Pollen Tube Growth and Seed Set of Banksia coccinea R.Br. (Proteaceae)

Controlled hand pollinations and field observations were usedin conjunction with fluorescence and scanning electron microscopyand fruit and seed set to investigate the timing of stigma receptivity,pollen tube growth and self-incompatibility in relation to fertilityin B. coccinea. The species showed both protandry and partialself-incompatibility. Peak stigma receptivity as measured bypollen germination was recorded at 3 d after anthesis and maximumproduction of stigmatic exudate at 6 d. Pollen tubes reachedthe base of the style by 6 d after pollination. A 5 x 5 diallelexperiment was conducted and the results measured by pollentube growth. Self-pollinations generally resulted in poorertube growth than crosses and there was significant specificand general combining ability as well as reciprocal effects.Cross-pollination resulted in improved fruit set and seed toflower ratio over both selfing and open pollination. Spatiallimitations to fertility due to infructescence size were alsorecorded, but the combination of outcrossing mechanisms andspatial limitation did not entirely account for the low fertility.It is suggested that environmental conditions and the availabilityof resources may also exert an influence. Banksia coccinea R.Br., scarlet banksia, Proteaceae, pollination, stigma receptivity, pollen tube growth, self-incompatibility, breeding system, seed set     

Structural analysis of stigma development in relation with pollen–stigma interaction in sunflower

Structural analysis of stigma development in sunflower highlights the secretory role of papillae due to its semi-dry nature. Production of lipid-rich secretions is initiated at the staminate stage of the flowers in stigma development and increases at the receptive stage, coinciding with an extensive development of elaioplasts and endoplasmic reticulum network in the basal region of the papillae. Transfer cells, earlier identified only in the wet type of stigma, are also present in the transmitting tissue of the sunflower stigma. Attainment of physiological maturity by the stigmatic tissue, accompanying development from bud to pistillate stage, appears to affect the initial steps of pollen–stigma interaction. The nature of self-incompatibility in Helianthus has also been investigated in relation with pollen adhesion, hydration and germination. Pollen adhesion to the stigma is a rapid process in sunflower and stigma papillae exhibit greater affinity for pollen during cross pollination as compared to self-pollination. Components of the pollen coat and the pellicle on the surface of stigmatic papillae are critical for the initial phase of pollen–stigma interaction (adhesion and hydration). The lipidic components of pollen coat and the proteinaceous and lipidic components from the surface of the papillae coalesce during adhesion, leading to the movement of water from stigma to the pollen, thereby causing pollen hydration and its subsequent germination. Pollen germination (both in self-and cross-pollen) on the stigma surface and the growth of the pollen tube characterize the flexibility of self-incompatibility in sunflower. Compatible pollen grains germinate and the pollen tube penetrates the stigma surface to enter the nutrient-rich transmitting tissue. The pollen tube from incompatible pollen germination, however, fails to penetrate the stigmatic tissue and it grows parallel to the papillae. Present findings provide new insights into structural and functional relationships during stigma development and pollen–stigma interaction.     

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.     

Pollination and Pollen-pistil Interaction in Oil Palm, Elaeis guineensis

Structural and cytochemical aspects of the pistil and detailsof pollination and pollen-pistil interaction were investigatedin the African oil palm (Elaeis guineensis Jacq.), an importantperennial oil crop. The stigma is trilobed, wet and papillate.The branched papillae are confined to a narrow linear zone oneach stigmatic lobe. Each stigmatic lobe harbours a deep stigmaticgroove, which runs adaxially along the surface. The stigmaticgroove is bordered by a well-defined layer of glandular cells,each of which has a pectinaceous cap on the inner tangentialwall. The style is hollow. The canal cells show thickeningson the inner tangential wall. The stigmatic groove and stylarcanal contain an extracellular matrix secreted by the canalcells which is rich in proteins, acidic polysaccharides andpectins. The canal cells at the base of the style are papillateand loosely fill the stylar canal. The stigma becomes receptivewhen the stigmatic lobes separate, and remains so for 24 h.Pollination is mediated by weevils as well as by the wind. Undernatural conditions the pollination efficiency was 100%. Pollinationinduces additional secretion in the stigmatic groove and stylarcanal. During post-pollination secretion, the pectinaceous capsof the cells lining the stigmatic groove are degraded. Pollengrains germinate on the stigmatic papillae and tubes grow onthe surface of the papillae, entering the stigmatic groove andadvancing along it into the stylar canal to eventually gainaccess to the locules. Pollen tubes are seen in the ovules 18–20h after pollination. Copyright 2001 Annals of Botany Company Arecaceae, Elaeis guineensis, African oil palm, pollination, stigmatic grove, stylar canal, Tenera hybrid, weevil     

