Analysis of pollen-style interactions in Petunia hybrida; the determination of variance in male reproductive success

Summary A factorial cross design was used to evaluate the influence of differences among pollen donors and recipients on variation in stylar attrition of pollen tubes in self-fertile plants of Petunia hybrida. Pollinations to flower buds were used to reduce the inhibitory ability of the style and these crosses were compared with flower pollinations to assess the degree of stylar influence on pollen fertilization ability. There was less pollen tube attrition after bud pollinations than after flower pollinations, indicating that styles of buds were less able to inhibit pollen tubes. The variance component for plants acting as pollen donors tended to be greater after flower pollinations than after bud pollinations. The lower variance in male success after bud pollinations indicates that differences among pollen donors after flower pollinations were due to stylar inhibition of pollen rather than differences in pollen vigor. Since the level of variation in pollen growth after pollination to flowers was greater among clones than among ramets within clones, the differences in pollen fertilization ability are probably genetically based.     

Flowering and male reproductive functions of chickpea (Cicer arietinum L.) genotypes as affected by salinity

The influence of salinity, given at germination (stage I) or 75 d after sowing (stage II), on flower production and characteristic features of male reproductive structures was studied in three promising genotypes of chickpea (Cicer arietinum L. cv. ICCV 88102, H 82-2 and C-235). In ICCV 88102 and H 82-2 salinity 20 meq 1^-1 increased the number of flowers when applied at both stages whereas in C-235 only when applied at the later stage. The salinity also delayed flowering; the higher salinity the greater was delay in flowering. In H 82-2 and C-235 salinity treatment given at stage II (when few flower buds appeared) hastened the flowering. The salinity curtailed pollen production; the reduction being minimum in C-235 and maximum in H 82-2. Germination was not significantly affected in C-235 pollen collected from plants grown under salinity conditions upto 60 meq 1^-1 applied at stage I but the tube elongation was inhibited. The inhibition of tube elongation was greatest in C-235. Salinity treatment administered at stage II did not affect significantly pollen germination excepting C-235 in which a consistent decline with increasing salinity was evident. Salinity stimulated tube growth in ICCV 88102. Na₂SO₄ in the germination medium was more detrimental for both pollen germination and tube growth than NaCl.     

Temperatures during flower bud development affect pollen germination,self‐incompatibility reaction and early fruit development of clementine (Citrus clementina Hort. ex Tan.)

• One of the key environmental factors affecting plant reproductive systems is temperature. Characterising such effects is especially relevant for some commercially important genera such as Citrus. In this genus, failure of fertilisation results in parthenocarpic fruit development and seedlessness, which is a much‐prized character. Here, we characterise the effects of temperature on flower and ovary development, and on pollen–pistil interactions in ‘Comune’ clementine (Citrus clementina Hort. ex Tan.).
• We examine flower bud development, in vitro pollen germination and pollen–pistil interaction at different temperatures (15, 20, 25 or 30 °C). These temperatures span the range from ‘cold’ to ‘hot’ weather during the flowering season in many citrus‐growing regions.
• Temperature had a strong effect on flower and ovary development, pollen germination, and pollen tube growth kinetics. In particular, parthenocarpic fruit development (indicated by juice vesicle growth) was initiated early if flowers were exposed to warmer temperatures during anthesis.
• Exposure to different temperatures during flower bud development also alters expression of the self‐incompatibility reaction. This affects the point in the pistil at which pollen tube growth is arrested and confirms the role of sub‐ and supra‐optimal temperatures in determining the numbers of pollen tubes reaching the ovary.

     

Effects of air pollutants on pollen

In vivo and in vitro pollen germination and pollen tube growth are affected by air pollutants. The stimulation and inhibition of these pollen characteristics depend on the pollen species as well as on the pollutant and its concentration, the exposure time, the relative humidity during exposure and some other factors. Mitosis, gas exchange, size and chemistry of pollen can also be influenced by air contaminants. Pollen germination and tube elongation differ in their response to air pollution stress, and both are more susceptible in vitro compared to the in vivo situation. Some possible mechanisms of action are reported in this review study. The effects of air contaminants on pollen are discussed in relation to sexual reproduction, gametophytic selection and adaptation to stress. Pollen may be used as sensitive biological indicators of pollution     

Stigma development and receptivity in almond (Prunus dulcis)

