Rat- and bat-pollination of Mucuna championii (Fabaceae) in Hong Kong

Mucuna (Fabaceae) species possess gullet-type flowers that open explosively and which are thought to be specifically adapted for bat- or bird-pollination. However, recent studies have shown that non-flying mammals are also important pollinators of this genus in Asia. Here we report on the pollination system of Mucuna championii (endemic in southeast China) in Hong Kong. As is typical for the genus, explosive opening is essential for fruit set, but flowers are unable to open in the absence of manipulation by an effective pollinator. Camera trap surveys of three individuals revealed both chestnut spiny rats (Niviventer fulvescens) and short-nosed fruit bats (Cynopterus sphinx) to be capable of triggering explosive opening. The number of flowers opened by each species did not differ significantly, and both removed most pollen grains from the flowers they visited, but either species visited different individuals. Sucrose-rich nectar was secreted by flowers throughout the day. Our results reveal that M. championii can be pollinated by both rats and bats, with this representing only the second report of rat-pollination in tropical Asia. The sympatric M. birdwoodiana often occurs in close proximity to M. championii and has an overlapping flowering season, suggesting that pollinator segregation may have played a role in shaping the evolutionary ecology of these two species.     

The pollination system of the widely distributed mammal‐pollinated Mucuna macrocarpa (Fabaceae) in the tropics

Although the pollinators of some plant species differ across regions, only a few mammal‐pollinated plant species have regional pollinator differences in Asia. Mucuna macrocarpa (Fabaceae) is pollinated by squirrels, flying foxes, and macaques in subtropical and temperate islands. In this study, the pollination system of M. macrocarpa was identified in tropical Asia, where the genus originally diversified. This species requires “explosive opening” of the flower, where the wing petals must be pressed down and the banner petal pushed upward to fully expose the stamens and pistil. A bagging experiment showed that fruits did not develop in inflorescences (n = 66) with unopened flowers, whereas fruits developed in 68.7% of inflorescences (n = 131) with opened flowers. This indicated that the explosive opening is needed for the species to reproduce. Four potential pollinator mammals were identified by a video camera‐trap survey, and 78.3% and 60.1% of monitored inflorescences (n = 138) were opened by gray‐bellied squirrels (Callosciurus caniceps) and Finlayson's squirrels (C. finlaysonii), respectively, even though more than 10 mammal species visited flowers. Nectar was surrounded by the calyx, and the volume and sugar concentration of secreted nectar did not change during the day. This nectar secretion pattern is similar to those reported by previous studies in other regions. These results showed that the main pollinators of M. macrocarpa in the tropics are squirrels. However, the species' nectar secretion pattern is not specifically adapted to this particular pollinator. Pollinators of M. macrocarpa differ throughout the distribution range based on the fauna present, but there might not have been no distinctive changes in the attractive traits that accompanied these changes in pollinators.     

Civet pollination in Mucuna birdwoodiana (Fabaceae: Papilionoideae)

Plant Ecology - Inferring pollinators from floral traits is problematic because many flowers do not conform to a prescribed phenotype by which they may be assigned to any one particular pollination...     

Floral traits of mammal‐pollinated Mucuna macrocarpa (Fabaceae): Implications for generalist‐like pollination systems

Floral traits are adapted by plants to attract pollinators. Some of those plants that have different pollinators in different regions adapt to each pollinator in each region to maximize their pollination success. Mucuna macrocarpa (Fabaceae) limits the pollinators using its floral structure and is pollinated by different mammals in different regions. Here, we examine the relationships between floral traits of M. macrocarpa and the external morphology of mammalian pollinators in different regions of its distribution. Field surveys were conducted on Kyushu and Okinawajima Island in Japan, and in Taiwan, where the main pollinators are the Japanese macaque Macaca fuscata, Ryukyu flying fox Pteropus dasymallus, and red‐bellied squirrel Callosciurus erythraeus, respectively. We measured the floral shapes, nectar secretion patterns, sugar components, and external morphology of the pollinators. Results showed that floral shape was slightly different among regions and that flower sizes were not correlated with the external morphology of the pollinators. Volume and sugar rate of nectar were not significantly different among the three regions and did not change throughout the day in any of the regions. However, nectar concentration was higher in Kyushu than in the other two regions. These results suggest that the floral traits of M. macrocarpa are not adapted to each pollinator in each region. Although this plant limits the number of pollinators using its flower structure, it has not adapted to specific mammals and may attract several species of mammals. Such generalist‐like pollination system might have evolved in the Old World.     

