Morph-ratio variation, population size and female reproductive success in distylous Pulmonaria officinalis (Boraginaceae)

Theory predicts that morph ratios in heterostylous populations are governed by negative frequency-dependent selection typically resulting in equal morph ratios at equilibrium. Previous work on the distylous perennial herb Pulmonaria officinalis, however, showed asymmetric mating between floral morphs and a weak self-incompatibility system, with the long-styled morph (L-morph) producing significantly higher seed set following intramorph crosses and even selfing than the short-styled morph (S-morph), two aspects thought to affect female fecundity and morph-ratio variation. Here, we evaluated morph ratios and population size of all known P. officinalis populations in the northern part of Belgium. Morph ratios deviated significantly from 1:1 (range 0.09-1 L-morph frequency, mean = 0.58). Relative fecundity of the S-morph (i.e. mean seed set of the S-morph/mean seed set of the L-morph) was on average 0.73, was positively related to the frequency of the L-morph, and reached 1 (similar levels of female fecundity) at an average L-morph frequency of 0.66 in the population. As some small populations had the S-morph in majority, our results suggest that local morph ratios are influenced both by the relative fecundity of L- and S-morph individuals and by stochastic processes in small populations.     

Floral display size and spatial distribution of potential mates affect pollen deposition and female reproductive success in distylous Pulmonaria officinalis (Boraginaceae)

In animal‐pollinated plants, both the spatial distribution of flowering individuals and the number of flowers that an individual displays affect pollen deposition rates and female reproductive success. Heterostylous species are likely to be particularly sensitive to the contingencies of spatial distribution, as they are reproductively subdivided into distinct mating groups, which usually exhibit self‐ and intra‐morph incompatibility and differ in floral morphology. In this paper, we explore the joint effects of both spatial distribution of potential mates and floral display size on morph‐specific pollen deposition rates and seed set patterns in two natural populations of Pulmonaria officinalis, a distylous species with a weak self‐incompatibility system. Both total stigmatic pollen load and the proportion of legitimate pollen decreased with increasing spatial isolation. Legitimate (intermorph) pollen transfer was, however, asymmetric and decreased more rapidly with decreasing proximity to a compatible legitimate mating partner in the S‐morph than in the L‐morph. Total stigmatic pollen loads per flower increased with increasing floral display size, indicating that large plants are disproportionately more visited than smaller individuals. However, because legitimate pollen deposition decreased with increasing floral display size, these results also suggest that larger numbers of flowers increase the degree of geitonogamous pollination. In both the L‐ and S‐morph, seed set significantly decreased with increasing isolation from a legitimate mating partner, but in the L‐morph seed set was less dependent on the spatial distribution of the S‐morph. In addition, seed set significantly increased with floral display size in the L‐morph, but not in the S‐morph. These findings indicate that the spatial distribution of potential mates and variation in floral display size may cause morph‐specific differences in pollen deposition rates and female reproductive success.     

The control of incompatibility in distylous Pulmonaria affinis Jordan (Boraginaceae)

In P. affinis, pin pollen is shorter on average than thrum pollen. Pins have more coronal hooks on stigmatic papillae than thrums, but gaps between papillae are relatively smaller for thrums. Pin stigmas receive more pollen than thrum stigmas. Thrum stigmas receive more (dissortive) pin pollen, but pin stigmas are assortively pollinated. Pollen only germinates when trapped below papilla coronas. On thrum stigmas, most trapped pollen is pin. Pollen germination is better on thrum stigmas than pin stigmas, and thrum stigmas show a close relationship between numbers of legitimate pollen grains, numbers of germinating grains, and numbers of pollen tubes in the style. There is no inhibition of illegitimate pollen germination. Illegitimate pollen tubes are inhibited in the style. Incompatibility operates by a combination of dissortive pollination, dissortive pollen trapping, and stylar pollen tube inhibition. All heteromorphic features differing between pins and thrums are implicated in the inhibition of within-morph fertilization in thrums.     

