Reproductive significance of indirect pollen-tube growth in Dalechampia (Euphorbiaceae)

Most species of Dalechampia have expanded stigmatic surfaces that extend from the stylar tip part way down the sides of the elongated styles. Pollen grains landing at the stylar tip grow tubes directly down the style to the ovary. Pollen grains landing on the lateral stigmatic surfaces, however, exhibit indirect pollen-tube growth: the tubes grow first to the stylar tip, bend 180°, and then grow to the ovary. Indirect pollen-tube growth appears to be reproductively important: more than half the arriving pollen lands on the lateral stigmatic surfaces, and virtually all of these grains produce tubes that grow indirectly to the ovary; pollen on the lateral surface fertilizes seeds about as effectively as pollen at the tip (with direct tube growth). Indirect pollen-tube growth may be the result of a correlated response to selection for expanded stigmatic surfaces in Dalechampia, or it may be an adaptation to increase the intensity of pollen competition.     

The control of incompatibility in distylous Pulmonaria affinis Jordan (Boraginaceae)

In P. affinis, pin pollen is shorter on average than thrum pollen. Pins have more coronal hooks on stigmatic papillae than thrums, but gaps between papillae are relatively smaller for thrums. Pin stigmas receive more pollen than thrum stigmas. Thrum stigmas receive more (dissortive) pin pollen, but pin stigmas are assortively pollinated. Pollen only germinates when trapped below papilla coronas. On thrum stigmas, most trapped pollen is pin. Pollen germination is better on thrum stigmas than pin stigmas, and thrum stigmas show a close relationship between numbers of legitimate pollen grains, numbers of germinating grains, and numbers of pollen tubes in the style. There is no inhibition of illegitimate pollen germination. Illegitimate pollen tubes are inhibited in the style. Incompatibility operates by a combination of dissortive pollination, dissortive pollen trapping, and stylar pollen tube inhibition. All heteromorphic features differing between pins and thrums are implicated in the inhibition of within-morph fertilization in thrums.     

Reproductive biology of Nymphaea capensis Thunb. var. zanxibariensis (Casp.) Verdc. (Nymphaeaceae)

Anthesis in Nymphaea capensis var. zanzibariensis is diurnal with flowers opening and closing for three consecutive days. On the first day of anthesis, the stigmatic papillae secrete fluid and the outermost anthers are dehiscent. On the second day of anthesis the stamens form a cone above the dry stigmatic cup. The middle stamens open and turn outward. On the third day of flowering, all the stamens open and the dry stigmatic cup is exposed. The flowers are homogamous and not protogynous as the other Nymphaea. The gynoecium of the self-compatible N. capensis var. zanzibariensis , is characterized by a wet papillate stigma, a short hollow style, and secretory cells on the ventral surface of the ovary. The pollen is released on the receptive stigma. Following initial growth in intercellular spaces in the transmitting tract of the stigma, pollen tubes travel through the stylar canal and into the ovary.     

Gametophyte–Sporophyte Interactions in the Pollen–Pistil System: 3. Hormonal Status at the Progamic Phase of Fertilization

The content of hormones, IAA, ABA, and cytokinins, as well as the rate of ethylene production in petunia (Petunia hybrida L.) pistils and their parts (stigma, style, and ovary) were determined over 8 h after compatible pollination. At the progamic phase of fertilization in the pollen–pistil system, the phytohormones were virtually absent from the ovary but were present in various proportions in stigma and style. The stigma was the main site of ethylene synthesis and contained 90% of ABA while the style contained 80% of cytokinins of their contents in the whole pollinated pistil. Stigma and style did not differ in their IAA levels. The interaction of the male gametophyte with the stigmatic tissues was accompanied by a threefold increase in the ethylene production and a 1.5-fold increase in the IAA content in the pollen–pistil system within 0–4 h. Growth of pollen tubes in the stylar tissues (4–8 h) was accompanied by a further increase in IAA content and a decrease in the ethylene production by stigmatic tissues, as well as by a decrease in the cytokinin content in the stylar tissues. The ethylene/auxin status of the stigma may be suggested to control the processes of adhesion, hydration, and germination of pollen grains during pollination, while the auxin/cytokinin status of the style controls the pollen tube growth.     

