Unpollinated Flowers Research Articles

Programme of senescence in petals and carpels of Pisum sativum L. flowers and its control by ethylene.

The role of ethylene in the control of senescence of both petals and unpollinated carpels of pea was investigated. An increase in ethylene production accompanied senescence, and the inhibitors of ethylene action were effective in delaying senescence symptoms in different flower verticils. Pollination did not seem to trigger the senescence syndrome in the corolla as deduced from the observation that petals from pollinated and unpollinated flowers and from flowers whose carpels had been removed senesced at the same time. A cDNA clone encoding a putative ethylene-response sensor (psERS) was isolated from pea flowers, and the pattern of expression of its mRNA was studied during development and senescence of different flower tissues. The levels of psERS mRNA paralleled ethylene production (and also levels of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) mRNA) in both petals and styles. Silver thiosulfate treatments were efficient at preventing ACO and psERS mRNA induction in petals. However, the same inhibitor showed no ability to modify expression patterns in pea carpels around the anthesis stage, suggesting different controls for ethylene synthesis and sensitivity in different flower organs.

The roles of abscisic acid and ethylene in the abscission and senescence of cocoa flowers

Cocoa flowers have a limited period of longevity; more than 90% of unpollinated flowers abscised within 32 h after anthesis. Abscisic acid (ABA) levels increased significantly prior to abscission. By 21 h after anthesis, ABA levels had increased almost 10-fold, and by 32 h flowers had 20-fold higher levels of ABA than at anthesis. Fluridone completely inhibited both the increase in ABA, the formation of an abscission zone, and the abscission and senescence of flowers. In contrast, ethylene production increased only slightly 21 h after anthesis and was only 2-fold higher after 32 h. Aminoethoxyvinylglycine (AVG) delayed but did not prevent abscission. In cocoa flowers, ABA is the primary regulator of abscission; ethylene accelerates abscission but only in the presence of ABA. Naphthalene acetic acid (NAA) treatment of flowers at anthesis prevented abscission zone formation and flower abscission, but did not induce fruit set. All parts of the NAA-treated flower except the pedicel senesced after 6 days. NAA+AVG treatment only delayed, whereas fluridone treatment completely prevented flower senescence.

Does self‐pollination provide reproductive Assurance in Aquilegia Canadensis (Ranunculaceae)?

The ability to produce seeds when pollinators or potential mates are scarce is thought to be one of the main advantages of self-fertilization in flowering plants. However, whether autonomous selfing increases seed set in natural populations has seldom been tested, and even fewer studies have evaluated the advantage of selfing across a gradient of pollen availability. This study examines the fertility consequences of autonomous selfing in Aquilegia canadensis (Ranunculaceae), a short-lived, spring-flowering perennial typically found in small, patchy populations on rock outcrops. We used a pollinator exclusion experiment to confirm reports that A. canadensis has a well-developed capacity for autonomous selfing resulting from incomplete protogyny and close proximity of stigmas and anthers during dehiscence. Flowers excluded from pollinators set 87% as many seeds per carpel (X +/- 1 SE = 7.1 +/- 1.4 seeds) as hand-pollinated flowers (8.1 +/- 1.3 seeds), and seed production in unpollinated flowers correlated negatively with the distance between stigmas and anthers (r = -0.46). Autonomous selfing could be potentially valuable in providing reproductive assurance because only 2.7 +/- 0.5 pollen grains were deposited on each stigma before anther dehiscence, compared to 134.1 +/- 17.9 pollen grains by the end of anther dehiscence. However, prevention of autonomous selfing by anther removal before dehiscence did not decrease seed set, even for plants at low plant densities where outcross pollen may have been in short supply. Emasculated flowers set as many seeds per carpel (9.3 +/- 0.9) as intact flowers (8.4 +/- 1.1), indicating that sufficient cross pollen is deposited for full seed set. These results do not support the hypothesis that autonomous selfing by A. canadensis has been selected because it provides reproductive assurance.

