Pollen Tube Growth Rate Research Articles

Diurnal pollen tube growth rate is slowed by high temperature in field-grown Gossypium hirsutum pistils

For Gossypium hirsutum pollination, germination, and pollen tube growth must occur in a highly concerted fashion on the day of flowering for fertilization to occur. Because reproductive success could be influenced by the photosynthetic activity of major source leaves, we hypothesized that increased temperatures under field conditions would limit fertilization by inhibiting diurnal pollen tube growth through the style and decreasing subtending leaf photosynthesis. To address this hypothesis, G. hirsutum seeds were sown on different dates to obtain flowers exposed to contrasting ambient temperatures while at the same developmental stage (node 8 above the cotyledons). Collection and measurement were conducted at 06:00, 09:00, 12:00, 15:00, and 18:00 h on August 4 (34.6 °C maximum air temperature) and 14, 2009 (29.9 °C maximum air temperature). Microclimate measurements included photosynthetically active radiation, relative humidity, and air temperature. Pistil measurements included pistil surface temperature, pollen germination, pollen tube growth through the style, fertilization efficiency, fertilized ovule number, and total number of ovules per ovary. Subtending leaf measurements included leaf temperature, photosynthesis, and stomatal conductance. Under high temperatures the first measurable pollen tube growth through the style was observed earlier in the day (12:00 h) than under cooler conditions (15:00 h). Also, high temperature resulted in slower pollen tube growth through the style (2.05 mm h −1) relative to cooler conditions (3.35 mm h −1), but there were no differences in fertilization efficiency, number of fertilized ovules, or ovule number. There was no effect of sampling date on diurnal photosynthetic patterns, where the maximum photosynthetic rate was observed at 12:00 h on both dates. It is concluded that, of the measured physiological and reproductive processes, pollen tube growth rate showed the greatest sensitivity to high temperature under field conditions.

Pollen density on the stigma affects endogenous gibberellin metabolism, seed and fruit set, and fruit quality in Pyrus pyrifolia

To clarify the relationship between pollen density and gametophytic competition in Pyrus pyrifolia, gametophytic performance, gibberellin metabolism, fruit set, and fruit quality were investigated by modifying P. pyrifolia pollen grain number and density with Lycopodium spores. Higher levels of pollen density improved seed viability, fruit set, and fruit quality. Treatments with the highest pollen density showed a significantly increased fruit growth rate and larger fruit at harvest. High pollen density increased germination rate and gave a faster pollen tube growth, both in vivo and in vitro. Endogenous gibberellin (GA) concentrations increased in pollen tubes soon after germination and the concentration of two growth-active GAs, GA3, and GA4, was positively correlated to final fruit size, cell numbers in the mesocarp, and pollen tube growth rate. These two GAs appear to be biosynthesized de novo in pollen tube and are the main pollen-derived bioactive GAs found after pollen germination. GA1 levels in the pollen tube appear to be related to a pollen–style interaction that occurred after the pollen grains landed on the stigma.

Cryptic self‐incompatibility and distyly in Hedyotis acutangula Champ. (Rubiaceae)

Distyly, floral polymorphism frequently associated with reciprocal herkogamy, self- and intramorph incompatibility and secondary dimorphism, constitutes an important sexual system in the Rubiaceae. Here we report an unusual kind of distyly associated with self- and/or intramorph compatibility in a perennial herb, Hedyotis acutangula. Floral morphology, ancillary dimorphisms and compatibility of the two morphs were studied. H. acutangula did not exhibit precise reciprocal herkogamy, but this did not affect the equality of floral morphs in the population, as usually found in distylous plants. Both pin and thrum pollen retained relatively high viability for 8 h. The pollen to ovule ratio was 72.5 in pin flowers and 54.4 in thrum flowers. Pistils of pin flowers remained receptive for longer than those of thrum flowers. No apparent difference in the germination rate of pin and thrum pollen grains was observed when cultured in vitro, although growth of thrum pollen tubes was much faster than that of pin pollen tubes. Artificial pollination revealed that pollen tube growth in legitimate intermorph crosses was faster than in either intramorph crosses or self-pollination, suggesting the occurrence of cryptic self-incompatibility in this species. Cryptic self-incompatibility functioned differently in the two morphs, with pollen tube growth rates after legitimate and illegitimate pollination much more highly differentiated in pin flowers than in thrum flowers. No fruit was produced in emasculated netted flowers, suggesting the absence of apomixis. Our results indicate that H. acutangula is distylous, with a cryptic self-incompatibility breeding system.

