Effects of pollen competitive environment on pollen performance in Mirabilis jalapa (Nyctaginaceae)

 We examined the influence of pollen competitive environment on pollen performance in Mirabilis jalapa. We used the number of pollen grains and the number of pollen tubes per pistil as measures of pollen competition. Pollen germination, pollen tube penetration into the style, and pollen tube growth rates were used as measures of pollen performance. All three measures of pollen performance were affected by the competitive environment. Pollen germination was greatest at intermediate pollen load sizes. The percentage of germinated pollen grains that penetrated the stigma and grew into the style decreased with pollen load size. Pollen tube growth rate in the style was greater and more variable with larger numbers of pollen tubes in the style. Controlling for the degree of selection at the stigma indicated that pollen-pollen or pollen-style interactions were the likely causes of increased growth rates. Received: 28 October 1996 / Revision accepted: 24 January 1997     

Interspecific crosses in the genus Tulipa L.: identification of pre-fertilization barriers

 Pollen tube growth in the pistil and pollen tube penetration of ovules have both been studied in crosses between cultivars from Tulipa gesneriana L. and 12 tulip species from all eight sections of the genus Tulipa to identify pre-fertilization barriers. Depending on the cross, pollen tubes grew as far as the stigma or the style or continued growing down into the ovary. Pollen tubes penetrated none or only a few percent of the ovules of some crosses, despite the presence of many pollen tubes in the ovary. In other crosses, from which no or only a few hybrids have been obtained after seed maturation on the plant, pollen tube penetration was found in up to 79% of the ovules. Apparently, various kinds of barriers preventing fertilization or normal embryogenesis occur in interspecific tulip crosses. Received: 26 July 1996 / Revision accepted: 31 January 1997     

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.     

荞麦亲和与不亲和授粉后柱头和花粉管中ATP酶的超微细胞化学定位

利用磷酸铅淀淀技术对荞麦（Fagopyrum esculentum Monch.)pin型植株分别进行亲和授粉和不亲和授粉后的柱头、花粉粒、花粉管进行了ATPase的超微细胞化学定位。结果表明（1）亲和授粉和不亲和授粉后0.5h,柱头细胞的ATPase活性反应水平较低或基于无酶活性;柱头表面、柱头上附着的花粉粒内ATPase活性在不亲和授粉时较低,亲和授粉时较高,花粉粒内ATPase主要定位于线粒体和精子细胞;（2）授粉后1.5h,不亲和授粉的柱头细胞及花粉管的ATPase活性均较低,花粉管停止生长,细胞质开始解体;而亲和授粉的柱头细胞及花粉管的ATPase活性均较高,ATPase主要定位于柱头细胞的质膜、胞基质以及花粉管的壁、质体的膜、高尔基体、线粒体上。根据不同时期不同部位ATPase活性的差异,我们认为荞麦发生自交不亲和时,花粉管在花柱中停止生长不仅是因此花粉管得不到花柱中的营养物质而引起的,可能也与花粉管自身物质代谢发生障碍有关。     

荞麦亲和与不亲和授粉后柱头和花粉管中ATP酶的超微细胞化学定位

利用磷酸铅沉淀技术对荞麦(Fagopyrum esculentum Month.)pin型植株分别进行亲和授粉和不亲和授粉后的柱头、花粉粒、花粉管进行了ATPase的超微细胞化学定位。结果表明(1)亲和授粉和不亲和授粉后0.5h,柱头细胞的ATPase活性反应水平较低或基本无酶活性;柱头表面、柱头上附着的花粉粒内ATPase活性在不亲和授粉时较低,亲和授粉时较高,花粉粒内ATPase主要定位于线粒体和精子细胞;(2)授粉后1.5h,不亲和授粉的柱头细胞及花粉管的ATPase活性均较低,花粉管停止生长,细胞质开始解体;而亲和授粉的柱头细胞及花粉管的ATPase活性均较高,ATPase主要定位于柱头细胞的质膜、胞基质以及花粉管的壁、质体的膜、高尔基体、线粒体上。根据不同时期不同部位ATPase活性的差异,我们认为荞麦发生自交不亲和时,花粉管在花柱中停止生长不仅是因为花粉管得不到花柱中的营养物质而引起的,可能也与花粉管自身物质代谢发生障碍有关。     

