Genetic Variation in Sorghum for the Inhibition of Maize Pollen Tube Growth

Aniline blue fluorescence was used to study the growth of maizepollen tubes in the stigmas of 13 diverse sorghum accessions.In 12, only short maize pollen tubes were formed, but in thesingle exception (Sorghum nervosum Nr481) maize pollen tubesgrew at least as far as the base of the style. The S. bicolorgenotypes S9B and CMS (a cytoplasmic male sterile line) werehybridized with Nr481, and analysis of maize pollen tube growthin F₁ plants, and BC₁ plants using Nr481 as the recurrent parent,suggested that differences in inhibition of pollen tube growthwere due to variation at a single locus, which we propose todesignate lap (Inhibition of alien pollen tubes). AccessionNr481 appears to be homozygous for a recessive allele permittingmaize pollen tube growth. Attempts were made to produce sorghumx maize hybrids using Nr481 and CMS derivatives which were knownto allow maize pollen tube growth to the base of the style.A putative hybrid endosperm was obtained in one Nr481 x Seneca60 maize cross, but this was not repeatable and no hybrid plantswere produced. A fundamental problem may be the large size ofthe maize pollen tube, which could have difficulty growing throughthe sorghum ovary and in entering the micropyle. Sorghum bicolor spp. bicolor (L.) Moench, Zea mays L, sorghum, maize, pollen tube growth, hybridization barriers     

Pistil Exudate Production and Pollen Tube Growth in Lilium longiflorum Thunb.

Exudate production in the pistil of Lilium longiflorum was studiedin relation to pollen tube growth, using scanning electron microscopy(SEM), transmission electron microscopy and light microscopy.In contrast with conventional fixation for SEM, during whichthe exudate of L. longiflorum largely washes away, the exudateremains present through freezing in case of cryo-SEM. Usingthe latter method we observed that exudate production on thestigma and in the style started before anthesis. Just underneaththe stigma the exudate was first accumulated at the top of eachsecretory cell, followed by a merging of those accumulationsas exudate production proceeded. Exudate is also produced bythe placenta. It was however not possible to determine whetherany of this fluid originated from the micropyle. Apart fromthe cell shape and the cuticle present in between the secretorycells, the ultrastructure of the secretory cells covering theplacenta was comparable to those of the stylar canal. The transferwall of the secretory cells of the placenta originated fromfusing Golgi vesicles but the endoplasmic reticulum seemed tohave an important role as well. After pollination the pollen tubes grew across the stigma andentered the style through one of the slits in the three stigmalobes. The pollen tubes grew straight downward through the styleand were covered by exudate. As the pollen tubes approachedthe ovary their growth was restricted to the areas with secretorycells. In the cavity the pollen tubes formed a bundle and theybent from this bundle in between the ovules towards the micropylarside. There they bent again to stay close to the secretory cells.After bud pollination the pollen tube growth was retarded. Laterarriving pollen tubes had a tendency to grow close to the secretorycells of the style, which resulted in a growth between thesecells and preceding pollen tubes. If there was still a littleexudate produced, it resulted in a lifting up of the pollentubes, out of the exudate. The relationship between exudateproduction and pollen tube growth is discussed. Both the speedand the guidance of the pollen tube seemed determined by theproperties of the exudate.Copyright 1994, 1999 Academic Press Cryo-scanning electron microscopy, exudate, Lilium longiflorum, lily, ovary, pollination, pollen tube growth, secretory cell, stigma, style     

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     

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

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

Mechanism of Pollination in Gladiolus: Roles of the Stigma and Pollen-tube Guide

Gladiolus has a dry type of stigma. Compatible pollen grainsalight and germinate on the receptive surface of the papillae,penetrate the cuticle and grow towards the style through a sub-cuticularpollen-tube guide of mucilage. This is secreted from epidermalcells of the stylodium and style canal. The cuticle, which coversthe pollen tube guide mucilage, is continuous through the stylecanal to the ovary. The wet stigma of Lilium also has cuticulartissue running through the style canal, covering the mucilage.     

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     

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.     

Successful Fertilization and Seed Set from Pollination on Immature Non-dehisced Flowers of Eucalyptus globulus

To minimize the number of visits required to achieve seed-setfrom hand pollination whilst maintaining full control of thereproductive process in Eucalyptus globulus, we investigatedthe importance of style maturation on the progamic phase ofreproduction. Controlled pollinations were performed in: (1)fully receptive styles; (2) immature cut styles at or afterflower dehiscence; and (3) immature cut styles before flowerdehiscence. We show here that pollen germination and tube growthto the ovule occurred in all three treatments, and all producedviable seeds. Remarkably, tubes growing on immature sliced stylesof non-dehisced flowers reached the ovules by day 7, 1–2d faster than tubes growing on normal receptive stigmas. Moreover,more tubes entered the ovary in this kind of pollination and,consequently, more viable seeds were produced. We establishedhistochemically that tube growth in immature sliced styles isconcomitant with the presence of lipid secretion. These resultsfully support the efficiency of the so-called ‘one-steppollination’ method in Eucalyptus globulus, which we nowprove can be applied more successfully to immature non-dehiscedflowers. General application of this procedure is likely tohave beneficial impacts on the planning and efficiency of large-scalegenetic improvement programs. Copyright 2001 Annals of BotanyCompany Eucalyptus globulus, one-step pollination, lipids, pollen, fertilization     

Regulation of LAT52 promoter activity during pollen tube growth through the pistil of Nicotiana alata.

