Pollen and Resource Limitation in Veratrum nigrum L. （Liliaceae）, an Andromonoecious Herb

Pollen limitation and resource limitation were invoked to account for the pattern that flowering plants produce more flowers and ovules than fruits and seeds. This study aimed to determine their relative importance in Veratrum nigrum, a self-compatible, perennial, andromonoecious herb. In order to determine whether female production was limited by pollen grains on stigmas or by available resources, we performed supplemental hand pollination in three populations, male-flower-bud removal in three other populations, and emasculation of hermaphroditic flowers in still another population, resulting in a total of seven populations experimentally manipulated. Across the three populations, supplemental hand pollination did not significantly increase fruit set, seed number per fruit, and total seed production per individual, nor did emasculation of hermaphroditic flowers. Taken together, our results suggest that pollen grains deposited on stigmas were abundant enough to fertilize all the ovules. Male-flower-bud removal significantly increased the mean size of hermaphroditic flowers in all three populations. Female reproductive success was increased in one population, but not in the other two populations possibly due to heavy flower/seed predation. We concluded that the female reproductive success of V. nigrum was not limited by pollen grains but by available resources, which is consistent with Bateman＇s principle. Furthermore, the female reproduction increase of male-flower-bud removal individuals might suggest a trade-off between male and female sexual functions.     

Airborne Pollen Grains of Bozuyuk （Bilecik, Turkey）

Airborne pollen grains in the atmosphere of Bozuyuk were investigated over a 2 yr period from 2000 to 2001 using a Durham sampler. A total number of pollen grains of 5 170 pollen grains belonging to 32 taxa were identified and recorded along with some unidentified pollen grains. Of all the pollen grains, 78.66% were arboreal, 19.20% were non-arboreal, and 2.12% were unidentified. The majority of pollen grains investigated were Pinus, Platanus, Quercus, Cupressaceae, Poaceae, Fagus, Salix, Rosaceae, Urticaceae, Asteraceae, and Chenopodiaceae. The maximum number of pollen grains was recorded in May.     

Evidence of Dichogamy in Santalum album L.

Flowering, fruit set, embryological development, and pollination trials were Investigated In Santslum album L. Each ovary may have three to four ovules. Mlcrosporogenesls and megasporogenesls In the same flower were synchronized at the earlier stages of flower development. However, at anthesls, when pollen was mature, the magaspore had developed only to the stage of a one- to two-nucleus embryo sac. As the eight-nucleus embryo sac developed, some mamelon cells began to undergo programmed cell death, forming holes Into which the eight-nucleus embryo sacs extended, becoming ＂N＂ or ＂S＂ shaped. The development from a two-nucleus embryo sac to a matured eight-nucleus embryo sac lasted up to 10 d. Fruit-set from open pollination was less than 2%. The endosperm develops prior to division of the zygotic embryo and one to three embryos and endosperms were formed In the same fruit. A mature seed usually germinates to produce one seedling; however, two and three seedlings from one seed were also observed, albeit at a low frequency. Pollination trials showed that no seed sets when Inflorescences were covered with s bag; however, artificial pollination could Improve fruit set. Our pollinaUon trials and embryological studies proved that the flower of S. album Is dlchogamous and fruit set has high heterozygosity.     

苏门白酒草的繁殖特征与入侵性的关系以及某些进化含义

Conyza sumatrensis （Retz.） E. Walker, a member of Asteraceae, is a highly invasive species. However, its reproduction biology remains poorly known. To understand the role of reproductive traits in successful invasion of the species, we studied several traits of its reproductive system： the miniature capitulum and gynomonoecious sexual system, the biology and phenology of capitula and florets, pollen/ovule ratio, the mating system （selfcompatibility）, flower visitors, physical traits and dispersal potential of achenes, germination potential of achenes from manually pollinated capitula, and the association of these traits with invasiveness. Our study showed that the reproductive traits of autonomous seed production, versatile mating system of self- and cross-pollination, and generalized pollination system might contribute to the species＇ successful invasive capability. The invasiveness was further enhanced by the high and rapid production of achenes, as well as the high percentage, rapid germination rate and high dispersal capability of achenes. It was concluded that in annual or winter-annual weeds, autonomous seed production contributed significantly to the invasiveness of the species.     