Pollination partners of Mucuna macrocarpa (Fabaceae) at the northern limit of its range

Mucuna macrocarpa is a plant found in tropical and subtropical regions that requires an “explosive opening.” Explosive opening is the process that exposes the stamen and pistil from the opening of the carina. This process is needed for cross pollination; however, the plant cannot open itself and opening by an animal is needed. The most common opener of Mucuna flowers is several nectar‐eating bats (e.g., Syconycteris), but the flying fox, Pteropus dasymallus, is the only opener of M. macrocarpa on the subtropical island of Okinawajima. Here, we present the explosive openers and possible pollinators in the northernmost and temperate Kamae region, Kyushu, Japan, where nectar‐eating bats are absent. The Japanese macaque, Macaca fuscata, and the Japanese marten, Martes melampus, were the explosive openers observed during our survey in Kamae. Martens opened flowers using their snout in a manner similar to that of the flying fox, whereas macaques opened flowers using their hands. This is the first time that an animal has been observed opening these flowers with its hands rather than snout. In total, 97% (n = 283) of explosively opened flowers were opened by macaques, and the macaque largely contributed to the overall flower opening. Because many pollen grains become attached to the explosive openers, they are considered to be primary pollinators. Furthermore, two bee species, Apis cerana japonica and Bombus ardens ardens, also visited opened flowers and collected pollen, and they were possibly secondary pollinators.     

First record of non‐flying mammalian contributors to pollination in a tropical montane forest in Asia

This study aims to identify the flower visitors of Mucuna thailandica (Fabaceae), endemic plant species in montane forests in Thailand, to determine their potential pollinators. The genus Mucuna produces papilionaceous flowers and has an explosive flower‐opening step. Explosive opening rapidly exposes stamens and pistil from keel petals and releases pollen. The flower of this species depends completely on animals to perform this step, essential for pollination success. Using a camera trap survey, we revealed that non‐flying mammals, such as squirrels (Callosciurus sp.) and masked palm civets (Paguma larvata), opened flowers explosively. Thus, these mammals contribute to the pollination of M. thailandica. This is the first report of non‐flying mammals contributing to pollination in montane forests in tropical Asia.     

Evidence of rodent pollination inCajophora coronata (Loasaceae)

Vegetative and floral features ofCajophora coronata (Loasaceae) suggest adaptations to pollination by rodents: (1) mammal guard represented by a covering of stinging hairs; (2) geoflory; (3) white corolla; (4) open flowers with copious low concentration nectar; (5) abundant pollen; (6) maximum pollen and nectar presentation in the afternoon hours and in the night. Palynological analysis revealed pollen loads ofC. coronata on the nostrils and whiskers of captured rodents (Graomys griseoflavus, fam.Muridae). Pollen and anther remains were also found in faeces sampled in the surroundings. Additional evidence includes rodent footprints obtained by placing smoked plates beneath the flowers, which revealed flower visitation during the night. These observations are to our knowledge the first evidence of flower visitation by rodents in South America and the first in the New World outside the range of flower bats and bat flowers.     

Study on the Chemical Constituens of Mucuna pruriens var. utilis (With Black seed Coat)

Five compounds were first isolated from the legumes of Mucuna pruriens (L.) DC. var. utilis (Wall. ex Wight) Baker ex Burck (M. atrocarpum Metcalf) with black seed coat. They were identified as L-dopa, stizolamine, D-erythro-neopterin, 6-hydroxymethylpterin and isoxanthopterin.     