BACKGROUND AND AIMS: Fertilization is essential in almond production, and pollination can be limiting in production areas. This study investigated stigma receptivity under defined developmental stages to clarify the relationship between stigma morphology, pollen germination, tube growth and fruit set. METHODS: Light and scanning electron microscopy were employed to examine stigma development at seven stages of flower development ranging from buds that were swollen to flowers in which petals were abscising. Flowers at different stages were hand pollinated and pollen germination and tube growth assessed. Artificial pollinations in the field were conducted to determine the effect of flower age on fruit set. KEY RESULTS: Later stages of flower development exhibited greater stigma receptivity, i.e. higher percentages of pollen germination and more extensive tube growth occurred in older (those opened to the flat petal stage or exhibiting petal fall) than younger flowers. Enhanced stigma receptivity was associated with elongation of stigmatic papillae and increased amounts of stigmatic exudate that inundated papillae at later developmental stages. Field pollinations indicated that the stigma was still receptive and nut set was maintained in older flowers. CONCLUSIONS: Stigma receptivity in almond does not become optimal until flowers are past the fully open stage. The stigma is still receptive and fruit set is maintained in flowers even at the stage when petals are abscising. Strategies to enhance pollination and crop yield, including the timing and placement of honey bees, should consider the effectiveness of developmentally advanced flowers.     

Sodium thioglycollate enhances pollen germination and pollen tube elongation in cruciferous species

Sodium thioglycollate is a reducing agent used in microbiological growth media to enhance the growth of anerobic, microaerophilic, and facultative organisms, and in eukaryotic tissue extraction buffers to inhibit damaging oxidative reactions. Sodium thioglycollate was added to a semi-solid pollen germination medium to evaluate its effects on in vitro pollen germination and pollen tube elongation, based on the assumption that conditions within stylar tissues are less aerobic than in ambient conditions. We observed significant increases in the percent germination and pollen tube elongation of both crop and weedy mustard family species, on a medium containing 2.2 mM sodium thioglycollate. This suggests that sodium thioglycollate may be a useful amendment to semi-solid media and to solutions that are used to study pollen vigor, physiology, or gene expression, and to bioassay sensitivities of different species or genotypes to diverse physical and chemical factors.     

Chasmogamous and Cleistogamous Pollination in Salpiglossis sinuata

Pollen from chasmogamous flowers of Salpiglossis sinuata L. could not be induced to germinate in vitro unless stigmatal extract was applied to the culture medium. The substance that induces pollen germination in the stigmatal extract is water-soluble and heat-stable. Crosses could not be achieved between chasmogamous and cleistogamous flowers because of structural incompatibility. Pollinated pistils of chasmogamous flowers release a large amount of ethylene. The burst of ethylene release is due to an interaction between pollen tubes and stylar tissue and is directly proportional to the quantity of pollen placed on the stigma. Cleistogamous flower buds also produce a burst of ethylene at the time of pollination within the closed flower. The ethylene release may be a cause of reduced corolla development associated with cleistogamous flowers.     

Flower bud differentiation and development in fruiting and non-fruiting shoots in relation to fruit set in apricot (Prunus armeniaca L.)

Situations of high flower bud drop and low fruit set without apparent causes are common in fruit trees. The term flower quality has been coined to explain differences among flowers in their capacity to set fruit, but the causes underpinning these differences are largely unknown. This lack of knowledge is based on the fact that these differences are established a posteriori and there are no criteria to determine a priori what will make a flower to set a fruit or to drop. In this work, we profit from the empirical knowledge that there are fruiting and non-fruiting shoots to explore to which extent flower bud differentiation and bud development will affect the subsequent fruit set. For this purpose, the processes from flower bud differentiation to fruit set were sequentially analyzed in both types of shoots, over 2 years. More than half of the buds from long shoots aborted development and dropped before flowering. At anthesis, most of the remaining flowers showed underdeveloped pistils that failed to sustain pollen germination or pollen tube growth along the pistil. This unsuccessful development resulted in clear differences in fruit set between both types of branches. These results highlight that flower bud differentiation and development play an important role for fruit set and that developmental timing appears critical to reach anthesis with a fully developed pistil.     