The pollen-specific promoter of the LAT52 gene is known to direct expression of marker proteins during the last stages of pollen maturation and in very early pollen tube growth.We have examined the expression of LAT52-GUS during later stages of pollen tube growth in style and ovary of the relatively long-styled species Nicotiana alata. GUS activity was detected histochemically and found to be present in germinating pollen grains of N. alata and in tubes growing through the upper part of the style. No GUS activity was detected in 99% of the pollen tubes growing through the lower part of the style, but activity was present in tubes within the ovary. This finding indicates that the LAT52 promoter is regulated in growing pollen tubes, and is most active during the earliest and latest stages of pollen tube growth. GUS activity was also detected in some ovules, where it presumably marked the release of pollen tube cytoplasm into the ovule. The distribution of ovules with GUS activity within the ovary is not consistent with high-precision pollen tube guidance to the ovule. Received: 16 August 1999 / Revision accepted: 20 December 1999     

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     

Fertilizing Pollen Tubes Parasitise Ovary Wall in Grevillea

Tissue-invasive pollen tube branching occurs in an Australianshrub, Grevillea banksii R.Br. (Proteaceae). Parenchyma andphloem of the ovary wall are soon infiltrated after fertilization,by intercellular branches and the lumina of xylem elements areentered. Ovular tissues are not affected. Grevillea banksii, pollen, haustoria, ovary wall     

The Significance of the Obturator in the Control of Pollen Tube Entry into the Ovary in Peach (Prunus persica)

Changes in the obturator of the peach (Prunus persica) havebeen investigated and related to pollen tube growth in thisregion. At anthesis, the cells of the obturator are active andrich in starch reserves. Twelve days after anthesis these cellsproduce a secretion that stains for carbohydrates and for proteins.As the secretion is produced, starch vanishes from these cellsand they degenerate and collapse as callose is accumulated.Secretion is independent of pollination as it takes place ina similar fashion both in pollinated and in unpollinated flowers. Pollen tube growth along the obturator surface depends on thissecretion for, although pollen tubes reach the base of the styleseven days after pollination, they cannot grow on the obturatoruntil five days later, when the secretion is produced. Thisdiscontinuous secretion taking place at the obturator may providea mechanism that controls the entrance of pollen tubes intothe ovary in the peach. Prunus persica, peach, obturator, pollen tube     

A novel pollen tube growth assay utilizing a transmitting tract-ablated Nicotiana tabacum style

Sexual plant reproduction requires multiple pollen–pistil interactions from the stigma (pollen adhesion, hydration, and germination) to the ovary (fertilization). Understanding the factors that regulate pollen tube growth is critical to understanding the processes essential to sexual reproduction. Many pollen tube growth assays (PTGAs) have shorter and slower pollen tube growth when compared to pollen tube growth through the style. The identification and study of factors that regulate pollen tube growth have been impeded by a lack of an efficient and reproducible PTGA. The objective of this research is to develop a robust assay for Nicotiana tabacum pollen tube growth in an environment that supports sustained and normal growth yet is amenable to testing the effects of specific factors. In this paper, we introduce a novel PTGA, which uses pistils from N. tabacum that lack a mature transmitting tract (TT) due to tissue-specific ablation. The TT-ablated style supports normal pollen tube growth and the hollow structure of the style allows modification of the growth environment by direct injection of test material. This PTGA is robust and allows for rapid and accurate measurement of pollen tube length and pollen tube morphology, supporting pollen tube growth from 20 to 35°C and at pH ranging from 4.8 to 7.6. Use of the ablated style for a PTGA is a novel method for the culture of pollen tubes with sustained growth in vivo while permitting the application of treatments to the growing pollen tubes.     

Polyethylene glycol improves the in vitro growth of Brassica pollen tubes without loss in germination

The culture media for the in vitro pollen assay of Brassicaspecies have so far shown good percentage germination, but limitedpollen tube growth. It is reported here that by lowering sucroseconcentration from 20% to 5 or 10 % and by adding polyethyleneglycol 4000 in the culture medium, high percentage germinationand high tube growth (10 times that in the standard media) canbe achieved in three species of cultivated Brassica. This improvedmedium should be useful in fundamental and applied studies onthe pollen biology of Brassica species. Key words: Brassica, pollen, pollen germination, pollen tube growth, polyethylene glycol     

Relationships of pollen size,pistil length and pollen tube growth rates in Rhododendron and their influence on hybridization