Pollen Tube Growth and Fertilization Efficiency in Salpiglossis sinuata: Implications for the Involvement of Chemotropic Factors

The possible involvement of chemotropic factors influencingpollen tube growth in the style and ovary of Salpiglossis wasstudied. Pollen deposited at the base of excised styles germinatedreadily, with pollen tubes penetrating the transmitting tissueand growing towards the stigma, implying that no chemotropicrequirement exists for pollen tube growth in the style. However,the position and high efficiency of seed set following limited-pollinationof intact pistils would support the involvement of localizedchemotropism operating within the ovary. Chemotropism, limited pollination, fertilization efficiency, Salpiglossis sinuata     

INFLUENCE OF THE PISTIL ON POLLEN TUBE KINETICS IN PEACH (PRUNUS PERSICA)

Pollen tube growth has been studied in peach and has been related to changes in the pistil structures which the pollen tube has to traverse in its way from the stigma down to the ovule. Growth of the pollen tubes along the pistil is not continuous. While pollen tubes reach the base of the style 7 days after pollination, fertilization does not take place until 12 days later. Pollen tubes stop for 5 days at the top of the obturator and they further stop for 3 days before entering the ovule. The pollen tube growth is heterotrophic; starch, present all along the pistilar tract at anthesis, vanishes as the pollen tubes pass by. Discontinuous pollen tube growth appears to be controlled by the pistil. At anthesis the pistil is not fully matured. Maturation of the pistil implies a number of secretory processes that occur in a basipetal way starting from the stigma down to the style and ending in the ovule. Some of these secretions at the stigma and the style are triggered by pollination; others appear to be a maturative stage of the pistil and are produced in a discrete way. The fact that the pollen tube depends on these secretions together with the fact that these secretions are not continuously produced confer upon the pistil a role of controlling pollen tube kinetics and point out that, for a successful fertilization, male gametophyte development and pistil maturation need to by synchronized.     

Pistil Receptivity and Pollen Tube Growth in Relation to the Breeding System of Eucalyptus woodwardii (Symphyomyrtus: Myrtaceae)

Pistil structure, stigma receptivity and pollen tube growthwere investigated in relation to seed set of Eucalyptus woodwardii.Self-pollination resulted in reduced capsule retention and seeddevelopment as compared with cross-pollination. The pistil consistedof an ovary with five locules, a long style with a canal extendingfor two-thirds of its length, and a papillate stigma. Therewas no change in style length with time after anthesis, butboth stigma secretion and ability to support pollen germinationand tube growth increased to reach a peak at 7 d. Pollen germinatedon the stigma surface and in the stylar canal, but most tubegrowth occurred intercellularly in the transmitting tissue surroundingthe canal. At the base of the style the pollen tubes split intofive groups following the transmitting tissue strands to theovary. Each group grew through a septum dividing two loculesand entered the placenta. The tubes then emerged from the placentato penetrate the ovules at between 10 and 20 d after pollination.Fewer ovules were penetrated following self- than cross-pollination. Eucalyptus woodwardii Maiden, Lemon-flowered gum, Pistil receptivity, Pollen tube growth, Breeding system, Self-incompatibility     

POLLEN-POLLEN AND POLLEN-STYLE INTERACTIONS DURING POLLEN TUBE GROWTH IN ERYTHRONIUM GRANDIFLORUM (LILIACEAE)

Using pairs of pollen donors, I analyzed the growth of pollen tubes growing in different channels of the same style in Erythronium grandiflorum. After 24 hr the length of pollen tubes of randomly selected pollen donors was affected by the presence of other donors. The mean pollen tube lengths of donors did not differ when taken across all of the donor pairings, but in individual pairs, pollen from one donor was often significantly longer than pollen from the other donor when tested across several recipients. Pollen tube lengths were also consistently longer for pollen paired with pollen from the same donor than when paired with pollen from other donors, apparently because of mutual stimulation between the pollen populations from the same plant. In a second experiment, the amount of pollen tube attrition after five days of growth of pollen from a donor growing near (within 3 m) the recipient depended upon the source of other pollen growing in the same style. Local pollen experienced more attrition paired with self pollen than when paired with outcross pollen. Pollen from different outcross distances also modified the attrition of local pollen, but local pollen usually outcompeted pollen from greater distances. Since the growth of local pollen was modified depending upon the source of other pollen growing in the same style, it is probable that recipient styles are selectively inhibiting pollen tubes to produce the patterns of pollen tube interaction observed. The results from these two experiments indicate that the amount of attrition for pollen can be dependent on the composition of the pollen pool. Both direct pollen tube interactions and mediation by the stylar tissue appear to affect the growth rate and attrition of pollen in Erythronium.     