Plasticity of flower longevity in Corydalis ambigua

We investigated the longevity of individual flowers of Corydalis ambigua Cham. et Schlecht. during different periods of pollinator activity and at different temperatures. To measure potential (unpollinated) flower longevity of C. ambigua, this study was conducted at forest islands where pollinator visits were scarce. The longevity of individual flowers of C. ambigua indicated high plasticity. The longevity of unpollinated flowers in natural pollination ranged from 2 to 25 days and continuously decreased with the date of flower opening. The temperature increased as the flowering season progressed. Furthermore, the greenhouse transplanting experiment showed that higher temperatures shorten the life span of flowers. The longevity of pollinated flowers subjected to hand pollination of newly opened flowers was shorter than that of unpollinated flowers in natural pollination regardless of the date of flower opening. These results showed that not only high temperature but also pollination shortens flower longevity. We discuss the role of plasticity in flower longevity for C. ambigua in relation to pollination success and reduction in the maintenance cost of the flowers.

Influence of intraovular reserves on ovule fate in apricot ( Prunus armeniaca L.)

In many plant species with multiovulate ovaries, a considerable reduction in the number of ovules takes place. However, the underlying physiological causes are not clear. In Prunus spp., although flowers present two ovules, usually only one seed is produced. We have followed the development and degeneration of the two ovules in apricot (Prunus armeniaca L.) and examined the extent to which carbohydrates within the ovule might be involved in determining the fate of the ovule. While the primary ovule grows in the days following anthesis, growth of the secondary ovule is arrested. Starch distribution along the different ovular tissues exhibits several changes that are different in the two ovules. Primary ovule growth is inversely related to starch content and this growth takes place independently of pollination since it occurs in the same way in pollinated and unpollinated flowers. In the secondary ovule, starch disappears simultaneously from all ovular structures and callose is layered at the chalazal end of the nucellus. The size of the secondary ovule does not change significantly from anthesis to degeneration, and callose starts to accumulate 5 days after anthesis. Likewise, this process occurs independently of pollination. These results are discussed in terms of the implications of the starch content of ovules in fertilization success and ovule fate.

Fruit and seed production in relation to pollination and resources in bluebell, Hyacinthoides non-scripta.

This paper explores spatial and temporal patterns in the interaction of pollination and resources as constraints on the initiation and maturation of fruits and seeds in bluebell (Hyacinthoides non-scripta, Liliaceae). Field and laboratory experiments showed a degree of self-incompatibility (cross-pollination gave greater seed and fruit numbers than self-pollination), and pollinator limitation (hand cross-pollination gave greater seed and fruit numbers than open-pollination) as well as␣resource limitation. Bud removal, designed to relax interfruit competition for resources in the remaining flowers, increased percent fruit maturation in unpollinated flowers and increased the percent of initiated seeds that matured in cross-pollinated flowers. Clear position effects were also found. In an ovary, ovules near the stylar end showed a higher percent seed maturation than basal ovules. On a raceme, terminal flowers showed lower percent fruit initiation and maturation and lower seed numbers than basal flowers. The balance between resource limitation and pollinator limitation changed with position on the raceme. Temporal patterns involved raceme death and abortion of flowers, fruits or seeds. A lattice diagram schematises the hypothetical modulation by pollination of resource-dependent serial adjustment of maternal investment.

Variation of Floral Scent Emission and Postpollination Changes in Individual Flowers of Ophrys sphegodes Subsp. sphegodes

We investigated the scent composition of individual flowers of Ophrys sphegodes, its alteration following pollination, and of picked flowers by day and at night. Odor samples were collected by headspace sorption and analyzed by gas chromatography and mass spectrometry. To evaluate the function of postpollination odor changes, we carried out behavioral tests on the pollinator Andrena nigroaenea with pollinated and unpollinated flowers. We identified 27 volatiles in the flower scents. Aldehydes and alkanes were most frequently found. Aldehydes were the most abundant class of compounds (40–50%). When flowers were picked, they emitted significantly lower total amounts of volatiles than unpicked flowers, and their odor bouquets were significantly different. Comparison of scents released by day and at night showed no decrease in scent emission during nighttime, but the odor bouquets were significantly different. Pollinated flowers produced significantly different odor bouquets, and the total amount of scent emitted two to four days after pollination was significantly lower compared with unpollinated flowers. In addition, behavioral tests with A. nigroaenea males showed that flowers were significantly less attractive three days after pollination. This reduced attractiveness is hypothesized to guide pollinators to the unpollinated flowers within an inflorescence, and thus increase the reproductive success of the plant.

