Orchid diversity: an evolutionary consequence of deception?

The Orchidaceae are one of the most species-rich plant families and their floral diversity and pollination biology have long intrigued evolutionary biologists. About one-third of the estimated 18,500 species are thought to be pollinated by deceit. To date, the focus has been on how such pollination evolved, how the different types of deception work, and how it is maintained, but little progress has been made in understanding its evolutionary consequences. To address this issue, we discuss here how deception affects orchid mating systems, the evolution of reproductive isolation, speciation processes and neutral genetic divergence among species. We argue that pollination by deceit is one of the keys to orchid floral and species diversity. A better understanding of its evolutionary consequences could help evolutionary biologists to unravel the reasons for the evolutionary success of orchids.     

ISSR and RAPD Markers Assessed Genetic Variation of Aerides maculosum — an Epiphytic Orchid from Goa, India

Aerides maculosum Lindl is one of the most important orchids valued for its beautiful inflorescence/flowers. The present study aimed to understand the level of genetic variation among the populations of A. maculosum using RAPD and ISSR markers. Among the 35 primers tested, 13 RAPD and 6 ISSR primers were selected for the analysis. Total of 101 RAPD and 40 ISSR fragments were generated. High level of polymorphism was recorded in RAPD (90.45%) compared to ISSR (72.85 %). Nei’s average genetic identity values for different populations of A. maculosum- ranged from 0.465 to 0.762 (RAPD), while for ISSR it ranged from 0.475 to 0.975. The present study provides important insights about genetic variation in A. masculosum and may facilitates the conservation and management of this species.     

Orchid Bee (Apidae: Euglossini) Communities in Atlantic Forest Remnants and Restored Areas in Paraná State,Brazil

In this study, we compare orchid bee communities surveyed in four forest remnants of the Atlantic Forest and four reforested areas characterized by seasonal semi-deciduous forest vegetation in different successional stages (mature and secondary vegetation), located in southern Brazil. The sizes of forest remnants and reforested areas varied from 32.1 to 583.9 ha and from 11.3 to 33.3 ha, respectively. All reforested areas were located near one forest remnant. During samplings, totaling nine per study area, euglossine males were attracted to eight scent baits and captured with bait trap and entomological nets. Each forest remnant and its respective reforested area were sampled simultaneously by two collectors. We collected 435 males belonging to nine species of orchid bees distributed in four genera. The number of individuals and species did not differ significantly between different areas, except for a reforested area (size 33.3 ha), which was located far from its respective forest remnant. Our findings also revealed an apparent association between an orchid bee species (Euglossa annectans Dressler 1982) and the most preserved area surveyed in our study, suggesting that this bee is a potential indicator of good habitat quality in recuperating or preserved areas. Our results suggest that reforested habitats located near forest remnants have a higher probability of having reinstated their euglossine communities.     

Mapping of the Interaction Between Agrobacterium tumefaciens and Vanda Kasem’s Delight Orchid Protocorm-Like Bodies

Physical contact between A. tumefaciens and the target plant cell walls is essential to transfer and integrate the transgene to introduce a novel trait. Chemotaxis response and attachment of Agrobacterium towards Vanda Kasem’s Delight (VKD) protocorm-like bodies (PLBs) were studied to analyse the interaction between Agrobacterium and PLB during the transformation event. The study shows that initially A. tumefaciens reversibly attached to PLB surface via polar and lateral mode of adherence followed by the irreversible attachment which involved the production of cellulosic fibril by A. tumefaciens. Cellulosic fibril allows formation of biofilm at the tip of trichome. Contrarily, attachment mutant Escherichia coli strain DH5α was significantly deficient in the attachment process. Spectrophotometric GUS assay showed the mean value of attachment by A. tumefaciens was 8.72 % compared to the negative control E. coli strain DH5α that produced 0.16 %. A. tumefaciens swarmed with sharper and brighter edge when severe wounding was applied to the PLBs producing the highest swarming ratio of 1.46 demonstrating the positive effect of the plant exudates on bacterial movement. The study shows that VKD’s PLBs are the suitable explants for Agrobacterium-mediated transformation since the bacteria expressed higher competency rate.

Electronic supplementary material

The online version of this article (doi:10.1007/s12088-015-0519-7) contains supplementary material, which is available to authorized users.     