Directional Guidance of Nicotiana alata Pollen Tubes in Vitro and on the Stigma

Pollen tubes navigate the route from stigma to ovule with great accuracy, but the cues that guide them along this route are not known. We reproduced the environment on the stigma of Nicotiana alata by immersing pollen in stigma exudate or oil close to an interface with an aqueous medium. The growth of pollen in this culture system mimicked growth on stigmas: pollen grains hydrated and germinated, and pollen tubes grew toward the aqueous medium. The rate-limiting step in pollen germination was the movement of water through the surrounding exudate or oil. By elimination of other potential guidance cues, we conclude that the directional supply of water probably determined the axis of polarity of pollen tubes and resulted in growth toward the interface. We propose that a gradient of water in exudate is a guidance cue for pollen tubes on the stigma and that the composition of the exudate must be such that it is permeable enough for pollen hydration to occur but not so permeable that the supply of water becomes nondirectional. Pollen tube penetration of the stigma may be the most frequently occurring hydrotropic response of higher plants.     

PROTANDRY,INCOMPATIBILITY, AND SECONDARY POLLEN PRESENTATION IN CEPHALANTHUS OCCIDENTALS (RUBIACEAE)

Cephalanthus occidentalis L. is protandrous and presents pollen secondarily on the stigma surface. Because self-pollen is present on the stigma, the degree of selling vs. outcrossing in this species will depend on 1) the phenology of pollen presentation and stigma receptivity; 2) whether the species is self-incompatible; and 3) the rates of self vs. crossed pollen tube growth. This study describes floral morphology and phenology, self-incompatibility, and pollen tube growth rates in self- and crosspollinations of C. occidentalis. Light and scanning electron microscopy were used to study stigma morphology after flower opening, while controlled pollinations tested for incompatibility. Stigmas were unreceptive initially but became receptive by the second day after flower opening. Ninety-two percent of cross-pollinated flowers set fruit, compared to 12% fruit set in self-pollinations. Pollen tubes from selfed and out-crossed pollen initially had similar growth rates. Out-crossed pollen tubes began to grow rapidly ca. 5 hr after pollination of a receptive stigma, whereas selfed pollen tubes ceased growth or grew slowly after this time. Pollen tubes from out-crossed pollen grew the length of the style within 24 hr after pollination, while selfed pollen tubes were inhibited at the stigma-style junction. Our results indicate that C. occidentalis has selfincompatibility, in addition to protandry and secondary pollen presentation. Protandry allows removal of self-pollen from the unreceptive stigma, while self-incompatibility prevents fertilization by unremoved self-pollen.     

Pistil Structure and Pollen Tube Pathways in Leptospermum myrsinoides and L. continentale (Myrtaceae)

Pistil structure and pollen tube growth were investigated inLeptospermum myrsinoides and L. continentale (Myrtaceae). BothL. myrsinoides and L. continentale pistils consist of an ovarywith five locules, a style and a five-lobed dry, papillate stigma.A centrally located stigmatic cleft is present but extends onlyto the base of the stigma. Pollen germinates and grows intercellularlythrough the stigma into the central transmitting tissue of thestyle. Pollen tubes do not grow down the stigmatic cleft. Atthe base of the style the transmitting tissue separates intofive, each tract leading through the placenta to one of thefive locules. The pollen tubes continue to grow intercellularlythrough these five tracts entering the locules between the lobesof the placenta. Pollen tubes are smooth-walled and straightwhilst in the transmitting tissue of the style but produce shortlateral branches at regular intervals when in the locules. Branchingcontinues until pollen tubes enter ovules. It is suggested thatthe observed branching in the locules is a result of pollentubes following a chemotropic or thigmotropic pathway to theovules. This behaviour was consistent in all pistils examinedand no difference was observed in the behaviour of self- orcross-pollen tubes in the style or ovary.Copyright 1994, 1999Academic Press Leptospermum myrsinoides Schldl., Leptospermum continentale J. Thompson, pistil structure, pollen tube pathway, pollen tube branching     