Gender specialization and distyly in hoary puccoon,LITHOSPERMUM CROCEUM (Boraginaceae)

This 2-yr study of gender and its relation to the sizes of pin and thrum plants in Lithospermum croceum was performed in central Michigan. The population was composed of 51-52% thrums, 42-43% pins, and ≈6% homostyles in 1988 and 1989. Pins produced more flowers and fruits than thrums, and a greater proportion of their ovaries produced more than one nutlet. However, the number of fruits produced by pins and thrums in 1988 did not differ when plant size is a covariate in ANCOVA. The number of seeds produced by plants in 1989 is also unaffected by floral morph when size is a covariate. Instead, plant size is the most important predictor of fecundity in both years. The average percent fruit sets (fruits/ovaries) of the morphs did not differ in either year. Herbivory was more common in 1988 than in 1989, but there was no difference in the frequency with which pins and thrums were attacked. Furthermore, the effect of herbivory on fecundity did not differ for the morphs. The fecundity of neither morph was limited by pollen in 1989. Apparent gender specialization was a consequence of subtle differences in the vegetative sizes of the morphs.     

Pollen morphology of the tribe Cynoglosseae of Boraginoideae (Boraginaceae) in China

Pollen morphology of 16 Chinese species representing 7 genera in the tribe Cynoglosseae of Boraginoideae(Boraginaceae) was examined under LM and SEM, and 5 species under TEM. Pollen grains are cocoon-shaped, rarely subprolate, prolate or ovoid, very small, 7～ 15.7μm× 3.5 ~ 13.9 μm in size, P/E = 1.6～2.02; 3-colporate apertures alternate with 3-pseudocolpi, with equatorial endocingulus except those in Bothriospermum; exine surface is usually smooth, with or without perforations in two poles, rarely with tuberculate ornamentation; exine is rather thin and includes ectexine and endexine, while ectexine consists of imperforate tectum, columellae and foot-layer. The tribe Cynoglosseae has many common characters of pollen grains, but there are some differences among genera. A key to the genera is given based on pollen morphology. Bothriospermum, without endocingulus, may be a primitive genus in this tribe, and its pollen morphology is more similar to that of Eritrichieae than to Cynoglosseae, thus it seems more reasonable to put it into Eritrichieae. Solenanthus may be the most advanced genus in the tribe Cynoglosseae for its pollen grains of sub-isopolar and ovoid shape.     

Cleistogamy in the rare high Andean perennial herb Cryptantha capituliflora (Boraginaceae)

 Axial and apical flowers of Cryptantha capituliflora were analyzed with regard to morphology and pollen tube growth to assess the occurrence of cleistogamy. Although intermediate floral forms do occur, cleistogamous flowers were significantly smaller than chasmogamous flowers, had fewer anthers, and showed a distinctive stigmatic surface. Chasmogamous flowers can be cross-pollinated. Nevertheless, the growth of self-pollen tubes in few chasmogamous buds jointly with flower characters suggests that these flowers can probably produce fruits through autonomous selfing. The mean seed number per fruit did not differ between fruits from chasmogamous and cleistogamous flowers. Cleistogamous flowers were only observed in axial inflorescences, which are completely covered by the leaf. Other species of section Cryptantha also show the same trend, with cleistogamous flowers located in the lower half of the stems. This pattern is discussed in relation to dissimilarities in the outcrossing opportunities between flower types within the plant. Received May 22, 2002; accepted November 14, 2002 Published online: March 20, 2003     

Size-dependent pollination efficiency in Anchusa officinalis (Boraginaceae): causes and consequences

Summary Bumblebees foraging on the self-incompatible Anchusa officinalis fly between near neighbour plants and between near neighbour inflorescences within plants. Although many-flowered plants attracted most bumblebees these plants received fewer visits on a per flower basis than smaller plants, and each bumblebee visited a smaller proportion of the flowers. The calculated effective visitation rate per flower was highest on plants of an intermediate size. If pollen-carryover was assumed to be limited the most efficient plant was predicted to be smaller since the proportion of fertilized flowers per bumblebee visit is expected to decrease further on the largest plants in relation to the total flower number. These predictions were tested by measuring fruit-set in the field. The percentage fruit-set decreased with plant size at all sizes that were investigated. That the most efficient plant was small indicates that pollen-carryover was indeed limited. However, the low percentage fruit-set associated with large size did not present a serious problem since the total estimated seed production per plant still increased with size. Selection favoring smaller plants may be low or absent in Anchusa.     

