Modification of the Pollen-Stigma Interaction in Brassica oleracea by Water

The presence of a film of distilled water on the stigma surfaceof freshly opened flowers results in complete inhibition ofpollen following both incompatible and compatible pollinationsin self-incompatible (SI) genotypes of Brassica oleracea, SIgenotypes of B. campestris and one self-compatible (SC) genotypeof B. campestris. The application of water to the stigmas afterpollination also resulted in a marked reduction in pollen germinationand tube penetration. An increase in the time intervals betweenthe application of pollen onto the stigma and the water treatmentprogressively reduced this inhibition. Pollen germination wasalso completely inhibited when stigmas from freshly-opened flowersof SI B. campestris and B. oleracea genotypes were washed inwater, dried and pollinated with pollen grains of either compatibility.The ability of stigmas to induce pollen germination and tubegrowth was restored over a period, the length of which was dependenton the incompatibility (S) genotype. Stigmas of B. napus (SC)and SC mutants of SI B. campestris were found to be affectedby washing, but stigmas of a SC variety of B. campestris andthe immature stigmas from buds of B. oleracea were found tobe considerably less affected. Microscopic examination of pollenplaced on washed stigmas reveals that grains, irrespective oftheir compatibility, fail to hydrate normally. When inducedto hydrate by raising atmospheric humidity, pollen grains onwashed stigmas did germinate, but most of the tubes failed topenetrate the papillar wall and very few entered the style.It is proposed that the water treatment mobilises componentsof the pellicle which reorganize to block the activity of molecules,present in both SC and SI individuals, responsible for establishingfull contact between the pellicle and pollen grain coating. Brassica, pellicle, pollen, recognition, self-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.     

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     

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     

The Site of the Self-incompatibility Mechanism in Tradescantia pallida

Cytochemical and ultrastructural investigations were carriedout on unpollinated, self-pollinated and cross-pollinated pistilsof Tradescantia pallida. A protein secretion was found to occuron the unicellular papillac of the stigma in increasing amountstowards the base of the cells. The active cytoplasm of the cellsseemed to be responsible for this secretion. Compatible cross-pollinationresulted in pollen tube growth through the secretion and downthe stylar canal, whereas self-pollination resulted in pollentube growth being halted at the base of the stigma. It is suggestedthat the secretion may play an important role in the self-incompatibilityresponse. Tradescantia pallida, self-incompatibility, self-pollination, cross-pollination     

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     

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     

Overcoming Self-incompatibility through the Use of Lectins and Sugars in Petunia and Eruca

Treatment of stigma with a lectin (Con A/PHA) before pollinationwas effective in overcoming self-incompatibility in Petuniahybrida, a gametophytic self-incompatible system, and Erucasativa, a sporophytic self-incompatible system. Treatment ofpollen with glucose/N-acetyl-D-galactosamine (tested only withPetunia) was also effective. These results suggest the involvementof pollen lectins and specific sugar components of the pistilin self-incompatibility recognition. Petunia hybrida, Eruca sativa, self-incompatibility, pollen recognition, lectins     

Reproductive Biology of Plantago: Shift from Cross- to Self-pollination

The breeding systems of an obligate outbreeder (Plantago lanceolata)and an obligate inbreeder (P. patagonica) were compared. Outbreedingin the former and inbreeding in the latter species is achievedby self-incompatibility and preanthesis cleistogamy, respectively.The difference in breeding system is accompanied by a differencein the pollen output, pollen to ovule ratio, stigma and anthersize, and seed weight. P. ovata, a third species known onlyin cultivated form, is intermediate. This species has two typesof plants, some of which are protogynous and others where stigmareceptivity and anther dehiscence synchronize. Although thedifferences between the two intraspecific variants are not large,they nevertheless suggest that the species is in evolutionaryflux. Domestication is known to have modified the breeding systemin many crop plants and there is evidence that, in P. ovata,a start in this direction has already been made. Plantago lanceolata, P. ovata, P. patagonica, protogyny, anther, stigma, pollen to ovule ratio     

Variations in and Inheritance of NS-Glycoprotein in Self-Incompatible Brassica campestris L.