两种姜科花柱卷曲性植物柱头的位置与其可授性的关系

花柱卷曲性是一种见于姜科山姜属(Alpinia)和砂仁属(Amomum)植物中的独特的性二态现象, 具有这一性系统的植物, 其居群包括上举型和下垂型两种表型, 所有个体的雌雄性别功能在时间和空间上分离。本研究通过操控授粉和花粉管生长两项实验, 探讨花柱卷曲性植物个体两性功能的隔离方式及其适应意义。云南草蔻(Alpinia blepharocalyx)操控授粉实验表明, 自花花粉对其异交率影响不显著(P > 0.05), 但可能由于自花花粉沉降导致胚珠贴现进而引起近交衰退, 操控条件下每果结籽数显著减少(P < 0.01)。云南草蔻和红豆蔻(Alpinia galanga)花粉管生长实验都显示无论对两种表型进行怎样的授粉处理, 花粉粒萌发和花粉管生长的速率仅在柱头处于可授位置时表现出最大值, 与自交和异交授粉方式无关; 上举型植株上午花粉囊虽未裂但其内花粉已成熟。研究结果表明即使不考虑花柱运动, 山姜属植物也具有异型雌雄异熟的特性。这一结果证实了花柱卷曲运动机制是通过互补式雌雄异位和异型雌雄异熟相结合形成的花部二态性, 异型雌雄异熟促进了异交, 而花柱运动的功能可能在于避免雌雄干扰。     

Heterostyly inPrimula. 2. Sites of pollen inhibition,and effects of pistil constituents on compatible and incompatible pollen-tube growth

Summary In incompatible (intramorph) pollinations of the heterostylousPrimula vulgaris, pollen germination or tube growth may be partially inhibited in several sites associated with the stigma or style. Blockage may occur, a) on the stigma surface through the failure of germination or of pollen tube penetration after germination, b) in the stigma head during the passage of the tube through the specialized transmitting tissue of the head, or c) in the transmitting tract of the style. None of the barriers is complete, and the prohibition of selfing or intramorph crossing depends upon the cumulative screening effect of one following upon the other. In both morphs, the germination of incompatible pollen on the stigma is enhanced in high ambient relative humidity, but many tubes still fail to penetrate the stigma. Those that do are retarded or blocked in their growth in the transmitting tissues of the stigma head and style. Crude extracts from the tissues of the stigma head and style show some differential effect on the growth of pollen tubesin vitro, and dialysates of extracts containing high molecular weight fractions show a consistent differential effect, those from thrum tissues retarding thrum tubes while having a lesser effect on pin tubes, and those from pin tissues retarding pin tubes while having lesser effect on thrum. It is suggested that the factors influencing tube growth are present in the intercellular secretions of the transmitting tract.     

Expression of stylar incompatibility in the Andean clonal tuber crop oca (Oxalis tuberosa Mol., Oxalidaceae)

Long-, mid-, and short-styled clonal accessions of oca (Oxalis tuberosa) were intercrossed in a complete diallelic design. Pollen tube growth in styles was monitored in all diallelic crosses. Pollen fertility was estimated by two tests: staining of pollen grains with aceto-carmine and detection of β-galactosidase activity by the substrate X-Gal. The two methods of pollen fertility estimation were equally useful to detect fertility levels. Pollen originating from short stamens had the highest fertility (85%) and pollen from long stamens had the lowest fertility (70%). Pollen fertility was high throughout, but its degree varied with the stylar morph on which the pollen was formed. Long-styled accessions had the highest rates of fertile pollen. Differences in pollen fertility at different anther levels in the same style morph were also apparent. Pollen grain diameter of the six morph-anther level combinations was inversely correlated with pollen fertility. Pollen grains from long stamens were the largest and pollen grains from short stamens were the smallest. Neither pollen fertility nor pollen grain size had an influence on pollen tube growth in the style or on fruit and seed set. Pollen tubes growing within the styles were inhibited at a different level for each of the 18 cross combinations in the diallel. Although legitimate crosses had greatest pollen tube growth, some of the illegitimate inter- and intramorph crosses had equally high scores. Of all illegitimate crosses, mid-styled seed parents had the lowest level of stylar incompatibility. Fruit and seed set were highly correlated with the extent of pollen tube growth in the style. The number of pollen tubes entering ovules in a flower was in good agreement with the number of seeds produced per fruit. Therefore, it is concluded that stylar incompatibility is the major determinant of limited seed formation in oca even in the most successful legitimate cross combinations. Received: 1 September 1999 / Revision accepted: 4 April 2000     

