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.     

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     

POLLEN TUBE DISTRIBUTIONS IN NICOTIANA GLAUCA: EVIDENCE FOR DENSITY DEPENDENT GROWTH

I hand sectioned styles of Nicotiana glauca at intervals along their length and counted the number of pollen tubes in each section using fluorescence microscopy. Evidence of density dependent growth was found for three stages of pollen growth. Pollen germination on the stigma increased with increasing pollen population size. Pollen tube penetration in the stigma was unaffected by increasing density from low to moderate levels but was reduced at high densities. Pollen tube penetration in the style was enhanced by increasing density. This enhanced growth in the style was apparently confounded by interference among pollen tubes growing at high densities. In particular, the area of tissue able to support pollen tube growth decreases from the stigma into the lower style, which could cause overcrowding of pollen tubes growing at high densities. Enhanced pollen tube penetration with increasing density combined with interference among pollen tubes growing at high densities resulted in greater mean pollen tube lengths for populations with moderate densities. The shift from density independent growth in the stigma to positively density dependent growth in the style may represent a shift from autotrophic to heterotrophic growth stages of pollen.     

Effects of pollen competitive environment on pollen performance in Mirabilis jalapa (Nyctaginaceae)

 We examined the influence of pollen competitive environment on pollen performance in Mirabilis jalapa. We used the number of pollen grains and the number of pollen tubes per pistil as measures of pollen competition. Pollen germination, pollen tube penetration into the style, and pollen tube growth rates were used as measures of pollen performance. All three measures of pollen performance were affected by the competitive environment. Pollen germination was greatest at intermediate pollen load sizes. The percentage of germinated pollen grains that penetrated the stigma and grew into the style decreased with pollen load size. Pollen tube growth rate in the style was greater and more variable with larger numbers of pollen tubes in the style. Controlling for the degree of selection at the stigma indicated that pollen-pollen or pollen-style interactions were the likely causes of increased growth rates. Received: 28 October 1996 / Revision accepted: 24 January 1997     

INFLUENCE OF THE PISTIL ON POLLEN TUBE KINETICS IN PEACH (PRUNUS PERSICA)

Pollen tube growth has been studied in peach and has been related to changes in the pistil structures which the pollen tube has to traverse in its way from the stigma down to the ovule. Growth of the pollen tubes along the pistil is not continuous. While pollen tubes reach the base of the style 7 days after pollination, fertilization does not take place until 12 days later. Pollen tubes stop for 5 days at the top of the obturator and they further stop for 3 days before entering the ovule. The pollen tube growth is heterotrophic; starch, present all along the pistilar tract at anthesis, vanishes as the pollen tubes pass by. Discontinuous pollen tube growth appears to be controlled by the pistil. At anthesis the pistil is not fully matured. Maturation of the pistil implies a number of secretory processes that occur in a basipetal way starting from the stigma down to the style and ending in the ovule. Some of these secretions at the stigma and the style are triggered by pollination; others appear to be a maturative stage of the pistil and are produced in a discrete way. The fact that the pollen tube depends on these secretions together with the fact that these secretions are not continuously produced confer upon the pistil a role of controlling pollen tube kinetics and point out that, for a successful fertilization, male gametophyte development and pistil maturation need to by synchronized.     

POLLEN LONGEVITY AND STIGMA PRE-EMPTION IN CLARKIA

The ability of pollen to effect seed set decreases with age when stored at room temperature or on the plant, but not when stored at 5 C. Seed set declines rapidly with pollen stored 10 days or more at room temperature. Seed set declines rapidly with pollen stored 4 days or more on the plant. The degree of pollen germination on the stigma and the number of pollen tubes in the style correlate positively with seed set. Pollen germination on the stigma and pollen tube growth in the style are strongly correlated with seed set for pollen stored at room temperature, but not, or only weakly, correlated for pollen stored at 5 C. In the latter case, seed sets are lower than expected, indicating a possible viability reduction associated with a post-pollination function. Inviable pollen grains temporarily interfere with normal pollen germination on the stigma. Essentially no seeds are produced when in viable pollen, immediately followed by viable pollen, is placed on a fresh stigma. However 27 to 65 seeds are produced when in viable pollen, followed in 1 to 6 days by viable pollen, is placed on fresh stigmas. New stigma surface is exposed at the center as expanding stigma surfaces carry the interfering inviable pollen toward the edge of the stigma.     

