Pollen digestion by flower-feeding Scarabaeidae: protea beetles (Cetoniini) and monkey beetles (Hopliini)

Pollen protoplasm is very nutritious, but the hard and highly resistant outer wall (exine) of the pollen grain presents an obstacle that pollen-feeders must overcome to benefit from the valuable protoplasm. Pollen digestion in three pollen-eating scarab beetles, the green protea beetle Trichostetha fascicularis and two monkey beetle species, Peritrichia cinerea and Pachynema flavolineata, was investigated. Adult beetles were collected for observations of feeding mechanisms, scanning electron microscopy of mouthparts, and histological examination of gut contents. Serial sections of different regions along the gut were stained for different nutrients and examined to determine the appearance of the pollen grains and the removal of carbohydrates, proteins and lipids from the pollen. During feeding, beetles used dense brushes of setae on the tips of their maxillae to mop up nectar and sweep pollen into their mouths. Grains were ingested intact. Nutrients were effectively removed from the grains as they passed along the gut and most pollen grains in the hindgut were empty, with broken exines. All the beetles excreted the undigested exine. Comparison with other studies on pollen digestion by a variety of insect and vertebrate pollen-feeders suggests that digestive enzyme penetration of ingested pollen grains and exudation of the products of digestion is the most likely method used by these beetles.     

Pollen digestion by nectarivorous and frugivorous Antillean bats

The ability to extract pollen contents may be related to the extent to which animals use this item as a regular part of their diet. In spite of the broad diversity of taxa that feed on pollen, comparative studies to test this hypothesis are scarce. We compared the extraction efficiency of pollen grains of Blue Mahoe (Talipariti elatum) by Antillean nectar bats (Brachyphylla nana) and Jamaican fruit bats (Artibeus jamaicensis). Antillean nectar bats extracted the contents of a higher percentage of pollen grains than Jamaican fruit bats, even though processing time in the gut was lower in the nectarivorous bats. Pollen extraction efficiency increased with time spent in the gut in each species. The gastrointestinal tract in both species resembled the functioning of a continuous stirred-tank reactor (CSTR) equivalent to the stomach, in series with a plug-flow reactor (PFR) equivalent to the intestine with varying degrees of longitudinal mixing. Accordingly, pollen grains flowed continuously out of the stomach and moved out through the intestine where they were mixed longitudinally. Our results support previous findings of higher extraction efficiencies in nectarivorous bats than their frugivorous relatives, and suggest that these differences may be the result of differences in the level of activity of the enzymes responsible for pollen wall degradation. Identification of enzymatic mechanisms of pollen degradation would allow a direct test of this hypothesis.     

Adaptations to a diet of nectar and pollen in the marsupial Tarsipes rostratus (Marsupialia: Tarsipedidae)

The mouse-sized marsupial Tarsipes rostratus , endemic to south-western Australia, feeds almost exclusively on nectar and pollen. Its tongue has long filiform papillae at the tip and shorter compound papillae over much of the upper surface. These collect nectar and pollen when the long tongue, stiffened by a keratinized keel, is protruded into flowers or over pollen presenters. Pollen is scraped from the papillae by a series of combs on the roof of the mouth. A large diverticulum, off the main chamber of the stomach, may store nectar in times of surplus. Pollen is not digested in the stomach but during passage down the simple intestine, which does not have a caecum. In captive animals, pollen passed through the gut in about six hours and the percentage of grains voided which had lost their contents was related to time spent in the gut. The digestion process remains unresolved but probably takes place through the apertures in the shells of pollen grains. Tarsipes ingests large numbers of pollen grains whose contents probably provide necessary nutrients absent from nectar.     

Use of maize pollen by adult Chrysoperla carnea (Neuroptera: Chrysopidae) and fate of Cry proteins in Bt-transgenic varieties

We investigated the use of maize pollen as food by adult Chrysoperla carnea under laboratory and field conditions. Exposure of the insects to insecticidal Cry proteins from Bacillus thuringiensis (Bt) contained in pollen of transgenic maize was also assessed. Female C. carnea were most abundant in a maize field when the majority of plants were flowering and fresh pollen was abundant. Field-collected females contained an average of approximately 5000 maize pollen grains in their gut at the peak of pollen shedding. Comparable numbers were found in females fed ad libitum maize pollen in the laboratory. Maize pollen is readily used by C. carnea adults. When provided with a carbohydrate source, it allowed the insects to reach their full reproductive potential. Maize pollen was digested mainly in the insect's mid- and hindgut. When Bt maize pollen passed though the gut of C. carnea, 61% of Cry1Ab (event Bt176) and 79% of Cry3Bb1 (event MON 88017) was digested. The results demonstrate that maize pollen is a suitable food source for C. carnea. Even though the pollen grains are not fully digested, the insects are exposed to transgenic insecticidal proteins that are contained in the pollen.     

