Electrophoretic profiling and immunocytochemical detection of pectins and arabinogalactan proteins in olive pollen during germination and pollen tube growth

Background and Aims

Cell wall pectins and arabinogalactan proteins (AGPs) are important for pollen tube growth. The aim of this work was to study the temporal and spatial dynamics of these compounds in olive pollen during germination.

Methods

Immunoblot profiling analyses combined with confocal and transmission electron microscopy immunocytochemical detection techniques were carried out using four anti-pectin (JIM7, JIM5, LM5 and LM6) and two anti-AGP (JIM13 and JIM14) monoclonal antibodies.

Key Results

Pectin and AGP levels increased during olive pollen in vitro germination. (1 → 4)-β-d-Galactans localized in the cytoplasm of the vegetative cell, the pollen wall and the apertural intine. After the pollen tube emerged, galactans localized in the pollen tube wall, particularly at the tip, and formed a collar-like structure around the germinative aperture. (1 → 5)-α-l-Arabinans were mainly present in the pollen tube cell wall, forming characteristic ring-shaped deposits at regular intervals in the sub-apical zone. As expected, the pollen tube wall was rich in highly esterified pectic compounds at the apex, while the cell wall mainly contained de-esterified pectins in the shank. The wall of the generative cell was specifically labelled with arabinans, highly methyl-esterified homogalacturonans and JIM13 epitopes. In addition, the extracellular material that coated the outer exine layer was rich in arabinans, de-esterified pectins and JIM13 epitopes.

Conclusions

Pectins and AGPs are newly synthesized in the pollen tube during pollen germination. The synthesis and secretion of these compounds are temporally and spatially regulated. Galactans might provide mechanical stability to the pollen tube, reinforcing those regions that are particularly sensitive to tension stress (the pollen tube–pollen grain joint site) and mechanical damage (the tip). Arabinans and AGPs might be important in recognition and adhesion phenomena of the pollen tube and the stylar transmitting cells, as well as the egg and sperm cells.     

Holocene pollen diagrams from Greece

Pollen diagrams are presented from three sites in Greece. Two cores from the Strymon Delta in Macedonia are thought to span parts of the period from Classical to Turkish times. A dated core from Kleonai in the Peloponnese has good pollen preservation in the early Neolithic, Bronze Age and Roman periods. A peat bog at Asi Gonia in the White Mountains of Crete has a continuous pollen record for the last 1500 years. The palynological evidence for human impact in various historic periods is assessed in the light of archaeological evidence. The problems of interpretation of pollen cores from Greece are discussed and the complex inter‐relationship between human impact and environmental factors is emphasised.     

Generalized food deception: colour signals and efficient pollen transfer in bee‐pollinated species of Eulophia (Orchidaceae)

Non‐rewarding plants use a variety of ruses to attract their pollinators. One of the least understood of these is generalized food deception, in which flowers exploit non‐specific food‐seeking responses in their pollinators. Available evidence suggests that colour signals, scent and phenology may all play key roles in this form of deception. Here we investigate the pollination systems of five Eulophia spp. (Orchidaceae) lacking floral rewards. These species are pollinated by bees, notably Xylocopa (Anthophorinae, Apidae) or Megachile (Megachilidae) for the large‐flowered species and anthophorid (Anthophorinae, Apidae) or halictid (Halictidae) bees for the small‐flowered species. Spectra of the lateral petals and ultraviolet‐absorbing patches on the labella are strongly contrasting in a bee visual system, which may falsely signal the presence of pollen to bees. All five species possess pollinarium‐bending mechanisms that are likely to limit pollinator‐mediated self‐pollination. Flowering times extend over 3–4 months and the onset of flowering was not associated with the emergence of pollinators, some of which fly year round. Despite sharing pollinators with other plants and lacking rewards that would encourage fidelity, the Eulophia spp. exhibited relatively high levels of pollen transfer efficiency compared with other rewarding and deceptive orchids. We conclude that the study species employ generalized food deception and exploit food‐seeking bees. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013 , 171 , 713–729.     

The influence of the progamic phase for fruiting in the apple tree

Final fruit production is the result of a number of processes, over which several environmental circumstances interact. But it is often difficult to disentangle the part played by each of these factors in the final crop. The aim of this work is to evaluate the influence of the progamic phase for fruiting in the apple tree. For this purpose we track back the process that goes from flower to fruit, identifying the inflection points where the final crop is reduced. We evaluate early versus late fruit development, pollination versus non‐pollination, and the effect of the progamic phase that goes from pollination to fertilization. From flowers to fruits 15 weeks elapsed, but the final fruit set settled 8 weeks after flowering, and the main flower–fruit drop occurred 3–4 weeks after flowering. Differences between dropped fruits and those that remained in the tree emerged earlier, and the onset of fruiting started 7 days after pollination. This time was coincident with the time lapse of the progamic phase. These results show that fruiting gets established well ahead of cropping, but also that the progamic phase is the main determinant of the final fruit set in apple trees.     