Pollen Tube Growth and Fertilization Efficiency in Salpiglossis sinuata: Implications for the Involvement of Chemotropic Factors

The possible involvement of chemotropic factors influencingpollen tube growth in the style and ovary of Salpiglossis wasstudied. Pollen deposited at the base of excised styles germinatedreadily, with pollen tubes penetrating the transmitting tissueand growing towards the stigma, implying that no chemotropicrequirement exists for pollen tube growth in the style. However,the position and high efficiency of seed set following limited-pollinationof intact pistils would support the involvement of localizedchemotropism operating within the ovary. Chemotropism, limited pollination, fertilization efficiency, Salpiglossis sinuata     

Interspecific crosses in the genus Tulipa L.: identification of pre-fertilization barriers

 Pollen tube growth in the pistil and pollen tube penetration of ovules have both been studied in crosses between cultivars from Tulipa gesneriana L. and 12 tulip species from all eight sections of the genus Tulipa to identify pre-fertilization barriers. Depending on the cross, pollen tubes grew as far as the stigma or the style or continued growing down into the ovary. Pollen tubes penetrated none or only a few percent of the ovules of some crosses, despite the presence of many pollen tubes in the ovary. In other crosses, from which no or only a few hybrids have been obtained after seed maturation on the plant, pollen tube penetration was found in up to 79% of the ovules. Apparently, various kinds of barriers preventing fertilization or normal embryogenesis occur in interspecific tulip crosses. Received: 26 July 1996 / Revision accepted: 31 January 1997     

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     

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     

不同葡萄品种柱头、花柱发育与种子形成的关系

对‘京秀’、‘香妃’(二倍体)和‘黑奥林’、‘巨峰’(四倍体)葡萄品种的柱头、花柱发生发育以及花粉管在雌蕊中生长与坐果和果实中种子形成的关系进行了研究。结果显示:‘京秀’、‘香妃’的坐果率和果实中的种子数明显高于‘黑奥林’和‘巨峰’,而‘巨峰’葡萄的坐果率和种子数最低。开花期‘京秀’和‘香妃’的花柱直径略小于‘黑奥林’和‘巨峰’,但花柱中部引导组织的直径和引导组织所占花柱的比例却大于‘黑奥林’和‘巨峰’;半实心花柱的比例也高于‘黑奥林’和‘巨峰’,说明花柱中引导组织的结构和发育状况可能与坐果和果实中种子形成有关;人工授粉24 h后,在‘京秀’和‘香妃’雌蕊中花粉管的生长速度快于‘黑奥林’和‘巨峰’,授粉后48 h‘京秀’和‘香妃’雌蕊中到达各部位的花粉管数要多于‘黑奥林’和‘巨峰’,授粉后72 h各品种到达子房各部位的花粉管数开始减少。表明‘京秀’和‘香妃’花柱中引导组织较为发达,而且半实心花柱比例也较高,有利于花粉管在雌蕊中的生长和完成授粉受精,使‘京秀’和‘香妃’的坐果率和果实中种子的形成都较高。     

异叶苦竹花粉管生长及双受精过程

以异叶苦竹为材料,采用扫描电镜、荧光显微镜技术及传统的石蜡制片技术,解剖观察其花粉管生长途径及双受精过程。结果表明:(1)授粉后,花粉在柱头上吸水膨胀,约30 min即可萌发。(2)授粉1～2 h后花粉管可达到花粉长度的5～10倍,花粉管在柱头分支中进一步伸长,并开始伸入花柱中生长。(3)授粉后5 h,大量花粉管沿引导组织进入花柱基部与子房顶部之间的子房壁,有少量花粉管在子房壁与外珠被之间的缝隙中生长。(4)授粉后8 h,少量花粉管到达珠孔端。(5)授粉后15～18 h,精核与极核融合,形成初生胚乳核;精、卵核融合,形成合子。(6)授粉后20～30 h,仍可在花柱中见到大量呈束状的花粉管。(7)授粉后48 h,子房内的大部分花粉管出现解体,大多数花粉死亡。研究认为,精细胞到达胚珠的时间为8 h。     

Expression of stylar incompatibility in the Andean clonal tuber crop oca (Oxalis tuberosa Mol., Oxalidaceae)