Mortality of pollen grains may result from errors of meiosis: study of pollen tetrads in Typha latifolia L

In the cattail Typha latifolia the four haploid products of meiosis remain attached and form the flat tetrad of pollen grains. Gametophytic lethals arisen de novo in diploid cells of sporophyte must manifest themselves as pollen tetrads with two dead grains. This could allow to estimate the rate of recessive lethals arresting pollen grain development. We studied pollen samples collected from 44 sprouts in two populations in the vicinity of Novosibirsk. The anomalous tetrads T1, T2, T3, and T4 carrying one, two, three, and four dead grains, respectively, were detected in each sampled individual. The mean frequency of all anomalous tetrads in the two populations was 3.4% and 8.7%. The frequencies of tetrad classes varied widely among the individuals with correlation coefficient up to 0.94, but their ratios remained nearly constant. The majority of anomalous tetrads were presented by T1 and T2 classes (their sum comprising 72.7 and 74.0% in two populations), T1 being a little more abundant. The observed pattern of frequencies of tetrads with dead grains can be explained by errors of male meiosis such as chromosome non-disjunction in both meiotic divisions. The tetrads with two dead pollen grains may result mostly from non-disjunction in anaphase I, and those with one pollen grain from non-disjunction in anaphase II, thus making tetrad analysis ineffective for estimating the rate of gametophytic lethals.     

Herbivory in Spiders: The Importance of Pollen for Orb-Weavers

Orb-weaving spiders (Araneidae) are commonly regarded as generalist insect predators but resources provided by plants such as pollen may be an important dietary supplementation. Their webs snare insect prey, but can also trap aerial plankton like pollen and fungal spores. When recycling their orb webs, the spiders may therefore also feed on adhering pollen grains or fungal spores via extraoral digestion. In this study we measured stable isotope ratios in the bodies of two araneid species (Aculepeira ceropegia and Araneus diadematus), their potential prey and pollen to determine the relative contribution of pollen to their diet. We found that about 25% of juvenile orb-weaving spiders’ diet consisted of pollen, the other 75% of flying insects, mainly small dipterans and hymenopterans. The pollen grains in our study were too large to be taken up accidentally by the spiders and had first to be digested extraorally by enzymes in an active act of consumption. Therefore, pollen can be seen as a substantial component of the spiders’ diet. This finding suggests that these spiders need to be classified as omnivores rather than pure carnivores.     

油菜和玉米花粉粒中元素分布的研究

以油菜和玉米花粉为材料,应用扫描电镜及能谱仪技术对花粉中内外元素分布进行了初步研究。结果表明：花粉粒萌发沟区与非萌发沟区,花粉壁与原生质内外的不同部位,Fe、Zn、Si、Al、Ca、P、S、Mg、K、Cl、Cu、Mn、Co及Ni 14种元素组成有很大差异;两种花粉的萌发沟区都未检测出Fe元素,而非萌发沟区的Fe元素相对含量为1.12%～368%;花粉壁中比花粉原生质中更富含Si,Ca,K,Cl等元素。本项研究为蜜源植物花粉的开发和利用提供了有价值的资料。     

Isoenzyme Polymorphism in Flowering Plants I. Diffusion of Enzymes Out of Intact Pollen Grains

Esterases, leucine aminopeptidases, catalases, amylases and acid phosphatases diffuse out of intact and ungerminated pollen grains of Oenothera organensis, whether suspended in 1 % sodium chloride or in pollen medium. A total of 15 esterase isozymes are recorded; 5 of them appear within 5 minutes, 8 within 30 minutes, 9 within 2 hours, and 13 within 19 hours. Pollen grains suspended for 19 hours gave much stronger isozyme bands than macerated pollen grains. However, one esterase hand was consistently missing from the 19 hour suspensions, although present in all others. It is suggested as a working hypothesis that the early growth of pollen tubes and possibly even germination of pollen grains may be influenced by the metabolic products of pistillate tissues caused by the diffused pollen enzymes, and that inactivation of these enzymes by stigmatic or stylar components could lead to incompatibility reactions.     

