Significance of flower exploding pollination on the reproduction of the Scotch broom, <Emphasis Type="Italic">Cytisus scoparius</Emphasis> (Leguminosae)

The significance of single flower visits by pollinators on the reproductive success of the Scotch broom, Cytisus scoparius, with explosive flowers was investigated. Unexploded flowers (intact flowers) produced no fruit, implying that autonomous selfing and apomixis do not occur and that the explosion of flowers by insects is necessary for the fertilization of flowers. The fruit set in the natural condition was intermediate between that in artificial selfing or exploding and that in artificial outcrossing. Seed set and the ratio of seeds to ovules did not differ significantly between the natural condition and artificial outcrossing. The fruit set by a single flower visit from Apis mellifera was similar to natural conditions. Apis mellifera was the most frequent visitor and the number of flowers tripped open per hour was the highest among all the flower visitors. Approximately 75% of pollen on a flower disappeared in a single flower visit by A.mellifera. The small amount of pollen remaining on already-exploded flowers appears to be the reason why A.mellifera do not frequently visit already-exploded flowers. The higher seed set by outcrossing was believed to contribute to the seed set in both natural conditions and single flower visits by A.mellifera, and did not differ between them. Therefore, at least in Japan, A.mellifera appears to be the most important pollinator for C.scoparius, resulting in a high outcrossing rate with each single flower visit.     

Cytokinins promotion of flowering in Cymbidium ensifolium var. misericors in vitro

Callus-derived rhizomes of Cymbidium ensifolium var. misericors produced flowers precociously on a defined basal medium (1/2MS) containing of NAA with thidiazuron (TDZ), N⁶-(2-isopentenyl) adenine (2iP) or N⁶-benzyladenine (BA) within 100 d of culturing. Among eight cytokinins tested, TDZ at 3.3–10 µM or 2iP at 10–33 µM combined with 1.5 µM NAA were the most effective combinations for achieving flower induction in vitro. These undersized flowers were physically normal and bloomed for two weeks in vitro.     

Bees assess pollen returns while sonicating Solanum flowers

Summary Can bees accurately gauge accumulating bodily pollen as they harvest pollen from flowers? Several recent reports conclude that bees fail to assess pollen harvest rates when foraging for nectar and pollen. A native nightshade (Solanum elaeagnifolium Cavanilles) that is visited exclusively for pollen by both solitary and social bees (eg. Ptiloglossa and Bombus) was studied in SE Arizona and SW New Mexico. The flowers have no nectaries. Two experiments were deployed that eliminated pollen feedback to the bees by experimentally manipulating flowers prior to bee visits. The two methods were 1) plugging poricidal anthers with glue and 2) emptying anthers of pollen by vibration prior to bee visitation. Both experiments demonstrated that bees directly assess pollen harvest on a flower-by-flower basis, and significantly tailor their handling times, number of vibratile buzzes per flower and grooming bouts according to the ongoing harvest on a given flower. In comparison to experimental flowers, floral handling times were extended for both Bombus and Ptiloglossa on virgin flowers. Greater numbers of intrafloral buzzes and numbers of times bees groomed pollen and packed it into their scopae while still on the flower were also more frequent at virgin versus experimental flowers. Flowers with glued andreocia received uniformly brief visits from Bombus and Ptiloglossa with fewer sonications and virtually no bouts of grooming. Curtailed handling with few buzzes and grooms also characterized visits to our manually harvested flowers wherein pollen was artificially depleted. Sonicating bees respond positively to pollen-feedback while harvesting from individual flowers, and therefore we expect them to adjust their harvesting tempo according to the currency of available pollen (standing crop) within Solanum floral patches.     