Hormonal Changes after Compatible and Incompatible Pollination in Theobroma cacao L.

In order to characterize the self-incompatibility system in Theobroma cacao, the levels of ethylene, indole-3-acetic acid (IAA), and abscisic acid (ABA) were determined after pollination with compatible and incompatible pollen and in unpollinated flowers. Pollen tube growth rates after incompatible and compatible pollinations were identical, and the majority of the pollen tubes reached the ovules between 12 and 20 hours after pollination. ABA levels rose in incompatibly pollinated flowers, and fell in compatibly pollinated flowers, prior to pollen tube—ovule contact. Ethylene evolution remained stable in compatibly pollinated flowers and rose in incompatibly pollinated flowers. IAA concentrations increased in compatibly pollinated flowers, and remained stable in incompatibly pollinated flowers after pollination and subsequent to pollen tube—ovule contact.

Pollination Requirements of Three Highbush Blueberry (Vaccinium corymbosum L.) Cultivars

The effects of pollination treatments on fruit set and five berry characteristics [mass, diameter, number of apparently viable seeds (well-developed, plump with dark seed coat), total seed number (includes apparently viable and partially developed seeds), and harvest date] were examined on three highbush blueberry cultivars. Pollination treatments included unpollinated, open pollinated, emasculated, and three hand pollinations that used pollen from the same flower, from the same cultivar, or from a different cultivar. Berries matured earliest and were smallest with the most apparently viable seeds in `Northland', `Patriot' had the greatest fruit set and smallest seed number, and `Bluecrop' matured the latest. Fruit set was greater, berry size larger, seed number smaller, and maturation later in 1990 than 1991. For all three cultivars, berries were generally smallest, latest maturing, and had the fewest seeds when pollination was prevented and were largest with the most seeds and earliest maturing in open visitation. Emasculation resulted in berries similar to those from unpollinated flowers. For berry characteristics, cross-pollination was of benefit for `Patriot' and possibly `Northland' but not `Bluecrop'. Thus, commercial highbush blueberry planting designs must be based on the pollination requirements of the particular cultivar. `Northland' berries almost always had seeds, while `Patriot' showed high levels and `Bluecrop' low levels of parthenocarpy.

A leucine-rich repeat containing receptor-like kinase marks somatic plant cells competent to form embryos.

The first somatic single cells of carrot hypocotyl explants having the competence to form embryos in the presence of 2,4-dichlorophenoxyacetic acid (2,4-D) were identified using semi-automatic cell tracking. These competent cells are present as a small subpopulation of enlarged and vacuolated cells derived from cytoplasm-rich and rapidly proliferating non-embryogenic cells that originate from the provascular elements of the hypocotyl. A search for marker genes to monitor the transition of somatic into competent and embryogenic cells in established suspension cell cultures resulted in the identification of a gene transiently expressed in a small subpopulation of the same enlarged single cells that are formed during the initiation of the embryogenic cultures from hypocotyl explants. The predicted amino acid sequence and in vitro kinase assays show that this gene encodes a leucine-rich repeat containing receptor-like kinase protein, designated Somatic Embryogenesis Receptor-like Kinase (SERK). Somatic embryos formed from cells expressing a SERK promoter-luciferase reporter gene. During somatic embryogenesis, SERK expression ceased after the globular stage. In plants, SERK mRNA could only be detected transiently in the zygotic embryo up to the early globular stage but not in unpollinated flowers nor in any other plant tissue. These results suggest that somatic cells competent to form embryos and early globular somatic embryos share a highly specific signal transduction chain with the zygotic embryo from shortly after fertilization to the early globular embryo.