Pollen-feeding in thrips (Insecta: Thysanoptera)

Eight species of thrips in three families fed readily on individual grains of pollen. Feeding behaviour is described. Thrips fed on the contents of the grains, but left the pollen wall. In Thrips fuscipennis Halid., the feeding time per grain increased in direct proportion to grain volume up to a grain volume of about 60,000 μm'. but then began to level off. Large grains and tetrads were usually abandoned before all the contents were removed. Mean feeding times ranged from 3 s to 120 s per grain.
Thrips appeared to have alternate periods of feeding and non-feeding. Drops of fluid were sometimes deposited from the end of the abdomen after non-feeding periods. Large amounts of pollen can be consumed; one thrips fed on more than 100 pollen grains in less than 15 min. Pollen-feeding appears to be widespread among flower thrips. The implications for pollination ecology are discussed.     

六个不同类型荞麦花花粉粒形态的电镜观察比较研究

对二倍体甜荞长花柱类型 (ES2 s)、二倍体甜荞短花柱类型 (ES2 S)、四倍体甜荞长花柱类型 (ES4s)、四倍体甜荞短花柱类型 (ES4S)、四倍体有翅细野荞 (GR4HI)、四倍体无翅细野荞 (GR4HO)等 6个不同荞麦类型花的花粉形态学进行了电镜观察。结果表明 :这些荞麦类型的花粉粒都是椭圆形 ,都有网状纹饰、3孔沟等特征。二倍体甜荞染色体加倍后形成的四倍体甜荞 ,相对于二倍体甜荞而言 ,花粉粒显著增大和明显变圆。无论二倍体还是四倍体甜荞 ,其长花柱型花的花粉粒都比其对应短花柱型花的花粉粒要小。尽管四倍体甜荞和四倍体细野荞在染色体数目和倍性上一样 ,但是它们在花粉粒的大小上有显著差异。四倍体细野荞有翅类型和无翅类型的花粉粒大小和形态差异不显著     

Protection of pollen germination from adverse temperatures: a possible role for proline

Abstract After germination, pollen grains of Lilium longiflorum became very sensitive to short periods of heat stress as shown by the greatly reduced germination percentages upon subsequent incubation at the optimal temperature. Addition of proline to the incubation medium made pollen more resistant to heat. It was demonstrated that in a short time a large amount of proline was taken up by the cell. Germination and metabolic functions were completely or partially protected from heat damage by proline treatment. As well, it was shown that proline treatment at least partially protected pollen grains from cold stress. These results suggest that the high proline concentrations found in pollen of many species may confer resistance to germinating pollen grains at unfavourable temperatures thereby enhancing the chances of successful fertilization.     

Sugar exudation by roots of kallar grass [<Emphasis Type="Italic">Leptochloa fusca</Emphasis> (L.) Kunth] is strongly affected by the nitrogen source

Exudation of sugars (glucose, fructose and sucrose) and that of cations and anions from intact roots of kallar grass [Leptochloa fusca (L.) Kunth] grown hydroponically with ammonium or nitrate (3 mM) as N source was investigated. In different experiments, plants grown on ammonium had slightly higher sugar contents than nitrate-grown plants, but their total sugar exudation during a 2-h period was up to 79-fold higher than under nitrate nutrition. Relative root exudation of inorganic anions and cations and that of amino acids (as a percentage of the internal contents exuded per time) was either similar or slightly higher from ammonium-grown than from nitrate-grown plants. Analysis of root architectural parameters revealed that ammonium-grown plants had a higher number of root tips/side roots per gram root fresh weight than nitrate-grown plants, whereas other root parameters, viz. length, diameter, volume and surface area were similar under the two N sources. A majority of the fine roots having diameter up to 0.4 mm represented up to 86% of the total root length, 64% of the total root surface area, and 35% of the total root volume; the root length and surface area per root system of that major root population were similar in ammonium- and nitrate-grown plants. Apparently, root architecture was not responsible for the different exudation rates. Within 12-24 h after shifting ammonium-grown plants to nitrate nutrition, root sugar levels and visible root architecture remained unchanged, yet the sugar exudation rate was reduced 30-fold. Short-term uptake of [14C]glucose (10 microM) from the rooting medium was similar for ammonium- and nitrate-grown plants. Thus, the very different sugar exudation rates were neither related to internal root sugar concentration, nor to the different root architecture, nor to differential resorption of sugars by ammonium- versus nitrate-grown plants. Increased external Ca2+ did not alter sugar exudation, and decreased external pH (4.5) only slightly increased sugar exudation from roots of nitrate-grown plants kept at pH 6.5. It is suggested that the much higher sugar exudation in response to ammonium may facilitate the ecologically and economically important association of diazotrophs with kallar grass roots.     