Analysis of Airborne Pollen Fall in Edirne, Turkey

In the atmosphere of Edirne 12691 pollen grains belonging to 42 taxa were identified by using of Durham sampler in 2000 and 2001. A total of 6 189 pollen grains per cm~2 were recorded in 2000 and a total of 6502 pollen grains per cm~2 in 2001. Total pollen grains consisted of 71.81% grains from arboreal plants, 25.88% grains from non-arboreal plants and 2.31% unidentified pollen grains. Pollen from the following taxa were also found to be prevalent in the atmosphere of Edirne: Gramineae, Pinus sp., Quercus sp., Cupressaceae/Taxaceae, Platanus sp., Salix sp., Morus sp., Populus sp., Carpinus sp., Juglans sp., Chenopodiaceae/Amaranthaceae, Fraxinus sp., Fagus sp., Ulmus sp., Ailanthus sp., Alnus sp., Ostrya sp., Helianthus sp. The season of maximum pollen fall was from April to June, with a prevalence of arboreal pollen in the first month, and of pollen from non-arboreal plants in the last months of the year.     

Phenotypic Characterization of a Female Sterile Mutant in Rice

A female sterile mutant, derived from a spontaneous mutation, was first discovered In rlce （Oryzs satlvs L. sep. Indlca） restorer llne 202R. Wlth normal fiowerlng, the mutant exhlblts an extremely low seed-sattlng rate. When the mutant Is crossed as a pollen donor, the seeds set normally; whereas when It Is used as a pollen recelver, no seeds are obtalned even wlth mlxed pollen gralns of dlfferent varletles sprlnkled over the atlgmas. The fioret of the mutant, conslatlng of slx stamens and one platll, looks the same as that of the wlld type In the malefemale organs, except that less than 10% of the mutant florets have three atlgmas on the ovary. Although the mutant has a low seed-setting rate, Its pollen fertility Is approximately 87.1%, which Is equal to that of the wild type. In addition, more than 90% of the mature embryo sacs of the mutant have complete Inner structures. At every stage after pollination, the sperm, embryo, and endosperm are not found In the mutant embryo sac, whereas the disintegration of the egg cell that does not accomplish fertilization Is visible. Through observetlons with a fluorescence microscope, we have found that the pollen grains germinate normally, whereas the pollen tube abnormally elongates in the style-transmitting tissue. The mutant pollen tubes display various defects In the style, such as slower elongation, conversed elongation, distorted elongation, swollen tips, or branched tips. As a result, the growth of the pollen tubes ceases In the style, and, therefore, the pollen tubes cannot reach the embryo sac and the process of double fertilization Is blocked. Based on these observations, we conclude that this mutant, designated as fs-202R, Is a novel type of female sterile mutation In rice, which causes the arrest of the elongation of the pollen tube.     

GNOM-LIKE 2, Encoding an Adenosine Diphosphate-Ribosylation Factor-Guanine Nucleotide Exchange Factor Protein Homologous to GNOM and GNL1, is Essential for Pollen Germination in Arabidopsis

In flowering plants, male gametes are delivered to female gametophytes by pollen tubes. Although it is important for sexual plant reproduction, little is known about the genetic mechanism that controls pollen germination and pollen tube growth. Here we report the identification and characterization of two novel mutants, gnom-like 2-1 （gnl2-1） and gn12-2 in Arabidopsis thaliana, in which the pollen grains failed to germinate in vitro and in vivo. GNL2 encodes a protein homologous to the adenosine diphosphate-ribosylation factor-guanine nucleotide exchange factors, GNOM and GNL1 that are involved in endosomal recycling and endoplasmic reticulum-Golgi vesicular trafficking. It was prolifically expressed in pollen grains and pollen tubes. The results of the present study suggest that GNL2 plays an important role in pollen germination.     

Composition and seasonal dynamics of seed rain in broad-leaved Korean pine (Pinus koraiensis) mixed forest in Changbai Mountain, China

To explore the composition and spatio-temporal dynamics of seed rain in broad-leaved Korean pine (Pinus koraiensis) mixed forest, 150 seed traps were set up in a 25 hm2 plot in Changbai Mountain. Seeds, fruits, anthotaxy and others in seed traps were collected, identified and divided into 4 types. From 2005 to 2006, we collected 47 different types. Total number of seeds and fruits was 121291, including 23147 mature seeds and fruits (19.1% of the total). Tilia amurensis and Fraxinus mandshurica, with the most seeds and fruits, accounted for 90% of the total. The analysis on seasonal dynamics of seed rain showed that there were the largest number of seeds and fruits between July and October, which were composed of immature seeds and fruits. In mid-October, mature seeds and fruits reached their peak, but immature seeds and fruits still accounted for high proportion. There were 91 traps that contained 100–200 mature seeds and fruits, and one trap without any mature seed or fruit. The largest number of species found in a trap was 7, and usually 3 or 4 species were found in most of the traps. There were obvious relationships between spatial patterns of mature seeds and fruits and their parent trees, indicating that their mature seeds and fruits were not dispersed far from their parent trees.     