The stigma of Brassica species contain NS-glycoproteins thatexhibit a high degree of structural homology to the S-glycoproteinsof self-incompatibility. Inheritance of and variations in theNS-glycoprotein were studied with reference to self-incompatibility.The detection of NS-glycoproteins was performed by cross-reactionwith an antiserum raised against a purified NS-glycoprotein.In B. campestris, four isoforms of the NS-glycoprotein weredifferentiated by their pI values, but their molecular weightswere identical to one another. The genes for these isoformsof NS-glycoprotein were controlled by alleles at a single locus,tentatively named the NS allele, which was independent of Salleles at both the protein and the DNA level. Segregation ofF₂ plants with respect to the self-incompatibility behaviorof pollen tubes can be explained by the S allele model, butit appears not to be affected by the NS alleles. NS-glycoproteinswere found in all 21 species of Brassica and its allies examinedto date. The pI values of these glycoproteins varied among differentspecies. In addition to the isoforms of the NS alleles, maturestigmas contained other groups of proteins that reacted weaklywith the antiserum against the NS-glycoprotein. (Received July 30, 1991; Accepted February 21, 1992)     

The Anatomy, Histochemistry and Ultrastructure of Stigmas and Styles in Commelinaceae

The anatomy and ultrastructure of stigmas in 37 species of 13genera of Commelinaceae are described. The stigmas are papillate,papillae forming a dense fringe of cells around the mouth ofthe stylar canal in most species. The papillar cell wall iscovered by an unstructured cuticle of variable thickness andis of variable thickness because of small wall ingrowths. Thecuticle and the external surface of the papillar cell wall arevariably disrupted, particularly in the mid and basal regionsof the cell. This was not found in species of the genus Aploleiaor Callisia. The cell cytoplasm possesses all major organellesexcept chloroplasts and each cell is vacuolate. In all species except Aploleia mulitiflora the style comprisesan epidermis, a cortex and a hollow, tripartite canal whichis continuous into the ovary cavity. The three vascular strandsare positioned at the apex of each canal lobe. The canal cellsare elongate and tabular and the wall abutting the canal hasingrowths. The style in Aploleia is solid and the transmittingtissue comprises cells whose walls are electron opaque. Thecytoplasms of both types of cell are similar in content althoughthere is a single, large vacuole in canal cells and many smallvacuoles in transmitting tissue. The morphology, position and histochemistry of stigmatic andstylar exudate was similar in all ‘wet’ stigmas.Most of the exudate originates from the stylar canal althoughsignificant contributions are made by the papillae in stigmasof Coleotrype, Dichorisandra and Thyrsanthemum. There is no apparent relationship between stigma structure andthe presence of self-incompatibility. Stigma papillae, stylar canal, transmitting tissue, Commelinaceae     

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.     

The Rapid Identification of Self-incompatibility of Cabbage by Amino Acid Analysis

Amino acid analysis was employed in the study of protein in the stigma and pollen of self-incompatible and self-compatible lines of cabbage ( Brassica oleracea L. var. capitata ); from which distinct differences in the contents of amino acids were found between the two lines. The analysis of variance indicated that the contents of 9 kinds of amino acids in the stigma and 6 kinds in the pollen were comparatively much higher in the self-incompatible ones. Cluster analysis was per-formed in order to delete the factors related to the gene type variance. Unassured variances were abandoned by accurate examination. The threonine and tyrosine contents in the stigma and the glycine and alanine content in the pollen were considered as standards to identify self-incompatibility. In the self-incompatible lines, the above-mentioned amino acid contents were higher than 0.223%, 0.358%, 1.593% and 1.464%, respectively; whereas in the self-compatible lines, they were less than 0.185% ,0.164%, 1.470% and 1.006% correspondingly.     

Loss of callose in the stigma papillae does not affect the Brassica self-incompatibility phenotype

As part of the Brassicaceae self-incompatibility response, callose is deposited in the stigma papillar cells. To determine if callose plays an important role in the rejection of incompatible pollen by the stigma, transgenic Brassica napus. L. plants were produced which express the tobacco β-1,3-glucanase cDNA (the enzyme which degrades callose) in the stigma papillae. Using aniline blue fluorescence, little or no callose was detected in the papillar cells of transgenic stigmas. However, the self-incompatibility system appeared to be unaffected based on the lack of pollen tube growth and the subsequent lack of seed set. The transgene had no effect on compatible pollinations. Thus, while callose deposition is associated with the B. napus self-incompatibility response, it is not required for the rejection of incompatible pollen. Received: 14 March 1997 / Accepted: 15 April 1997     