中国紫草科紫草亚科琉璃草族花粉形态的研究

对中国紫草科Boraginaceae 紫草亚科Boraginoideae琉璃草族Trib．Cynoglosseae 7属16种的花粉形态进行了光学显微镜和扫描电镜观察,并对其中5种的花粉外壁超微结构进行了透射电镜观察。该族花粉为茧形,少数为近长球形、长球形或卵球形;体积很小,大小为7～15.7μm×3.5～13.9μm,P／E=1.6～2.02;三孔沟和三假沟相间排列;除斑种草属Bothriospermum Bge．外,均有赤道环带;外壁常光滑,两极具或不具穿孔,少数具小瘤状纹饰;外壁较薄,包括外壁外层和外壁内层,外壁外层又由完全覆盖层、柱状层和基层组成。以上表明该族花粉有很多共同特征,但属之间还是有区别。文中作了花粉分属检索表。在该族中,斑种草属的花粉没有赤道环带,表明它是比较原始的类群。它和齿缘草族的花粉更相似,因此把它放在齿缘草族Trib．Eritrichieae比放在琉璃草族似乎更合适。长蕊琉璃草属SolenanthusLedeb．由于花粉卵球形、亚等极,可认为是琉璃草族中最进化的属。     

Pollen Morphology of Microula Benth. and Allied Genera (Boraginaceae)

Pollen grains of 16 species of Microula Benth. and six species of three related genera were examined under LM and SEM, and four of them also under TEM. Pollen grains of Microula and three related genera are dumb-bell-shaped, 3-colporate apertures alternate with three pseudocolpi. Pollen grains are very small,ranging from 12.18 x 7.13 μm to 6.36 x 3.66μm. In general, colpi with os are wider and shorter, rhomboid, but sometimes they are equal to pseudocolpi in length. Colpus margins are regularly or irregularly tooth-like. The surface of colpi is psilate or processed. Ora are circular or lalongated in outline, protruded or not; surface of os membrane is smooth or scabrid. The exine is usually indistinctly layered under LM. The exine surface is psilate, and more or less perforate. TEM examination shows that the pollen wall is differentiated into exine and intine: the exine includes ectexine and endexine, while the ectexine consists of tectum, columellae and foot-layer. However, there are differences in constriction of equatorial area, apertural characters, ornamentation and exinous ultrastructure between these genera. Pollen morphology indicates that the genus Microula Benth. is primitive, directly related to the genus Actinocarya Benth .; the genus Asperugo L. Is more advanced. The genus Eritrichium Schrad. which has two ora or one os and is anisopolar, represents the most advanced group among them. Noteworthily, the diorate phenomenon is found for the first time not onlyin the genus but in the family Boraginaceae.     

Pollen morphology in theBoragineae (Boraginaceae) in relation to the taxonomy of the tribe

The results of a comparative survey on pollen morphology in theBoragineae by means of light, scanning and transmission electron microscopy are presented and discussed in relation to the taxonomy of the tribe. Multivariate analyses lead to the recognition of 15 pollen morphotypes for which a discriminant key is proposed. The discriminatory characters concern mainly the stereostructural and ultrastructural features of the grains, such as tectum sculpture and aperture morphology, while the number of apertures appears variable within genera and even single species. Seven out of the 12 investigated genera, as currently circumscribed, are matched by palynological data:Anchusella, Borago, Brunnera, Elizaldia, Lithodora, Symphytum andTrachystemon. On account of aperture shape,Lithodora is however closer to theLithospermeae than to theBoragineae. Other genera, and in particularNonea, show a wide variation in tectum ornamentation, shape of grains and number of apertures. Palynological data do not support a broad concept of the genusAnchusa, and point to the autonomy of the satellite generaHormuzakia, Gastrocotyle, Phyllocara, Pentaglottis andCynoglottis.     

Pollen wall ultrastructure and ontogeny in Heliotropium europaeum L. (Boraginaceae)

The pollen grains of Heliotropium europaeum are heterocolpate, with alternation of 3 colpori and 3 pseudocolpi. The exine is characterized by a scabrate and thick tectum, massive columellae with a granular appearance and a thick nexine. The thickening of the intine at the apertural level makes the interpretation of this zone difficult. The ontogenetic study helped to understand the ultrastructure of the exine and the apertures. The different steps are as follows. The primexine matrix is formed during the beginning of the tetrad stage; it consists of an outer thick and electron dense zone and an inner one, less dense to electrons. The tectum and the infratectum begin to form in the outer zone of the matrix, towards the middle of the tetrad stage. The infratectum consists of a network of columellae variable in thickness and oriented in different directions. The foot layer is lacking. The endexine is formed on a lamella system during the callose loss and microspore separation. The endexine becomes compact very early on its inner part. The apertures are initiated during the tetrad stage; a granulo-fibrillar oncus develops. At the free microspore stage, the oncus gets fibrillar and is bordered by endexine lamellae on its outer side and by endexine granulations on its inner one and laterally. The intine is set at the end of this stage. At the vacuolated microspore stage, the intine shows three layers: two thin, clear and homogeneous layers, one outside and the other inside, and a thick middle layer that forms the zwischenkörper, crossed by trabecula, in the apertural areas.     