STIGMA,STYLE, AND OBTURATOR OF SOYBEAN,GLYCINE MAX (L.) MERR. (LEGUMINOSAE) AND THEIR FUNCTION IN THE REPRODUCTIVE PROCESS

Soybeans have a wet stigma overtopped by a pellicle that originates from the cuticle. There are numerous exudate-filled, axially oriented channels between cells of the transmitting tissue in the stigma and style. Pollen tubes grow in these channels and receive nutrition and mechanical guidance. Transmitting-tissue cells of the obturator are secretory also, but the obturator in soybean does not appear to control direction of pollen tube growth mechanically. The significant function of transmitting tissue in soybeans is to provide nutrition and to control direction of pollen tube growth.     

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.     

Observation of Pollen Tube Behaviour and Early Embryogenesis Following Interspecies Pollination Between Actinidia deliciosa and A. arguta

The pollen tube behaviour in the style and early embryogenesis following interspecies pollination between Actinidia deliciosa No. 26 and A. arguta were observed by means of fluorescence and light microscopy. Pollen grains germinated on the papillate stigma and pollen tubes grew along the V-shaped open-type style. Pollen tubes showed slower growth and reached the ovules 50--60 hours later than those of the control. Several abnormalities of pollen tubes have been observed at the base of the style, including wave-like pollen tubes, pollen tubes with swollen or pointed tips, with variable diameters, and a few with irregular growth. Random deposition of callose along pollen tube wall and even the whole wall was observed. About 26.74 % of the ovules were successfully fertilized and developed into seeds, among them 68.50% of the seeds were normal and 31.50% were abortive. About 11.41% were empty seeds without embryo and endosperm. Unfertilized small ovule was 61.45 %. Normal seed and its embryo were smaller than those of the control. The development of embryo was of the Soland type. The endosperm was cellular. The zygote remained quiescent for about 12-15 days before it started to divide, eventually forming a cotyledonary embryo 50 days after pollination.     

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.     

DYNAMICS OF POLLEN TUBE GROWTH IN THE WILD RADISH RAPHANUS RAPHANISTRUM (BRASSICACEAE). II. MORPHOLOGY,CYTOCHEMISTRY AND ULTRASTRUCTURE OF TRANSMITTING TISSUES,AND PATH OF POLLEN TUBE GROWTH

The relative importance of prezygotic mechanisms of gametophytic competition and selection are often unclear due to an absence of observations on the gynoecium and pollen tube growth in vivo. We used LM, SEM, and TEM to study the structure of the gynoecium and the path of pollen tube growth in Raphanus raphanistrum, a sporophytically self-incompatible annual. Wild radish has a papillate stigma and a solid style. A septum, which is characteristic of cruciferous gynoecia, is present in the ovary. After germination on the stigma, pollen tubes grow in the secretion of the transmitting tract of the style. The stylar secretion stains positive for acidic polysaccharides and insoluble carbohydrates, and negative for lipids and protein. In the ovary, the transmitting tissue is contained inside the septum. The secretion in the ovary stains positive only for acidic polysaccharides. Pollen tubes travel inside the septum as they enter the ovary and must exit to the surface of this tissue before ovule fertilization can occur. Pollen tube growth on the septum tracks the intercellular junctions of the septum epidermis where the secretion leaks out through a torn cuticle. Tubes must grow across the obturator before reaching the micropyle of an ovule. The temporal pattern with which tubes growing into the ovary exit the septum can contribute to the previously observed nonrandom patterns of fertilization (Hill and Lord, 1986).