Structure and Development of Stigmatic Branches and Style and Their Relation to Pollen Tube Growth in Wheat

The style of wheat divides into 2 branches, separated from its base and covered with a large number of slender stigmatic branches. The stigma is of dry type. The style is solid. There is no transmitting tissue differentiated in the style. Young stylar cells appear polygonal in transverse sections and elongated in longitudinal sections with an increase in length of the cells from periphery towards center. In transverse sections, mature stylar cells look extremely irregular. They are contorted and mosaicked with one another. During their development, stylar cells elongated vigorously with intrusive growth. The wall of stylar cells is thin, except at the corners where cells connect, that slight thickening of the cell wall occurs. Stylar cells start vacuolation at the earlier stages and gradually become highly vacuolated, but still remain rich in organelles, such as mitochondria, endoplasmic reticulum, dictyosomes and chloroplasts, the amount of which varied with the development stages of the style. Stigmatic branches are differentiated from the stylar epidermal cells, composed of 4 files of cells which link end to end with one another. Not long before anthesis, wall material in the intercellular corners becomes loose and porous. After pollination, pollen tubes grow along the intercellular spaces among the 4 files of cells in the stigmatic branches and then enter the style. Pollen tubes may pass through any intercellular corner throughout the 2 branches of the style, except for the lateral-outer portion which is composed of larger stylar cells. Eventually, pollen tubes enter the ovary.     

Dynamics of pollen tube growth under different competition regimes

Pollen tube dynamics following different competition regimes were studied in sweet cherry (Prunus avium L.). In the process from pollination to fertilization, a constant reduction in the number of pollen tubes that travel along the style is observed. There could be two main causes of this reduction. One is a physical or physiological constraint consisting of the progressive decrease in the reserves and space available for pollen tube growth along the transmitting tissue of the style, and the other is genetic interaction both among the male gametophytes and between the male gametophytes and the female tissues of the flower. To evaluate the roles that these two forces play in reducing the number of pollen tubes that travel along the style, pistils were subjected to various pollen competition regimes by applying different mixtures of live and dead pollen onto the stigmata. The results obtained were similar when the experiment was repeated with different genotypes over 2 years, both in the laboratory and in the field. The role of stylar constriction is important, but it is not the only cause of pollen tube attrition because with low pollen loads fewer pollen tubes reach the different parts of the style than could fit therein. The fact that under different pollen competition regimes the number of pollen tubes is reduced by the same proportion in each stylar level indicates that genetic interactions play an important role in the control of pollen tube attrition.     

Self-incompatibility inCrocus vernus subsp.vernus (Iridaceae)

Outcross, self- and mixed pollinations were performed inCrocus vernus subsp.vernus, a species with bicellular pollen, dry stigmas and hollow style. No differences were noted among the above pollinations concerning the germination of pollen and the growth of pollen tubes until the top of ovary. Within 45 min after pollinations 62% of pollen grains germinated. Pollen tubes penetrated the papilla cuticle extending along the papilla wall; on entry into stigmatic lobes they continued growth in the stylar secretion to ovarian locules. Here, however, self-pollen tubes failed to reach or to enter the ovule micropyle; while pollen tubes from either outcross- or mixed pollinations grew until fertilizing ovules. These observations gave evidence of a self-incompatibility system inCrocus, which appeared to be neutralized by mentor effect. The ovary as site of incompatibility response is discussed.     

