Differential localization of enzymes in the stigmatic exudates ofPrimula obconica

Summary The stigmatic dimorphisms in the distylous speciesPrimula obconica, which has been dealt with in an earlier paper, has been described further. An ultrastructural localization of various enzymes in the exudates is hereby revealed, and evidence is given for intermorph differences. Esterases are confined to the lipid phase of the exudates, including the lipid globuli in the papillae walls of both morphs, but they are not found in the pellicle of the dry thrum stigma. However, this pellicle exhibits acid phosphatase activity, as does the lumen of the blisters in the viscous exudate on pin stigmas. The blisters are presumed to hold the watery phase of the pin secretion. From the present findings and from previous results with LM cytochemical methods it is suggested that the sites of peroxidase activity resemble those of acid phophatases.     

Physiology and biochemistry of the stigmatic fluid of Petunia hybrida

Summary Production of stigma exudate per flower of Petunia hybrida is about 200 g. The effect of light, temperature, metabolic poison and emasculation on the production of the exudate at different ages of the bud has been studied. The presence of a thin film of water below the stigmatic exudate has been demonstrated. Physical properties of the exudate such as relative viscosity and surface tension have also been determined. Chemical analysis of the stigmatic fluid showed that it consists primarily of an oil, sugars and amino acids. No protein could be detected. It also contains no acid phosphatase.Behaviour of the pollen from its deposition on the stigmatic fluid until it germinates on the stigma surface has been studied in vivo and also with the aid of an artificial stigma.The role of the stigmatic fluid in pollination has been determined.

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Zusammenfassung Die Narbenflüssigkeit, welche von zahlreichen Pflanzen ausgeschieden wird zu einem Zeitpunkt, da die Blüte reif ist zur Bestäubung, spielt eine wichtige Rolle bei der Fixierung des Pollens. Um so überraschender ist die Tatsache, daß hinsichtlich der Physiologie und Biochemie des Narbenschleimes so wenig detaillierte Informationen zur Verfügung stehen.Bei Petunia wird je Blüte im Mittel 200 g Narben-Flüssigkeit produziert. Die Intensität der Narbenschleimproduktion ist abhängig von der Temperatur und der Länge der Lichtperiode. Hingegen wird die Ausscheidung der Narbenflüssigkeit durch Hemmung der Atmung und durch partielle Kastration nicht beeinflußt.Die physikalischen Eigenschaften (Dichte, relative Viscosität und Oberflächenspannung) werden bestimmt. Bei der chemischen Analyse des Narbenschleimes ergab sich, daß dieser hauptsächlich aus einem öligen Fett besteht, dessen Fettsäure-Zusammensetzung ermittelt wurde. Hingegen werden nur sehr geringe Spuren von Zuckern und Aminosäuren gefunden. Der Narbenschleim ist frei von Proteinen und zeigt keine enzymatische Aktivität.Das Verhalten des Pollens bei der Landung in einem Tropfen der Narbenflüssigkeit wird in vivo und mit Hilfe einer künstlichen Modell-Narbe untersucht.Die Bedeutung der Narbenflüssigkeit im Zusammenhang mit der Bestäubung und den einleitenden Stadien der Pollenkeimung wird diskutiert.

     

Fungicide sprays can injure the stigmatic surface during receptivity in almond flowers