Asymmetrical conspecific seed-siring advantage between Silene latifolia and S. dioica.

Silene dioica and S. latifolia experience only limited introgression despite overlapping flowering phenologies, geographical distributions, and some pollinator sharing. Conspecific pollen precedence and other reproductive barriers operating between pollination and seed germination may limit hybridization. This study investigates whether barriers at this stage contribute to reproductive isolation between these species and, if so, which mechanisms are responsible. Pollen-tube lengths for pollen of both species in styles of both species were compared. Additionally, both species were pollinated with majority S. latifolia and majority S. dioica pollen mixes; then seed set, seed germination rates and hybridity of the resulting seedlings were determined using species-specific molecular markers. The longest pollen tubes were significantly longer for conspecific than heterospecific pollen in both species, indicating conspecific pollen precedence. Seed set but not seed germination was lower for flowers pollinated with pure heterospecific versus pure conspecific pollen. Mixed-species pollinations resulted in disproportionately high representation of nonhybrid offspring for pollinations of S. latifolia but not S. dioica flowers. The finding of conspecific pollen precedence for pollen-tube growth but not seed siring in S. dioica flowers may be explained by variation in pollen-tube growth rates, either at different locations in the style or between leading and trailing pollen tubes. Additionally, this study finds a barrier to hybridization operating between pollination and seed germination against S. dioica but not S. latifolia pollen. The results are consistent with the underlying cause of this barrier being attrition of S. dioica pollen tubes or reduced success of heterospecifically fertilized ovules, rather than time-variant mechanisms. Post-pollination, pre-germination barriers to hybridization thus play a partial role in limiting introgression between these species.

Pollen behaviour and fertililization impairment in Bambara groundnut (Vigna subterrenea [L.] Verdc.)

Two field experiments were conducted in April and August, 2007 cropping season at the Department of Crop Science research farm, University of Nigeria, Nsukka, to evaluate the pollen germination potentials and the pollen tube growth of thirteen bambara groundnut cultivars. The harvested pollen grains were exposed for 0, 5, 10 and 15 min durations to ambient conditions before in vitro germination. Pollen germination and pollen tube growth were tested using a medium containing 10 g sucrose, 100 mg/l boric acid and 300 mg/l calcium nitrate made up to 100 ml with deionized water. The results showed that cultivars, had significant effect on the pollen germination only at the late planting. The early and late planting results showed that pollens incubated immediately after harvest had the highest germination percentage,while pollen exposed for five minutes prior to germination showed very poorgermination. Pollens exposed beyond five minutes after harvest did not germinate. The cultivars significantly (P < 0.05) affected the pollen tube growth at both early and late planting dates. The pollen tube growth decreased drastically with increase in duration of pollen exposure. The pollen tube had an exponential growth rate at the on set of the pollen tube growth, followed by lag and stationary growth phases in the pollen tube growth curves. The exponential, logistic and Gomperzt growth models were used to estimate the best fit model for the pollen tube growth in bambara groundnut cultivars. In the early planting, the average straightness (R2) value of these models for growth estimate were 98.4, 98.6 and 98.5%, respectively. During the late planting, the average R2 values were 91.5, 96.6 and 96.6%, respectively. The three models are therefore considered suitable for the computation of the pollen tube growth rates. The principal component and cluster analyses were used to groupthe cultivars in relation to the levels of pollen survival under ambient conditions. At early planting, cultivar Bg-01 had moderate surviving pollen grains while Bg-08, Bg-09, Bg-10 and Bg-11 were found to have poor surviving pollens. At late planting, cultivar Bg-04 and Bg-07 had high survival pollens while Bg-01 had poor pollen survival. Key words: Pollen behaviour, pollination, fertilization, Bambara groundnut, Vigna subterranea (L.) Verdc.