叶下珠科花粉组织化学、花粉数和胚珠数及其与传粉者关系的研究

植物花粉中营养物质的储存形式以及单花的花粉数与胚珠数被认为与其传粉系统有一定的联系。本文研究了叶下珠科部分种类的花粉组织化学、花粉数和胚珠数, 以及它们与传粉者之间的关系。结果显示: 在叶下珠科内, 植物花粉所含营养成分和传粉者之间存在相关性: 蛾类传粉的类群主要属“淀粉型”花粉, 蝇类和蜂类传粉的类群主要属“非淀粉型”花粉。蝇类传粉和蛾类传粉的植物花粉数没有一定规律。蝇类传粉的类群比蛾类传粉的类群胚珠数少, 这可能是由于蝇类携带花粉能力及传粉精确性均较小, 导致植物以减少胚珠数来适应的结果。对同一属内不同生活型植物的花粉数比较, 发现乔木的单花花粉数高于灌木, 灌木的单花花粉数明显高于草本。这可能是由于不同生活型的植株, 其花朵大小不同, 导致花粉数出现明显差别。另外, 通过扫描电子显微镜对花粉形态的观察, 发现蝇类传粉的类群和蛾类传粉的类群间的花粉表面纹饰存在显著差异。     

Organelle Movement in the Germinated Plooen Grains and Pollen Tubes of Oenothera odorata

The movement of organelles in the germinated pollen of Oenothera odorata was studied in detail by video microscopy. The image of the organelle movement was processed by computergation. The pollen grain of Oenothera odorata is large and easy to germinate in vitro and is suitable for the study of organelle movement . The motion of organelles in the germinated pollen grains and pollen tubes is very vigorous. But the movement of organelles in the pollen tubes is more vigorous than that in the germinating pollen grains. Some of their motion is saltatory. A kind of fibrils was observed in the germinated pollen grains. They are supposed to be made up of actin filaments. Some of the fibrils have one end connected to the plasma membrane and others have both ends linked to the plasma membrane , forming a network. Organelles move along the fibrils continuosely and the speed changes constantly . The speed of movement of organelles is not related to their dimensions. Cytochalasin B can inhibit the movement of organelles. Our results suggest that the movement of organelles is independent of the cytoplasmic streaming in the germinated pollen grains and pollen tubes.     

Digestion of maize and sunflower pollen by the spotted maize beetle Astylus atromaculatus (Melyridae): is there a role for osmotic shock?

We investigated the mechanism and efficiency of digestion of two types of pollen, maize, Zea mays, and sunflower, Helianthus annuus, by the spotted maize beetle, Astylus atromaculatus (Melyridae). We found similar and high extraction efficiencies, but different mechanisms of digestion. Osmotic shock was apparently involved in digestion of the large and thin-walled maize pollen grains. In the anterior midgut most maize pollen grains were already ruptured, in contrast with the intact exines of sunflower pollen, which suggests another mechanism of digestion for the latter, such as enzymatic action. We investigated the effect of osmotic shock on maize pollen in vitro by looking at the behavior of pollen grains in varying osmotic concentrations. Maize pollen grains burst in both distilled water and sugar solutions of various concentrations, and the amount of rupturing decreased with an increase in sugar concentration. Digestion of maize pollen was much slower in honeybees than in spotted maize beetles. Maize pollen bursts early in the midgut of maize beetles, but remains intact in honeybees: this suggests that osmotic shock may not be as important for honeybees as previously suggested.     