Pollen and stigma size changes during the transition from tristyly to distyly in Oxalis alpina (Oxalidaceae)

• Pollen and stigma size have the potential to influence male fitness of hermaphroditic plants, particularly in species presenting floral polymorphisms characterised by marked differences in these traits among floral morphs. In this study, we take advantage of the evolutionary transition from tristyly to distyly experienced by Oxalis alpina (Oxalidaceae), and examined whether modifications in the ancillary traits (pollen and stigma size) respond to allometric changes in other floral traits. Also, we tested whether these modifications are in accordance with what would be expected under the hypothesis that novel competitive scenarios (as in distylous‐derived reproductive system) exert morph‐ and whorl‐specific selective pressures to match the available stigmas.
• We measure pollen and stigma size in five populations of O. alpina representing the tristyly–distyly transition.
• A general reduction in pollen and stigma size occurred along the tristyly–distyly transition, and pollen size from the two anther levels within each morph converged to a similar size that was characterised by whorl‐specific changes (increases or decreases) in pollen size of different anthers in each floral type.
• Overall, results from this study show that the evolution of distyly in this species is characterised not only by changes in sexual organ position and flower size, but also by morph‐specific changes in pollen and stigma size. This evidence supports the importance of selection on pollen and stigma size, which increase fitness of remaining morphs following the evolution of distyly, and raises questions to explore on the functional value of pollen size in heterostylous systems under pollen competition.

     

Pollen flow, population composition, and the adaptive significance of distyly in Linum tenuifolium L. (Linaceae)

The function of distyly as a device promoting inter-morph pollination was proposed by Darwin over a century ago, though only recently have experimental studies been undertaken to test this hypothesis. These studies are unrealistic since they assume regular spatial distribution of morphs within a population and fail to consider intra-flower pollination in assessing distyly's effects. Pollen flow in two populations of Linum tenuifolium L. was investigated using a fluorescent-dye marker technique. One population was dimorphic and self-incompatible; the other was monomorphic and self-compatible. In both populations pollen dispersal was leptokurtic with over 75% of grains detected within 5 m from the source. Intra-flower pollinations comprised a large proportion of individual stigma loads (42% for the dimorphic form; 49% for the monomorphic form). Total stigmatic loads for the dimorphic population showed equal proportions of pin and thrum pollen on both long-style and short-style stigmas, though variation in individual loads was marked. Values ranging from 34.2 to 61.5% legitimate pollen were recorded. The irregular distribution of morphs observed at the dimorphic site will reduce the efficiency of heterostyly as a device promoting intermorph pollination. The reduction in intra-flower pollination suggests adaptive significance of this floral dimorphism in increasing the efficiency of pollen dispersal to the individual.     

Pollen morphology of the tribes Eritrichieae and Cynoglosseae (Boraginaceae) in the Iberian Peninsula and its taxonomic significance

DÍEZ, M. J. & VALDÉS, B., 1991. Pollen morphology of the tribes Eritrichieae and Cynoglosseae (Boraginaceae) in the Iberian Peninsula and its taxonomic significance.
The pollen morphology of 33 species of Boraginaceae from the Iberian Peninsula belonging to the tribes Eritrichieae and Cynoglosseae has been studied by light and scanning electron microscopy. The results confirm the eurypalinous character of this family, in which a large number of species can be recognized by their pollen characters.
The pollen of these two tribes is basically heterocolpate, a type which is very uncommon in the angiosperms, with 6, 8 or 10 apertures, sometimes with ectoapertures anastomosed at the equator (ectocingulum), or diorate, minute or small in size, outline elliptic or rectangular-elliptic in equatorial view, with or without a constriction at the equator, exine 0.5-1.0 urn thick and the tectum psilate with granules around the colpi. The similarity in shape and apertural system found in several genera such as Asperugo, Gyrocarion and Omphalodes, and some species of Myosotis, does not permit the two tribes to be distinguished on pollen characters. The generic limits of Cynoglossum and the infrageneric classification of Myosotis are discussed.     