The role of pollen in the changing environmental conditions of Scots pine

Rapid climate warming poses a challenge to forest trees adapted to cooler conditions. However, a genetic safety net may exist for wind pollinated tree species in the form of long distance gene flow mediated by pollen containing alleles that are (pre)adapted to a warmer climate. In order to examine long distance gene flow in Scots pine (Pinus sylvestris L.), we monitored flowering phenology in central, northern, and northernmost Finland and measured the amount and viability of airborne pollen over four years. Viable airborne pollen grains were detected during female flowering periods and prior to local pollen shedding in all study sites. Depending on the year and study site, female flowers were exposed only to pollen originating elsewhere for between one and four days. Foreign airborne pollen represented 2.3% of all viable grains detected and varied between years and study sites. Artificial pollination trials were conducted to evaluate the effect of pollen origin on seed siring success. Pollen genotypes originating from southern Finland sired 76% and 48% of seeds in competition experiments where pollen from both populations was introduced simultaneously into the female strobilus. Furthermore, we investigated the importance of pollen arrival order by introducing pollen of northern and southern genotypes in to the strobilus at two-hourly intervals. Northern genotypes sired 76% of the analysed seeds when injected first but when southern genotypes were supplied first, both pollen types enjoyed equal success. The first pollen grain in the pollen chamber did not always fertilize the ovum. Instead there is likely a more subtle and complex form of competition between genotypes and their pollen grains. To examine chemically-mediated pollen interactions we conducted an in vitro germination experiment where different genotypes were in chemical but not physical contact. Both positive and negative interactions were found. We found highly negative effects in the germinability of northern pollen grains when in contact with southern pollen, and an increase in the germinability of southern pollen when in contact with northern pollen. There was no variation in the size of dry pollen grains between sources, but after hydration and germination, northern pollen grains were larger than southern pollen. Genotypes with high hydration rates had low germinability and slow pollen tube growth rates. This suggests that early germination and growth of the pollen tube is more dependent on pollen storage materials and less dependent on water intake. Pollen source affected the outcome of competition experiments and therefore is an important factor in long distance gene flow. However, genotype also plays a role and must be taken into account.

Vacuolar sorting receptors (VSRs) and secretory carrier membrane proteins (SCAMPs) are essential for pollen tube growth

Vacuolar sorting receptors (VSRs) are type-I integral membrane proteins that mediate biosynthetic protein traffic in the secretory pathway to the vacuole, whereas secretory carrier membrane proteins (SCAMPs) are type-IV membrane proteins localizing to the plasma membrane and early endosome (EE) or trans-Golgi network (TGN) in the plant endocytic pathway. As pollen tube growth is an extremely polarized and highly dynamic process, with intense anterograde and retrograde membrane trafficking, we have studied the dynamics and functional roles of VSR and SCAMP in pollen tube growth using lily (Lilium longiflorum) pollen as a model. Using newly cloned lily VSR and SCAMP cDNA (termed LIVSR and LISCAMP, respectively), as well as specific antibodies against VSR and SCAMP1 as tools, we have demonstrated that in growing lily pollen tubes: (i) transiently expressed GFP-VSR/GFP-LIVSR is located throughout the pollen tubes, excepting the apical clear-zone region, whereas GFP-LISCAMP is mainly concentrated in the tip region; (ii) VSRs are localized to the multivesicular body (MVB) and vacuole, whereas SCAMPs are localized to apical endocytic vesicles, TGN and vacuole; and (iii) microinjection of VSR or SCAMP antibodies and LlVSR small interfering RNAs (siRNAs) significantly reduced the growth rate of the lily pollen tubes. Taken together, both VSR and SCAMP are required for pollen tube growth, probably working together in regulating protein trafficking in the secretory and endocytic pathways, which need to be coordinated in order to support pollen tube elongation.