Localization and release of allergens from tapetum and pollen grains ofBetula pendula

Summary Although intact pollen grains are assumed to be the primary carrier of pollen allergens, specific immunoreactive components have been found in other aerosol fractions, e.g., starch grains and remains of tapetal cells Cryo-scanning-electron-microscopy results demonstrate the presence of a clear network of strands connecting the tapetum with the microspores. The distribution of protein in tapetal orbicules, pollen wall, and pollen cytoplasm was tested by histochemical stains for light microscopy and transmission electron microscopy. The protein is mainly localized at the apertures and starch grains in the cytoplasm of pollen and in the core and on the surface of tapetal orbicules. Monoclonal antibodies Bv-10, BIP3, and BIP4 have been used to locate the cellular sites of pollen and tapetal allergens inBetula pendula (syn.B. verrucosa). The application of rapid-freeze fixation prevented relocation of allergens from their native sites. The allergens are predominantly found in the starch grains and to lesser extent in the exine. We also tested interactions between mature birch pollen and human fluids: saliva, nostrils fluid, and eyes solution. The aim was to mimic more closely the in vivo situation during allergenic response. In all cases we observed several pollen grains that were burst and had released their cytoplasmic contents. In the nose the allergens are released from the pollen within minutes. In rhinitis, nasal pH is increased from the normal pH 6.0 to 8.0. When we used nasal fluid at pH 8.0, the number of ruptured pollen grains increased. The mechanism that might induce formation of small allergen-bearing particles from living plant cells is discussed.     

To Repeat the Effects of Hormones on the Early Microspore Developments of Wheat in Vitro in Anther Culture

1. The normal development of pollen cells can be transformed by the exoision itself of anther culture: The second mitotic division of pollen grains has been prevented; The frequency of anomalous division of pollen grains was higher than that present in anthers in vive; The generative nuclei after the first mitosis were more or less globular in form and in their subsequent developments most of them do not become spindly-shape which is particular to the generative cells in vive. In the meantime, they show a weak staining reaction with Feulgen reagent. 2. The higher concentrations of hormones were found to enhance the frequency of abnormal division obviously. Of anthers cultured on the four N6 media added with various concentration ratios of IAA to Kinetin 2:10, 10:2, 2:12, and 12:2 mg/l. The mean percentages of abnormal pollen grains were 34.02%, 35.28%, 34.27% and 36.65% respectively. 3. The higher hormone level may promote the formation of multicellular pollen grains obviously. When the IAA concentration was raised up to 12 mg/l, the mean multieellular pollen grain yields per anther increased to 13.3 unit, while the control without hormone was only 4 unit.     

Gametophyte–Sporophyte Interactions in the Pollen–Pistil System: 3. Hormonal Status at the Progamic Phase of Fertilization

The content of hormones, IAA, ABA, and cytokinins, as well as the rate of ethylene production in petunia (Petunia hybrida L.) pistils and their parts (stigma, style, and ovary) were determined over 8 h after compatible pollination. At the progamic phase of fertilization in the pollen–pistil system, the phytohormones were virtually absent from the ovary but were present in various proportions in stigma and style. The stigma was the main site of ethylene synthesis and contained 90% of ABA while the style contained 80% of cytokinins of their contents in the whole pollinated pistil. Stigma and style did not differ in their IAA levels. The interaction of the male gametophyte with the stigmatic tissues was accompanied by a threefold increase in the ethylene production and a 1.5-fold increase in the IAA content in the pollen–pistil system within 0–4 h. Growth of pollen tubes in the stylar tissues (4–8 h) was accompanied by a further increase in IAA content and a decrease in the ethylene production by stigmatic tissues, as well as by a decrease in the cytokinin content in the stylar tissues. The ethylene/auxin status of the stigma may be suggested to control the processes of adhesion, hydration, and germination of pollen grains during pollination, while the auxin/cytokinin status of the style controls the pollen tube growth.     

An analysis of cedar of Lebanon (Cedrus libani A. Rich.) pollen grain development in cultural plantings]

A cytological study of the Cedrus libani mature pollen from 3 culture areas (Italy, France, USSR) has shown that 69-71% of pollen grains have two-celled protallium and antheridial cell. About 5% of pollen grains are characterized by accelerated of delayed development, otherwise apparently normal. The pollen sterility (up to 30% of grains) is due to the abortive spore development. Anomalous cenocyte and multinuclear pollen grains were found thus suggesting that multicellular haploid structures capable of further growth and development may arise in the course of natural anther development.