Pollen tubes enter neighbouring ovules by way of receptacle tissue,resulting in increased fruit-set in Sagittaria potamogetifolia Merr

Using fluorescence microscopy, deposition of pollen on stigmas and pollen tube growth in the gynoecium of Sagittaria potamogetifolia Merr., a monoecious species with an apocarpous gynoecium, were observed. The maximum rate of pollination averaged 83.9 +/- 4.7 %, and the number of pollen grains per stigma ranged from zero to 30. Pollen tubes grew through one stigma to the base of the ovary at almost the same speed, but generally only one of the pollen tubes then turned towards the ovule and finally entered the nucellus through the micropyle. The other pollen tubes grew through the ovary base and the receptacle tissue into ovules of adjacent carpels whose stigmas were not pollinated or which had been pollinated later. This phenomenon is termed pollen tube 'reallocation' by the authors. To verify the direct effect of the phenomenon on fruit set, artificial pollination experiments were conducted in which two or more pollen grains were placed onto only one stigma in each gynoecium; frequently more than one fruitlet was obtained from each flower treated. The reallocation of pollen tubes among pistils in the gynoecium could effect fertilization of ovules of unpollinated pistils and lead to an increase in sexual reproduction efficiency. It would, to some extent, also increase pollen tube competition among pistils of the whole gynoecium.     

CHEMOTROPIC ACTIVITY AND THE PATHWAY OF THE POLLEN TUBE IN LILY,

A study of the pollen tube pathway in Lilium leucanthum var. centifolium and in L. regale reveals that the entire pathway from stigma to ovule is lined with cytologically unique stigmatoid cells. Assays for chemotropic activity of tissues and exudates along the pathway of pollinated or unpollinated pistils showed that onset of chemotropic activity progressed basipetally (and, when pollinated, in advance of the pollen tubes), commencing at the stigma 3-5 days before anthesis and appearing in the ovules 1-2 days after anthesis. Activity persists about 10 days in ovules of pollinated pistils and for 14-16 days in ovules of non-pollinated pistils. Attempts to localize the source of the chemotropic factor showed that gynoecial tissues bearing stigmatoid cells are chemo-tropically active while slices of style or ovary wall lacking stigmatoid cells are inactive. When ovules were sliced transversely and the micropylar and chalazal halves assayed, only the micropylar half showed activity. We suggest that the ovules and the stigmatoid tissue along the pollen tube pathway are the sources of the chemotropic factor responsible for the directional growth of the pollen tube.     

The breeding system of Lycium cestroides: a Solanaceae with ovarian self-incompatibility

To determine the breeding system of Lycium cestroides,several treatments were performed: self-, cross-, and geitonogamous pollinations, autonomous self-pollination, and a control (flowers exposed to natural pollinators). Production and quality of fruits and seeds as well as pollen tube growth were evaluated for each treatment. Experimental pollinations indicate that L. cestroides is self-incompatible since fruits were obtained only under cross-, and open-pollination treatments. However, in self- and geitonogamous hand pollinations, as well as in autonomous self-pollination, pollen tubes developed successfully and reached the ovules. The speed of pollen tube growth did not differ significantly among the different hand-pollinated flowers (cross-, self-, and geitonogamous). These facts indicate the presence of an ovarian self-incompatibility system. Significant differences were observed in fruit set, fruit size, and seed number per fruit between cross-pollinated flowers and open pollination (control). These results could be explained in terms of quantity and quality of pollination in each case. Received: 2 November 2000 / Revision accepted: 26 January 2001     