The joint evolution of mating system and pollen performance: Predictions regarding male gametophytic evolution in selfers vs. outcrossers

Abstract Studies of sexual selection in plants historically have focused on pollinator attraction, pollen transfer, gametophytic competition, and post-fertilization discrimination by maternal plants. Pollen performance (the speeds of germination and pollen tube growth) in particular is thought to be strongly subject to intrasexual selection, but the effect of mating system on this process has not been rigorously evaluated. Here we propose four predictions derived from the logic that pollen performance should evolve with mating system as an adaptive response to: (1) the competitive environment among pollen genotypes and (2) variation among female genotypes regularly encountered by a given pollen genotype. First, as previously predicted, due to the higher potential for intense selection among diverse pollen genotypes in outcrossing relative to selfing taxa, pollen should evolve to germinate and/or to grow more rapidly in outcrossers than in selfers. Second, due to stronger selection on pollen performance in outcrossing than in selfing taxa, heritable variation in pollen tube growth rate is more likely to be purged in outcrossers. In selfers, by contrast, genetic variation in pollen tube growth rates may readily accumulate because selfing reduces the number of genetically distinct male gametophytes likely to be deposited on any given stigma, thereby relaxing selection on male gametophytic traits. A summary of published studies presented here provides preliminary support for this prediction. Third, due to the high probability that the pollen of outcrossing individuals will be exposed to multiple pistil genotypes, we predict that the pollen of habitually outcrossing taxa will evolve to perform more consistently across female genotypes than the pollen of selfing taxa. Fourth, we predict that epistatic interactions between pollen and pistil genotypes are more likely to evolve in selfers than in outcrossers. We suggest several empirical approaches that may be used to test these predictions.

Incidence and post-pollination mechanisms of nonrandom mating in Arabidopsis thaliana

Compatible pollinations from many different taxa display nonrandom mating. Here we describe a system for examining questions of nonrandom mating in Arabidopsis thaliana. Using this system, we demonstrate that Arabidopsis thaliana displays nonrandom mating between distinct accessions. Statistical analysis of these data demonstrates aspects of both pollen competition and male-female complementarity in these matings. Cytological experiments implicate pollen germination and pollen tube growth rates as possible causal factors in these nonrandom mating efficiencies.

Effects of Temperature and the Combination of Liquid Lime Sulfur and Fish Oil on Pollen Germination, Pollen Tube Growth, and Fruit Set in Apples

Effects of temperature and the combination of liquid lime sulfur (LLS) and fish oil (FO) applied during bloom on pollen germination and pollen tube growth in flowers and fruit set were examined in apples (Malus ×domestica Borkh.). Percent germination of pollen of ‘Manchurian’ crabapples and ‘Golden Delicious’ apple flowers on the stigmatic surface of ‘Golden Delicious’ pistils increased with increasing temperature from 13 to 29 °C in the first 24 and 48 h after pollination, respectively, but not thereafter. Pollen tube growth rate in the style increased quadratically with increasing temperature from 13 to 29 °C. ‘Manchurian’ was a more effective pollenizer of ‘Golden Delicious’ than was ‘Golden Delicious’ pollen. For example, at 24 or 29 °C, some ‘Manchurian’ pollen tubes grew to the base of ‘Golden Delicious’ styles by 24 h after pollination. On the other hand, no ‘Golden Delicious’ pollen tube grew to the base of a ‘Golden Delicious’ style regardless of temperature and time. Pollen tube growth rate in the style increased with increasing day/night temperature from 7/0 to 24/7 °C. The time required for pollen tubes to grow to the base of styles decreased with increasing day/night temperature from 13/2 to 24/7 °C. Only ≈36 h was required for pollen tubes to grow to the base of style at 24/7 °C, whereas pollen tubes grew very slowly and no pollen tubes grew to the base of style at 7/0 °C regardless of pollen source. LLS + FO, applied 4 or 24 h after pollination, inhibited pollen germination, pollen tube growth in the style, fertilization, and fruit set, but it had no effect when applied 48 h after pollination. These results suggest that LLS + FO applied at this bloom stage causes flower or fruit abscission most likely by inhibiting pollen germination, pollen tube growth in the style, and fertilization.