Donkey Orchid Symptomless Virus: A Viral ‘Platypus’ from Australian Terrestrial Orchids

Complete and partial genome sequences of two isolates of an unusual new plant virus, designated Donkey orchid symptomless virus (DOSV) were identified using a high-throughput sequencing approach. The virus was identified from asymptomatic plants of Australian terrestrial orchid Diuris longifolia (Common donkey orchid) growing in a remnant forest patch near Perth, western Australia. DOSV was identified from two D. longifolia plants of 264 tested, and from at least one plant of 129 Caladenia latifolia (pink fairy orchid) plants tested. Phylogenetic analysis of the genome revealed open reading frames (ORF) encoding seven putative proteins of apparently disparate origins. A 69-kDa protein (ORF1) that overlapped the replicase shared low identity with MPs of plant tymoviruses (Tymoviridae). A 157-kDa replicase (ORF2) and 22-kDa coat protein (ORF4) shared 32% and 40% amino acid identity, respectively, with homologous proteins encoded by members of the plant virus family Alphaflexiviridae. A 44-kDa protein (ORF3) shared low identity with myosin and an autophagy protein from Squirrelpox virus. A 27-kDa protein (ORF5) shared no identity with described proteins. A 14-kDa protein (ORF6) shared limited sequence identity (26%) over a limited region of the envelope glycoprotein precursor of mammal-infecting Crimea-Congo hemorrhagic fever virus (Bunyaviridae). The putative 25-kDa movement protein (MP) (ORF7) shared limited (27%) identity with 3A-like MPs of members of the plant-infecting Tombusviridae and Virgaviridae. Transmissibility was shown when DOSV systemically infected Nicotiana benthamiana plants. Structure and organization of the domains within the putative replicase of DOSV suggests a common evolutionary origin with ‘potexvirus-like’ replicases of viruses within the Alphaflexiviridae and Tymoviridae, and the CP appears to be ancestral to CPs of allexiviruses (Alphaflexiviridae). The MP shares an evolutionary history with MPs of dianthoviruses, but the other putative proteins are distant from plant viruses. DOSV is not readily classified in current lower order virus taxa.     

Spatial–Temporal Variation in Orchid Bee Communities (Hymenoptera: Apidae) in Remnants of Arboreal Caatinga in the Chapada Diamantina Region, State of Bahia, Brazil

The spatial and temporal distribution of organisms is a fundamental aspect of biological communities. The present study focused on three remnants of arboreal Caatinga in northeastern Brazil between May, 2009 and April, 2010. A total of 627 euglossine males were captured in traps baited with artificial aromatic compounds. The specimens belonged to 14 species and four genera: Euglossa Latreille, Eulaema Lepeletier, Eufriesea Cockerell, and Exaerete Hoffmannsegg. Eulaema nigrita Lepeletier (41.6), Euglossa carolina Nemésio (15.3%), Eulaema marcii Nemésio (13.6%), and Euglossa melanotricha Moure (12.8%) were the most common species sampled. The distribution of collected specimens per fragment was as follows: Braúna (280?ha)??259 individuals belonging to 14 species; Cambuí (179?ha)??161 individuals from eight species; and Pindoba (100?ha)??207 individuals represented by seven species. Braúna had the highest diversity (H???=?1.91) and estimated species richness. The largest fragment was the main source of the observed variation in species richness and abundance, indicating a non-random pattern of spatial distribution. The analysis of environmental factors indicated that seasonal variation in these factors was the principal determinant of species occurrence and abundance.     

Friends or Relatives? Phylogenetics and Species Delimitation in the Controversial European Orchid Genus Ophrys

