Stigma-surface Esterase Activity and Stigma Receptivity in Some Taxa Characterized by Wet Stigmas

Stigma-surface esterase activity and stigma receptivity througha sequence of developmental stages of the pistil have been studiedin four taxa characterized by having wet stigmas — Petuniahybrida, Nicotiana tabacum, Crinum defixum and Amaryllis vittata.The style is solid in the first two and hollow in the lattertwo taxa. In all the taxa, stigma—surface esterase couldbe detected in a thin surface layer (pellicle) from a very earlystage of pistil development, irrespective of the presence orabsence of the exudate. However, the taxa showed variation instigma receptivity. In Petunia and Nicotiana, stigmas from pistilsof all the stages supported pollen germination and tube growth.In Amaryllis and Crinum, stigmas of only the mature pistils,when the exudate is present on the stigma, supported normalpollen germination and tube growth. It is inferred that in taxacharacterized by a wet stigma and solid style, the factors requiredfor pollen germination are present from an early stage of pistildevelopment and the exudate per se is not involved in pollengermination. In taxa characterized by a wet stigma and hollowstyle, however, the pellicle does not carry the factors requiredfor pollen germination and tube growth; they appear to be presentin the exudate. Petunia hybrida Vilm, Nicotiana tabacum L., Crinum defixum, Ker-Gawl, Amaryllis vittata Ait., tobacco, pollination, pollen germination, stigmatic exudate, stigma receptivity, stigma-surface esterase, esterase activity     

Physiology of Post-Pollination Exudate Production in Acacia

Marginson, R., Sedgley, M. and Knox, R. B. 1985. Physiologyof post-pollination exudate production in Acacia—J. exp.Bot. 36: 1660–1668. Stigmas of A. wattsiana produced 1260 nl of secretion by 90min after pollination. The optimum temperature for post-pollinationsecretion in A. baileyana, A. brownii and A. iteaphylla was20°C. Both self and cross intraspecific, interspecific andintergeneric pollinations produced a similar positive responsein A. iteaphylla and A. baileyana. In all cases aged pollenwas as effective as fresh pollen despite reduced fluorescencein the fluorescein diacetate test. Live yeast cells, fixed chickenerythrocytes, glass beads, talc and Biogel P150 did not stimulatesecretion, nor did pollen which had been washed in water orethanol despite high retention of viability in some cases Pollendiffusates dried on to glass beads produced a positive responsein A. iteaphylla and A. gracifolia. Intraspecific and interspecificpollinations involving fresh Acacia pollen resulted in pollengermination on the stigma and pollen tube growth in the styleof A. iteaphylla and A. baileyana. In contrast washed pollenfailed to germinate and pollen germination and tube growth werereduced at 35 °C as compared with 20 °C. Key words: Acacia, pollination, secretion     

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     

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.     

The extracellular lipase EXL4 is required for efficient hydration of Arabidopsis pollen

Pollination in species with dry stigmas begins with the hydration of desiccated pollen grains on the stigma, a highly regulated process involving the proteins and lipids of the pollen coat and stigma cuticle. Self-incompatible species of the Brassicaceae block pollen hydration, and while the early signaling steps of the self-incompatibility response are well studied, the precise mechanisms controlling pollen hydration are poorly understood. Both lipids and proteins are important for hydration; loss of pollen coat lipids and proteins results in defective or delayed hydration on the stigma surface. Here, we examine the role of the pollen coat protein extracellular lipase 4 (EXL4), in the initial steps of pollination, namely hydration on the stigma. We identify a mutant allele, exl4-1, that shows a reduced rate of pollen hydration. exl4-1 pollen is normal with respect to pollen morphology and the downstream steps in pollination, including pollen tube germination, growth, and fertilization of ovules. However, owing to the delay in hydration, exl4-1 pollen is at a disadvantage when competed with wild-type pollen. EXL4 also functions in combination with GRP17 to promote the initiation of hydration. EXL4 is similar to GDSL lipases, and we show that it functions in hydrolyzing ester bonds. We report a previously unknown function for EXL4, an abundant pollen coat protein, in promoting pollen hydration on the stigma. Our results indicate that changes in lipid composition at the pollen–stigma interface, possibly mediated by EXLs, are required for efficient pollination in species with dry stigmas.     