Long-, mid-, and short-styled clonal accessions of oca (Oxalis tuberosa) were intercrossed in a complete diallelic design. Pollen tube growth in styles was monitored in all diallelic crosses. Pollen fertility was estimated by two tests: staining of pollen grains with aceto-carmine and detection of β-galactosidase activity by the substrate X-Gal. The two methods of pollen fertility estimation were equally useful to detect fertility levels. Pollen originating from short stamens had the highest fertility (85%) and pollen from long stamens had the lowest fertility (70%). Pollen fertility was high throughout, but its degree varied with the stylar morph on which the pollen was formed. Long-styled accessions had the highest rates of fertile pollen. Differences in pollen fertility at different anther levels in the same style morph were also apparent. Pollen grain diameter of the six morph-anther level combinations was inversely correlated with pollen fertility. Pollen grains from long stamens were the largest and pollen grains from short stamens were the smallest. Neither pollen fertility nor pollen grain size had an influence on pollen tube growth in the style or on fruit and seed set. Pollen tubes growing within the styles were inhibited at a different level for each of the 18 cross combinations in the diallel. Although legitimate crosses had greatest pollen tube growth, some of the illegitimate inter- and intramorph crosses had equally high scores. Of all illegitimate crosses, mid-styled seed parents had the lowest level of stylar incompatibility. Fruit and seed set were highly correlated with the extent of pollen tube growth in the style. The number of pollen tubes entering ovules in a flower was in good agreement with the number of seeds produced per fruit. Therefore, it is concluded that stylar incompatibility is the major determinant of limited seed formation in oca even in the most successful legitimate cross combinations. Received: 1 September 1999 / Revision accepted: 4 April 2000     

Pollen tube dynamics following half- and fully-compatible pollinations in self-compatible almond cultivars

. Pollen tube growth dynamics along the style at different times after self- and cross-pollination of four heterozygous self-compatible (SfSx) almond cultivars were studied by means of fluorescence microscopy. Results showed a reduction of pollen tube number along the style following both self-pollination (half-compatible reaction) and cross-pollination (fully-compatible). An important decrease in the percentage of pollen tubes present took place in the upper section of the style in both crosses. As expected in a gametophytic incompatibility system, a higher number of pollen tubes were stopped along the style in the case of the half-compatible crosses; however, the percentage of pollen tubes reaching the base of the style was similar in both cases. These results reveal the existence of other mechanisms controlling pollen tube growth along the style.     

Observation of Pollen Tube Behaviour and Early Embryogenesis Following Interspecies Pollination Between Actinidia deliciosa and A. arguta

The pollen tube behaviour in the style and early embryogenesis following interspecies pollination between Actinidia deliciosa No. 26 and A. arguta were observed by means of fluorescence and light microscopy. Pollen grains germinated on the papillate stigma and pollen tubes grew along the V-shaped open-type style. Pollen tubes showed slower growth and reached the ovules 50--60 hours later than those of the control. Several abnormalities of pollen tubes have been observed at the base of the style, including wave-like pollen tubes, pollen tubes with swollen or pointed tips, with variable diameters, and a few with irregular growth. Random deposition of callose along pollen tube wall and even the whole wall was observed. About 26.74 % of the ovules were successfully fertilized and developed into seeds, among them 68.50% of the seeds were normal and 31.50% were abortive. About 11.41% were empty seeds without embryo and endosperm. Unfertilized small ovule was 61.45 %. Normal seed and its embryo were smaller than those of the control. The development of embryo was of the Soland type. The endosperm was cellular. The zygote remained quiescent for about 12-15 days before it started to divide, eventually forming a cotyledonary embryo 50 days after pollination.     

Effects of pollen quality and quantity on pollen limitation in Crataegus monogyna (Rosaceae) in NW Spain

Pollen limitation occurs when plants produce less fruits and/or seeds than they would with adequate pollen receipt. If the addition of cross-pollen to stigmas increases fruit/seed production, it is interpreted as an evidence of pollen limitation. Much of the limitation may be associated with the quality rather than quantity of pollen; however, most studies do not discriminate between the two, which may lead to misinterpretation of the results. We studied the effects of quality and quantity of pollen on the reproduction of a northern Spanish population of Crataegus monogyna. The treatments included self- and cross-pollination, and supplementation to open and bagged flowers. The response variables considered were number of pollen grains per stigma, pollen tubes per style, and initial and final fruit set. In the Cantabrian range, C. monogyna requires insect pollinators to set fruit and is partially self-incompatible. We found that the number of pollen tubes did not differ between cross- and self-pollination treatments; however, self-pollinated flowers set less fruits than flowers that received pure cross-pollen or were supplemented with both cross- and self-pollen. The experimental design allowed us to infer qualitative rather than quantitative pollen limitation. Comparison of the number of pollen grains and tubes, and initial and final fruit set among pollination treatments suggested post-zygotic embryo selection against selfed progeny.     