BACKGROUND AND AIMS: Highly variable, yet possibly convergent, morphology and lack of sequence variation have severely hindered production of a robust phylogenetic framework for the genus Ophrys. The aim of this study is to produce this framework as a basis for more rigorous species delimitation and conservation recommendations. METHODS: Nuclear and plastid DNA sequencing and amplified fragment length polymorphism (AFLP) were performed on 85 accessions of Ophrys, spanning the full range of species aggregates currently recognized. Data were analysed using a combination of parsimony and Bayesian tree-building techniques and by principal co-ordinates analysis. KEY RESULTS: Complementary phylogenetic analyses and ordinations using nuclear, plastid and AFLP datasets identify ten genetically distinct groups (six robust) within the genus that may in turn be grouped into three sections (treated as subgenera by some authors). Additionally, genetic evidence is provided for a close relationship between the O. tenthredinifera, O. bombyliflora and O. speculum groups. The combination of these analytical techniques provides new insights into Ophrys systematics, notably recognition of the novel O. umbilicata group. CONCLUSIONS: Heterogeneous copies of the nuclear ITS region show that some putative Ophrys species arose through hybridization rather than divergent speciation. The supposedly highly specific pseudocopulatory pollination syndrome of Ophrys is demonstrably 'leaky', suggesting that the genus has been substantially over-divided at the species level.     

Why Do Floral Perfumes Become Different? Region-Specific Selection on Floral Scent in a Terrestrial Orchid

Geographically structured phenotypic selection can lead to adaptive divergence. However, in flowering plants, such divergent selection has rarely been shown, and selection on floral signals is generally little understood. In this study, we measured phenotypic selection on display size, floral color, and floral scent in four lowland and four mountain populations of the nectar-rewarding terrestrial orchid Gymnadenia odoratissima in two years. We also quantified population differences in these traits and pollinator community composition. Our results show positive selection on display size and positive, negative, or absence of selection on different scent compounds and floral color. Selection on the main scent compounds was consistently stronger in the lowlands than in the mountains, and lowland plants emitted higher amounts of most of these compounds. Pollinator community composition also differed between regions, suggesting different pollinators select for differences in floral volatiles. Overall, our study is the first to document consistent regional differences in selection on floral scent, suggesting this pattern of selection is one of the evolutionary forces contributing to regional divergence in floral chemical signaling.     

Studies of Mycorrhizal Fungi of Chinese Orchids and Their Role in Orchid Conservation in China—A Review

China has over 1,200 species of native orchids in nearly 173 genera. About one fourth of native species are of horticultural merit. Some species are of Chinese medicinal value. In fact, the demand on orchid species with high Chinese medicinal values such as Gastrodia elata, Dendrobium offcinale, along with demands on species of cultural importance, such as those in the genus of Cymbidium, is a major factor causing wild populations to diminish and in some cases, drive wild populations to the brink of extinction. These market demands have also driven studies on the role of mycorrhizal fungi in orchid seed germination, seedling and adult growth, and reproduction. Most of these mycorrhizal studies of Chinese orchids, however, are published in Chinese, some in medical journals, and thus overlooked by the mainstream orchid mycorrhizal publications. Yet some of these studies contained interesting discoveries on the nature of the mycorrhizal relationships between orchids and fungi. We present a review of some of these neglected publications. The most important discovery comes from the mycorrhizal studies on G. elata, in which the researchers concluded that those fungi species required to stimulate seed germination are different from those that facilitate the growth of G. elata beyond seedling stages. In addition, presence of the mycorrhizal fungi associated with vegetative growth of post-seedling G. elata hindered the germination of seeds. These phenomena were unreported prior to these studies. Furthermore, orchid mycorrhizal studies in China differ from the mainstream orchid studies in that many epiphytic species (in the genus of Dendrobium, as medicinal herbs) were investigated as well as terrestrial orchids (mostly in the genus Cymbidium, as traditional horticultural species). The different responses between epiphytic and terrestrial orchid seeds to fungi derived from roots suggest that epiphytic orchids may have a more general mycorrhizal relationship with fungi than do terrestrial orchid species during the seed germination stage. To date, orchid mycorrhizal research in China has had a strongly commercial purpose. We suggest that this continuing research on orchid mycorrhizal relationships are a solid foundation for further research that includes more rare and endangered taxa, and more in-situ studies to assist conservation and restoration of the endangered orchids. Knowledge on the identities and roles of mycorrhizal fungi of orchids holds one of the keys to successful restoration and sustainable use of Chinese orchids.     