Factors influencing the growth of micropropagated shoots and in vitro flowering of gentian

About 70% of the shoots developed from nodal explants ofGentiana triflora flowered in vitroondouble strength WPM medium containing 3% (w/v) sucrose, 0.5mg/l BA after 12 weeks of culture in a growth room at 22°Cwith continuous illumination (PPFD=60molm^–2 s^–1). The influences oninvitro shoot development and flowering of several factors includingthe position of the explant, requirements for sucrose, cytokinin orGA₃, variations of pH and photosynthetic photon flux density (PPFD)were investigated. In vitro flowering but not shootdevelopment of G. triflora decreased notably withincreaseddistance from the apex of the shoot, indicating the presence of a floralgradient in the micropropagated shoots. Conversely, as little as 0.01mg l^–1 GA₃ in the medium promotedshootdevelopment but even up to 0.2 mg l^–1GA₃ did not induce in vitro flowering.Even though BA could substitute GA₃ for a high level of shootdevelopment, it also promoted a high level of in vitroflowering at the PPFD of 60 molm^–2 s^–1. Sucrose was required for shootdevelopment and flowering in vitro and higher levels ofPPFD could not compensate effectively for the omission of the sugar from themedium. In general, the effects of different concentrations of BA in the mediumor variations of pH on shoot development and flowering invitro were found to be influenced by PPFD. A novel observation isthat precocious flowering of micropropagated gentian shoots did not occur ifthey were first cultured for 5 weeks in the dark before transfer to the lightcondition.     

Seed set inCichorium intybus L. by pollination of flowers developedin vitro

Formation of viable seeds ofCichorium intybus L. was achieved in anin vitro system. Flower formation, pollination, fertilization, embryogenesis and seed development occurredin vitro on chicory root explants on culture medium lacking plant growth regulators. After flower induction under a 24-h daylength treatment, the explants were transferred to a 16-h daylength at 40 E m^-2s^-1 irradiance for pollination and further seed development. Negative results were obtained when root explants were maintained continuously under a 24-h daylength during the whole culture period. Lower seed set was obtained when the cultures were at low irradiance. The need of a dark period and adequate level of irradiance are suggested as important factors to obtain viable seeds. The developedin vitro system can be used as a model to study the factors controlling the reproductive processes, and for the study of self-incompatibility in chicory.     

Notes on the pollination mechanisms ofMoraea inclinata andM. brevistyla (Iridaceae)

Individual flowers ofMoraea inclinata are nectariferous and last about six hours. They appear to be pollinated largely by bees in the familyHalictidae (Lasioglossum spp.,Nomia spp.,Zonalictus) and to a lesser extent by bees in the familyAnthophoridae (Amegilla). The mechanism of bee-pollination inM. inclinata is the Iris type; i.e., each flower consists of three pollination units (an outer tepal, a partly exserted anther, and the opposed style branch which terminates in a pair of petal-like crests). Bees rarely visit more than one pollination unit per flower. Transferral of pollen to the bee is passive and nototribic although all bees collected on the flowers were female and 55% of the bees carried pollen loads with 2–5 pollen taxa in their scopae.Moraea brevistyla flowers are nectariferous but lack scent and last two days. They are visited infrequently by bees and only one femaleLasioglossum spec. carried the pollen ofM. brevistyla. Unlike flowers ofM. inclinata those ofM. brevistyla deposit pollen only on the head and thorax. Bee-mediated autogamy in both species is avoided due to the erratic foraging patterns of the bees and the flexibility of each stigma lobe as the bee backs out of the flower. Approximately 2–4 flowers in the inflorescences of both species (6–8 flowers/infloresence) develop into capsules.     

Stem elongation and floral initiation on in vitro chicory root explants: influence of photoperiod

Chicory root explants (Cichorium intybus L.) were cultured in vitro under different photoperiods. In complete darkness, strong stem elongation, but no flowering induction was observed. We suggest that this stem elongation could be homologous to the pit growth in chicory heads in vivo. Under a photoperiod of 12 h (LI=±40 E m^–2 s^–1), only vegetative growth was observed. Photoperiods of 16 h or more light a day induced the in vitro explants to develop stems bearing flower buds. When the in vitro cultures were kept in the dark for different durations starting from the first day of culture and afterwards transferred to long-day conditions, 4 days dark were sufficient to cause a decrease in flowering induction. We suggest that during the dark culture, a flowering inhibitory process was started.     