The transition to gender dimorphism on an evolutionary background of self-incompatibility: an example from Lycium (Solanaceae)

Populations of three North American species of Lycium (Solanaceae) are morphologically gynodioecious and consist of male-sterile (i.e., female) and hermaphroditic plants. Marked individuals were consistent in sexual expression across years and male sterility was present throughout much of the species' ranges. Controlled pollinations reveal that L. californicum, L. exsertum, and L. fremontii are functionally dioecious. Fruit production in females ranged from 36 to 63%, whereas hermaphrodites functioned essentially as males. Though hermaphrodites were mostly male, investigation of pollen tube growth reveals that hermaphrodites of all dimorphic species were self-compatible. Self-fertilization and consequent inbreeding depression are commonly invoked as important selective forces promoting the invasion of male-sterile mutants into cosexual populations. A corollary prediction of these models is that gender dimorphism evolves from self-compatible ancestors. However, fruit production, seed production, and pollen tube number following outcross pollination were significantly higher than following self-pollination for three diploid, cosexual species that are closely related to the dimorphic species. The data presented here on incompatibility systems are consistent with the hypothesis that polyploidy disrupted the self-incompatibility system in the gynodioecious species leading to the evolution of gender dimorphism.     

The Proteaceous Pistil: Morphological and Anatomical Aspects of the Pollen Presenter and Style of Eight Species Across Five Genera

The morphology and anatomy of pollen presenters, styles andpollen of species ofBanksia, Dryandra, Hakea, IsopogonandMacadamiawerestudied. Serial sections of pistils and SEM images of pollenwere quantified to determine whether the low fertility observedin the Proteaceae has a structural basis. Pollen access to thestigma was investigated. There were three types of stigmaticcavity. A groove in which the stigmatic papillae were enclosedwas present inDryandra, BanksiaandHakea. Macadamiahad a groovewith protruding papillae, andIsopogonhad a tube which enclosedthe papillae. Anatomical studies showed the pollen presenterto be structurally complex but overall to have similar internalanatomy across the species studied. The species could be groupedaccording to presence or absence of transfer tissue and presenceor absence of sclerenchyma, but these groups were not mutuallyexclusive. In the pistil there were three structural filtersto pollen tube passage. The first was at the stigma, where pollengrain access could be limited by the diameter or length of thestigmatic groove or the capacity of the pollination chamberin relation to pollen volume. The second and third related toa narrowing of the transmitting tissue tract within the pollenpresenter and in the lower style which could influence pollentube passage to the ovule.Copyright 1999 Annals of Botany Company Proteaceae,Banksia coccinea, Banksia ericifolia, Dryandra formosa, Dryandra nana, Dryandra quercifolia, Hakea bucculenta, Isopogon cuneatus, Macadamia integrifolia,stigmatic cavity, fertility, pollen presenter, structural limitation, stigma, pollen grain, image analysis, transfer tissue.     

INDIVIDUAL VARIATION IN THE VIGOR OF SELF POLLEN AND SELFED PROGENY IN HIBISCUS MOSCHEUTOS (MALVACEAE)

In some self-compatible species, self pollen tubes grow more slowly than outcross pollen, presumably leading to low selfing rates when mixtures of self and outcross pollen reach the stigma simultaneously. Here we show that the competitive ability of self pollen differed among individuals of Hibiscus moscheutos. Self pollen tubes grew slower than outcross pollen in three plants, faster than outcross pollen in four plants, and showed no difference in five other plants (based on rates of callose plug formation). Levels of inbreeding depression were examined by comparing progeny from self and outcross pollinations in seven maternal families. Self pollination led to reduced seed number in only one maternal family, and a slight decrease in seed size was seen in two maternal families. Considerable inbreeding depression occurred later in the life cycle, and the degree of inbreeding depression varied among maternal families of 6-week-old plants. Our results demonstrate the potential for unpredictable effects of pollen competition on individual selfing rates, which in turn may affect progeny vigor. This complex situation contrasts with previous reports of species in which outcross pollen consistently outcompetes self pollen (cryptic self-incompatibility).     

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     

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.