Post-pollination mechanisms in Nicotiana longiflora and N. plumbaginifolia: pollen tube growth rate, offspring paternity and hybridization

In natural populations where interfertile species coexist, conspecific and heterospecific pollen can be delivered to the stigmas. Post-pollination mechanisms might determine the seed siring success of different pollen donors within species as well as the chances for hybridization between species. Nicotiana longiflora and N. plumbaginifolia occur in sympatry in Northwest Argentina, where they have overlapping flowering seasons and share floral visitors. We explored (1) pollen tube growth rates for outcross versus self pollen in single-donor pollinations; (2) siring success of self versus outcross pollen donors in competitive pollinations, and (3) possibilities for hybridization by performing two- (outcross conspecific vs. heterospecific) and three-pollen donor (self vs. outcross vs. heterospecific) crosses. In N. longiflora, both pollen tube growth rate and siring success favored outcross pollen over self pollen and strong rejection of heterospecific pollen. In N. plumbaginifolia, pollen tube growth rate was similar for self and outcross pollen, self pollen sired similar numbers of offspring than outcross pollen and heterospecific pollen sired roughly the same number of progeny than self pollen. Results suggest that in natural sympatric populations, interspecific crosses would likely lead to unidirectional hybridization with N. plumbaginifolia as the seed parent.

H +-ATPase in the plasma membrane of Arabidopsis pollen cells is involved in extracellular calmodulin-promoted pollen germination

In this paper, the role of the plasma membrane (PM) H +-ATPase in extracellular calmodulin (CaM)-promoted pollen germination and in tube growth of Arabidopsis thaliana was investigated. Pollen germination, pollen tube growth rate, free cytosolic Ca 2+ concentration ([Ca 2+] cyt) and Ca 2+ channel activity in the PM of pollen cells were measured. In response to fusicoccin or CaM treatment, in vitro pollen germination and pollen tube growth rate, [Ca 2+] cyt and activity of a hyperpolarization-activated Ca 2+-permeable channel increased. Sodium vanadate inhibited the promotion of these parameters by extracellular CaM. The results suggest that H +-ATPase may be involved in extracellular CaM-regulated pollen germination and in tube growth by modulation of the hyperpolarization-activated Ca 2+ channel in the PM of A. thaliana pollen cells.

Analysis of the tip-to-base gradient of CaM in pollen tube pulsant growth using in vivo CaM–GFP system

Ca(2+)-CaM signaling is involved in pollen tube development. However, the distribution and function of CaM and the downstream components of Ca(2+)-CaM signal in pollen tube development still need more exploration. Here we obtained the CaM-GFP fusion protein transgenic line of Nicotiana tobacum SRI, which allowed us to monitor CaM distribution pattern in vivo and provided a useful tool to observe CaM response to various exogenous stimulations and afforded solid evidences of the essential functions of CaM in pollen tube growth. CaM-GFP fusion gene was constructed under the control of Lat52-7 pollen-specific promoter and transformed into Nicotiana tobacum SRI. High level of CaM-GFP fluorescence was detected at the germinal pores and the tip-to-base gradient of fluorescence was observed in developing pollen tubes. The distribution of CaM at apical dome had close relationship with the pulsant growth mode of pollen tubes: when CaM aggregated at the apical dome, pollen tubes stepped into growth state; When CaM showed non-polarized distribution, pollen tubes stopped growing. In addition, after affording exogenous Ca(2+), calmidazolium (antagonism of CaM) or Brefeldin A (an inhibitor of membrane trafficking), CaM turned to a uniform distribution at the apical dome and pollen tube growth was held back. Taken together, our results showed that CaM played a vital role in pollen tube elongation and growth rate, and the oscillation of tip-to-base gradient of CaM was required for the normal pulsant growth of pollen tube.

Consequences of Pollination Syndrome Evolution for Postpollination Biology in an Ancient Angiosperm Family

Evolutionary shifts from insect to wind pollination involve a host of modifications to floral structure and phenology, but little is known about how floral modifications that facilitate pollination might affect the fertilization process. Within the water lily family Cabombaceae, there is evidence that wind pollination arose recently in Brasenia, whereas the sister genus Cabomba became specialized for fly pollination. Both species have an apomorphic stylar extension, which in Brasenia became greatly elongated to produce a much larger stigmatic surface. Consequently, pollen tubes in Brasenia must travel much farther to reach ovules, and because mean pollen tube growth rates are similar (750–950 μm/h), fertilization occurs ∼4 h later in Brasenia than in Cabomba. In both genera, pollen tubes grow between cells of the substigmatic ground tissue and then within an open, secretion‐filled stylar canal and ovarian cavity. In Brasenia, early pollen tube development is slower than in Cabomba, which may be a result o...

