Impairment in reproductive development is a major factor limiting yield of black gram under zinc deficiency

Black gram [Vigna mungo (L.) Hepper] cv. IPU 94 plants grown in sand culture with deficient zinc (0.1 μM Zn) nutrition and those deprived of normal (1 μM) Zn supply at the initiation of flowering, showed decrease in dry matter production and especially seed yield. These plants showed a decrease in the size of anthers and stigmatic heads, pollen producing capacity of the anthers and stigmatic exudations. Zn deficiency caused structural alterations in exine and retarded germination of pollen grains and tube growth. The pollen extracts and stigmatic exudates of the Zn-deficient plants showed increase in activity of acid phosphatase isoforms and inhibition of esterase isoforms. Zn deficiency led to decrease in number of pods, seeds per pod and seed mass, altered seed coat topography and reduced seeds germinability. Low seed yield under Zn deficiency is attributed to a role of Zn in pollen function, as also in pollen-pistil interaction conducive to fertilization and development of seeds.     

The effects of pollen load size and donor diversity on pollen performance, selective abortion, and progeny vigor in Mirabilis jalapa

The influence of pollen competitive environment on pollen performance (pollen germination, stigmatic penetration, and pollen tube growth rate), the maturation or abortion of initiated fruit, seed size, and seedling vigor was explored by manipulating the size and diversity of stigmatic pollen loads on Mirabilis jalapa. All aspects of pollen performance significantly increased with the number of pollen grains on a stigma or pollen tubes in a style, but was not influenced by the diversity of pollen donors. Plants tended to mature single-ovulate fruits that came from flowers where pollen load size and diversity were greatest and aborted those where these were lowest. No plants from seeds resulting from pollinations with a single pollen grain survived, but other fitness measures were mostly determined by maternal plant. The data suggest that pollen performance is influenced by pollen competitive environment, and both the genetic diversity of the pollen load and number of competing pollen tubes are important determinants of seed/fruit abortion.     

Stigmatic fluid aids self-pollination in Roscoea debilis (Zingiberaceae): a new delayed selfing mechanism

Background and Aims

Delayed selfing is the predominant mode of autonomous self-pollination in flowering plants. However, few delayed selfing mechanisms have been documented. This research aims to explore a new delayed selfing mechanism induced by stigmatic fluid in Roscoea debilis, a small perennial ginger.

Methods

Floral biology and flower visitors were surveyed. The capacity of autonomous selfing was evaluated by pollinator exclusion. The timing of autonomous selfing was estimated by emasculation at different flowering stages. The number of seeds produced from insect-pollination was assessed by emasculation and exposure to pollinators in the natural population. The breeding system was also tested by pollination manipulations.

Key Results

Autonomous self-pollination occurred after flowers wilted. The stigmatic fluid formed a globule on the stigma on the third day of flowering. The enlarged globule seeped into the nearby pollen grains on the fourth flowering day, thus inducing pollen germination. Pollen tubes then elongated and penetrated the stigma. Hand-selfed flowers produced as many seeds as hand-crossed flowers. There was no significant difference in seed production between pollinator-excluded flowers and hand-selfed flowers. When emasculated flowers were exposed to pollinators, they produced significantly fewer seeds than intact flowers. Visits by effective pollinators were rare.

Conclusions

This study describes a new form of delayed autonomous self-pollination. As the predominant mechanism of sexual reproduction in R. debilis, delayed self-pollination ensures reproduction when pollinators are scarce.     

Pollen morphology and germination inNarcissus

Pollen grains of several species and varieties ofNarcissus were examined with a light microscope and a scanning electron microscope. Normal pollen grains were kidney- or spindle-shaped with a germination furrow and a reticulate structure similar to that of the pollen grains ofAmaryllis. Pollen grains germinated within 2 to 3 hr. Percentage of germination was dependent upon temperature and treatment. Pollen tubes grew in length up to 1,000 μm and branched occasionally or behaved in strange fashion. Fresh pollen grains germinated more in distilled water at lower temperature than in sucrose-aqueous medium. Both in the presence and in the absence of stigmatic exudate calcium increased the percentage of germination. Gibberellic acid, abscisic acid and coumarin inhibited the pollen germination. Plasmoptysis occurred in all species and in all media tested except in a medium containing coumarin without stigmatic exudates. Plasmoptysis did not seem to be induced by hypotonic medium alone. Pollen of high germination capacity showed a high percentage of plasmoptysis. Based on the results obtained, evolution and sterility of theNarcissus plant was discussed.     