Fungicides can be detrimental to flower development, pollen function and fruit set in a number of crops. Almond is a self-incompatible nut crop that has a fruit set of only approx. 30 % of the total number of flowers. Thus, interference of pollination and fertilization by fungicide sprays is of concern, and identification of chemicals having the least detrimental effects would be desirable. The objective of this study was to evaluate the effect of fungicide sprays on stigma morphology in almond using a laboratory spray apparatus that simulated field applications. Four fungicides (azoxystrobin, myclobutanil, iprodione and cyprodinil) were applied, and fresh, unfixed stigmatic surfaces were observed using a scanning electron microscope at 4 and 24 h after spraying. Increased exudate accumulation was induced by azoxystrobin at both time periods, and localized damage and collapse of stigmatic cells were observed after 24 h. Damaged stigmatic papillae exhibited wrinkling, surface distortion or collapse. Likewise, myclobutanil caused significant damage to and collapse of papillae; these were more extensive at later observations. Iprodione had no effect on exudate accumulation but caused marked and severe collapse of stigmatic papillae which was pronounced at 24 h. Cyprodinil promoted a copious increase in exudate secretion and caused the most severe collapse of stigmatic cells of all the fungicides evaluated. Damage was somewhat localized at 4 h but more global at 24 h. This study has verified that certain fungicide sprays have direct detrimental effects on stigma morphology and enhance exudate production in almond flowers.     

Composition of the stigmatic secretions of long and medium styled flowers ofEichhornia

Like wet stigmas, the stigmatic secretions of dry stigmas ofEichhornia also have proteins, sugars, amino acids, phenols and fatty acids. No significant difference could be observed in the composition of stigmatic secretions of long and medium styled flowers except that the components were marginally higher in concentration in the former than the latter. The UV absorption profiles of the stigmatic secretions also showed no difference. Stigma surface esterases could be detected in both the stigmas from a very early stage of development.     

Polysaccharide component in the stigmatic exudate from Lilium longiflorum

The polysaccharide component of the stigmatic exudate from Lilium longiflorum has the composition, arabinose (26%), rhamnose (6%), galactose (57%) and glucuronic acid (11%). The highly branched polysaccharide bears a striking resemblance to the acidic polysaccharide exudate from Araucaria bidwillii in belonging to the galactan group and in carrying outer chains terminated by arabinofuranose, rhamnopyranose, galactopyranose and glucuronic acid residues. Both polysaccharides contain the sequence O-rhamnopyranosyl-(1→4)-glucopyranosyluronic acid-(1→6)-galactopyranose in some of the outer chains.     

Two novel reports of semidry stigmatic surface in Asteraceae

Research documents related to the morphology and function of style branches and stigmatic surface of Asteraceae are still rather few, and the literature reports are thus controversial. We report in the present study that the stigmatic surfaces of two non-related species of Asteraceae (Lessingianthus grandiflorus and Lucilia lycopodioides) have features of semidry stigmas. Sporodermis of both species was also analyzed so that we could understand how the stigmatic surface works during pollen deposition and rehydration. Stylar branches and pollen grains (sporodermis) were studied using scanning and transmission electron microscopy (SEM and TEM) and histochemistry techniques. The inner and marginal bands of stylar branches in these species display intermediary features between the dry and wet types of stigma: the cuticle characterizes the dry stigma and cells with secretory activity characterize the wet stigma; these showed differences from what has been described to the Asteraceae family, where stigmatic surface of species from several tribes is considered dry. Pollen grains are medium-size to large with exine ornamentation (echinate and echinolophate) and abundant secretion which latter characterizes pollenkitt. We can assume that two processes might help pollen grain hydration on stigmatic surface in Lessingianthus grandiflorus and Lucilia lycopodioides: (1) the presence of pollenkitt, as observed in the secretory content inside exine cavities and around pollen grains; and (2) the secretory activity of stigmatic surface cells, whose secretion accumulates among intercellular and subcuticular spaces and leads to cuticle disruption during the floral receptive phase. Our results suggest that ultrastructural and histochemical studies should be considered when describing stigmatic surface and that the “semidry” feature within Asteraceae should be investigated still more in detail, so that the taxonomic or adaptation value of this trait in the family can be verified.     