Floral rewards and pollination in Cytiseae (Fabaceae)

 Cytiseae have been reported to be mostly nectar-lacking, yet some taxa secrete nectar from extrastaminal nectaries. We studied the pollination biology of four shrubby species of Cytiseae (Cytisophyllum sessilifolium (L.) Lang, Spartium junceum L., Genista radiata (L.) Scop., Genista cilentina Valsecchi) which differ for ecology, distribution and population size. All species resulted obliged xenogamous, insect visits being necessary for successful pollination. Bumblebees and solitary bees are the principal pollinators, but also many beetles, some hover-flies, and few bugs visit flowers. Pollinator specificity is low, and this may be the reason of the scarce seed set compared to the number of ovules. Pollen is the main reward, but traces of glucose were detected in all species, at the base of vexillum or on the reproductive column. Nectar production is irregular in time, and apparently unpredictable. We suppose that nectar may play a role in attracting pollinators determining their right position for a successful pollination. Received August 12, 2002; accepted November 25, 2002 Published online: June 2, 2003     

Spatiotemporal Evolution of Calophaca (Fabaceae) Reveals Multiple Dispersals in Central Asian Mountains

Background

The Central Asian flora plays a significant role in Eurasia and the Northern Hemisphere. Calophaca, a member of this flora, includes eight currently recognized species, and is centered in Central Asia, with some taxa extending into adjacent areas. A phylogenetic analysis of the genus utilizing nuclear ribosomal ITS and plastid trnS-trnG and rbcL sequences was carried out in order to confirm its taxonomic status and reconstruct its evolutionary history.

Methodology/Principal Finding

We employed BEAST Bayesian inference for dating, and S-DIVA and BBM for ancestral area reconstruction, to study its spatiotemporal evolution. Our results show that Calophacais monophyletic and nested within Caragana. The divergence time of Calophaca is estimated at ca. 8.0 Ma, most likely driven by global cooling and aridification, influenced by rapid uplift of the Qinghai Tibet Plateau margins.

Conclusions/Significance

According to ancestral area reconstructions, the genus most likely originated in the Pamir Mountains, a global biodiversity hotspot and hypothesized Tertiary refugium of many Central Asian plant lineages. Dispersals from this location are inferred to the western Tianshan Mountains, then northward to the Tarbagatai Range, eastward to East Asia, and westward to the Caucasus, Russia, and Europe. The spatiotemporal evolution of Calophaca provides a case contributing to an understanding of the flora and biodiversity of the Central Asian mountains and adjacent regions.     

Beyond the pollination syndrome: nectar ecology and the role of diurnal and nocturnal pollinators in the reproductive success of Inga sessilis (Fabaceae)

Inga species present brush‐type flower morphology allowing them to be visited by distinct groups of pollinators. Nectar features in relation to the main pollinators have seldom been studied in this genus. To test the hypothesis of floral adaptation to both diurnal and nocturnal pollinators, we studied the pollination ecology of Inga sessilis, with emphasis on the nectar secretion patterns, effects of sequential removals on nectar production, sugar composition and the role of diurnal and nocturnal pollinators in its reproductive success. Inga sessilis is self‐incompatible and pollinated by hummingbirds, hawkmoths and bats. Fruit set under natural conditions is very low despite the fact that most stigmas receive polyads with sufficient pollen to fertilise all ovules in a flower. Nectar secretion starts in the bud stage and flowers continually secreting nectar for a period of 8 h. Flowers actively reabsorbed the nectar a few hours before senescence. Sugar production increased after nectar removal, especially when flowers were drained during the night. Nectar sugar composition changed over flower life span, from sucrose‐dominant (just after flower opening, when hummingbirds were the main visitors) to hexose‐rich (throughout the night, when bats and hawkmoths were the main visitors). Diurnal pollinators contributed less than nocturnal ones to fruit production, but the former were more constant and reliable visitors through time. Our results indicate I. sessilis has floral adaptations, beyond the morphology, that encompass both diurnal and nocturnal pollinator requirements, suggesting a complementary and mixed pollination system.     

Evolution of floral morphology and pollination system in Bignonieae (Bignoniaceae)

The radiation of angiosperms is associated with shifts among pollination modes that are thought to have driven the diversification of floral forms. However, the exact sequence of evolutionary events that led to such great diversity in floral traits is unknown for most plant groups. Here, we characterize the patterns of evolution of individual floral traits and overall floral morphologies in the tribe Bignonieae (Bignoniaceae). We identified 12 discrete traits that are associated with seven floral types previously described for the group and used a penalized likelihood tree of the tribe to reconstruct the ancestral states of those traits at all nodes of the phylogeny of Bignonieae. In addition, evolutionary correlations among traits were conducted using a maximum likelihood approach to test whether the evolution of individual floral traits followed the correlated patterns of evolution expected under the "pollination syndrome" concept. The ancestral Bignonieae flower presented an Anemopaegma-type morphology, which was followed by several parallel shifts in floral morphologies. Those shifts occurred through intermediate stages resulting in mixed floral morphologies as well as directly from the Anemopaegma-type morphology to other floral types. Positive and negative evolutionary correlations among traits fit patterns expected under the pollination syndrome perspective, suggesting that interactions between Bignonieae flowers and pollinators likely played important roles in the diversification of the group as a whole.     