Pollen morphology and structure of Tropical and Subtropical American genera of the Piptadenia -group (Leguminosae: Mimosoideae)

A survey of the pollen morphology of the American taxa of the Piptadenia -group (Mimoseae) was done including all together 384 species of the following genera: Piptadenia , Parapiptadenia , Pseudopiptadenia , Piptadeniopsis , Microlobius (= Goldmania ), Stryphnodendron , Adenopodia , Anadenanthera and Mimosa . Because of a high morphological and structural variation the definition and characterization of pollen types turned out to be very difficult. Nevertheless the presence and durability of some pollen characters allow a provisional grouping: (1) always tetrads or polyads, (2) small size 6 (20) 40 mum, (3) unstable shape but currently ovoidal and, if biconvex, ellipsoidal in the outline, (4) number of pollen grains of the polyads very variable and unstable; the most frequent numbers are 8-12-16, (5) pollen grains irregularly united in polyads, with homomorphous exine and, (6) pores always located in the angles of the pollen grains in distal-subdistal position, rarely subdistal, (7) without or with pseudocolpi or subpseudocolpi (never colpi) in the distal face, (8) endexine forming an annulus around the endoaperture with ends of free lamellae with a central white line, (9) the tectal stratum above the pores forming a small vestibular space (except in Anadenanthera ). These palynological characters or most of them confirm the affinity among genera in the Piptadenia -group and its distinction from other taxa of the subfamily Misosoideae with tetrads or polyads.     

A melissopalynological study of artisanal honey produced in Catamarca (Argentina)

Spores belonging to 43 taxa of pteridophytes were observed from the ambient air of 11 localities in India. It is suggested that pteridophyte spores could be listed among viable particulate air pollutants, especially in the light of the earlier reports regarding their allergenicity.     

Pollen morphology of Justicia L. (Acanthaceae) from Thailand and its taxonomic value

The acetolysed pollen of 33 species of the genus Justicia in Thailand is investigated using both light and scanning electron microscopy. The pollen of the genus is characterised as being isopolar, bilaterally or radially symmetrical, with mostly prolate or prolate spheroidal shape. Based on characters with high factor loading in the cluster analysis and principal components analysis, the studied species are divided into two major groups; one with 3-colporate with six pseudocolpi and the other with 2-porate or 2–3-colporate with aperture areas. The pollen morphology of each group is described and illustrated. Pollen characters are useful for taxonomic delimitation and relationships among Thai species.     

Botanical affinity of pollen harvested by Apis mellifera L. in a semi-arid area from Bahia,Brazil

We analysed the botanical composition of pollen harvested by Apis mellifera L. in the Canudos Biological Station, Bahia, Brazil, and the influence of climatic factors on pollen sample composition was assessed. Forty-six pollen types were identified belonging to species occurring in the study area. The family Leguminosae was of significant importance amongst the samples, represented by ten pollen types. Diodia radula, Rhaphiodon echinus, and Mimosa misera pollen types occurred most constantly among the samples. We observed that isolated pollen class characterises samples analysed. It was also observed that pollen type richness is directly linked to rainfall, reflecting the strong influence of this climatic parameter on flowering intensity, and thus on the ability of the bees to obtain food resources.     

Analysis of protein content in pollen loads produced in north-west Spain

An analysis was made of the protein content of pollen loads produced by the bees in a hive situated in Viana do Bolo (Ourense, north-west Spain), to establish whether or not the relative quantity of protein in the pollen of each plant species influences the preference made by the bee of the flowers that supply pollen to the hive. This analysis was performed on all types of pollen that formed more than 5% of the pollen spectrum. Pollen load samples were collected directly from the hive from March to September. Pollen loads were separated by colour, and their specific homogeneity was confirmed microscopically. The Bradford method has been used for protein extraction and spectrophotometry was used for the determination of protein content. The results show that the different pollen loads have high protein content. Pollen of the plant species that reached relatively higher percentages in the pollen spectrum are also those that have the highest protein content. These were Cytisus scoparius type, uncultivated Poaceae, Quercus robur type, Sanguisorba minor, Salix fragilis and Spergularia rubra type. The pollen of the systematic units, which had pollen loads that could be identified at the level of species, maintained a constant value of protein content independently of the date the samples were obtained. The pollen of the systematic units, which had pollen loads that could be identified at the level of pollen type, has varied in protein content in the analyses performed on samples obtained on different dates. This result is due to the fact that the different species that integrate the pollen type flower on different dates, and thus have a pollenkitt with different characteristics.     

Pollen morphology in the genus Veronica L. (Plantaginaceae) and its systematic significance

Pollen grains of 30, mainly annual, species from 134 populations of the genus Veronica (Plantaginaceae, formerly Scrophulariaceae) from the Mediterranean Region, have been studied with light and scanning electron microscopy. Three pollen types are defined based on pollen size and exine surface sculpture. In some cases within the study group pollen size can be useful in species determination. Hypothetical relationships of pollen grain size and aperture number with ploidy level, style length and corolla diameter are discussed. The ornamentation of the exine in Veronica, although generally a conservative character within the genus, gives some additional support to the most recent infrageneric classification of Veronica.