Effects of floral thermogenesis on pollen function in Asian skunk cabbage Symplocarpus renifolius

The effects of temperature on pollen germination and pollen tube growth rate were measured in vitro in thermogenic skunk cabbage, Symplocarpus renifolius Schott ex Tzvelev, and related to floral temperatures in the field. This species has physiologically thermoregulatory spadices that maintain temperatures near 23 degrees C, even in sub-freezing air. Tests at 8, 13, 18, 23, 28 and 33 degrees C showed sharp optima at 23 degrees C for both variables, and practically no development at 8 degrees C. Thermogenesis is therefore a requirement for fertilization in early spring. The narrow temperature tolerance is probably related to a long period of evolution in flowers that thermoregulate within a narrow range.

Pollen-tube growth rates in Collinsia heterophylla (Plantaginaceae): one-donor crosses reveal heritability but no effect on sporophytic-offspring fitness

Evolutionary change in response to natural selection will occur only if a trait confers a selective advantage and there is heritable variation. Positive connections between pollen traits and fitness have been found, but few studies of heritability have been conducted, and they have yielded conflicting results. To understand better the evolutionary significance of pollen competition and its potential role in sexual selection, the heritability of pollen tube-growth rate and the relationship between this trait and sporophytic offspring fitness were investigated in Collinsia heterophylla. Because the question being asked was if female function benefited from obtaining genetically superior fathers by enhancing pollen competition, one-donor (per flower) crosses were used in order to exclude confounding effects of post-fertilization competition/allocation caused by multiple paternity. Each recipient plant was crossed with an average of five pollen donors. Pollen-tube growth rate and sporophytic traits were measured in both generations. Pollen-tube growth rate in vitro differed among donors, and the differences were correlated with in vivo growth rate averaged over two to four maternal plants. Pollen-tube growth rate showed significant narrow-sense heritability and evolvability in a father-offspring regression. However, this pollen trait did not correlate significantly with sporophytic-offspring fitness. These results suggest that pollen-tube growth rate can respond to selection via male function. The data presented here do not provide any support for the hypothesis that intense pollen competition enhances maternal plant fitness through increased paternity by higher-quality sporophytic fathers, although this advantage cannot be ruled out. These data are, however, consistent with the hypothesis that pollen competition is itself selectively advantageous, through both male and female function, by reducing the genetic load among successful gametophytic fathers (pollen), and reducing inbreeding depression associated with self-pollination in plants with mix-mating systems.

Amborella trichopoda (Amborellaceae) and the evolutionary developmental origins of the angiosperm progamic phase

A remarkable number of the defining features of flowering plants are expressed during the life history stage between pollination and fertilization. Hand pollinations of Amborella trichopoda (Amborellaceae) in New Caledonia show that when the stigma is first receptive, the female gametophyte is near maturity. Pollen germinates within 2 h, and pollen tubes with callose walls and plugs grow entirely within secretions from stigma to stylar canal and ovarian cavity. Pollen tubes enter the micropyle within 14 h, and double fertilization occurs within 24 h. Hundreds of pollen tubes grow to the base of the stigma, but few enter the open stylar canal. New data from Amborella, combined with a review of fertilization biology of other early-divergent angiosperms, show that an evolutionary transition from slow reproduction to rapid reproduction occurred early in angiosperm history. I identify increased pollen tube growth rates within novel secretory carpel tissues as the primary mechanism for such a shift. The opportunity for prezygotic selection through interactions with the stigma is also an important innovation. Pollen tube wall construction and substantial modifications of the ovule and its associated structures greatly facilitated a new kind of reproductive biology.

Ectopic expression of Arabidopsis thaliana plasma membrane intrinsic protein 2 aquaporins in lily pollen increases the plasma membrane water permeability of grain but not of tube protoplasts.