The pollination biology of two species of Helicteres (Malvaceae) with different mechanisms of pollen deposition

Species of Helicteres are pollinated mainly by hummingbirds and bats. Most species pollinated by hummingbirds have a mechanism of depositing pollen on the top of the pollinator's head such as is shown in this work for Helicteres sacarolha. H. brevispira has an unusual mechanism of depositing pollen grains under the tail or on the abdomen of the hummingbirds. The top of the birds’ head may be considered an efficient place to transport pollen grains, because it is plain, easily accessible for deposition and donation of pollen and not easily accessible for grooming, while the tail is movable and inclined. Thus, H. brevispira pollinators may carry or transfer fewer amounts of pollen grains than H. sacarolha pollinators from one flower to another. If a large amount of pollen grains is lost, a higher quantity of flower or pollen has to be produced to guarantee reproductive success. Plants of H. brevispira set higher number of flowers and pollen grains per flower than H. sacarolha and have also higher rates of fruit and flower abortion. Thus, pollination efficiency of H. brevispira may be reached by high pollen and flower production. Flower change mechanism presented in this species may be also involved with the optimization of pollinator feeding and pollination efficiency.     

Pollination of four sympatric species ofAngelonia (Scrophulariaceae) by oil-collecting bees in NE. Brazil

The manner whereby the oil-producing bisaccate flowers ofAngelonia (Scrophulariaceae) are pollinated by female oil-collecting bees is reported for the first time. Observations were made in the Caatinga formation of Pernambuco, NE. Brazil, on four synchronopatric species. These differ in sizes and structural details of the corolla, level of flower exposition, and habitat preferences. All legitimate visitors wereCentris spp. (Anthophoridae):Angelonia hirta was mainly pollinated byC. fuscata andA. pubescens byC. hyptidis; A. bisaccata andA. hookeriana shared an unidentified species. Several exomalopsine, tetrapediine and meliponid bees exploit the flowers less descriminately for oil or pollen, respectively, without regularly contacting anthers and stigma. The flowers are protandrous, and are self-incompatible except those of the annualA. pubescens. After alighting, theCentris bees introduce their front legs simultaneously into each of the pouches and start alternate collecting movements to gather the oil from the trichome elaiophores. While doing so, they are forced by projections of the corolla floor to press their head under the anthers and stigma, whereby pollen is transferred with their frons or clypeus. On account of their collector type and behaviour,C. fuscata andC. spec. are not specialized toAngelonia but may equally exploit other nonrelated taxa for oil, whereasC. hyptidis exhibits oligolecty onA. pubescens. It possesses relatively elongate forelegs with padlike collectors suitable for sweeping the lipids from the scattered glandular hairs inside the divergent spurs of its host. It is the only species that also collects pollen (by buzzing) from the oil host.A. hirta and relatives, provided with dense elaiophore carpets, are, for their part, adapted to scrapingCentris species with typical oil collectors. Flower and bee phenologies, although largely dependent on the irregular rainfalls, are not always coincident.     

TheCaMV 35S promoter is highly active on floral organs and pollen of transgenic strawberry plants

We have evaluated the expression of the reporter -glucuronidase (GUS) gene driven by the cauliflower mosaic virus 35S (CaMV 35S) promoter in flowers and pollen from 14 independent transgenic strawberry lines. Of the 14 lines evaluated, 13 (92.8%) showed GUS activity—as estimated by the histochemical GUS assay—in some floral organs, with expression being most common in the flower stem, sepals, petals, ovary and stigma. Ten of these thirteen transgenic lines (77%) showed GUS activity in pollen, although the percentages of positive pollen per flower varied greatly among the different lines. A study of the GUS expression during pollen maturation showed that the (CaMV 35S) promoter showed low expression in pollen from flower buds before anthesis but was activated in mature pollen following anther dehiscence. The percentages of pollen grains that showed GUS activity ranged from 2.1% to 46.3%. These percentages were similar or even higher when mature pollen was stored dry at room temperature for 2 weeks. After 5 weeks of storage, the percentages of GUS-positive pollen decreased in two of the six lines analysed but remained at similar values in the other four lines. GUS activity was also measured in protein extracts of mature pollen by means of the fluorometric GUS assay, with the values obtained ranging from 3.8 mol MU mg protein^–1 h^–1 to 0.26 mol MU mg protein^–1 h^–1. Contrary to the generally held view that the CaMV 35S promoter is virtually silent in pollen, we conclude that it is highly expressed in transgenic strawberry pollen.Abbreviations CaMV 35S Cauliflower mosaic virus promoter - GUS -Glucuronidase (EC 3.2.1.31) - MU 4-Methyl umbelliferone - nos Nopaline synthase promoter - nptII Neomycin phosphotransferase - X-Gluc 5-Bromo-4-chloro-3-indolyl--d-glucuronic acid     