Raising the Sugar Content – Orchid Bees Overcome the Constraints of Suction Feeding through Manipulation of Nectar and Pollen Provisions

Unlike most other bees, the long-tongued orchid bees ingest nectar using suction feeding. Although long tongues allow exploitation of flowers with deep spurs, the energy intake rate is optimal at 10–20% lower nectar sugar concentrations compared to that of lapping bees. This constraint might be compensated by a higher digestive throughput. Additionally, orchid bees might evaporate water from regurgitated droplets of crop contents. We found male Euglossa championi (n = 10) and Euglossa dodsoni (n = 12) to regularly regurgitate droplets of crop content to the base of their proboscis, generating a fluid film between the proximal parts of the galeae, glossa and labial palps. Rhythmic movements of the proboscis may help to increase convection. There was a significant change in sugar concentration between the initially imbibed solution and the resulting crop content (P<0.05) and the time individual bees had engaged in this liquid exposure behavior was positively correlated with the resulting crop sugar concentration. Female Euglossa townsendi and Euglossa viridissima showed the same behavior. Additionally, they manipulated their nectar-enriched pollen provisions for extensive periods of time before deposition in brood cells. The deposited pollen loads (n = 14) showed a significantly higher sugar concentration than the sugar-water available to the bees (P<0.001). Thus, both male and female euglossines show behaviors that promote evaporative water loss from nectar. We suggest that the behaviors have evolved in concert with suction feeding on dilute nectar from deep floral tubes.     

Orchid bees as bio-indicators for organic coffee farms in Costa Rica: does farm size affect their abundance?

The potential of Euglossini bees, especially Euglossa, as biological indicators of organic vs nonorganic coffee farms was studied in Atenas and San Isidro, Alajuela, Costa Rica using 1.8-cineole as lure. Observations were made for three days at each of four farms and complemented with data from a year of observations. Orchid bees were in greater abundance in the organic farms (t-Student test). However, lower abundances suggest that an organic farm may be negatively affected by the proximity of non-organic farms, depending on its size and distance. Orchid bees may be indicators of organic coffee farms.     

The Cold Awakening of <Emphasis Type="Italic">Doritaenopsis</Emphasis> ‘Tinny Tender’ Orchid Flowers: The Role of Leaves in Cold-induced Bud Dormancy Release

Massive flowering of tropical Phalaenopsis orchids is coordinated by the cold-induced release of reproductive bud dormancy. Light and temperature are the two key factors integrated by the dormancy mechanism to both stop and reactivate the meristem development of many other angiosperm species, including fruit trees and ornamental plants. It is well established that leaves and roots play a major role in inducing flower development; however, currently, knowledge of molecular events associated with reproductive bud dormancy release in organs other than the bud is limited. Using differential gene expression, we have shown that the leaves of a hybrid of Phalaenopsis species, Doritaenopsis ‘Tinny Tender’, undergo major metabolic modifications. These changes result in the production of sucrose and amino acids, both of which can sustain bud outgrowth, and auxin and ethylene, which may play important roles in awaking the dormant buds. Intake of abscisic acid and synthesis of the hormone jasmonate may also explain the inhibition of vegetative growth that coincides with bud growth. Interestingly, many genes that were upregulated by cold treatment are homologous for genes involved in flower induction and vernalization in Arabidopsis, indicating that processes regulating flowering induction and those regulating reproductive bud dormancy release may use similar pathways and effector molecules.     

Autophagosomes,phagosomes, autolysosomes,phagolysosomes, autophagolysosomes… Wait,I’m confused

When an autophagosome or an amphisome fuse with a lysosome, the resulting compartment is referred to as an autolysosome. Some people writing papers on the topic of autophagy use the terms “autolysosome” and “autophagolysosome” interchangeably. We contend that these words should be used to denote 2 different compartments, and that it is worthwhile maintaining this distinction—the autophagolysosome has a particular origin in the process of xenophagy that makes it distinct from an autolysosome.     

Autophagosomes,phagosomes, autolysosomes,phagolysosomes, autophagolysosomes… Wait,I’m confused

When an autophagosome or an amphisome fuse with a lysosome, the resulting compartment is referred to as an autolysosome. Some people writing papers on the topic of autophagy use the terms “autolysosome” and “autophagolysosome” interchangeably. We contend that these words should be used to denote 2 different compartments, and that it is worthwhile maintaining this distinction—the autophagolysosome has a particular origin in the process of xenophagy that makes it distinct from an autolysosome.     