Pollination-enhanced expression of a receptor-like protein kinase related gene in tobacco styles.

We have isolated a putative serine/threonine receptor kinase gene with an expression pattern indicating that it may play a role in the stylar response to pollination. Differential display PCR was used to select tobacco mRNAs with increased accumulation following pollination. NTS16, a cDNA identified by this method, is homologous to a ca. 2.4 kb mRNA primarily expressed in pistil tissues. Levels of this mRNA increase during floral development and are further increased by pollination reaching maximal accumulation 12-18 hours after pollination and then declining. mRNA levels can also be increased by the application of ethylene to unpollinated flowers. A polypeptide encoded by the NTS 16 open reading frame has sequence similarity to the catalytic domain of several receptor protein kinases from plants including the S-receptor kinases implicated in the rejection of self-pollen in Brassica species and the Pto gene product of tomato which confers resistance to a bacterial pathogen.

Effects of pollinia removal and insertion on flower longevity inChloraea alpina (Orchidaceae)

Although it is known that stigmatic pollen deposition may trigger early flower senescence, the existence of a similar plastic response of flower lifespan to pollen removal has been much less studied. Here we report on a factorial, manipulative experiment in which all 2 × 2 flower combinations of pollinia removal and stigmatic pollinia insertion were performed in inflorescences of the Patagonian ground orchidChloraea alpina. This experiment was conducted in the laboratory, in a population of cut inflorescences and in the field. We hypothesized that if expected fitness gains, through both the male and female functions, were weighed against the costs of flower maintenance, then early flower senescence should be triggered by either pollinia removal or insertion. The shortest flower lifespan would be expected in flowers where both processes occurred. Results showed that flower longevity was very strongly affected by pollinia insertion, reducing the flower lifespan by approximately 60%. The response of pollinia removal was much weaker. A significant reduction in flower longevity caused by pollinia removal was only detected in unpollinated flowers (i.e. no pollinia inserted). Within the racemose inflorescences, flowers in basal positions lived longer than flowers in terminal ones, which might be evidence of the importance of resource availability in determining maximum flower longevity. The observed responses of flower lifespan plasticity to pollinia manipulation only partially supported our expectations based on fitness benefit—cost relationships. Other factors that might explain these discrepancies are the different fitness gains that may indeed accrue to the processes of pollinia removal and insertion as they occur in nature, donor manipulation of the recipient flower lifespan associated with the evolution of pollen clustering into pollinia and physiological constraints in terms of the extent to which flower longevity may respond to pollen removal.

Pollen‐pistil interaction in kiwifruit (Actinidia deliciosa; Actinidiaceae)

The pollen‐pistil interaction has been examined in kiwifruit (Actinidia deliciosa). In this species a large number of seeds are produced in each fruit and a great many pollen grains germinate and grow to reach the ovules. This growth is assisted by an abundant secretion that is present all along the pistilar tract. At anthesis, the stigma is covered by a secretion where the pollen grains germinate and grow. The stylar transmitting tissue is initially rich in starch reserves, but the starch gradually disappears and, concomitantly, an abundant secretion that stains for carbohydrates appears in all of the intercellular spaces. Pollen tube growth relies on this secretion since it is depleted after pollen tube passage, while in unpollinated flowers it remains unaltered throughout the flower life‐span. In the ovary a similar situation occurs. The placental surface, where the pollen tubes grow before reaching the ovules, is covered by a number of obturators. At anthesis, these obturators are rich in starch reserves and have an abundant secretion on their outer surface. As time passes, starch disappears while the secretion increases. It is in this secretion that the pollen tubes grow on their way toward the ovules. These observations are discussed in terms of the support given by the pistil to pollen tube growth to achieve the highly successful reproductive performance of this species.