Pistil anatomy and pollen tube development in Polygala vayredae Costa (Polygalaceae)

Low seed ovule ratios have been observed in natural populations of Polygala vayredae Costa, a narrowly endemic species from the oriental pre-Pyrenees. To evaluate physical and nutritional constraints and pollen tube attrition in this endemic species, stigma and style anatomy, as well as pollen tube development along the pistil were investigated using light and fluorescence microscopy. The structural morphology of the stigmatic region was also examined with scanning electron microscopy. Pollen grains that reached the stigmatic papillae came into contact with a lipid-rich exudate and germinated easily. Although a large number of pollen grains reach the stigmatic papillae, few pollen tubes were able to grow into the style towards the ovary. The style was hollow, with the stylar channel beginning a few cells below the stigmatic papillae. Initially, the stylar channel area was small compared to other levels of the style, and was surrounded by lipid-rich, highly metabolic active cells. Furthermore, lipid-rich mucilage was detected inside the stylar channel. At subsequent style levels towards the ovary, no major reserves were detected histochemically. The reduced intercellular spaces below the stigmatic papillae and the reduced area of the stylar channel at its commencement are suggested to physically constrain the number of pollen tubes that can develop. In subsequent levels of the style, the stylar channel could physically support a larger number of pollen tubes, but the lack of nutritional reserves cannot be disregarded as a cause of pollen tube attrition. Finally, the number of pollen tubes entering the ovary was greater than the number of ovules, suggesting that interactions occurring at this level play a major role in the final reproductive outcome in this species.     

Stigmatic Self-Incompatibility and Mating Patterns in Trillium grandiflorum and Trillium erectum(Melanthiaceae)

Post-pollination processes governing mating patterns in Trillium,a well-known genus of insect-pollinated woodland herbs, arepoorly understood. Mechanisms influencing outcrossing were investigatedin T. grandiflorum and T. erectum, two widespread species nativeto eastern North America. In southern Ontario, Canada, the twospecies are often sympatric; they flower in early May, and arepollinated by different assemblages of insects. Controlled cross-and self-pollinations and structural observations of pollengermination and pollen tube growth were conducted to determinewhether the two species possess a self-incompatibility (SI)system and, if so, the specific site(s) of self-rejection. Controlledpollinations indicated that both species set significantly moreseeds from cross-pollination than self-pollination, implicatingthe action of SI. This was confirmed by structural studies whichdemonstrated that self-recognition and rejection reactions occurredon dry-type stigmatic papillae. Observations of pollen hydrationrevealed that self-rejection was rapid, being initiated within10 min of pollination and prior to pollen tube emergence. Finalself-rejection resulted in failure of pollen tube growth atthe base of stigmatic papillae. SI was expressed more weaklyin T. erectum and thereby resulted in considerable self-seedset in some individuals . Estimates of outcrossing rates usingallozyme markers indicated that T. erectum displayed a mixed-matingsystem whereas T. grandiflorum was more highly outcrossed. Structuralstudies of pollen traits indicated that the two species differedwith respect to the size of grains and their aggregation withimplications for pollen dispersal and mating. The ecologicaland evolutionary implications of the variable expression ofSI in Trillium are discussed. Copyright 2001 Annals of BotanyCompany Trillium grandiflorum, Trillium erectum, self-incompatibility, mating     

Exudation of carboxylates in Australian Proteaceae: chemical composition

Roots of a wide range of plant species exude carboxylates, such as citrate, into the rhizosphere. In the present study, seedlings of a range of Australian Banksia, Hakea and Dryandra species (Proteaceae) were assayed for their exudation of carboxylates. All of these species (Hakea prostrata, Hakea undulata, Hakea petiolaris, Hakea baxteri, Banksia grandis, Banksia prionotes, Banksia occidentalis and Dryandra sessilis) form cluster roots when grown in nutrient solution with a low phosphate concentration. Exudation of carboxylates was studied for cluster roots and non‐cluster roots separately, and for the entire root system. Cluster roots of these Proteaceae exuded malate, malonate, lactate, acetate, maleate, citrate, fumarate, cis‐ and trans‐aconitate. The relative contributions of each of these carboxylates differed between species. Malate, malonate, lactate, citrate and trans‐aconitate, however, were invariably present in large proportions of total carboxylate exudation. Non‐cluster roots of H. prostrata exuded a spectrum of carboxylates (mainly malonate, lactate and citrate), which differed somewhat from the exudation pattern of cluster roots (mainly malate, malonate, lactate and citrate). The rate of exudation for cluster roots of the seven species was approximately 1·6 nmol g^?1 FM s^?1, which is considerably higher than that reported for a variety of crop and native species that do or do not form cluster roots. Contrary to what occurs in the cluster roots of Lupinus albus, which release carboxylates accompanied by protons so that the rhizosphere is acidified, the present Proteaceae exude the carboxylates as anions without concomitant proton release. The role of carboxylates in the mobilization of phosphate and other nutrients from soil is discussed.     