Microgametophyte population sizes and plant reproductive output in the insect-pollinated Prunella grandiflora (Lamiaceae)

Intraspecific variation in pollen deposition and number of pollen tubes per style is rarely quantified, but is essential for assessing the occurrence of pollen limitation and pollen competition and their evolutionary implications. Moreover, pollen deposition, pollen tube growth, and the fate of fertilized ovules are rarely distinguished in field studies. Here we present such a study in eight natural populations of Prunella grandiflora. We quantified microgametophyte population sizes and inferred pollen limitation when the number of fertilizable ovules exceeded pollen tubes, and assessed seed set and fate after open pollination. Two and three populations had on average significantly fewer pollen grains and pollen tubes per flower, respectively, than the fixed number of fertilizable ovules per fruit, while one population experienced significant pollen competition. Style length was positively correlated with the number of pollen tubes. While pollen availability was very variable, seed abortion was significantly less frequent in denser populations, and in one population the proportion of well-developed seeds was significantly, positively correlated with the number of pollen tubes in the style. Less pollen deposition, lower numbers of pollen tubes reaching the base of the style, lower pollen quality and therefore increased abortion of fertilized ovules can all reduce seed set in natural P. grandiflora stands. Substantial intraspecific variability implies that microgametophyte competition also occurs in this species. Finally, style morphology may affect pollen receipt.     

The relationship of the female reproductive success of Eucalyptus globulus to the endogenous properties of the flower

Low capsule and seed set is a major factor limiting seed production in Eucalyptus globulus seed orchards. Controlled pollination studies showed that the reproductive success (number of seeds produced per flower pollinated) was primarily determined by the female. We aimed to identify the factors contributing to the differences in reproductive success between female genotypes in terms of the physical and anatomical properties of the flower. We studied pairs of genotypes of high and low reproductive success from each of three races (Furneaux Group, Strzelecki Ranges and Western Otways) growing in a seed orchard. Controlled pollinations were performed on six females and along with flower physical measurements, pollen tube growth and seed set were assessed. Overall tree reproductive success was positively correlated with flower size, ovule numbers, style size, cross-sectional area of conductive tissue within the style (all of which were inter-correlated) and the proportion of pollen tubes reaching the bottom of the style. Significant positive correlations of reproductive success and flower physical properties between different ramets of the same genotypes across seasons suggests a genetic basis to the variation observed. The majority of pollen tube attrition occurred within the first millimetre of the cut style and appeared to be associated with differences in style physiology. When examined as pairs within races the difference in reproductive success for the Western Otways pair was simply explained by differences in flower size and the number of ovules per flower. Physical features did not differ significantly for the Strzelecki Ranges pair, but the proportion of pollen tubes reaching the bottom of the style was lower in the less reproductively successful genotype, suggesting an endogenous physiological constraint to pollen tube growth. The difference in reproductive success between the females from the Furneaux Group was associated with a combination of these factors.     

Pollen-pistil interactions in Hermodactylus tuberosus Mill. (Iridaceae)

ABSTRACT

Hermodactylus tuberosus Mill. (Iridaceae) is a species with a very short style and a tricarpellar unilocular ovary. For ornamental purposes it is usually reproduced by means of tubers, although both vegetative and seed reproduction occur in nature. However, its capsules do not contain many seeds in comparison to the high number of ovules inside the ovary. To understand this conflicting process a study on H. tuberosus pollen-pistil interaction was carried out. Pollen viability was evaluated by different techniques. Germination rate was tested in vitro and in vivo after stigma, intrastylar and intraovarian pollinations. A high percentage of the pollen grains were found to be viable resulting in a high percentage of in vitro germination and a low rate of germination on the stigma. Indeed, many pollen tubes developed after intrastylar and intraovarian pollinations. Several anomalies characterized pollen germination and pollen tube growth in vitro and in vivo. Observations from field trials indicated that capsules were only produced from open or hand, cross- pollinated flowers, confirming that the plant is self- incompatible and not apomictic. In addition, all the mature capsules contained only a few mature seeds suggesting the existence of pre- and postzygotic seed set inhibition.     

荞麦亲和与不亲和授粉后柱头和花粉管中ATP酶的超微细胞化学定位

利用磷酸铅淀淀技术对荞麦（Fagopyrum esculentum Monch.)pin型植株分别进行亲和授粉和不亲和授粉后的柱头、花粉粒、花粉管进行了ATPase的超微细胞化学定位。结果表明（1）亲和授粉和不亲和授粉后0.5h,柱头细胞的ATPase活性反应水平较低或基于无酶活性;柱头表面、柱头上附着的花粉粒内ATPase活性在不亲和授粉时较低,亲和授粉时较高,花粉粒内ATPase主要定位于线粒体和精子细胞;（2）授粉后1.5h,不亲和授粉的柱头细胞及花粉管的ATPase活性均较低,花粉管停止生长,细胞质开始解体;而亲和授粉的柱头细胞及花粉管的ATPase活性均较高,ATPase主要定位于柱头细胞的质膜、胞基质以及花粉管的壁、质体的膜、高尔基体、线粒体上。根据不同时期不同部位ATPase活性的差异,我们认为荞麦发生自交不亲和时,花粉管在花柱中停止生长不仅是因此花粉管得不到花柱中的营养物质而引起的,可能也与花粉管自身物质代谢发生障碍有关。