The possible role of embryo sac degeneration in yield reduction of summer turnip rape cultivar Candle

When pollination occurred within 24 h after anthesis, 69.2% of the pods of Brassica rapa cultivar Candle contained at least 50% fertilized ovules. If pollination occurred later than 4 d after anthesis, an occasional ovule near the base of the pistil was fertilized; however, no significant seed set was observed. Pollen tubes were observed entering the locules of the ovary from pollinations carried out as late as 5 d after anthesis. At 3 d after anthesis, in unpollinated flowers, a general deterioration of the embryo-sac contents began and gradually increased to include integumentary cells by 6 d after anthesis. Some ovules swelled as though they were fertilized, but no embryos were present. The results suggest that yield reductions in this species may occur because of the reduction of fertilization potential through rapid degeneration of embryo sacs following anthesis. Key words:Brassica rapa, embryo sac, yield

The ponticulus: a structure bridging pollen tube access to the ovule in Pistacia vera

The path of the pollen tube has been examined in pistachio (Pistacia vera), a chalazogamous species where the pollen tube penetrate the ovule via the chalaza. Special attention was paid to the way the pollen tube gains access to the ovule. A single anatropous ovule with a big funiculus occupies the entire ovary cavity. At anthesis, a physical gap exists between the ovule and the base of the style. However, upon pollen tube arrival a protuberance, the ponticulus, develops in the uppermost area of the funiculus between the style and ovule. This structure appears to facilitate access to the ovule by the pollen tube. The pollen tube penetrates the ovule via this ponticulus. Upon penetration, callose develops in the ponticulus cells surrounding the pollen tube. After pollen tube passage, the upper layer of the ponticulus lignifies and isolates the ovule from the style. This separation is further enlarged 2 weeks later when the ovary starts to develop without expansion of the ovule and a large gap develops separating the ovule from the style. Except for the induction of callose formation by the pollen tube in the funiculus, this process is independent of pollination and appears to be developmentally regulated since it occurs in the same way and at the same time in pollinated and unpollinated flowers. The ponticulus, although by a different mechanism, appears to be playing the role of an obturator regulating access of the pollen tube to the ovule. Furthermore, this access is restricted to a particular time during the development of the ovule.

An increase in ethylene sensitivity following pollination is the initial event triggering an increase in ethylene production and enhanced senescence of Phalaenopsis orchid flowers

Cut Phalaenopsis (Phalaenopsis hybrid, cv. Herbert Hager) flowers usually last about 2 weeks. Following pollination however, there is a rapid acceleration of the wilting process, which is completed within 2 days. The first event detected following pollination was an increase in ethylene sensitivity. This increased sensitivity began about 4 h after pollination and peaked 6 h later. A subsequent increase in ethylene production could only be detected 12 to 14 h after pollination. Treatment of the flowers with silver thiosulfate or 1‐methylcyclopropene, both inhibitors of ethylene action, completely inhibited the pollination‐induced increase in ethylene production and the enhanced senescence of the flowers. This indicates that the pollination‐induced increase in ethylene production is a response to the existing ethylene. Treatment of flowers with calcium and its ionophore A23187, which increased ethylene sensitivity and protein phosphorylation, also promoted ethylene production and senescence of unpollinated flowers, EGTA, a calcium chelator, decreased the sensitivity of pollinated flowers to ethylene and delayed and decreased the pollination‐induced increase in ethylene production. We suggest that the pollination‐induced increase in ethylene sensitivity is the initial pollination‐induced event triggering the enhancement of ethylene production, which leads to enhanced senescence of Phalaenopsis flowers.

Pollen tube penetration and fertilization inLilium longiflorum (Liliaceae)

The ultrastructure of the embryo sac, nucellus, and parts of the micropyle of Lilium longiflorum were studied both before and after pollen tube penetration to examine the interactions between ovule and pollen tube, using transmission electron microscopy and light microscopy. Before pollen tube penetration the egg cell and two synergids are similar. No filiform apparatus was detected and no synergid degeneration occurs prior to pollen tube penetration. The polar nuclei do not fuse until fertilization. No differences in embryo sac ultrastructure were detected between pollinated ovules unpenetrated by pollen tubes and unpollinated flowers of a comparable age. Shortly after the discharge of the pollen tube two enucleated cytoplasmic bodies with different ribosome densities were observed in the degenerated cytoplasm. These structures border both on the central cell and the egg cell as well as each other and are interpreted as remains of sperm cytoplasm after transmission of sperm nuclei. In the central cell both the sperm nucleus and the polar nuclei are associated with endoplasmic reticulum (ER). ER is thought to be a transport mechanism to achieve contact between the haploid polar nuclei and the sperm nucleus. In the egg cell sperm nucleus alignment is not visibly achieved by ER. The persistent cells of the egg apparatus and the central cell appear to become more metabolically active after pollen tube penetration. Pollen tube penetration already occurs despite the absence of a filiform apparatus and a low level of differences between the cells of the egg apparatus.