Evolution of secondary metabolites in legumes (Fabaceae)

Legumes produce a high diversity of secondary metabolites which serve as defence compounds against herbivores and microbes, but also as signal compounds to attract pollinating and fruit-dispersing animals. As nitrogen-fixing organisms, legumes produce more nitrogen containing secondary metabolites than other plant families. Compounds with nitrogen include alkaloids and amines (quinolizidine, pyrrolizidine, indolizidine, piperidine, pyridine, pyrrolidine, simple indole, Erythrina, simple isoquinoline, and imidazole alkaloids; polyamines, phenylethylamine, tyramine, and tryptamine derivatives), non-protein amino acids (NPAA), cyanogenic glucosides, and peptides (lectins, trypsin inhibitors, antimicrobial peptides, cyclotides). Secondary metabolites without nitrogen are phenolics (phenylpropanoids, flavonoids, isoflavones, catechins, anthocyanins, tannins, lignans, coumarins and furanocoumarins), polyketides (anthraquinones), and terpenoids (especially triterpenoid, steroidal saponins, tetraterpenes). While some secondary metabolites have a wide distribution (flavonoids, triterpenes, pinitol), however, others occur in a limited number of taxa. The distributions of secondary metabolites with an irregular occurrence are mapped on a molecular phylogeny of the Fabaceae, reconstructed from a combined data set of nucleotide sequences from rbcL, matK and ITS genes. In most cases, the distribution patterns of secondary metabolites do not agree with the phylogeny of the plants producing them. In contrary, the distribution of many secondary metabolites is patchy and irregular. Thus, the use of phytochemical data to reconstruct a phylogeny of plants is often not informative and can be misleading. The patchy distribution may be due to convergent evolution, a contribution of endophytic fungi or more likely, to an early acquisition of the key genes of secondary metabolism in the evolution of land plants among others by horizontal gene transfer from bacteria. Thus it would be a matter of gene regulation whether these genes are active in some but not all taxa.     

Evolution,pollination, and systematics of the tribeNeottieae (Orchidaceae)

The various classifications of the orchid tribeNeottieae are reviewed and a new classification is proposed that divides the tribe into three subtribes,Neottiinae, Limodorinae, andCephalantherinae, based primarily on characters of the column (gynostemium). A cladistic analysis illustrates that these three subtribes are more closely related to one another than either is to any other group in subfam.Neottioideae, although there are very few apomorphic characters for the tribe. Pollination biology is also discussed showing links between breeding systems and distribution. There is also a possible role between column and labellum morphology and the emergence of a deceptive pollination syndrome from one of reward.     

Specialized pollination by carpenter bees in Calanthe striata (Orchidaceae), with a review of carpenter bee pollination in orchids

Calanthe striata has nectarless flowers that are self‐compatible, but pollinator dependent. Field observations showed that the flowers were pollinated exclusively by the carpenter bee Xylocopa appendiculata circumvolans, although the bees occasionally wasted pollen by delivering to the stigmatic surface pollinaria that retained their anther caps. Fruit set ratios at the population level varied spatiotemporally, but were generally low (8.3–17.3%). Calanthe striata blooms in spring when post‐overwintering carpenter bees have not yet started foraging for brood production. It can therefore exploit an abundance of opportunistic/naïve foragers. This timing may also increase the possibility of pollinator visits, because no rewarding co‐flowering plants are available in the orchid habitats. A literature review of Orchidaceae pollinated by carpenter bees revealed that at least 14 species of Orchidoideae and Epidendroideae have evolved flowers specialized for carpenter bee pollination. They typically have shallow pink/magenta flowers with a foothold for pollinators; pollinaria are attached to the head, ventral thorax or base of the middle legs of carpenter bees when they insert their heads and/or proboscises into flowers; pollination success is generally low, a probable consequence of the deceptive pollination systems. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013 , 171 , 730–743.     