Pollen walls of Araceae,with special reference to their fossilization potential

The pollen grains, the pistil, growth of the pollen tube and its pathway are described in Borago officinalis and Heliotropium europaeum. The exine is thick in both taxa but it is covered with dense gemmae in Borago. The intine seems to be thicker and contains more proteins in Borago than in Heliotropium. Starch is very abundant in the latter while it is lacking in the former. The style is hollow in Borago with a stylar canal running from the stigma to the ovary, while in Heliotropium it is broadly cone‐shaped with papillae located at the base of the cone (the “stigmatic ring") and not at the top of the style as usual. In Borago stigmatic papillae are unicellular, skittle‐shaped and have a thick pectocellulosic wall and an equally thick cuticular layer, while in Heliotropium the stigmatic papillae are unicellular, elongated cone‐shaped with a thin pecto‐cellulosic wall and have an apparently reduced cuticular layer. The stigmatic exudate is very abundant on the stigmas of Heliotropium even before anthesis while it is absent on those of Borago except when allo‐pollination occurs. Pollen tube growth has been followed from stigma to ovules in both taxa.     

POLLEN TUBE ATTRITION IN ERYTHRONIUM GRANDIFLORUM

Seed set after selfing in E. grandiflorum is often reduced relative to seed set after crossing; however, the compatibility patterns seen are not due to genes of major effect (i.e., S alleles). There is quantitative variation in the proportion of pollen tubes reaching the base of the style after both self- and cross-pollinations. Pollen tubes require between 24 and 72 hr to reach the ovary, but pollen tube growth ceases after 72 hr. When styles were removed from the ovaries 5 days after pollination, between 10 and 80% of the pollen tubes in the stigma had not reached the base of the style. The number of pollen tubes at the base of the style is a much better predictor of seed set than is the number of pollen tubes in the stigma. Pollen tube attrition is not affected by the age of the recipient flower or by the number of pollen donors contributing to the stigmatic pollen pool. The number of pollen tubes reaching the base of the style is dependent upon the source of the pollen and appears to be a decelerating function of the number of pollen tubes present in the stigma.     

不同葡萄品种柱头、花柱发育与种子形成的关系

对‘京秀’、‘香妃’(二倍体)和‘黑奥林’、‘巨峰’(四倍体)葡萄品种的柱头、花柱发生发育以及花粉管在雌蕊中生长与坐果和果实中种子形成的关系进行了研究。结果显示:‘京秀’、‘香妃’的坐果率和果实中的种子数明显高于‘黑奥林’和‘巨峰’,而‘巨峰’葡萄的坐果率和种子数最低。开花期‘京秀’和‘香妃’的花柱直径略小于‘黑奥林’和‘巨峰’,但花柱中部引导组织的直径和引导组织所占花柱的比例却大于‘黑奥林’和‘巨峰’;半实心花柱的比例也高于‘黑奥林’和‘巨峰’,说明花柱中引导组织的结构和发育状况可能与坐果和果实中种子形成有关;人工授粉24 h后,在‘京秀’和‘香妃’雌蕊中花粉管的生长速度快于‘黑奥林’和‘巨峰’,授粉后48 h‘京秀’和‘香妃’雌蕊中到达各部位的花粉管数要多于‘黑奥林’和‘巨峰’,授粉后72 h各品种到达子房各部位的花粉管数开始减少。表明‘京秀’和‘香妃’花柱中引导组织较为发达,而且半实心花柱比例也较高,有利于花粉管在雌蕊中的生长和完成授粉受精,使‘京秀’和‘香妃’的坐果率和果实中种子的形成都较高。     

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

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

INCOMPATIBILITY IN AMSINCKIA GRANDIFLORA (BORAGINACEAE): DISTRIBUTION OF CALLOSE PLUGS AND POLLEN TUBES FOLLOWING INTER- AND INTRAMORPH CROSSES

The distribution of callose plugs and pollen tubes was investigated following inter- and intramorph crosses of Amsinckia grandiflora (Boraginaceae), a distylous species possessing cryptic self-incompatibility. Callose plug distribution provided a good indication of the distribution of pollen tubes. Compared to intramorph crosses, many more callose plugs and pollen tubes were found in basal stylar regions following intermorph crosses, indicating that differential pollen tube growth is a likely cause of cryptic self-incompatibility. The incompatibility response differed for the floral morphs: in the pin (long-styled) morph pollen tubes were most likely to cease growth in the midstylar region, while inhibition was more likely to occur in the upper stylar region of the thrum (short-styled) morph. There was no evidence of stigmatic inhibition of pollen tubes for either morph, although the incompatibility response in the Boraginaceae is normally located in the stigmatic region.     