The spatial dynamics of pollen beetles in relation to inflorescence growth stage of oilseed rape: implications for trap crop strategies

Semi‐field‐scale arrays of oilseed rape (Brassica napus L.) (Brassicaceae) plants were used to observe the development of distributions of pollen beetles (Meligethes aeneus Fabricius) (Coleoptera: Nitidulidae) in a simulated trap crop system where inflorescence growth stage alone was used to manipulate the pest. Over two successive years, pairs of 1 m spaced square arrays of 100 glasshouse‐grown plants were placed 40 m apart in the field in May, and were subject to natural infestation by pollen beetles. The test plot of each pair had a simulated trap crop, with an outer row of plants at early flowering stage intended to protect more susceptible inner plants at late bud stage, and the control plot had all plants at the late‐bud stage, simulating a standard crop situation. Pollen beetles were counted daily on each plant for 10–13 days. The spatio‐temporal development of plot infestation was analysed in relation to the distribution of racemes in bud and raceme in flowers using Spatial Analysis by Distance IndicEs (SADIE), and tests of edge and centre distribution. Inflorescence growth stage characteristics were shown to be important in determining the spatial distributions of pollen beetles. In control plots, the numbers of racemes in bud and in flower were never edge or centre distributed. In test plots, racemes in flower were always edge distributed, and racemes in bud began edge distributed and became centre distributed. Pollen beetle numbers were usually spatially associated with the abundance of racemes in bud and/or in flower. In control plots, pollen beetles were neither edge nor centre distributed, but in test plots they maintained a significant edge distribution for 7–10 days. At the end of the experiments, females were more centre distributed in the test plots than males, and were more closely associated with racemes with buds, whereas males were more associated with racemes with flowers. In early flowering stage plants, the number of racemes in flowers were a good indicator of the abundance of racemes in buds, but this relationship was lost as flowering progressed. Although flowering racemes provide strong cues for immigrating pollen beetles, the abundance of buds may be a more important determinant of residence time, particularly for females, and is therefore a critical determinant of trap crop effectiveness.     

Sunny-side up: flower heliotropism as a source of parental environmental effects on pollen quality and performance in the snow buttercup, Ranunculus adoneus (Ranunculaceae)

Floral traits affect mating success via their influence on the microenvironment in which sexual reproduction occurs as well as their impact on pollinator attraction. Here we investigate the importance of flower heliotropism as a source of parental environmental effects on pollen quality and performance. Flowers of the snow buttercup, Ranunculus adoneus, closely track the sun's rays. We experimentally restrained flowers to test for effects of heliotropism on pollen quality and performance after pollination. When equivalent amounts of pollen were transferred to recipient pistils, pollen from solar-tracking donor flowers exhibited a 32% advantage in germination compared to pollen from stationary (tethered) donor flowers. By the end of anthesis, pistils of tracking flowers contained 40% more germinating pollen grains and 44% more pollen tubes midway down the style than pistils of stationary ones. Solar tracking had no direct effect on pollen tube growth. The greater amount of germinating pollen in tracking flowers accounted for the treatment effect on pollen tube density. A survey of pollen receipt and pollen germination in naturally tracking flowers indicated that solar tracking primarily affects pollen tube density by promoting pollen germination rather than pollen deposition. We conclude that flower heliotropism, by enhancing the paternal environment for pollen development and the maternal environment for pollen germination, represents a source of positive parental environmental effects on pollen performance in snow buttercups.     

In vitro germination and transient GFP expression of American chestnut (Castanea dentata) pollen

The development of the male reproductive structures of American chestnut (Castanea dentata) is described to advance our understanding of its reproductive behavior. This information has been vital in the development of a strategy to collect pollen grains from male catkins suitable for in vitro germination and transformation experiments. Cutting male catkins into small segments and rolling them over a culture plate resulted in evenly dispersed and large amounts of pollen with minimal unwanted accessory floral parts. To optimize pollen viability, the effect of various storage conditions on in vitro germination was examined. Our results showed that initial storage at 4°C for 2 weeks significantly increased percent germination as compared to freshly collected pollen and those stored directly at −20°C or −80°C. This also means that for long-term storage of American chestnut pollen, the catkins should first be kept at 4°C for a couple of weeks and then at −80°C. The use of pollen grains with high viability is necessary for the transformation of American chestnut pollen. To optimize pollen transformation via particle bombardment, the effects of target distance, target pressure, and pollen developmental stage were examined. Statistical analysis showed that bombardment of ungerminated pollen at 1,100 psi resulted in the highest percent transient GFP expression (4.1%).     