濒危植物安徽羽叶报春两种花型的繁育特性及其适应进化

二型花柱的维持机制和自然选择压力多年来一直是生态学和进化学研究领域的热点之一。通过实验室栽培和野外观察统计相结合的方法,对安徽羽叶报春两种花型(长柱花和短柱花)的形态特征、花粉活力、柱头可授性、花粉/胚珠比、自然授粉及结籽能力、自交亲和性等繁育特性进行了比较研究。结果表明:长、短柱花的花冠直径和裂片宽无明显差异,而花冠筒、雌蕊和雄蕊高、花粉数目及大小、P/O比均有显著差异。在自然条件下,长柱花所接受的总花粉数要明显高于短柱花的总花粉数,但所接受的异型花花粉数和平均每果结籽数两者无显著差异。长柱花和短柱花的花粉和柱头活力相似,均能在较长时间内维持较高活力,仅在开花末期显著下降。两种花型的花在自花授粉、同型异花授粉、异型花授粉条件下均能结籽,但异型花授粉的结籽数均明显高于自花授粉和同型异花授粉结籽数。在长柱花各种授粉方式中,花粉萌发率无明显差异,但异型花花粉管的生长速度明显比同型异花花粉和自花花粉的快,而在短柱花柱头上表现为异型花授粉的萌发率最高,但只要萌发后在花柱中的生长速度无显明差异。此外,综合上述结果,对二种花型花部综合征的维持机制及自然选择压力进行了探讨。     

Pollen tubes lacking a pair of K+ transporters fail to target ovules in Arabidopsis

Flowering plant reproduction requires precise delivery of the sperm cells to the ovule by a pollen tube. Guidance signals from female cells are being identified; however, how pollen responds to those cues is largely unknown. Here, we show that two predicted cation/proton exchangers (CHX) in Arabidopsis thaliana, CHX21 and CHX23, are essential for pollen tube guidance. Male fertility was unchanged in single chx21 or chx23 mutants. However, fertility was impaired in chx21 chx23 double mutant pollen. Wild-type pistils pollinated with a limited number of single and double mutant pollen producing 62% fewer seeds than those pollinated with chx23 single mutant pollen, indicating that chx21 chx23 pollen is severely compromised. Double mutant pollen grains germinated and grew tubes down the transmitting tract, but the tubes failed to turn toward ovules. Furthermore, chx21 chx23 pollen tubes failed to enter the micropyle of excised ovules. Green fluorescent protein-tagged CHX23 driven by its native promoter was localized to the endoplasmic reticulum of pollen tubes. CHX23 mediated K(+) transport, as CHX23 expression in Escherichia coli increased K(+) uptake and growth in a pH-dependent manner. We propose that by modifying localized cation balance and pH, these transporters could affect steps in signal reception and/or transduction that are critical to shifting the axis of polarity and directing pollen growth toward the ovule.     

Seed abortion in the sexual counterpart of <Emphasis Type="Italic">Brachiaria brizantha</Emphasis> apomicts (Poaceae)

Gametophytic organization, fertilization and reproductive success are described for the fertile diploid Brachiaria brizantha accession BRA-002747 which is being raised for use in Brachiaria breeding programs, as well as to understand and control of apomixis in this genus. The current paper reports on reproductive biology and analysis of seed set in field experiments during three consecutive years. Unsuccessful seed production in this plant is believed to correlate with early inbreeding depression, based on the reproductive features analyzed. Caryopsis development was observed using differential interference contrast microscopy with seed set determined by the number of self- and open-pollinated pistils that fully developed into viable seeds. Developing and mature female and male gametophytes were observed in the context of flower phenology, morphology and anthesis patterns. Pollen viability was determined by acetocarmine staining and by observation of germination in vivo, which was also used to observe pollen tube/pistil interaction. Although normal development was observed in floral structures, anthesis and gametophytes, seed set was low, with 2 and 6% in self- and open-pollination, respectively, producing seed. Variations observed in the female organs, such as the presence of a hermaphrodite flower in 50% of the inferior floscules and the presence of multiple embryo sacs of the Polygonum type within the same ovule in 15% of the pistils, are not related to low fertility. The majority of pollen grains are viable, in spite of the reduced number of pollen tubes within the style and ovary carpel, and a developing caryopsis was observed in 70% of self-pollinated pistils, indicating successful double fertilization from 2 days after anthesis (DAA). Nevertheless, abortion gradually increased from 2 until 7 DAA and remains elevated until 12 DAA, when caryopsis maturity is achieved. These data confirm low seed set in this accession and indicate that low fertility is not a consequence of abnormalities, either in the floral or gamete structures, or pollen tube rejection, but most likely a consequence of inbreeding depression.     