Der Einfluß endogener Faktoren auf den Asche-, Kalzium-, Magnesium-, Kalium-, Natrium-, Phosphor-, Zink-, Eisen-, Kupfer- und Mangangehalt der Schweineborste

The objective of the present study was to investigate physiological effects of a marginal copper and iron supply on pigs. Therefore an experiment was conducted with 4 × 12 growing pigs of the crossbreed Pietrain × Deutsche Landrasse. The animals were fed for a period of 119 days with a diet poor of copper (1.5 mg Cu/kg diet) and/or poor of iron (35 mg Fe/kg diet). Control animals were supplied adequately with copper (4.8 mg Cu/kg diet) and iron (85 mg Fe/kg diet). The diet was given according to weight. After reaching an average weight of 102.6 ± 3.5 kg the animals were slaughtered. Due to the activity of the coerulplasmin and katalase enzyme and the haematological parameters, the supply of copper and iron could be classified as marginal. There was no interaction between copper deficiency and iron metabolism. The protein metabolism was unchanged. Low copper intake reduced the copper concentrations in serum, liver, muscle and backfat, and low iron intake reduced the iron concentration in serum, liver and muscle. Marginal copper and iron supply had no relevant effect on either food intake and growth performance or carcass characteristics and meat quality.     

Der Mengen- und Spurenelementgehalt des Rinderhaares als Indikator der Calcium-, Magnesium-, Phosphor-, Kalium-, Natrium-, Eisen-, Zink-, Mangan-, Kupfer-, Molybdän- und Kobaltversorgung

Beyond the energy requirement of maintenance, the assimilated energy, occurring in bioproducts, is linearly proportional to the intake of metabolizable energy in non‐underfed conditions. In contrast, resting metabolic rate is differing between individuals within a population of an animal species. As adaptability to changed environmental conditions may play a role, young bulls were exposed to thermoneutral (18°C) and low (4°C) ambient temperatures and were fed at two feeding levels (1.0 and 1.6 times energy requirement in maintenance) to produce metabolic rate differences, using the same animals, metabolic rate was altered by reducing the sympathetic outflow in each case. Expression of sulfonylurea receptors in circulating mononuclear leukocytes and cells from skeletal muscle (m. semitendinosus) was studied by flow cytom‐etry. Changes of metabolic rate at rest corresponded to the portion of cells with sulfonylurea receptors expression. The data from reducing the sympathetic outflow and those from sulfonylurea receptors expression are useful to explain metabolic rate differences among individuals of an animal population.     

Der Mengen- und Spurenelementgehalt des Rinderhaares als Indikator der Calcium-, Magnesium-, Phosphor-, Kalium-, Natrium-, Eisen-, Zink-, Mangan-, Kupfer-, Molybdän- und Kobaltversorgung

Many health effects can be attributed to the Mediterranean herb oregano (Origanum vulgare L.) and several studies demonstrated the improving effect on performance, changes in blood count, antibacterial, antifungal and immunmodulating abilities. The majority of these investigations were carried out with processed essential oil, while whole plant material was only used in a few studies. Thus, the aim of the present experiment was to test the effect of increasing proportions of dried oregano in piglet feed on health and performance, with a special focus on immune modulation. A total of 80 male castrated weaned piglets (body weight [BW] 7.9 kg ±1.0 kg) were used in a feeding experiment lasting 5 weeks. They were assigned to 4 experimental groups: a control diet, and three diets with an oregano supplementation at 2 g, 4 g and 8 g per kg feed, respectively, corresponding to 23.5 mg, 46.9 mg and 93.9 mg carvacrol/kg DM. After 3 weeks, half of each group was challenged with 5 µg lipopolysaccharides (LPS) per kg BW. Blood samples were collected 2 h after LPS stimulation and analysed for T-cell phenotypes, granulocyte activity, clinical-chemistry as well as white and red blood count. The results indicate no effects of oregano on performance. In contrast, oregano altered the lymphocyte proportion and the ratio of CD4⁺ and CD8⁺ T-cells as well as the triglyceride concentration in the serum of non-stimulated and in LPS-stimulated piglets. In conclusion, whole plant supplementation of oregano to piglet feed altered immune-related parameters, but did not modulate the acute inflammatory response induced by LPS stimulation.