Papillar integrity as an indicator of stigmatic receptivity in kiwifruit (Actinidia deliciosa)

Stigmatic receptivity is a major factor limiting fruit set inkiwifruit (Actinidia deliciosa). The aim of this work was toknow what determines the cessation of stigmatic receptivityin this species. Stigmatic receptivity has been analysed inkiwifruit through the capacity of the stigma to sustain pollengermination and has been related to stigmatic development anddegeneration.The stigma of kiwifruit flowers has a papillatesurface which from anthesis is covered with an abundant exudate.Papillae are unicellular and contain a number of phenolic deposits.During the lifetime of the flower these papillae gradually loseturgidity, while stigmatic secretion increases until 5 d afteranthesis when papillae start rupturing and the papillar contentis liberated into the germination medium. Stigmatic receptivityis high at anthesis and lasts for the next 4 d. However, itdrastically decreases 5 d after anthesis and it is nil 2 d later.The pattern of stigmatic receptivity closely fits that of papillarintegrity, indicating that stigmatic receptivity relies on thisintegrity. Since papillar integrity can easily be evaluatedin stigmatic arms, this criterion can be used as a quick methodto estimate stigmatic receptivity. Key words: Actinidia deliciosa, kiwifruit, stigma, receptivity, 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     

The Pollen Tube Pathway in the Pistil of Lycopersicon peruvianum

The pollen tube pathway has been studied in unpollinated andpollinated pistils of Lycopersicon peruvianum using histochemicalstains for detection of proteins, lipids and arabinogalactansby bright-field microscopy, and decolourized aniline blue fordetection of pollen tubes by epifluoresence microscopy. Thepollen tube pathway is a continuous tract of mucilage from thestigma surface to the ovule micropyles, and is associated witha continuous tract of specialized, protein-rich transmittingcells comprising the stigmatic papillae, vertical files of stylartransmitting cells and the placental epithelium within the ovary.The superficial exudate of the stigma is hydrophobic and richin lipids. The mucilage of the style and ovary is hydrophilicand rich in arabinogalactans but low in proteins. Pollen tubesgrow between cells through the mucilage of the stigma and stylartransmitting tract, and across the surface of the placenta inthe ovarian mucilage. The structure of the junction of the stylartransmitting tract with the top of the ovary placenta assistseffective distribution of pollen tubes within the ovary. Lycopersicon, solanaceae, fertilization, pistil, pollen tube, pollination     

Heterostyly and Pollen-tube Interactions in Luculia gratissima (Rubiaceae)

Observations on the floral biology of Luculia gratissima (Rubiaceae)showed that this species is distylous with complementary positioningof anthers and stigmas in the two floral forms. Unusual featuresof distyly in this species include the larger size of the corolla,the stigma surface and the stigmatic papillae in the thrum flowerscompared to the pin ones. Stigmatic surfaces have similar secretionsbut they appear more copious in thrum than pin. The floral dimorphismwas accompanied by a very effective self-incompatibility systemand no seed was set on selfing. Seed number per capsule on crossingwas significantly greater in thrum flowers compared to pin.Incompatible pollen tubes were inhibited within 24 h at thebase of the stigma/top of the style in both morphs. Amputationof this region of the gynoecium removed the self-incompatibilityreaction in thrum but not pin flowers. Pollination with a mixtureof compatible and incompatible pollen and sequential pollinationwith self followed by cross pollen showed that there were interactionsbetween the two types of pollen tube. The presence of compatibletubes was found to cause the excessive swelling of the pollen-tubetip of the incompatible ones. The incompatible tubes did notappear to have any effect on the growth of compatible ones. Luculia gratissima, distyly, floral biology, self-incompatibility, pollen-tube interactions     