Pollen tube reallocation in two preanthesis cleistogamous species, Ranalisma rostratum and Sagittaria guyanensis ssp. lappula (Alismataceae)

Pollen deposition on stigmas and pollen tube growth in two apocarpous species, Ranalisma rostratum and Sagittaria guyanensis ssp. lappula (Alismataceae), were examined with fluorescence microscopy. The reallocation of pollen tubes among pistils was observed in both species. The percentage of pollinated stigmas per flower was only 22.0% in R. rostratum and 51.0% in S. guyanensis, though the seed/ovule ratios are higher than 65% in both species. The number of pollen grains on each single stigma ranged from 0 to 96 in R. rostratum, and from 0 to 125 in S. guyanensis. When more than one pollen grain deposited on a stigma, all pollen tubes grew to the ovary, but only one of them turned towards the ovule and finally entered the nucleus. The other tubes grew through the receptacle tissue into ovules of adjacent carpels whose stigmas were unpollinated or pollinated later. The intercarpellary growth of pollen tubes could be a mechanism to increase the efficiency of sexual reproduction in an apocarpous gynoecium with low pollination on the pistils.     

Pollination intensity and potential seed set in Passiflora vitifolia

Summary Initial seed set and fruit set were pollen-limited in a Costa Rican population of Passiflora vitifolia, a self-incompatible species with 200–350 ovules per flower. Pollination intensity was measured by counting the number of allogamous pollen grains on stigmas of the large one-day flowers. Hand-pollinations demonstrated that 25–50 pollen grains are required for fruit set, and >450 are needed for maximum seed set, with a pollen:seed ratio of about 1.6:1.0. Hummingbirds (Phaethornis superciliosus) delivered sufficient allogamous pollen for maximum seed set to only 28% of the flowers examined. Naturally pollinated flowers yielded fewer fruits and fewer seeds per fruit than those pollinated by hand. Most pollen transferred by humming-birds was self-incompatible; emasculated flowers yielded higher seed set than flowers with intact anthers. Visitation rates did not provide a good index of effective pollination.There were significant differences in ovule number, maximum seed set, and maximum per cent seed set among individual vines. More than half of an individual's flowers failed to set fruit, whether pollinated by birds or by hand. In this population, maximum reproductive potential may be limited by maternal resources for fruit development, but seed set varies with pollination intensity. Pollen-limited seed set may be a disadvantage of self-incompatibility, especially in species with many-seeded fruits.     

The signals to trigger the initiation of ovule enlargement are from the pollen tubes: The direct evidence

In angiosperms, initiation of ovule enlargement represents the start of seed development, the molecular mechanism of which is not yet elucidated.It was previously reported that pollen tube contents,rather than double fertilization, can trigger ovule enlargement. However, it remains unclear whether the signal(s) to trigger the initiation of ovule enlargement are from the sperm cells or from the pollen tubes.Recently, we identified a mutant drop1- drop2-, which produces pollen tubes with no sperm cells. Taking advantage of this special genetic material, we conducted pollination assays, and found that the ovules pollinated with drop1- drop2- pollen could initiate the enlargement and exhibited significant enlarged sizes at 36 h after pollination in comparison with those unpollinated ovules. However, the sizes of the ovules pollinated with drop1- drop2- pollen are significantly smaller than those of the ovules pollinated with wildtype pollen. These results demonstrate that the pollen tube, rather than the sperm cells, release the signal to trigger the initiation of ovule enlargement, and that double fertilization is required for further enlargement of the seeds.     

Variation of pollination and resource limitation in a low seed-set tree, Liriodendron chinense (Magnoliaceae)

Pollen limitation and resource limitation have been documented as the major factors responsible for plants commonly producing more ovules than seeds, but few studies have examined pollen deposition directly in natural populations at different sites and times. We investigated the causes of low seed set in four populations of Liriodendron chinense (Magnoliaceae), an insect‐pollinated endangered tree endemic to southern China, over 2–3 years. One pistil potentially produces two ovules. The number of pistils per flower varies among populations, but in three of the four populations the variation in a given population was not significantly different among years. Overall, populations with higher pistil numbers tend to set more seeds per flower, but a positive correlation between pistil numbers and seed production per flower was observed in only one of the four populations. The numbers of pollen grains deposited per stigma varied from 0 to 60. The proportion of pollinated stigmas per flower ranged from 44% to 88% among populations and years. The numbers of pollen grains deposited per stigma and the percentages of pollinated stigmas were significantly different between populations, and two populations showed significant differences between years. A positive correlation between stigmatic pollen load and seed set was sought in ten population‐by‐year combinations but, in a given population, high stigmatic pollen loads did not always result in high seed set. Examination of pollen deposition, pistil and seed production over several sites and years showed that in addition to pollination, other factors such as resource or genetic loads were likely to limit the (lower than 10%) seed set in L. chinense. It appears that small, isolated populations experience severe pollination limitation; one population studied had seed/ovule ratios of 0.84% and 1.88% in 1995 and 1996. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 140 , 31–38.