Temperature and water stress affect plant–pollinator interactions in Borago officinalis (Boraginaceae)

Climate change alters the abiotic constraints faced by plants, including increasing temperature and water stress. These changes may affect flower development and production of flower rewards, thus altering plant–pollinator interactions. Here, we investigated the consequences of increased temperature and water stress on plant growth, floral biology, flower‐reward production, and insect visitation of a widespread bee‐visited species, Borago officinalis. Plants were grown for 5 weeks under three temperature regimes (21, 24, and 27°C) and two watering regimes (well‐watered and water‐stressed). Plant growth was more affected by temperature rise than water stress, and the reproductive growth was affected by both stresses. Vegetative traits were stimulated at 24°C, but impaired at 27°C. Flower development was mainly affected by water stress, which decreased flower number (15 ± 2 flowers/plant in well‐watered plants vs. 8 ± 1 flowers/plant under water stress). Flowers had a reduced corolla surface under temperature rise and water stress (3.8 ± 0.5 cm² in well‐watered plants at 21°C vs. 2.2 ± 0.1 cm² in water‐stressed plants at 27°C). Both constraints reduced flower‐reward production. Nectar sugar content decreased from 3.9 ± 0.3 mg/flower in the well‐watered plants at 21°C to 1.3 ± 0.4 mg/flower in the water‐stressed plants at 27°C. Total pollen quantity was not affected, but pollen viability decreased from 79 ± 4% in the well‐watered plants at 21°C to 25 ± 9% in the water‐stressed plants at 27°C. Flowers in the well‐watered plants at 21°C received at least twice as many bumblebee visits compared with the other treatments. In conclusion, floral modifications induced by abiotic stresses related to climate change affect insect behavior and alter plant–pollinator interactions.     

The influence of enhanced UV-B radiation on the spring geophyte Pulmonaria officinalis

Pulmonaria officinalis is an understorey spring geophyte, which starts its vegetative period before full foliation of the tree storey. During its early growth phase it is exposed to full solar radiation, therefore the enhanced UV-B radiation could present a threat to this species. An outdoor experiment in which potted plants were exposed to below ambient, ambient, and above ambient (corresponding to 17% ozone reduction) UV-B radiation, was conducted in order to evaluate the radiation effects. The amount of photosynthetic pigments and photochemical efficiency of PSII were not affected, but the amount of UV-B absorbing compounds was lower in plants grown under reduced UV-B. This change was measurable after only fourteen days in reproductive shoots, while in the vegetative shoots, it was not detectable until after three months. The leaves of P. officinalis are variegated and the light green spots became less transparent to PAR under enhanced UV-B. The results reveal that under simulated 17% ozone depletion the harmful effects of UV-B on the measured parameters were negligible.     

Discovery of distyly in Narcissus (Amaryllidaceae)

There has been disagreement concerning the nature of sexual polymorphisms in Narcissus, a genus of insect-pollinated geophytes native to the Mediterranean. The existence of trimorphic heterostyly in the genus has recently been confirmed, but the occurrence of distyly remains enigmatic. All sexually dimorphic species previously investigated possess two distinct style lengths but anthers of similar height. Stigma-height dimorphism does not qualify as true distyly because of the absence of a reciprocal correspondence in stigma and anther position in the floral morphs. Such reciprocal herkogamy is generally regarded as the defining feature of heterostyly. Here we report on distyly in N. albimarginatus (section Apodanthae), a rare species confined to a single mountain in northwestern Morocco. A population composed of equal numbers of long- and short-styled plants exhibited reciprocal herkogamy with lower anthers of the long-styled morph and upper anthers of the short-styled morph corresponding in height to stigmas of short- and long-styled plants, respectively. The presence of both stigma-height dimorphism and distyly in Narcissus is of general significance to theoretical models of the evolution of heterostyly.     

中国紫草科破布木属花粉形态和外壁超微结构

为了深入探讨紫草科（Boraginaceae）的分类问题,用光学显微镜和扫描电子显微镜观察了该科破布木属（CordiaL.) 10种植物的花粉形态和外壁超微结构。发现该属花粉具三孔、三孔沟、三拟孔沟和三合沟4种萌发孔类型。外壁表面具微刺状纹饰、刺状纹饰、网状纹饰和不规则的条纹网状纹状。破布木属的花粉特征表明,该属花粉在紫草科中既是独特的分类群,又是比较原始的属种。