To investigate the role of aquaporin-mediated water transport during pollen grain germination and tube growth, Arabidopsis thaliana plasma membrane intrinsic proteins (PIPs) were expressed in pollen of Lilium longiflorum (lily). Successful expression of AtPIPs in particle-bombarded lily pollen grains was monitored by co-expression with fluorescent proteins and single-cell RT-PCR, and by measuring the water permeability coefficient (P(os)) in swelling assays using protoplasts prepared from transformed pollen grains and tubes. Expression of AtPIP1;1 and AtPIP1;2 in pollen grains resulted in P(os) values similar to those measured in nontransformed pollen grain protoplasts (6.65 +/- 2.41 microm s(-1)), whereas expression of AtPIP2 significantly increased P(os) (AtPIP2;1, 13.79 +/- 6.38; AtPIP2;2, 10.16 +/- 3.30 microm s(-1)). Transformation with combinations of AtPIP1 and AtPIP2 did not further enhance P(os). Native pollen tube protoplasts showed higher P(os) values (13.23 +/- 4.14 microm s(-1)) than pollen grain protoplasts but expression of AtPIP2;1 (18.85 +/- 7.60 microm s(-1)) did not significantly increase their P(os) values. Expression of none of the tested PIPs had any effect on pollen tube growth rates. The ectopic expression of AtPIP2s in lily pollen increased the water permeability of the plasma membrane in pollen grains, but not in pollen tubes. The measured endogenous water permeability does not limit water uptake during tube growth, but has to be regulated to prevent tube bursting.

Spray pollination as a labor-saving pollination system in Japanese pear ( Pyrus pyrifolia (Burm.f.) Nakai): Development of the suspension medium

Effects of suspension media used for spray pollination on pollen grain viability were investigated in Japanese pear ( Pyrus pyrifolia (Burm.f.) Nakai ‘Kosui’). The suspension media tested in this study consisted of pectin methylesterase (PME) and polygalacturonase (PG) combined with either 0.1% agar or 0.04% xanthangum (XG). We also evaluated the influences of spray pollination on fruit set and fruit quality. Pollen grain viability, as measured by germination and pollen tube growth rates, was maintained in media supplemented with 0.1 mg L −1 PME or 0.1 mg L −1 PG. The level of fruit set after spray pollination using media containing PME or PG in combination with either 0.1% agar or 0.04% XG was almost the same as the level after artificial pollination by hand (hand pollination). The media containing XG combined with either PME or PG seemed to show better results for pollen grain viability and fruit set, although the results were variable from year to year. With regard to fruit size, shape and other parameters for fruit quality, spray pollination and hand pollination gave comparable results, irrespective of the medium composition. The time required for spray pollination was less than half of that required for hand pollination, and the amount of pollen grain required for spray pollination was one-third or less than the amount required for hand pollination. Thus, spray pollination may be a time- and labor-saving pollination system for the cultivation of Japanese pear.

The pollen receptor kinase LePRK2 mediates growth-promoting signals and positively regulates pollen germination and tube growth.

In flowering plants, the process of pollen germination and tube growth is required for successful fertilization. A pollen receptor kinase from tomato (Solanum lycopersicum), LePRK2, has been implicated in signaling during pollen germination and tube growth as well as in mediating pollen (tube)-pistil communication. Here we show that reduced expression of LePRK2 affects four aspects of pollen germination and tube growth. First, the percentage of pollen that germinates is reduced, and the time window for competence to germinate is also shorter. Second, the pollen tube growth rate is reduced both in vitro and in the pistil. Third, tip-localized superoxide production by pollen tubes cannot be increased by exogenous calcium ions. Fourth, pollen tubes have defects in responses to style extract component (STIL), an extracellular growth-promoting signal from the pistil. Pollen tubes transiently overexpressing LePRK2-fluorescent protein fusions had slightly wider tips, whereas pollen tubes coexpressing LePRK2 and its cytoplasmic partner protein KPP (a Rop-GEF) had much wider tips. Together these results show that LePRK2 positively regulates pollen germination and tube growth and is involved in transducing responses to extracellular growth-promoting signals.