Summary Pollen size and pistil length data have been collected for 93 species of Rhododendron (Ericaceae) belonging to a number of different subgeneric taxa. For a sample of eight species in section Vireya, pollen tube growth in the style after selfor interspecific pollination has been quantified. Pollen volume and the time taken for pollen tubes to reach the ovary were both related to pistil length. Pollen-tube growth rates were generally greater for species with longer pistils and larger pollen. Increasing temperature increased the rate of pollen-tube growth. There was no detectable effect of pollen tube density on tube growth rate in the style. After interspecific pollinations tube growth rates in foreign styles could be faster or slower than in self styles. A semisterile individual with two viable pollen grains per tetrad and a plant grafted as scion to a longer-styled stock both showed more rapid pollen-tube growth than expected on the basis of pistil size. Data collected for 26 species in section Vireya showed that where extreme disparity of pollen/pistil size causes failure of interspecific crosses, one or more bridging species with intermediate pollen/pistil size can generally be selected.     

Effects of Cross-pollination and Flower Removal on Fruit Set in Macadamia

Macadamia racemes were cross-pollinated and had flowers removedto determine whether manipulation of initial fruit set wouldaffect final fruit set or yield. Examination of flowers collected7 d after pollination indicated that the cross-pollination treatmentgenerally induced a small (13-19%) but significant increasein the number of pollen grains per stigma. All flowers possessedsome pollen grains, but in all cases cross-pollination approximatelydoubled the percentage of flowers with a pollen tube at thebase of the style (42-68% of flowers on cross-pollinated racemescompared with 21-35% of flowers on control racemes). Fruit setafter 14 d was always improved by cross-pollination, and thistranslated into increased final fruit numbers on two out offour occasions. In these two cases, kernel weights were increasedby 24 and 31% in the cross-pollinated treatments. This indicatesa possible xenic (paternal parent) effect in macadamia. Flowerremoval resulted in retention of a greater percentage of flowersas fruits, so that only severe reductions in flower number decreasedfinal fruit numbers. Improved efficiency of cross-pollinationin macadamia may increase both nut numbers and nut size.Copyright1994, 1999 Academic Press Macadamia integrifolia Maiden and Betche, Macadamia tetraphylla L. A. S. Johnson, Proteaceae, macadamia, pollination, pollen tube growth, flower number, fruit drop, fruit set, xenia     

The Importance of Light Intensity for Pollen Tube Growth and Embryo Survival in Wheat x Maize Crosses

The success of Triticum aestivumxZea mays crosses, used to producewheat doubled haploids, is influenced by light intensity. Toexamine the basis for this response, pollen tube growth, embryosurvival and indicators of photosynthetic rate were measuredin two wheat cultivars (‘Karamu’ and ‘Kotuku’)crossed with maize at two irradiance levels (250 or 750 µmolm^-2s^-1, PAR). Pollen tube growth was significantly affectedby light intensity in ‘Karamu’ plants but not in‘Kotuku’ plants, despite both cultivars being pollinatedby the same maize source. The percentage of pollen tubes reachingthe cavity between the ovarian wall and integuments, or in themicropyle of ‘Karamu’ plants at high light intensity(65%) was nearly three-times greater than that at low lightintensity (22%). Thus, either low light intensity can affectthe maternal wheat plant in a way that inhibits pollen tubegrowth and/or high light intensity may promote pollen tube growthin ‘Karamu’ plants. Significant differences in ratesof electron transport in plants grown at the two light intensitiesindicated that the rate of photosynthesis may also have an effecton pollen tube growth. These results have importance for improvingthe efficiency of wheat x maize crosses and other wide cerealcrosses. Copyright 2001 Annals of Botany Company Intergeneric hybridization, light intensity, pollen tube growth, embryo survival, Triticum aestivum, wheat,Zea mays , maize     

Semi in vivo pollen tube growth of Aechmea fasciata

With semi in vivo pollen tube growth assays, stigmas are pollinated in vivo and, after a fixed time interval, the styles are isolated from the ovary and placed on culture medium in vitro. Semi in vitro pollination includes isolation of the stigma and style complex, followed by pollination and placing the stylar end on nutrient medium. After semi in vivo pollination more and longer pollen tubes protruded from the cut end of the styles into medium, in comparison to semi in vitro pollination. Medium with 3 g l^–1 agar was better than that with 6 g l^–1 agar for pollen tube growth after the tubes emerged from the cut style. Semi in vitro pollination of the reversed style indicated that pollen tube growth was not influenced by the direction of the style. Fructose and glucose inhibited pollen tube growth compared to sucrose. Swollen tips characterized tube growth inhibition. After semi in vivo pollination all generative nuclei had divided to give two sperm nuclei. The average distance between the last sperm nucleus and the pollen tube tip as well as the distance between the two sperm nuclei diminished in growing pollen tubes between 24 and 48 h after pollination. The arrangements between the vegetative and the generative nuclei did not differ in semi in vivo and in vitro cultured pollen tubes of Aechmea fasciata. This information is important to explain why fertilization rate is low after placental pollination in comparison to placental grafted style pollination of Aechmea fasciata. The data may also contribute to the improvement of in vitro fertilization methods in Bromeliaceae and other higher plants.     

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.