Haplotype structure of the stigmatic self-incompatibility gene in natural populations of Arabidopsis lyrata

We describe analyses of almost full-length sequences (including both the kinase domain and the S-domain) of the putative SRK incompatibility gene of the self-incompatible plant Arabidopsis lyrata. In A. lyrata, the SRK S-domain controls the pistil recognition specificity, as in self-incompatible Brassica species. In alleles from plants derived from natural A. lyrata populations, nonsynonymous and synonymous site diversity values are very high in both domains; even in exons 3 to 7 of the kinase domain, which probably have no recognition functions, 39% of the amino acids are polymorphic. Within populations, diversity between alleles is high, as expected for an incompatibility locus, which should be under frequency-dependent selection within populations, whereas within the different putative allelic classes polymorphism is very low, as predicted from theoretical models when recombination is rare. Nonsynonymous site variability declines in the kinase domain with increasing distance from the S-domain border, although synonymous diversity remains high, and the introns are unalignable. A decline in nonsynonymous diversity is expected due to selective constraints in the kinase domain, in combination with recombination (allowing diversity to decrease at sites distant from those under balancing selection). However, it is unclear whether recombination occurs in the SRK locus, and interpretation of the observed diversity pattern is complicated by apparent gene conversion with a paralogous gene (or genes). Patterns of linkage disequilibrium in our SRK sequences do not support the conclusion that recombination occurs, which was suggested from previous analyses based on Brassica SLG sequences.     

Influence of the pH of the stigmatic exudate on male-female interaction in Rosa hybrida L

Summary The in vitro germination of rose pollen is influenced by the pH of the medium. Both germination percentage and length of emitted pollen tubes were maximal for in vitro germination and tube elongation at pH 5 and minimal at pH 3 and 9 (Rosa hybrida L. var P 30 pollen). Three varieties characterized by having a stigmatic exudate of pH = 5 and another three varieties having one of pH = 9 were pollinated with the same pollen. Pollination effectiveness, as indicated by hip set (number of hips/pollmated flowers × 100) and mean number of achenes per hip, were significantly different: it was much higher for the varieties with the stigmatic exudate of pH = 5. pH control on pollination efficiency and subsequent fecundation success is proposed and discussed.     

Arabinogalactan-protein secretion is associated with the acquisition of stigmatic receptivity in the apple flower

Background and Aims

Stigmatic receptivity plays a clear role in pollination dynamics; however, little is known about the factors that confer to a stigma the competence to be receptive for the germination of pollen grains. In this work, a developmental approach is used to evaluate the acquisition of stigmatic receptivity and its relationship with a possible change in arabinogalactan-proteins (AGPs).

Methods

Flowers of the domestic apple, Malus × domestica, were assessed for their capacity to support pollen germination at different developmental stages. Stigmas from these same stages were characterized morphologically and different AGP epitopes detected by immunocytochemistry.

Key Results

Acquisition of stigmatic receptivity and the secretion of classical AGPs from stigmatic cells occurred concurrently and following the same spatial distribution. While in unpollinated stigmas AGPs appeared unaltered, in cross-pollinated stigmas AGPs epitopes vanished as pollen tubes passed by.

Conclusions

The concurrent secretion of AGPs with the acquisition of stigmatic receptivity, together with the differential response in unpollinated and cross-pollinated pistils point out a role of AGPs in supporting pollen tube germination and strongly suggest that secretion of AGPs is associated with the acquisition of stigma receptivity.Key words: AGPs, arabinogalactan proteins, apple, Malus × domestica, pollen, pollen tube, stigma, stigmatic receptivity, flower receptivity     

Involvement of SLR1 genes in pollen adhesion to the stigmatic surface in Brassicaceae

 The S-locus-related gene SLR1 is highly conserved and highly expressed in several species of the Brassicaceae family. Its function has not been determined, although several features would suggest a fundamental role in pollination. A second related gene (SLR2) is conserved and expressed in a subset of Brassica genotypes. We analysed the stigmatic expression of SLR1 and SLR2 genes among 11 different plants from various species or genera of the Brassicaceae and examined the extent of the pollen-stigma interaction during intraspecific, interspecific and intergeneric pollinations between them. Appropriate statistical tests on these variables (pollen adhesion, germination, penetration into the stigma, style and ovary, and SLR gene expression) showed that expression of SLR1 (but not SLR2) may be a factor in pollen-stigma adhesion. This hypothesis was supported by the observation of reduced pollen-stigma adhesion in transgenic B. napus plants modified for SLR1 expression. Received: 25 February 1997/Revision accepted: 24 June 1997     