Floral ontogeny in Dipterygeae (Fabaceae) reveals new insights into one of the earliest branching tribes in papilionoid legumes

Flowers of Dipterygeae (Fabaceae, Papilionoideae) exhibit an unusual petaloid calyx. The two adaxial sepals are large and petaloid, and the three abaxial sepals form a three‐toothed lobe. The goal of this study was to elucidate the ontogenetic pathways of this peculiar calyx in light of the floral development of the three genera that comprise the tribe. Floral buds of Dipteryx alata, Pterodon pubescens and Taralea oppositifolia were analysed using scanning electron microscopy and light microscopy. The order of bracteole and sepal initiation varies among the species. The androecium is asymmetric. The carpel cleft is positioned to the right or to the left, and is opposite the adaxial antepetalous stamen. The peculiarity of the calyx becomes noticeable in the intermediate stages of floral development. It results from the differential growth of the sepal primordia, in which the abaxial and lateral primordia remain diminutive during floral development, compared with the adaxial ones that enlarge and elongate. Bracteoles, abaxial sepals, petals and anthers are appendiculate, except in T. oppositifolia, in which the appendices were not found in bracteoles or anthers. These appendices comprise secretory canals or cavities. Considering that the ontogenetic pathway for the formation of the petaloid calyx is similar and exclusive for Dipterygeae, it might be a potential synapomorphy for the group, with the presence of secretory canals in the appendices of abaxial and lateral sepals and petals. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174 , 529–550.     

Isolation and characterization of a Forssman antigen-binding lectin from velvet bean (Mucuna derringiana) seeds

A Forssman antigen (GalNAc1-3GalNAc1-3Gal1-4Gal1-4Glc1-1Cer)-binding lectin has been purified from velvet bean (Mucuna derringiana) seeds by a combination of affinity chromatography and reversed phase HPLC. This lectin agglutinates both native and trypsin-treated sheep erythrocytes as well as trypsinized rabbit erythrocytes, but neither native rabbit nor human erythrocytes, irrespective of blood group type. SDS-PAGE and gel filtration chromatography reveal the lectin to be a homodimer consisting of two 54 kDa subunits linked by non-covalent bonds. The results obtained by quantitative precipitation, haemagglutination inhibition and TLC overlay assays indicate that theMucuna lectin specifically recognizes Forssman antigen and Forssman disaccharide (GalNAc1-3GalNAc)-related structures. Abbreviations: The abbreviations and the trivial names used are: AH, 6-aminohexyl; BSA, bovine serum albumin; Cer, ceramide; HPLC, high performance liquid chromatography; PAGE, polyacrylamide gel electrophoresis; PBS, 10mm phosphate-buffered saline, pH 7,2, containing 0.15m NaCl; PMSF, phenyl methyl sulfonyl fluoride; SDS, sodium dodecyl sulphate; TFA, trifluoroacetic acid; TBS, 20mm tris-buffered saline, pH 7.2; TLC, thin-layer chromatography; A disaccharide, GalNAc1-3Gal; A trisaccharide, GalNAc1-3[Fuc1-2]Gal; Forssman disaccharide, GalNAc1-3GalNAc; CDH (ceramide dihexoside or lactosyl ceramide) Gal1-4Glc1-1Cer (LacCer); CTH (ceramide trihexoside or globotriosyl ceramide), Gal1-4Gal1-4Glc1-1Cer (GbOse₃Cer or Gb₃); globoside (globotetraosyl ceramide), GalNAc1-3Gal1-4Gal1-4Glc1-1Cer (GbOse₄Cer or Gb₄); Forssman antigen (globopentaosyl ceramide), GalNAc1-3GalNAc1-3Gal1-4Gal1-4Glc1-1Cer (GbOse₅Cer).     