POLLEN-PISTIL INTERACTIONS IN TRISTYLOUS PONTEDERIA SAGITTATA (PONTEDERIACEAE). I. FLORAL HETEROMORPHISM AND STRUCTURAL FEATURES OF THE POLLEN TUBE PATHWAY

Heteromorphic characters and structural features of the pollen tube pathway are described in tristylous Pontederia sagittata to assess their influence on the pollination process and in mediating self-incompatibility behavior. Heteromorphic characters that distinguish the floral morphs include style length, stigma depth, stigmatic papillae length, stylar parenchyma cell length, area of the stylar canal, stamen height, anther size, and pollen size. Unlike several distylous species that have been investigated, the exine of pollen in P. sagittata was not strongly differentiated among the pollen types, and no differences in stigma cytochemistry were apparent. Features common to the floral morphs were a wet stigma, a hollow trilobed stylar canal separating into two sterile and one fertile canal, and a single anatropous ovule with a highly elaborated integumentary obturator. The similarity in structural features of the pollen tube pathway of P. sagittata to those found in monocotyledonous taxa with gametophytic self-incompatibility suggests that phylogenetic constraints may have influenced the evolution of trimorphic incompatibility in the Pontederiaceae.     

Structure and Cytochemistry of the Stigma and Pollen--Pistil Interaction in Zephyranthes

The stigma in Zephyranthes candida and Z. citrina is of thedry type with a continuous cuticle—pellicle. In some papillae,however, the terminal portion of the cuticle—pellicleis lifted upwards and occasionally even disrupted by the accumulationof a secretion product below it. Both non-specific esterasesand acid phosphatases are present on the stigma surface. Thestyle is solid with a central core of transmitting tissue whichhas conspicuous intercellular spaces containing a matrix thatincludes proteins, polysaccharides and pectic substances. Zephyranthes citrina is self-compatible while Z. candida isself-incompatible. Followng incompatible pollination in Z. candida,pollen germination is normal but pollen tube growth is inhibitedat the junction of the stigma and style. Self-incompatibilitycan be overcome by bud pollination. Protein synthesis is necessaryfor pollen germination in both species. Concanavalin A bindsto the stigma surface of both species, but does not affect pollentube penetration in Z. candida. In crosses between the two speciestypical unilateral incompatibility is observed when Z. candidais used as the pistillate parent. Zephyranthes, stigma-surface enzymes, dry stigma, pollen-pistil interaction, self-incompatibility, unilateral-incompatibility     

Papillate Tepal Hoods and Delayed Pollen Germination inAlbucaL. (Liliaceae)

In the genusAlbuca, pollen germination is delayed until thetepals close around the stigma immediately before the onsetof floral senescence. At this time, papillae on the tepal apicesand the stigma swell and produce an exudate in which pollenrapidly germinates. No pollen germination occurs when the tepalsare artificially prevented from closing around the stigma.Copyright1998 Annals of Botany Company Albuca, Liliaceae, delayed pollen germination, gametophytic self-incompatibility, perianth contributing to pollen germination, pollen tube inhibition in ovary.     

Floral Structure, Stigma Receptivity and Pollen Viability in Relation to Protandry and Self-incompatibility in Silky Oak (Grevillea robusta A. Cunn.)

The reproductive biology of Grevillea robusta growing underexotic conditions in Kenya and Australia is reported. The speciesshowed both protandry and a self-incompatibility mechanism.The stigma was wet and papillate with a distinct groove in themiddle. The anthers dehisced prior to anthesis, when the perianthopened. Stigmatic receptivity began 1 d after anthesis, withthe greatest pollen germination rates and longest pollen tubesobtained 2 d after anthesis. Nectar secretion commenced withpollen dehiscence and was abundant at anthesis. Most stigmaticgrooves opened widely 1–2 d after anthesis and stigmasshowed taller papillae and abundant secretion. Controlled pollinationsgave a greater fruit set from cross-pollination (5.9% in Apriland 17.5% in July) than open-pollination (0.1% in April and3.3% in July). No fruit set from self-pollination was obtainedin April, and very few fruit set for geitonogamous (two outof 1622; 0.1%) or for autogamous (one out of 2707 flowers; 0.04%)pollination treatments in July. Following self-pollination,growth of pollen tubes was poorer than in other treatments,and was generally arrested in the upper style. Cross-pollinatedflowers produced normal and straight pollen tubes, while self-pollentubes had growth abnormalities. Most of the open-pollinatedflowers were found without pollen or with only self-pollen ontheir stigmas indicating that the amount of cross-pollen reachingthe stigma under open-pollination may be a factor limiting seedproduction. Flowers shed soon after the fertilization phasewere those with ungerminated pollen or no pollen. Although avery low rate of selfing may occur, G. robusta presents a self-incompatibilitysystem and allogamy is its primary breeding behaviour.Copyright2000 Annals of Botany Company Grevillea robusta, silky oak, Proteaceae, protandry, controlled pollinations, receptivity, pollen-tube growth, self-incompatibility, pollination     

