycorrhizal fungi (AMF). These plants grow in the biologically diverse but endangered Greater Everglades habitat. Roots from field-grown and potted plants were cleared and stained. All 27 species had AMF and include 14 species having an endangered or threatened status. The Paris-type colonization occurred in two species in the families Annonaceae and Smilacaceae. The Arum-type occurred in 22 species in the families Anacardiaceae, Arecaceae (Palmae), Boraginaceae, Cactaceae (questionable), Euphorbiaceae, Fabaceae, Lauraceae, Melastomataceae, Polygalaceae, Rubiaceae, Simaroubaceae, Ulmaceae, and Zamiaceae. Three species in the families Fabaceae, Lauraceae, and Simaroubaceae had a mix of Paris- and Arum-types. The results have implications for the restoration of these endangered plant communities in the Everglades.

Biodiversity↗

Wide-band tracheids in genera of Portulacaceae: novel, non-xylary tracheids possibly evolved as an adaptation to water stress.

Wide-band tracheids (WBTs) are novel tracheids with wide, lignified secondary walls that intrude deeply into the cell lumen when viewed in transverse sections. These tracheids are found in a few genera in related families in the order Caryophyllales: Aizoaceae, Cactaceae, and Portulacaceae. WBTs in these three families vary in (1) systematic occurrence (found in more highly derived genera in each family), (2) location in plant organs, and (3) structure and dimensions. In addition, an analysis was conducted of WBT cell walls to test the hypothesis that WBTs evolved as an adaptation to water stress (i.e., the wide secondary walls should prevent collapse of the primary wall during water stress). The cell wall data show that primary cell walls in WBTs cannot inwardly collapse to occlusion, thus providing support for the water stress hypothesis of WBT evolution. With consideration of their systematic occurrence, the molecular phylogenetic data, and data here showing support for a water stress adaptive origin, it is logical to assume that WBTs evolved in genera that were adapting to environments undergoing a rapid trend toward aridification.

Adaptation, Physiological↗

Purification and partial characterization of two lectins from the cactus Machaerocereus eruca.

Two lectins (MEAI and MEAII) were isolated from the cactus Machaerocereus eruca by affinity chromatography on mucin-Sepharose and partially characterized with respect to their biochemical and carbohydrate binding properties. Both are oligomeric glycoproteins consisting of 35 kDa monomers. Amino acid analysis indicates that both lectins have similar composition with high amounts of glycine, glutamic acid and serine. MEAI and MEAII contain approximately 36 and 24% (w/w) of carbohydrates, respectively. They agglutinate erythrocytes from several animal species. Binding specificity was directed to galactose-containing oligosaccharides and glycopeptides. The M. eruca lectins are the first lectins to be isolated from a species belonging to the plant family of Cactaceae.

Amino Acids↗

Comparative anti-hyperglycemic potentials of medicinal plants.

Validation of the ethnobotanical use of the leaves of Artemisia vulgaris Linn. (Compositae), Eucalyptus tereticornis Sm. (Myrtaceae), Solanum nigrum Linn. (Solanaceae), and Vitex negundo Linn. (Verbenaceae); stems of Nopalea cochinellifera (Linn.) Salm-Dyck (Cactaceae); roots of Imperata cylindrica Beauv. (Gramineae); dried bark of Syzygium cumini (Linn.) Skeels (Myrtaceae) as anti-diabetic agents using the oral glucose tolerance test showed that only the bark of Syzygium cumini and the leaves of Vitex negundo and Eucalyptus tereticornis exhibited anti-hyperglycemic activities when fed simultaneously with glucose. At the same dosages of 5 mg/20 g mouse, Syzygium cumini-treated mice showed a significant decrease in blood glucose levels (BGLs) at 30 min (alpha=0.10) and from 45 min onwards at alpha=0.05. Vitex negundo exhibited greater anti-hyperglycemic activity than Eucalyptus tereticornis. Both showed a significant decrease in BGLs at 60 min but at alpha=0.05 for Vitex negundo and at alpha=0.07 for Eucalyptus tereticornis. There was no significant lowering in BGLs for Imperata cylindrica and Solanum nigrum while there was even an increase in BGLs for Nopalea cochinellifera and Artemisia vulgaris.

Animals↗

Color, betalain pattern, and antioxidant properties of cactus pear (Opuntia spp.) clones.

Total phenolics, ascorbic acid, and betalain contents of differently colored cactus pear clones (nine Opuntia ficus-indica [L.] Mill. clones and one O. robusta Wendl. clone) were investigated and related to their respective antioxidant potential assessed by Trolox-equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays. TEAC and ORAC values were very highly correlated with each other and also with values for total phenolics, betalain contents, and ascorbic acid concentrations. Total phenolics had the greatest contribution to ORAC and TEAC values. High-performance liquid chromatography (HPLC)-diode array detector (DAD)-tandem mass spectrometry (MS/MS) measurements of cactus pear juices permitted the differentiation of the clones based on variations in pigment patterns and betalain concentrations. The red and yellow betalains were absent in lime green colored cactus fruits. The ratio and concentration of these pigments were responsible for the yellow, orange, red, and purple colors in the other clones. Progeny of purple and lime green colored parents were characterized by 12% and 88% of plants bearing lime green and purple fruit, respectively. This implies that the genes for betalain production were lacking in the lime green fruits but could be provided by a parent with a complete set of genes, that is, purple fruits. Besides known pigments typical of Cactaceae, two unexpected betalains were identified. Whereas gomphrenin I was found for the first time in tissues of cactus plants, methionine-betaxanthin has never been described before as a genuine betalain. In addition to their alleged health-promoting properties, various combinations of yellow betaxanthins and red-purple betacyanins may allow the development of new food products without using artificial colorants.