Effects of high humidity and temperature stress on pollen membrane integrity and pollen vigour in Nicotiana tabacum

Summary The prolonged exposure of pollen Nicotiana tabacum to high humidity at both room temperature and 38° C did not affect membrane integrity as revealed by the fluorochromatic reaction (FCR) test, but did affect pollen vigour. At room temperature germination was not affected, although tube growth was reduced; at 38° C, there was both a reduction in tube growth and delayed germination. When the pollen was subjected to 1 h hydration followed by 1 h desiccation (up to a maximum of four cycles) at room temperature, a reduction in the FCR, germination and tube length after each desiccation treatment was observed. Subsequent hydration fully restored the FCR, but only partially restored germination and tube growth. At 38° C, however, FCR, germination, and tube growth were drastically reduced. The implications of these results on the relationship between FCR and germinability, the responses of pollen exposed to humidity and temperature stress in the field, and on pollen storage are discussed.     

Attractive Spring-food for willow grouse (Lagopus lagopus subsp. lagopus L.) studied using plant macrofossils and pollen in faeces: a methodological discussion

While macrofossil remains of herbs and flowers have disintegrated beyond identification, pollen survives the digestive process and can be identified, sometimes to species level, and is hence an important tool in determining and reconstructing animal diets.

This study compares macroscopic plant parts and pollen remains in faeces from grouse. While macroscopic remains reflect the quantity of bark, twigs, berries and leaves in the birds' food, the pollen also reflects other food, such as catkins, flower buds with developed pollen, flowers and pollen attached to leaves.

Both techniques reflect that Betula and Salix are the most important food for grouse in early Spring. Pollen analysis shows that later in Spring, after snow-melt, the birds are attracted to different plants, firstly, it is taxa within the Ericales and their fruits from the previous year. The nutrient value of food in periods with more or less snow cover is based on the environmental conditions of the previous year. Later on, different herbs, including insect-pollinated plants rich in nectar/sugar, are recorded in the pollen but are not reflected in the macro study, and are therefore lost in most calculations and not discussed. Nutrient content calculations based on Spring and Summer flowers need to be compared with the environmental and climate conditions of the current year.     

Effects of simulated acid rain on the pollen germination and pollen tube growth of apple (Malus sylvestris Miller cv. Golden)

The pollens of apple flowers have been treated with simulated acid rain solutions in range of pHs 2.9 to 5.0 in order to determine the threshold proportion values that lead the observed symptoms of detriments of acid rain. Compared to controls (pH 6.5), pollen germination decreased by 41.75% at pH 3.3 and pollen tube elongation decreased by 24.3% at pH 3.4. Acid rain threshold proportion value was around pH 3.3 and 3.4 for apple pollen germination and pollen tube elongation, respectively. Furthermore, pollen tube elongation was determined to be more sensitive to acid rain than pollen germination. The pH values below 3.1 resulted in complete destruction of pollen tubes. Pollen germination entirely stopped at around pH 3.0. Finally, it has been shown that the acid rain has a blocking effect on pollen germination and pollen tube elongation in apple. The conclusion is that not only pH value but also the quantity of acid rain is important factor in germination. The results were found statistically significant through the LSD test at levels of p < 0.05 and p < 0.01.     

Adaptive function of duodichogamy: Why do chestnut trees have two pollen emission phases?

Premise

Intersexual mating facilitation in flowering plants has been largely underexplored. Duodichogamy is a rare flowering system in which individual plants flower in the sequence male-female-male. We studied the adaptive advantages of this flowering system using chestnuts (Castanea spp., Fagaceae) as models. These insect-pollinated trees produce many unisexual male catkins responsible for a first staminate phase and a few bisexual catkins responsible for a second staminate phase. We hypothesized that duodichogamy increases female mating success by facilitating pollen deposition on stigmas of the rewardless female flowers through their proximity with attractive male flowers responsible for the minor staminate phase.

Methods

We monitored insect visits to 11 chestnut trees during the entire flowering period and explored reproductive traits of all known duodichogamous species using published evidence.

Results

In chestnuts, insects visited trees more frequently during the first staminate phase but visited female flowers more frequently during the second staminate phase. All 21 animal-pollinated duodichogamous species identified are mass-flowering woody plants at high risk of self-pollination. In 20 of 21 cases, gynoecia (female flower parts) are located close to androecia (male flower parts), typically those responsible for the second minor staminate phase, whereas androecia are often distant from gynoecia.

Conclusions

Our results suggest that duodichogamy increases female mating success by facilitating pollen deposition on stigmas by means of the attractiveness of the associated male flowers while effectively limiting self-pollination.