A study on floral biology of seedlings in vitro in Orychophragmus violaceus: Induction of flowers in seedlings of O. violaceus cultured in vitro

Seedlings of O. violaceus were cultured on MS media and treated at low temperature. Cold treatment at 5–7 °C for more than 7 days was needed for flower induction of seedlings in vitro originated from germinated seeds. When cultured on MS media supplemented with 2.5 mg l⁻¹ zeatin and 2 mg l⁻¹ gibberellin (GA₃), seedlings in vitro did initiate flowers without cold treatment. When MS media was used with a reduced amount of NH₄NO₃, flower induction of seedlings in vitro could be accelerated. This revised version was published online in June 2006 with corrections to the Cover Date.     

Induction of stem elongation on in vitro chicory root explants under unfavourable photoperiodic conditions by gibberellin A3

Flowering stems are formed under long-day conditions on root explants of chicory (Cichorium intybus L.) cultured in vitro while under short days, only vegetative growth is observed. Under short-day conditions (12 h), stem elongation is induced by treating newly formed shoot apices with GA₃ (1 l, 10^–3M). No flower buds were formed on the GA₃-induced stems. Long days seem to be indispensable for the induction of flower buds on the elongated stem.     

In vitro pollination of angiosperm ovules with gymnosperm pollen grains

Summary Pollen grains of the gymnosperm species Ephedra distachya and Pinus wallichiana germinated abundantly on the in vitro cultured placentae of the angiosperm species Nicotiana tabacum, Melandrium album and Allium moly. Some P. wallichiana pollen tubes entered the micropyles of M. album. Embryological observations of cross-pollinated M. album ovules 2 or 3 d after cross-pollination revealed the presence of pollen tube remnants within the embryo sacs. Karyological disturbances in the two- and three-celled proembryos and endosperm nuclei indicated their probable hybrid origin. In some crosses, gynogenetic haploid proembryos were also noticed.     

Characterization of <Emphasis Type="Italic">HoMADS 1</Emphasis> and its induction by plant hormones during <Emphasis Type="Italic">in vitro</Emphasis> ovule development in <Emphasis Type="Italic">Hyacinthus orientalis</Emphasis> L.

To understand the molecular mechanism of ovule development, a MADS box gene,HoMADS 1, has been isolated from the ovule tissues of Hyacinthus. Sequence comparison showed that HoMADS 1 is highly homologous to both class C and D genes. Furthermore, phylogenetic analysis suggests that HoMADS 1 is most likely a class D MADS box gene. RNA hybridization revealed that HoMADS 1 was exclusively expressed in the ovules. Over-expressing HoMADS 1 in transgenic Arabidopsis plants produced ectopic carpelloid structures, including ovules, indicating that HoMADS 1 is involved in the determination of carpel and ovule identities. Interestingly, during in vitro flowering, no HoMADS 1 mRNA was detected in the floral tissues at high level hormones in the media. However, HoMADS 1 mRNA accumulated in the floral tissues when the regenerated flowers were transferred to the media containing low level hormones which could induce in vitro ovule formation. Our data suggest that the induction of HoMADS 1 by plant hormones may play important roles during ovule initiation and development in the regenerated flower. Whether HoMADS 1 expression is also regulated by cytokinin and auxin during ovule development in planta remains to be investigated.     