Asynchronous development of stigmatic receptivity in the pear (Pyrus communis; Rosaceae) flower

While stigma anatomy is well documented for a good number of species, little information is available on the acquisition and cessation of stigmatic receptivity. The aim of this work is to characterize the development of stigma receptivity, from anthesis to stigma degeneration, in the pentacarpellar pear (Pyrus communis) flower. Stigma development and stigmatic receptivity were monitored over two consecutive years, as the capacity of the stigmas to offer support for pollen germination and pollen tube growth. In an experiment where hand pollinations were delayed for specified times after anthesis, three different stigmatic developmental stages could be observed: (1) immature stigmas, which allow pollen adhesion but not hydration; (2) receptive stigmas, which allow proper pollen hydration and germination; and (3) degenerated stigmas, in which pollen hydrates and germinates properly, but pollen tube growth is impaired soon after germination. This developmental characterization showed that stigmas in different developmental stages coexist within a flower and that the acquisition and cessation of stigmatic receptivity by each carpel occur in a sequential manner. In this way, while the duration of stigmatic receptivity for each carpel is rather short, the flower has an expanded receptive period. This asynchronous period of receptivity for the different stigmas of a single flower is discussed as a strategy that could serve to maximize pollination resources under unreliable pollination conditions.     

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.     

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     

Self-incompatibility in Papaver rhoeas: there is no evidence for the involvement of stigmatic ribonuclease activity

Abstract. Ribonuclease assays have revealed, in contrast to the self-incompatibility (SI) system of Nicotiana alata , there is no detectable ribonuclease activity that correlates with the presence of the functional stigmatic S-gene product in Papaver rhoeas . Thus, we have shown that the inhibition of incompatible pollen tube growth in P. rhoeas is not associated with ribonuclease activity. Furthermore, the finding that pollen from P. rhoeas , unlike that from N. alata , is insensitive to purified bovine pancreatic ribonuclease A at very high concentrations, suggests that the involvement of ribonucleases in the inhibition reaction of the SI response in P. rhoeas is unlikely. In addition, the level of ribonuclease activity in mature stigmas of P. rhoeas is very much lower than that in N. alata and significantly, the level of ribonuclease activity did not rise in conjunction with the developmental expression of SI. Therefore, as a result of these studies, we believe that SI in P. rhoeas does not involve ribonuclease activity.     

Mating system and stigmatic behaviour during flowering of Alpinia kwangsiensis (Zingiberaceae)

 Little is known about the breeding systems of tropical perennial ginger species. In this paper, we provide information about the breeding system of Alpinia kwangsiensis in Yunnan, Southwest China, specifically self-compatibility, mechanisms promoting outcrossing, and the visitation characteristics of pollinators. Populations of A. kwangsiensis have two specific phenotypes that differ in flowering behaviour: 1) “cataflexistyle” individuals in which the stigma is held erect above the dehiscent anther when anthesis begins in the morning and becomes decurved under the anther at noon and 2) “anaflexistyle” individuals in which the receptive stigma is decurved under the indehiscent anther first and moves into a reflexed superior position above the anther as it begins to shed pollen at mid-day. The stigmatic movements in the two floral phenotypes, which occur in a ratio of 1:1 in natural populations, are synchronous and correlate with the foraging behaviour of floral visitors; pollination is effected only between floral forms. Field experiments indicate that A. kwangsiensis is self-compatible and dependent upon insects for fertilization. This newly reported floral mechanism, which we have named “flexistyly,” adds to the repertoire of devices that have evolved in flowering plants to insure outcrossing. Received November 29, 2001 Accepted January 8, 2002