Female flowers and inflorescences of Didymelaceae

 In molecular analyses Didymelaceae together with Buxaceae form a fairly well-supported clade among families near the base of eudicots. Only little is known, however, about the flowers and inflorescences of Didymelaceae. In this study, the structure of the female flowers and inflorescences of Didymeles integrifolia was studied. Flowers are unicarpellate and orientation of the carpel is slightly deflected abaxially as in Proteaceae. Otherwise, Didymelaceae share many features of the gynoecium with Buxaceae and some other basal eudicots: the carpels are ascidiate in the lower half; anthetic carpels are completely closed by postgenital fusion; stigma is double-crested and widely decurrent; stigmatic papillae are unicellular and pear-shaped; the pollen tube transmitting tract is extensive and prominently differentiated; fruits are fleshy drupes with persistent stigma and style. However, the exceedingly elongate base of the integuments of Didymelaceae is an unusual feature among basal eudicots and even angiosperms. Received October 31, 2002; accepted December 17, 2002 Published online: March 31, 2003     

High humidity partially rescues the Arabidopsis thaliana exo70A1 stigmatic defect for accepting compatible pollen

We have previously proposed that Exo70A1 is required in the Brassicaceae stigma to control the early stages of pollen hydration and pollen tube penetration through the stigmatic surface, following compatible pollination. However, recent work has raised questions regarding Arabidopsis thaliana Exo70A1’s expression in the stigma and its role in stigma receptivity to compatible pollen. Here, we verified the expression of Exo70A1 in stigmas from three Brassicaceae species and carefully re-examined Exo70A1’s function in the stigmatic papillae. With previous studies showing that high relative humidity can rescue some pollination defects, essentially bypassing the control of pollen hydration by the Brassicaceae dry stigma, the effect of high humidity was investigated on pollinations with the Arabidopsis exo70A1-1 mutant. Pollinations under low relative humidity resulted in a complete failure of wild-type compatible pollen acceptance by the exo70A1-1 mutant stigma as we had previously seen. However, high relative humidity resulted in a partial rescue of the exo70A1-1 stigmatic papillar defect resulting is some wild-type compatible pollen acceptance and seed set. Thus, these results reaffirmed Exo70A1’s proposed role in the stigma regulating compatible pollen hydration and pollen tube entry and demonstrate that high relative humidity can partially bypass these functions.     

The Structure and Cytochemistry of the Pistil of Sternbergia lutea (Amaryllidaceae)

Studies were carried out on structural and cytochemical aspectsof the pistil of Sternbergia lutea (L.) KerGawl. The stigmais of the wet papillate type; the papillae are unicellular andare arranged densely around the rim of a funnel-shaped stigma.The stigma exudate is limited and is confined to the bases ofthe papillae and the inner lining of the stigma. The papillaeare smooth in the distal part and are covered with intact cuticle-pelliclelining. The cuticle is disrupted at places towards the baseof the papillae releasing the exudate. The exudate is rich inpectins and other polysaccharides but poor in proteins and lipids.The papillae show dense cytoplasmic profiles with extensiveendoplasmic reticulum (ER), abundant mitochondria, polyribosomesand active dictyosomes. The style is hollow. The stylar cavityis surrounded by two to four layers of glandular cells. In theyoung pistil the canal is lined with a continuous cuticle, butin the mature pistil the cuticle becomes disrupted and the canalis filled with the secretion produced by the cells of the surroundingglandular tissue. Ultrastructurally, the cells of the glandulartissue are very similar to the stigmatic papillae. The innertangential wall of the cells bordering the canal is uniformlythicker than other walls. The secretion in the stylar canal,as well as the intercellular spaces of the glandular tissue,stain intensely for pectins and polysaccharides but poorly forproteins and lipids. Pollen tubes grow through the stylar canal.Structural and cytochemical details of the pistil of Sternbergiaare compared with other hollow-styled systems. Pistil, Sternbergia lutea (L.) Ker-Gawl., stigma and style, structure and cytochemistry