     

MARIS plays important roles in Arabidopsis pollen tube and root hair growth

In flowering plants, male gametes are delivered to female gametes for double fertilization through pollen tubes.Therefore, pollen tube growth is crucial for double fertilization. Despite its importance to sexual reproduction, genetic mechanisms of pollen tube growth remain poorly understood.In this study, we characterized the receptor-like cytoplasmic protein kinase(RLCK) gene, MARIS(MRI) that plays critical roles in pollen tube growth. MRI is preferentially expressed in pollen grains, pollen tubes and roots. Mutation in MRI by a Ds insertion led to a burst of pollen tubes after pollen germination. Pollen-rescue assay by pollen and pollen tubespecific expression of MRI in the mri-4 mutant showed that loss of MRI function also severely affected root hair elongation. MRI protein interacted with the protein kinase OXIDATIVE SIGNAL INDUCIBLE1(OXI1) in the in vitro and in vivo assays, which functions in plant defence and root hair development, and was phosphorylated by OXI1 in vitro. Our results suggest that MRI plays important roles in pollen tube growth and may function in root hair elongation through interaction with OXI1.     

Patterns and sources of variation in pollen deposition and pollen tube formation in flowers of the endemic monoecious shrub Cnidoscolus souzae (Euphorbiaceae)

Pollen deposition and pollen tube formation are key components of angiosperm reproduction but intraspecific variation in these has rarely been quantified. Documenting and partitioning (populations, plants and flowers) natural variation in these two aspects of plant reproduction can help uncover spatial mosaics of reproductive success and underlying causes. In this study, we assess variation in pollen deposition and pollen tube formation for the endemic monoecious shrub Cnidoscolus souzae throughout its distribution range in Mexico, and determine how this variation is structured among populations, plants and flowers. We also infer the relative importance of pollen quantity and quality in determining pollination success in this species. While we found no evidence suggesting that pollen receipt limits C. souzae reproduction across 19 populations, we did find extensive variation in pollen load size and pollen tube number per flower. Total variation in pollen receipt and pollen tube number was mostly explained by intra‐individual and among‐population variance. Furthermore, pollen load size had a stronger effect on the number of pollen tubes at the base of the style than pollen germination rate, suggesting that pollen quantity may be more important than quality for pollen tube success in C. souzae. Our results suggest that both small within‐plant flower differences and broad‐scale differences in community attributes can play an important role in determining pollination success. We emphasise the need to evaluate patterns and sources of variation in pollen deposition and pollen tube formation as a first step in understanding the causes of variation in pollination success over broad spatial scales.     

Nectarless flowers with deep corolla tubes in Pedicularis: does long pistil length provide an arena for male competition?

Pollinator‐mediated selection does not seem to have a direct influence on the evolution of a long corolla tube in a nectarless flower. We hypothesized that the long pistil length of the nectarless flower with a deep corolla tube provided an opportunity for male competition. Pedicularis siphonantha, a nectarless and partially self‐incompatible lousewort with substantial variation in corolla tube length, was used to test the hypothesis. We tested whether and how corolla tube length affected seed production per capsule and seed germination rate with different pollination treatments. Flowers were hand‐pollinated with pollen from one self donor and one outcross donor and mixed pollen grains consisting of equal amounts from the two donor types, respectively. Additionally, seeds from open‐pollinated flowers with different corolla tube lengths were collected separately for measurement of germination rate. Pollination treatment and corolla tube length significantly affected number of seeds per capsule. Moreover, a significant positive relationship between seeds per capsule and corolla tube length was found when mixed hand pollination was conducted. Seeds of self hand‐pollinated flowers had a lower germination rate than those from outcross‐pollinated flowers. Under open pollination, seeds from flowers with longer corolla tubes tended to have higher germination rate. In P. siphonantha, outcross pollen may have a higher probability of contributing to the next generation when transferred to flowers with longer corolla tubes. The pistil length, therefore, should be seen as a female choice mechanism, which provides an arena for male‐to‐male competition. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 526–532.     

β-Glucan synthetase II activity upon callose formation in the flower ofArabidopsis thaliana

Callose formation was observed in the pollens during flower development and pollen tube grown in the pistil ofA. thaliana. The accumulation of callose occurred in the tetrad in the flower bud and pollen tube. Therefore, the activity of β-glucan synthetase II (GS II), which is responsible for synthesizing the callose, was measured in the flowers on the same developmental stages. The enzyme activity was increased by about 10% while the level of callose contents was increased by about 70% in tetrads. Then, callose accumulation was increased during pollen tube growth by about 30% higher than the other stages and enzyme activity was detected, 30% more too. These results suggest that callose plays an important role in the growth of pollen and pollen tube by increasing GS II activity.