Pollen tube attrition as related to breeding systems inBrassicaceae

In order to estimate pollen tube attrition and to relate it to the breeding system, numbers of microgametophytes within the stigma and at the bottom of the style were scored in species of theBrassicaceae, 17 sporophytically self-incompatible, 11 predominant inbreeders, 4 facultative outcrossers. Attrition was evident in almost all species. The mean number of tubes at the bottom of the style, and mean number of stylar tubes per ovule were significantly lower in outbreeders. The degree of attrition, measured as attrition rate (the proportion of tubes in the stigma that do not reach the bottom of the style) was higher in outbreeders; it was positively correlated with the pollen : ovule ratios as estimates of the degree of outbreeding in the species involved. In outbreeders, sexual selection seems to be rather effective in inhibiting certain microgametophytic genotypes from reaching the ovules. Because of the low number of tubes per ovule in most cases, we assume that, in theBrassicaceae, female choice may be more effective in sexual selection and in ensuing pollen tube attrition.     

A lipid transfer-like protein is necessary for lily pollen tube adhesion to an in vitro stylar matrix

Flowering plants possess specialized extracellular matrices in the female organs of the flower that support pollen tube growth and sperm cell transfer along the transmitting tract of the gynoecium. Transport of the pollen tube cell and the sperm cells involves a cell adhesion and migration event in species such as lily that possess a transmitting tract epidermis in the stigma, style, and ovary. A bioassay for adhesion was used to isolate from the lily stigma/stylar exudate the components that are responsible for in vivo pollen tube adhesion. At least two stylar components are necessary for adhesion: a large molecule and a small (9 kD) protein. In combination, the two molecules induced adhesion of pollen tubes to an artificial stylar matrix in vitro. The 9-kD protein was purified, and its corresponding cDNA was cloned. This molecule shares some similarity with plant lipid transfer proteins. Immunolocalization data support its role in facilitating adhesion of pollen tubes to the stylar transmitting tract epidermis.     

The effect of varying the number of pollen grains used in fertilization

Summary The number of pollen grains placed upon a stigma influence both the development of pollen tubes and subsequently the progeny which result from fertilizations by gametes from these pollen tubes. The first influence is demonstrated by reduced pollen tube growth rates when pollen grains are few in number. This may indicate direct effects of pollen tubes upon the stylar tissues or perhaps more complex interactions between pollen and style. The second and potentially more important influence of limited pollination is upon the progeny. This was demonstrated with studies on three species. In each case, variation among the resultant plants was greater when pollen was limited than when normal, that is excessive, pollen was used. The mechanism of this phenomenon is not certain, but our data indicate that it is not simply an artefact of variation in seed size.     

Evidence for Stigmatic Self-incompatibility, Pollination Induced Ovule Enlargement and Transmitting Tissue Exudates in the Paleoherb, Saururus cernuus L. (Saururaceae)

Saururus cernuus, a species belonging to the primitive herbaceousangiosperm family Saururaceae, exhibits high rates of self-sterility.We investigated the structural and functional aspects of pollen-carpelinteractions following cross and self pollination to assessthe tissue specific site and timing of self-sterility and factorsimportant for successful cross pollen tube growth. Self-sterilitywas due to inhibition of self pollen germination at a dry stigma.Self pollination was associated with anomalous foot formation,reduced cell wall expansion and secretory activity of stigmaticpapillae, and callose production in stigmatic papillae. Followinggermination, cross compatible pollen tubes entered a solid coreof transmitting tissue and grew to the base of a short style.Entry of cross pollen tubes into the ovary was coincident withovule enlargement which placed the micropyle in the proximityof cross pollen tube tips. Ovule enlargement also occurred followingself pollination. Cross pollen tubes either entered an exudate-filledmicropyle directly from the style, or growth in the ovary waslocalized to the epidermis of the locule and outer integumentprior to entry into the micropyle. Prior to pollination, thetransmitting tract was void of secretions except for exudatein the micropyle. Growth of pollen tubes on the locule and integumentwas associated with exudate apparently arising from transmittingcells adjacent to growing pollen tubes. The present study providesthe first evidence in a primitive herbaceous species of stigmaticself-incompatibility (SI) in association with a dry stigma,pollination-induced signalling events affecting developmentof carpellary tissues, and micropylar exudates. Copyright 1999Annals of Botany Company SI evolution, dry stigma, exudates, pollen-carpel signalling, Saururaceae.     

