Tansley Review No. 110.

S UMMARY 367
I. I NTRODUCTION 367
II. N UMBER 368
III. S IZE 379
IV. A IR SPACE IN THE SEEDS 381
V. F LOATATION AND DISPERSAL 383
1. Air 383
(a) Physical considerations 383
(b) Dispersal 387
(c) Birds 415
2. Water 416
(a) Physical considerations 416
(b) Dispersal 416
VI. C ONCLUSIONS 417
Acknowledgements 417
References 418
Orchid seeds are very small, extremely light and produced in great numbers. Most range in length from c . 0.05 to 6.0 mm, with the difference between the longest and shortest known seeds in the family being 120-fold. The 'widest' seed at 0.9 mm is 90-fold wider than the 'thinnest' one, which measures 0.01 mm (because orchid seeds are tubular or balloon-like, 'wide' and 'thin' actually refer to diameter). Known seed weights extend from 0.31 lg to 24 μg (a 78-fold difference). Recorded numbers of seeds per fruit are as high as 4000000 and as low as 20–50 (80000–200000-fold difference). Testae are usually transparent, with outer cell walls that may be smooth or reticulated. Ultrasonic treatments enhance germination, which suggests that the testae can be restrictive. Embryos are even smaller: their volume is substantially smaller than that of the testa. As a result, orchid seeds have large internal air spaces that render them balloon-like. They can float in the air for long periods, a property that facilitates long-distance dispersal. The difficult-to-wet outer surfaces of the testa and large internal air spaces enable the seeds to float on water for prolonged periods. This facilitates distribution through tree effluates and/or small run-off rivulets that may follow rains. Due to their size and characteristics, orchid seeds may also be transported in and on land animals and birds (in fur, feathers or hair, mud on feet, and perhaps also following ingestion).     

Manipulation of trait expression and pollination regime reveals the adaptive significance of spur length

Understanding the mechanisms of adaptive population differentiation requires that both the functional and adaptive significance of divergent traits are characterized in contrasting environments. Here, we (a) determined the effects of floral spur length on pollen removal and receipt using plants with artificial spurs representing the species-wide variation in length, and (b) quantified pollinator-mediated selection on spur length and three traits contributing to floral display in two populations each of the short-spurred and the long-spurred ecotype of the orchid Platanthera bifolia. Both pollen receipt and removal reached a maximum at 28–29 mm long spurs in a short-spurred population visited by short-tongued moths. In contrast, pollen receipt increased linearly across the tested range (4–52 mm) and pollen removal was unrelated to spur length in a long-spurred population predominantly visited by a long-tongued moth. The experimentally documented effects on pollen transfer were not reflected in pollinator-mediated selection through female fitness or pollen removal indicating that the natural within-population variation in spur length was insufficient to result in detectable variation in pollen limitation. Our study illustrates how combining trait manipulation with analysis of causes and strength of phenotypic selection can illuminate the functional and adaptive significance of trait expression when trait variation is limited.     

Evaluating asymbiotic seed culture methods and establishing Disa (Orchidaceae) germinability in vitro: relationships, requirements and first-time reports

Many Disa species (80%) have not been germinated in vitro, fuelling conservation fears. Winter-rainfall species are typically germinable, whilst summer-rainfall species are exclusively intractable in vitro. We aimed to establish the in vitro germination requirements for previously ungerminated Disa species. Conventional asymbiotic seed culture protocols were reviewed and their efficacy in summer-rainfall Disa established. Mature seed was subjected to manipulations of media composition and viscosity, incubation temperature and illumination. Immature cultures were also established across four seed maturity classes. Four first-time germinations (D. cooperi, D. nervosa, D. pulchra and D. woodii) resulted from media with increased water availability. Germination rate (>12 weeks), percentage (<30%) and synchrony were not in accordance with values reported for winter-rainfall species (>80% germination in 8 weeks). Germination of immature seed under control conditions was similar to mature seed germination under modified conditions, but neither percentage approached the calculated germination potential (∼viability). Germination control in Disa is proposed as a trade-off between water availability and the presence of phyto-inhibitors in the environment of the embryo—two features typical of seeds exhibiting water-impermeable dormancy.     

Minituber production from immature seed suspension culture ofOrchis papilionacea

Summary Ripe and immature seeds ofOrchis papilionacea (method I and II, respectively) cultured on modified double strength Curtis medium were assayed for minituber production. Ripe seed germination both on solid and in liquid medium was low and the protocorms obtained developed into white calluses. Germination increased from, 9 to 33% when immature seed suspension culture was used. Protocorms obtained in suspension culture under light developed into minitubers, whereas those obtained on solid media developed into callus. A 30 s ultrasonication of immature seeds 1 wk after suspension, culture initiation further enhanced germination and minituber production. Minitubers had to be transferred and embedded in solid regeneration medium for normal growth.     