Effects of condensed tannins in white clover flowers on their digestion in vitro

Protein in white clover (Trifolium repens L.) is poorly utilised by ruminants because of its extensive degradation to ammonia in the rumen. However, white clover produces condensed tannins (CT) in its flowers, which can reduce rumen proteolysis. Effects of increasing proportions of clover dry matter (DM) as flowers (and therefore floral CT) on soluble protein, ammonia and volatile fatty acid (VFA) concentrations were determined with in vitro incubations. Minced mixtures of 0, 250, 500, 750 and 1000 g/kg of DM as white clover flower (F) with the remainder as white clover leaf, were incubated in vitro and sampled after 0, 2, 4, 8, 12 and 24 h. Treatments contained 0, 13, 26, 39 and 52 g CT/kg DM, respectively. A further treatment with 500 g/kg DM as flower and 500 g/kg DM as leaf had polyethylene glycol added to remove effects of CT. Increasing the proportion of white clover as flowers from 0 to 1000 g/kg DM reduced net conversion of plant N to ammonia N from 290 to 120 mM/M at least partly due to reduced solubility of the protein. Treatments with 750 g/kg DM or more as clover flowers reduced ammonia concentrations to levels likely to limit microbial growth. Total VFA production was not affected by flower content, although the proportion of acetate to propionate increased. The contribution of CT to treatment effects was small compared to effects attributed to difference in chemical composition between flowers and leaves.     

Insect-induced wind pollination of the palm Chamaedorea pinnatifrons and pollination in the related Wendlandiella sp.

In Peruvian Amazonia pollination ecology of two palms, Chamaedorea pinnatifrons (Jacq.) Oerst. and Wendlandiella sp. was studied from October 1988 to January 1990. Both palms are dioecious understory species that form locally dense populations.In C. pinnatifrons both sexes flower synchronously during the dry season. Prior to anthesis, the pendulous male inflorescence is inhabited by numerous thrips (Thysanoptera) and Ptiliidae (Coleoptera). Staminate flowers open by a small basal slit between the petals. At anthesis pollen is shed and the movements of the insects inside the flowers trigger pollen release in small clouds. Thus, the powdery pollen becomes airborne and finally air currents act as a vector, carrying pollen to the inconspicuous female plants, which usually are not visited by insects. The term insect induced wind pollination is suggested for this pollination mode. Wendlandiella flowers during four months in the dry season. Male and female plants were not visited by insects. The dry condition of the pollen indicates that anemophily is the pollination mode in Wendlandiella. Fructification is rare but the plants show intense vegetative propagation.The significance of anemophily in the tropical lowland rain forest is discussed.     

Morphological development of anthers induced by the dimorphic smut fungus<Emphasis Type="Italic"> Microbotryum violaceum</Emphasis> in female flowers of the dioecious plant<Emphasis Type="Italic"> Silene latifolia</Emphasis>

When inoculated with the dimorphic smut fungus Microbotryum violaceum (Pers.) G. Deml and Oberwinkler, the female flower of the dioecious plant Silene latifolia (Miller) E.H.L. Krause develops anther-like structures filled with spores instead of pollen grains. Using natural scanning electron microscopy, Nomarski interference microscopy, and fluorescence microscopy, we investigated the morphological modifications of the host plant resulting from this parasitism and the localization of smut hyphae in the flower bud. Flowers of infected plants lasted significantly longer than those of healthy plants, probably because the infection strengthened floral organs, such as the flower base and the anther filaments. Smut hyphae were observed throughout all organs of the young flower buds of infected plants, including sepals, petals, stamens, and pistil primordia. In healthy female flowers, anthers initiated sporogenous cell formation, but lacked parietal cell layers. By contrast, the parietal cell layers of infected female flowers differentiated into tapetal tissue, middle cell layers, and endothecial layers, as in the anthers of healthy male flowers. Smut spore formation in the infected anther was initiated in intercellular regions between the sporogenous cells, resulting in degeneration of premature sporogenous cells, tapetal tissue, and middle cell layers. The development of the endothecial layers and epidermis in the infected anther were morphologically normal.Abbreviations DAPI 4,6-diamidino-2-phenylidole - i infected - PMC pollen mother cell     