Transmitting tissue ECM distribution and composition, and pollen germinability in Sarcandra glabra and Chloranthus japonicus (Chloranthaceae)

BACKGROUND: and Aims Free-flowing surface exudates at the stigmatic (wet versus dry stigma) and adaxial epidermis at the site of angiospermy in carpels of Chloranthaceous species have been proposed to comprise a continuous extracellular matrix (ECM) operating in pollen tube transmission to the ovary. The aim of this research was to establish the spatial distribution and histo/immunochemical composition of the ECM involved in pollen tube growth in Sarcandra glabra and Chloranthus japonicus (Chloranthaceae). METHODS: Following confirmation of the pollen tube pathway, the histo/immunochemical make-up of the ECM was determined with histochemistry on fresh tissue to detect cuticle, esterase, proteins, pectins, and lipids and immunolocalization at the level of the TEM on sections from cryofixed/freeze-substituted tissue to detect molecules recognized by antibodies to homogalacturonans (JIM7, 5), arabinogalactan-proteins (JIM13) and cysteine-rich adhesion (SCA). KEY RESULTS: Pollen germinability is low in both species. When grains germinate, they do so on an ECM comprised of an esterase-positive cuticle proper (dry versus wet stigma). Pollen tubes do not track the surface ECM of stigma or adaxial epidermal cells at the site of angiospermy. Instead, tubes grow between stigmatic cells and subsequently along the inner tangential walls of the stigmatic and adaxial carpel cells at the site of angiospermy. Pollen tubes enter the ovary locule at the base of the funiculus. The stigmatic ECM is distinct by virtue of the presence of anti-JIM5 aggregates, lipids, and a protein recognized by anti-SCA. CONCLUSIONS: The Chloranthaceae joins a growing number of basal angiosperm taxa whereby pollen tubes germinate on a dry versus wet stigma to subsequently grow intercellularly en route to the ovary thereby challenging traditional views that the archetype pollen tube pathway was composed of the surface of stigma and adaxial epidermal cells covered with a free-flowing exudate.     

Pollen performance before and during the autotrophic-heterotrophic transition of pollen tube growth

For species with bicellular pollen, the attrition of pollen tubes is often greatest where the style narrows at the transition between stigmatic tissue and the transmitting tissue of the style. In this region, the tubes switch from predominantly autotrophic to predominantly heterotrophic growth, the generative cell divides, the first callose plugs are produced, and, in species with RNase-type self-incompatibility (SI), incompatible tubes are arrested. We review the literature and present new findings concerning the genetic, environmental and stylar influences on the performance of pollen before and during the autotrophic-heterotrophic transition of pollen tube growth. We found that the ability of the paternal sporophyte to provision its pollen during development significantly influences pollen performance during the autotrophic growth phase. Consequently, under conditions of pollen competition, pollen selection during the autotrophic phase is acting on the phenotype of the paternal sporophyte. In a field experiment, using Cucurbita pepo, we found broad-sense heritable variation for herbivore-pathogen resistance, and that the most resistant families produced larger and better performing pollen when the paternal sporophytes were not protected by insecticides, indicating that selection during the autotrophic phase can act on traits that are not expressed by the microgametophyte. In a study of a weedy SI species, Solanum carolinense, we found that the ability of the styles to arrest self-pollen tubes at the autotrophic-heterotrophic transition changes with floral age and the presence of developing fruits. These findings have important implications for selection at the level of the microgametophyte and the evolution of mating systems of plants.