Stigma Constriction in Pearl Millet, a Factor Influencing Reproduction and Disease

The sequence of events during pollination in pearl millet isdescribed. Four or five hours after pollination with compatiblepollen a constriction region located in the fused stylodia,consisting of a collar of parenchymatous cells, becomes apparent.The structural integrity of the constriction region is lostwithin 6 h of pollination, thereby eventually isolating theovary from other pollen and also from ovary pathogens whichnormally enter throught the stigma. The constriction developsin unpollinated gynoecia 4–5 d after stigma emergence,thereby contributing to out-crossing in the protogynous commercialhybrid commonly grown in the semi-arid tropics. The potentialsignificance of a stigma constriction character in a wider botanicalcontext is discussed, particularly as it is a hitherto unrecognizedincompatibility barrier. Pennisetum americanum (L.), pearl millet, Sorghum bicolor (L.), sorghum, pollination, stigma constriction, ergot, protogyny, incompatibility barrier     

Reproduction in Seagrasses: Nature of the Pollen and Receptive Surface of the Stigma in the Hydrocharitaceae

This paper describes the characteristics of the pollen and thereceptive surfaces of the stigmas in the three marine angiospermsincluded in the Hydrocharitaceae. The pollen in Enhalus acoroidesand Thalassia hemprichii is spherical and has an ornamentedexine. An exine layer is not found in Halophila stipulacea wherereniform pollen grains are contained within transparent moniliformtubes. Cytochemical tests show that the pollen wall in the threespecies contains acidic and neutral polysaccharides and acidhydrolase acitivity is detected in the intine of H. stipulaceaand T. hemprichii. In Thalassia, one of the intine enzymes,acid phosphatase, is unambiguously associated with cytoplasmicinclusions. Flowering in Thalassia is coincident with the spring tides andthe pollen is released as a mass suspended in a thecal slimewhich contains approximately 5 per cent by weight carbohydrate,the principal mono-saccharide being mannose. Electrophoreticanalysis of the pollen-free slime shows a single glycoproteincomponent. The stigmas of the three seagrasses are papillate and of the‘dry’ type possessing a continuous protein-aceouspellicle subtended by a cuticle. The stigma pellicle exhibitscytochemically detectable esterase activity and binds the lectinconcanavalin A. Acid phosphatase activity is localized beneaththe cuticle at the tips of the stigma papillae. The discoveries show that the characteristics of the pollenand stigmas in the seagrasses are comparable with those foundin terrestrial flowering plants. The similarity in enzymaticproperites of the pollen wall and stigma pellicle suggests that,intriguingly, a similar mechanism of cuticle erosion might wellfollow compatible pollination both on land and in the sea. Enhalus acoroides, Thalassia hemprichii, Halophila stipulacea, Halophila decipiens, seagrasses pollen wall, stigma surface, hydrolytic enzymes     

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     

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.     

Development and Histochemistry of Style and Stigma, and Pollen-Pistil Interaction in Madhuca indica (Sapotaceae)

The histological and histochemical changes during the developmentof the stigma and style of Madhuca indica of the family Sapotaceaehave been investigated. The stigma is of the wet type and thestyle is open. The stigma secretion, present from stage I onwardsis lipophilic. But the protein, polysaccharides and variousenzymes are secreted only by the third, receptive stage. Thestigma has a stellate cavity opening into the stylar canals.The exudate along with various constituents is present up tostage VI of flower maturity. The pollen—pistil interactionstudies show a strong self-incompatibility response. The pollentubes show coiling and clubbing in the stigmatic cavity andstylar canals, even in the case of compatible cross pollination.Various methods are successfully employed to overcome self-incompatibility. Madhuca indica, Mahuda, development, histochemistry, pollen—pistil interaction, self-incompatibility, style and stigma     

Effective Pollination Period and Nature of Pollen-collecting Apparatus in the Gymnosperm, Larix leptolepis

In controlled environment, the effective pollination periodin Larix leptolepis is characterized by its brevity, lastingfor less than 1 d. It is associated with the opening of thepapillae of the pollen collecting apparatus which possess surfaceesterase activity, a condition that has its parallel in thesurface pellicle of angiosperm stigmas. Larix leptolepis, ovule receptivity, morphometric analysis, pollination, stigma surface, esterase activity     

Pollination Ecophysiology of Mercurialis annua L. (Euphorbiaceae), an Anemophilous Species Flowering all Year Round

Mercurialis annua L. is a dioecious anemophilous species thatflowers all year round in central and southern Italy. The flowersof both sexes are dimorphic: the female flower has a vestigialcalyx; the male flower consists only of a calyx that opens atanthesis. The anthers always dehisce after anthesis. The anthesisof male flowers seems to be temperature dependent, whereas antherdehiscence is related to relative humidity. The pollen grainsvary in volume according to the season: they are smaller whenrelative humidity is low and vice versa. They always decreasein volume after anther dehiscence and have the capacity to varyin volume and reach equilibrium with a changing environment.Viability is high, but may drop suddenly during heavy rain orhail that damage the exposed male flowers. The number of pollengrains per stigma varies from 0 to 300. The data is discussedin relation to the type of pollination and environmental characteristics.Copyright1994, 1999 Academic Press Mercurialis annua, dioecism, anthesis, anther dehiscence, pollen volume, pollen viability, anemophilous pollination, pollination ecology     

Le Infezioni Con “Stubborn” Ottenute per Trauma E Il Grado di Recettività dei Tipi di Grano

Abstract

Dioecy ensures cross pollination but pollen must travel from male to female plants, sometimes covering long distances. The present study concerns the reproductive strategy of two Italian dioecious plants: Mercurialis annua L. and Bryonia dioica L. The former is anemophilous and the latter is entomophilous; the latter has nectar and pollen as rewards. The distribution of pollen grains on the stigma is very different in the two species, reflecting the different types of pollination. In both species there is a number-dependent mechanism of pollen germination on the stigma. This mechanism could enhance male gametophytic competition.     