Evidence for rodent pollination in Erica hanekomii (Ericaceae)

Pollination by rodents, originally discovered in Cape Proteaceae, is known from a handful of plant lineages and may be more widespread than current data suggest. Here, we demonstrate the occurrence of this mode of pollination in Erica, the largest plant genus in the Cape flora of South Africa. The localized endemic Erica hanekomii appears to be adapted for pollination by rodents, on account of its low, mat‐forming habit with many dull‐coloured, pendulous flower heads, large volumes (up to c. 30 µL per inflorescence) of sucrose‐dominated nectar, hook‐shaped styles and late winter flowering phenology. Rodents trapped in the vicinity of E. hanekomii plants had many thousands of Erica tetrads in their droppings. Nocturnal laboratory observation showed that Acomys subspinosus (Cape spiny mouse) is adept at manipulating E. hanekomii inflorescences and lapping nectar without destroying gynoecia. During terrarium experiments, pollen was deposited mainly on the rostrum, but also on the chest area and forelegs of A. subspinosus. No birds or insects were seen to visit E. hanekomii in the field. Whereas natural fruit set was 73%, inflorescences from which rodents, but not insects, were excluded using mesh cages, had fruit set of just 3.3%. Exclusion of all visitors resulted in zero seed production. Broken anther rings (a sign of flower visitation in Erica) were observed in 90.1% of flowers in the field and 92% of flowers exposed to rodents in the laboratory, but in virtually none of the flowers from which rodents were selectively excluded. This first record of a non‐flying mammal pollination syndrome in Ericaceae raises interesting new questions about the origins of the rodent pollination system and the selective factors that lead to its evolution. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 166 , 163–170.     

Temporal variation of flower scent in Silene otites (Caryophyllaceae): a species with a mixed pollination system

Floral scent composition and emission are often related to olfactory abilities, preferences and activity times of pollinating agents. However, most studies have focused on species with one pollinator type, and little is known about species with mixed pollination systems. We investigated the temporal variation in flower visitor activity and flower scent emission of Silene otites (Caryophyllaceae), a species regarded as being nocturnal and pollinated by moths and mosquitoes. We found, however, that S. otites is not only visited at night by moths and mosquitoes, but also by a wide range of day‐active insects. Furthermore, both the intensity and composition of floral scent change during the course of the day. Total scent emission reached its maximum in the early hours of the night when compounds known to be attractants for moths and/or mosquitoes, such as lilac aldehyde, benzaldehyde, phenylacetaldehyde and 2‐phenylethanol, dominated the scent. However, although less intensive, emission continued during the day. In the early morning, α‐pinene, hotrienol, 2‐phenylethanol and δ‐3‐carene were the dominant scent compounds. (E)‐β‐Ocimene, hotrienol and benzyl alcohol contributed proportions of at least 10% before midday when flies, such as syrphids, were abundant flower visitors, whereas linalool and hotrienol contributed to the scent with at least 10% each in the afternoon when bees also contributed to flower visits. We conclude that, in S. otites, the changing emission patterns are ‘in tune’ with the olfactory abilities, preferences and activity times of its day‐ and night‐active potentially pollinating flower visitors. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169 , 447–460.     

Flower visitors and pollination in the Oriental (Indomalayan) Region

Current knowledge of flower visitors and pollination in the Oriental Region is summarised. Much less is known about pollination than seed dispersal and the coverage of habitats and taxa in the region is very uneven. The available evidence suggests that pollination in lowland forests is dominated by highly social bees (mainly Trigona and Apis species), with beetles probably the next most important group, followed by other bees and flies. In comparison with the better-studied Neotropics, large solitary bees, moths, Lepidoptera and vertebrates are relatively less important. These differences are greatest in the canopy of the lowland dipterocarp forests of Southeast Asia, where they probably reflect the unique temporal pattern of floral resource availability resulting from 'general flowering' at supra-annual intervals. Apis bees (but not Trigona species) are also important in most montane, subtropical and non-forest habitats. Apart from the figs (Ficus spp.), there are few well-documented examples of plant species visited by a single potential pollinator and most plant-pollinator relationships in the region appear to be relatively generalised. The small sizes of most pollinators and the absence of direct human exploitation probably make pollination mutualisms less vulnerable to failure as a result of human impacts than dispersal mutualisms, but more subtle impacts, as a result of altered gene flows, are likely to be widespread. On current evidence, pollination systems in the Oriental Region do not require any specific conservation action, but this review reinforces arguments for making the preservation (or restoration) of habitat connectivity the major focus of Oriental conservation.