Floral sources used by the orchid bee Euglossa cordata (Linnaeus, 1758) (Apidae: Euglossini) in an urban area of south-eastern Brazil

Although orchid bees (Apidae: Euglossini) are known as key pollinators in tropical ecosystems, knowledge of their floral sources is still scarce, especially for those species commonly found in urban environments. We aimed to identify the pollen, nectar and resin sources used by the widespread species Euglossa cordata in an urban area in south-eastern Brazil. The residual pollen from 81 brood cells of nine nests reactivated between October 2013 and December 2014 was acetolysed and analysed. A total of 50 pollen types belonging to 20 botanical families were identified in the samples. Pollen sources included species from seven families; five of them were plants with poricidal anthers (Bixaceae, Commelinaceae, Fabaceae, Melastomataceae and Solanaceae). The mass-flowering trees Handroanthus chrysotrichus and H. heptaphyllus (Bignoniaceae) are firstly reported as important pollen sources to an orchid bee species. Nectar was collected primarily from plants with long, tubular corolla as Acanthaceae, Apocynaceae, Bignoniaceae and Convolvulaceae. The vine species Dalechampia stipulacea (Euphorbiaceae) acted as a floral resin source. All pollen sources consisted of native plants whereas some exotic plant species were visited for nectar collection. Although nesting in an area encompassed by a high proportion of invasive plant species, Euglossa cordata females preferred to visit native plants to gather floral resources.     

Floral induction and development in Phalaenopsis in vitro

Adventitious shoots formed from nodal sections of floral-stalks of Phalaenopsis Pink Leopard Petra that had been propagated on Hyponex medium with 6-benzyladenine (BA). After subculturing adventitious shoots on Vacin-Went medium with BA, more than 70% of the shoots formed floral buds, and 10% of the floral buds flowered in 9-month-old shoots. Floral buds were not formed on the adventitious shoots cultured on Vacin-Went medium lacking BA. A high concentration of total nitrogen inhibited the development of floral buds, and low-temperature treatments failed to induce the formation of floral buds in vitro.     

The effects of smoke derivatives on in vitro seed germination and development of the leopard orchid Ansellia africana

Plant‐derived smoke and smoke‐isolated compounds stimulate germination in seeds from over 80 genera. It has also been reported that smoke affects overall plant vigour and has a stimulatory effect on pollen growth. The effect of smoke on orchid seeds, however, has not been assessed. In South Africa, orchid seeds from several genera may be exposed to smoke when they are released from their seedpods. It is therefore possible that smoke may affect their germination and growth. Therefore, the effects of smoke [applied as smoke‐water (SW)] and two smoke‐derived compounds, karrikinolide (KAR₁) and trimethylbutenolide (TMB), were investigated on the germination and growth of orchid seeds in vitro. The effect of SW, KAR₁ and TMB were investigated on the endangered epiphytic orchid, Ansellia africana, which is indigenous to tropical areas of Africa. Smoke‐water, KAR₁ and TMB were infused in half‐strength MS medium. The number of germinated seeds and number of seeds and protocorm bodies to reach predetermined developmental stages were recorded on a weekly basis using a dissecting microscope for a 13‐week period. Infusing SW 1:250 (v:v) into half‐strength MS medium significantly increased the germination rate index (GRI) and the development rate index (DRI) of the A. africana seeds. All the SW treatments significantly increased the number of large protocorm bodies at the final stage of development. Infusing KAR₁ into the growing medium had no significant effect on germination or development of the seeds. The TMB treatment, however, significantly reduced the GRI and DRI of A. africana seeds.     

Evolution and diversity of floral scent chemistry in the euglossine bee-pollinated orchid genus Gongora

•Background and Aims Animal-pollinated angiosperms have evolved a variety of signalling mechanisms to attract pollinators. Floral scent is a key component of pollinator attraction, and its chemistry modulates both pollinator behaviour and the formation of plant–pollinator networks. The neotropical orchid genus Gongora exhibits specialized pollinator associations with male orchid bees (Euglossini). Male bees visit orchid flowers to collect volatile chemical compounds that they store in hind-leg pouches to use subsequently during courtship display. Hence, Gongora floral scent compounds simultaneously serve as signalling molecules and pollinator rewards. Furthermore, because floral scent acts as the predominant reproductive isolating barrier among lineages, it has been hypothesized that chemical traits are highly species specific. A comparative analysis of intra- and inter-specific variation of floral scent chemistry was conducted to investigate the evolutionary patterns across the genus.•Methods Gas chromatography–mass spectrometry (GC-MS) was used to analyse the floral scent of 78 individuals belonging to 28 different species of Gongora from two of the three major lineages sampled across the neotropical region. Multidimensional scaling and indicator value analyses were implemented to investigate the patterns of chemical diversity within and among taxonomic groups at various geographic scales. Additionally, pollinator observations were conducted on a sympatric community of Gongora orchids exhibiting distinct floral scent phenotypes.•Key Results A total of 83 floral volatiles, mainly terpenes and aromatic compounds, were detected. Many of the identified compounds are common across diverse angiosperm families (e.g. cineole, eugenol, β-ocimene, β-pinene and terpinen-4-ol), while others are relatively rare outside euglossine bee-pollinated orchid lineages. Additionally, 29 volatiles were identified that are known to attract and elicit collection behaviour in male bees. Floral scent traits were less variable within species than between species, and the analysis revealed exceptional levels of cryptic diversity. Gongora species were divided into 15 fragrance groups based on shared compounds. Fragrance groups indicate that floral scent variation is not predicted by taxonomic rank or biogeographic region.•Conclusions Gongora orchids emit a diverse array of scent molecules that are largely species specific, and closely related taxa exhibit qualitatively and quantitatively divergent chemical profiles. It is shown that within a community, Gongora scent chemotypes are correlated with near non-overlapping bee pollinator assemblies. The results lend support to the hypothesis that floral scent traits regulate the architecture of bee pollinator associations. Thus, Gongora provides unique opportunities to examine the interplay between floral traits and pollinator specialization in plant–pollinator mutualisms.