The Effect of Brassica oleracea Stigma Extracts on the Germination of B. oleracea Pollen in a Thin Layer Chromatographic Bioassay

Hodgkin, T. and Lyon, G. D 1986. The effect of Brassica oleraceastigma extracts on the germination of B. oleracea pollen ina thin layer chromatographic bioassay.—J. exp. Bot. 37:406–411. A procedure for germinating Brassica oleracea pollen on thinlayer chromatography plates pretreated with 20 mol m^–3tris(hydroxymethyl) methyl-aminopropanesulphonic acid (TAPS)buffer, pH 8·0 has been devised and used to detect pollengermination inhibitors in B. oleracea stigma extracts. Inhibitory zones in extracts of stigmas, unpollinated, or collected0·5, 4, 8 and 24 h after self- or cross-pollination,differed little in R_F values and sizes. Extracts of stigmascollected 1 h and 2 h after self-pollination gave a small additionalinhibitory zone which was not detected in 1 h and 2 h cross-pollinatedstigma extracts. The results showed some differences from thoseobtained using Petunia hybrida pollen germinated on T.L.C. platesthat were not pretreated with buffer. The nature of the differencesbetween the two bioassays is discussed and some possible reasonsfor them indicated. Key words: Pollen, germination inhibitors, self-incompatibility, Brassica oleracea     

The Pollen-stigma Interaction: Pollen-tube Penetration in Crocus

In a compatible pollination in Crocus, pollen tube tips enterthe stigma papillae after the enzymic erosion of the cuticle,and the tubes continue downward growth towards the ovary betweenthe cuticle and the underlying pectocellulosic wall. The cuticleof the receptive zone of the stigma papilla is chambered, thechambers containing a secretion accumulated during the maturationof the stigma. Pollen exudates contain various acid hydrolases,but are incapable alone of eroding stigma cutin. Furthermore,there is no penetration when the proteins of the wall-held stigmasecretions are degraded enzymically. These facts are taken toindicate that the pollen contributes a ‘cutinase’precursor which is activated by a factor or factors held inthe stigma secretion. Pollens of certain Cruciferae producetubes capable of penetrating the Crocus stigma cuticle, suggestingthat notwithstanding the taxonomic remoteness of Cruciferaeand Iridaceae the enzyme activation systems are quite similar.     

The Growth Activity and Fertilizing Capacity of Submersely Cultivated Pollen Tubes of Nicotiana tabacum L.

Tobacco pollen was cultivated in shaken suspension culturesand the growth estimated by weighing the mass of germinatedpollen separated from the nutrient solution. A formula for calculatingmean pollen tube length from the weight of the culture has beenderived for this pollen species. The growth of pollen tubes in vitro is shown to have a rhythmiccharacter. The rapid growth comparable to the mean growth ratein styles is limited to short time intervals alternating withprogressively extending periods of very depressed growth, whichceased entirely after 10–12 h of cultivation. When tested by placental pollination in vitro, the fertilizingcapacity of the pollen culture was found to increase duringthe first hour of cultivation but to decrease steadily thereafter.On the other hand, with the application of pollen tubes fromculture on stigma, a short precultivation period and even themere wetting of pollen had a negative effect on the seed set.With pollen cultivated longer than 4 h, no seed formation wasobserved.     

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.     

Flavanoids in Pollen and Stigma of Brassica oleracea and Their Effects on Pollen Germination In Vitro

Brassica oleracea pollen was applied to a basic medium of 1.5per cent agar and 15 per cent sucrose to which flavanoids wereadded at three concentrations. Two types of agar were used;with agar 1, quercetin at a concentration of 0.5 x 10^–3per cent gave an increase in percentage germinating grains.With agar 2, an increase in germination occurred with kaempferoland naringin at concentrations of 0.5 x 10^–3 and 0.5 x10^–1 per cent respectively. Increase in pollen tube lengthoccurred with agar 2 and quercetin at a concentration of 0.5x 10^–3 per cent. The stigma tissue of B. oleracea contains at least three andthe pollen at least one glycoside of quercetin. The sugars inthe glycosides were not identified. Pollen germination and pollentube extension were not stimulated exclusively by the flavanoidspresent in the stigma. The flavanoid composition of the stigmadid not vary amongst five different S-allele genotypes, indicatingthat flavanoids are probably not directly involved in the incompatibilityreaction of B. oleracea.