Antioxidants↗

Identification of new cactus alkaloids in Backebergia militaris by tandem mass spectrometry.

Tandem mass spectrometry, applied to simple extracts of Backebergia militaris, indicated the presence of a number of new alkaloids, including fully aromatic oxygenated isoquinolines, their di- and tetra-hydro analogs, and beta-phenethylamines. These conclusions were supported by separation using radial tlc and comparison with authentic compounds. Traces of seven alkaloids new to this cactus species were identified; four (3,4,8,11) were known previously from other cacti. Three novel cactus alkaloids were identified and confirmed by synthesis as 7,8-dimethoxy-3,4-dihydroisoquinoline (12, dehydrolemaireocereine), 6,7-dimethoxyisoquinoline (13, backebergine), and 7,8-dimethoxyisoquinoline (14, isobackebergine). The last two compounds are the first simple, fully aromatic, isoquinoline alkaloids to be reported from the Cactaceae. The sensitivity of this approach to new alkaloid discovery is emphasized; the entire project consumed only 10 g of dried plant material.

Alkaloids↗

Effects of temperature on the hill reaction and photophosphorylation in isolated cactus chloroplasts.

Chloroplasts isolated from Opuntia polyacantha Haw. (Cactaceae) are capable of noncyclic electron transport and ATP synthesis. Hill reaction rates, measured by O(2) evolution or by ferricyanide reduction, increase with increasing temperature to approximately 40 C. The temperature optimum of NADP reduction is 42 C while the optimum for noncyclic photophosphorylation is 35 C. NADP-linked phosphorylation exhibits a higher coupling ratio (P/e(2)) than ferricyanide-linked photophosphorylation. The temperature optima for photochemical energy production correlate with photosynthetic properties of Crassulacean acid metabolism (CAM) plants and are discussed in relation to the operation of CAM at high tissue temperature.

Journal Article↗

Water Relations, Diurnal Acidity Changes, and Productivity of a Cultivated Cactus, Opuntia ficus-indica.

Physiological responses of the Crassulacean acid metabolism (CAM) plant Opuntia ficus-indica (Cactaceae) were studied on a commercial plantation in central Chile. Young cladodes (flattened stems) and flower buds exhibited daytime stomatal opening, whereas mature cladodes and fruit exhibited the nocturnal stomatal opening characteristic of CAM plants. Severe water stress suppressed the nocturnal stomatal opening by mature cladodes, but their high water vapor conductance occurring near dawn was not affected. Nocturnal acidity increases were not as sensitive to water stress as was the nocturnal stomatal opening. The magnitude of the nocturnal acidity increases depended on the total daily photosynthetically active radiation (PAR), being 90% PAR-saturated at 27 moles per square meter per day for a mean nighttime air temperature of 5 degrees C and at 20 moles per square meter per day for 18 degrees C. Inasmuch as the PAR received on unshaded vertical surfaces averaged about 21 moles per square meter per day, nocturnal acidity increases by the cladodes were on the verge of being PAR-limited in the field. The net assimilation rate, which was positive throughout the year, annually averaged 3.4 grams per square meter per day for 1.0- and 2.0-year-old plants. Plants that were 5.4 years old had 7.2 square meters of cladode surface area (both sides) and an annual dry weight productivity of 13 megagrams (metric tons) per hectare per year when their ground cover was 32%. This substantial productivity for a CAM plant was accompanied by the highest nocturnal acidity increase so far observed in the field, 0.78 mole H(+) per square meter.

Journal Article↗

Day-Night Variations in Malate Concentration, Osmotic Pressure, and Hydrostatic Pressure in Cereus validus.

Malate concentration and stem osmotic pressure concomitantly increase during nighttime CO(2) fixation and then decrease during the daytime in the obligate Crassulacean acid metabolism (CAM) plant, Cereus validus (Cactaceae). Changes in malate osmotic pressure calculated using the Van't Hoff relation match the changes in stem osmotic pressure, indicating that changes in malate level affected the water relations of the succulent stems. In contrast to stem osmotic pressure, stem water potential showed little day-night changes, suggesting that changes in cellular hydrostatic pressure occurred. This was corroborated by direct measurements of hydrostatic pressure using the Jülich pressure probe where a small oil-filled micropipette is inserted directly into chlorenchyma cells, which indicated a 4-fold increase in hydrostatic pressure from dusk to dawn. A transient increase of hydrostatic pressure at the beginning of the dark period was correlated with a short period of stomatal closing between afternoon and nighttime CO(2) fixation, suggesting that the rather complex hydrostatic pressure patterns could be explained by an interplay between the effects of transpiration and malate levels. A second CAM plant, Agave deserti, showed similar day-night changes in hydrostatic pressure in its succulent leaves. It is concluded that, in addition to the inverted stomatal rhythm, the oscillations of malate markedly affect osmotic pressures and hence water relations of CAM plants.

Journal Article↗

Biogeography of the yeasts of ephemeral flowers and their insects.

We studied specific yeast communities vectored by beetles, drosophilids, and bees that visit ephemeral flowers, mostly in the genus Hibiscus and in the families Convolvulaceae and Cactaceae, in the Neotropical, Nearctic, and Australian biogeographic regions. The communities consist mostly of yeasts in four clades centered around the genera Metschnikowia, Kodamaea, Wickerhamiella, and Starmerella. The largest geographic discontinuity occurs as a function of the nitidulid beetle species that dominate the non-pollinator insect visitors of the flowers. This partitions the New World, where the dominant beetle is in the genus Conotelus, from the Australian biogeographic region, dominated by species of Aethina. Distinct but sympatric insects may also carry radically different yeast communities.

Animals↗