Gender variation in Actinidia deliciosa,the kiwifruit

Summary Flower and fruit characters were measured in ten female, five male and five fruiting male selections of A. deliciosa var deliciosa (A. Chev) Liang and Ferguson. Flowers from female vines had functional pistils, which contained many ovules. Stamens appeared to be fully developed but produced only empty pollen grains. Flowers from male vines had functional stamens that produced high percentages of pollen grains with stainable cytoplasmic contents. Pistils did not contain ovules and were generally small with vestigial styles. Fruiting male vines had both staminate and bisexual flowers. Staminate flowers were similar to those found on strictly male vines. Bisexual flowers produced ovules and stainable pollen. Pistils were smaller than in pistillate flowers. Although the three flower sexes differed in style length, ovary dimensions and ovules per carpel, staminate and bisexual flowers were similar in number of flowers per inflorescence, stamen filament length, pollen stainability, inflorescence rachis length and carpel number, and differed from pistillate flowers in these characters. The three flower sexes had similar sepal and petal numbers. The fruit of fruiting males were considerably smaller than those of females. Low ovule number appears to be the major factor limiting fruit size in the fruiting males studied. Prospects for developing hermaphroditic kiwifruit cultivars through breeding are discussed.     

The importance of non-pollen plant parts as food sources for the common blossom thrips, Frankliniella schultzei

Like other flower thrips, Frankliniella schultzei Trybom (Thysanoptera: Thripidae) feeds on pollen. However, the influence of a pollen diet on the life history of F. schultzei may not be as significant as reported for other thrips species. Frankliniella schultzei was reared successfully and with low mortalities (20%) on Wax Mallow, (Malvaviscus arboreus Cav.) plant part diets. Development times and fecundity on a petal diet were not significantly different from that on a pollen diet. Fecundity on a diet combining M. arboreus pollen, petal and leaf tissues was significantly higher than those individually containing these tissues. In laboratory choice tests, F. schultzei females encountered petal most often of the three plant parts. Pollen and leaf were encountered with similar frequencies. A significantly higher proportion of petal encounters (0.8) resulted in feeding than did pollen encounters (0.5). Few leaf encounters (0.1) were followed by feeding. Adult and larval F. schultzei were found in M. arboreus flowers but not on leaves. All parts of the flower were inhabited and not just the pollen-bearing petal apices and anthers. We propose that feeding on pollen within M. arboreus flowers is just one of many influences on the life history of F. schultzei and suggest that this may extend to other thrips species/host-plant combinations.     

Microgametophytes in flowers with and without fruits ofPhlox drummondii (Polemoniaceae)

Relationships between microgametophyte numbers per flower and fruit-set were sought inPhlox drummondii Hook. This was accomplished by counting fluorescentstained pollen tubes in flowers with and without fruits in 21 populations, and in flowers subject to either supplemental or sparse pollination, and then determining whether the flowers initiate fruits. There was a conspicuous variation in mean pollen tubes per flower (3–38) and in percent fruit-set (17%–92%) among populations. Neighboring populations often differed strikingly in these respects. Flowers with fruits had an average of 15.2 pollen tubes per stigma whereas those without had an average of 3 7 tubes. Over all flowers, there was a mean of 3.7 tubes per ovule. There was a strong correlation between mean tube number within populations and their percent fruit-set (r = 0.86). Supplemental pollination significantly enhanced fruit-set, thereby supporting the notion that fruit-set was pollen-limited in most populations. In both natural and artificial pollinations a portion of flowers without fruits still had several pollen tubes. The occurrence of fruitless flowers containing tubes is explained by attrition factors, like late-acting pollen-pistil incompatibility, and resource limitation. Fruit-set could be initiated with one pollen tube, but this rarely occurred in nature. Higher pollen loads were accompanied by higher percents of fruit-set and number of seeds per flower.