A reproductive calendar of Lilium longiflorum Thunb. cv. Gelria

The development and timing of different processes in the floral biology and sexual reproduction of Lilium longiflorum were studied and brought together into a chronological reproductive calendar. Care must be taken when emasculating flowers later than 4 days before anthesis (DBA), because the stomium of the anther was open from 8 DBA and the pollen grains appear to be mature from 3–2 DBA. The receptivity period of the pistil lasts for a rather long period, but before wilting of an unpollinated flower as determined by a change of the colour of the perianth from white into brown, the flower had already lost its receptivity. Because L. longiflorum pollen grains can easily be stored, the pistil receptivity should thus determine the pollination schedule. The relationships between pollen tubes and pistil are discussed with respect to plant breeding.

Strong synergistic effects of gibberellins with the synthetic cytokinin N-(2-chloro-4-pyridyl)-N-phenylurea on parthenocarpic fruit set and some other fruit characteristics of apple

The induction of parthenocarpic fruit set was investigated using the apple cvs. Golden Delicious and Jonagold. The gibberellins GA3, GA4, GA5 and GA7 and the synthetic phenylurea-type cytokinin CPPU (N-(2-chloro-4-pyridyl)-N-phenylurea), were applied alone and in combination to unpollinated flowers at the end of petal fall. Gibberellins induced only a marginal final set of parthenocarpic fruits. CPPU sprays were more effective, particularly in the first year. When applied in combination, CPPU and gibberellins had a positive synergistic effect on parthenocarpic fruit set and fruit size, but a negative effect on flower induction the next year. After CPPU + GA sprays, percent fruit set was similar, or greater, compared to natural pollinated trees. The parthenocarpic fruits induced by CPPU + GA had an increased length to diameter ratio. CPPU stimulated, and GA4 and GA7 reduced, the russeting of the parthenocarpic fruits. The internal quality of the fruits was hardly affected, but Ca-deficiency symptoms occurred more frequently in parthenocarpic fruits.

Functional Anatomy of the Ovule in Broad Bean, Vicia faba L. II. Ultrastructural Development up to Early Embryogenesis

Ovules of broad beans (Vicia faba) have been studied to analyze ultrastructural features for nutrient transport to the embryo sac at various ontogenetic stages up to 10 d after pollination. In unpollinated flowers a notable homogeneous or fibrillar material is deposited in the endostome, between the two integuments and on the nucellus. Osmiophilic globules accumulate at the plasmalemma and in the walls at the micropylar end of the inner integument. These globules increase in number after fertilization and appear also in other cells near the embryo sac. The central cell, which has some wall ingrowths typical for transfer cells, shows intrusive growth between cells of the nucellar cap. After fertilization wall thickenings occur in cells close to the embryo sac. At 10 d after pollination the inner integument has degenerated entirely. Also the nucellus, including the nucellar cap, is digested. In the endosperm free-nuclear divisions start and the cytoplasm increases in amount. Wall ingrowths are formed along the whole embryo sac boundary. The suspensor consists of two pairs of multinucleate cells: the pair adjacent to the embryo proper have rounded cells; the other pair have elongated ones. The suspensor cells that are attached to the embryo sac boundary become transfer cells. Their plastids have prolamellar bodies, and these structures are not seen anywhere else in the ovule. Our study confirms that transfer cells are common at junctions between the different generations in the ovules, that the transport to the embryo sac is apoplastic, and that symplastic transport is possible between endosperm and embryo and further between suspensor and embryo proper.