Transmitting tissue ECM distribution and composition, and pollen germinability in Sarcandra glabra and Chloranthus japonicus (Chloranthaceae)

BACKGROUND: and Aims Free-flowing surface exudates at the stigmatic (wet versus dry stigma) and adaxial epidermis at the site of angiospermy in carpels of Chloranthaceous species have been proposed to comprise a continuous extracellular matrix (ECM) operating in pollen tube transmission to the ovary. The aim of this research was to establish the spatial distribution and histo/immunochemical composition of the ECM involved in pollen tube growth in Sarcandra glabra and Chloranthus japonicus (Chloranthaceae). METHODS: Following confirmation of the pollen tube pathway, the histo/immunochemical make-up of the ECM was determined with histochemistry on fresh tissue to detect cuticle, esterase, proteins, pectins, and lipids and immunolocalization at the level of the TEM on sections from cryofixed/freeze-substituted tissue to detect molecules recognized by antibodies to homogalacturonans (JIM7, 5), arabinogalactan-proteins (JIM13) and cysteine-rich adhesion (SCA). KEY RESULTS: Pollen germinability is low in both species. When grains germinate, they do so on an ECM comprised of an esterase-positive cuticle proper (dry versus wet stigma). Pollen tubes do not track the surface ECM of stigma or adaxial epidermal cells at the site of angiospermy. Instead, tubes grow between stigmatic cells and subsequently along the inner tangential walls of the stigmatic and adaxial carpel cells at the site of angiospermy. Pollen tubes enter the ovary locule at the base of the funiculus. The stigmatic ECM is distinct by virtue of the presence of anti-JIM5 aggregates, lipids, and a protein recognized by anti-SCA. CONCLUSIONS: The Chloranthaceae joins a growing number of basal angiosperm taxa whereby pollen tubes germinate on a dry versus wet stigma to subsequently grow intercellularly en route to the ovary thereby challenging traditional views that the archetype pollen tube pathway was composed of the surface of stigma and adaxial epidermal cells covered with a free-flowing exudate.     

Regulation of seed number and female incitation of mate competition by a pH-dependent proteinaceous inhibitor of pollen grain germination in Leucaena leucocephala

Summary Leucaena leucocephala generally produces pods with more than 7–9 seeds. This is regulated by the stigmatic inhibition of pollen grain germination when the pollen grains are less than a critical number in the stigma. This number-dependent inhibition of pollen grain germination is effected by a pH-dependent proteinaceous inhibitor active at the stigmatic pH. Only when the pollen grains in the stigma exceed the critical number, they inactivate the inhibitor by collectively raising the stigmatic pH and thus overcoming the inhibition. The adaptive significance of such pre-fertilization mechanism for the female in inciting mate competition among the pollen grains is discussed. The evolution of en masse pollen grain dispersal units is explained as a sexual selection strategy by males in response to such stigmatic inhibition by females.     

Ultrastructural Studies on the Pollen-stigma Interaction of Intersectional Hybridization in Populus

The ultrastructural changes of pollen-stigma interaction were studied under artificial pollination in intersectional crosses of Populus lasiocarpa Oliv. × P. nigra L., P. lasiocarpa Oliv. × P. simonii Cart., P. lasiocarpa Oliv. × P. adenopoda Maxim. and P. lasiocarpa Oliv. X P. lasiocarpa Oliv. The results are as follows. Before pollination, the stigmatic cells of P. lasiocarpa containing large amounts of endo- plasmic reticulum, abund ant dictyosomes and well-developed mitochondria appear physiologi- cally active. Its outer wails are covered by a layer of cuticle with numerous branching channels toward the outer space. When the pollens of P. lasiocarpa and P. nigra are dusted, respectively, to the stigma of P. lasiocarpa, the degeneration of the stigmatic cells are rather slow. The cuticle of the stigma surface just adjacent to the alighting pollen is dissolved, but the cell contents still have large amounts of endoplasmic reticulum and the structure of the membrane system remains distinctly. When the pollens of P. simonii and P. adenopoda are forced, respectively, upon the stigma of P. lasiocarpa, the stigmatic cells degenerate rapidly. In addition to the disruption of the cuticle, heavy callose is deposited on the walls of the stigma cells adjoining the dropping pollens. Numerous vesicles are formed by the rough endoplasmic reticulum, and the electron density is increasing. The structure of the membrane system disappears gradually and becomes indistinct. Osmiophilic globuli occurs in the central vacuole. Plastids accumulate starch grains. The behavior of pollen tubes of P. nigra is similar to that of P. lasiocarpa. The pollen grains germinate rapidly and the pollen tubes penetrate into the stigma after the short-tube patten. Abnormal behavior is rare. A lot of minute exudates appear on the pollen walls. As for P. lasiocarpa × P. simonii and P. lasiocarpa × P. adenopoda, only half of the tubes can penetrate into the stigma after short-tube patten and is associated with callose deposition on the walls. The other half arresting on the stigma surface shortly after germination shows abnormal behavior; such as swelling, creeping, bursting, twisting and distortion with callose deposition on the tip of the pollen tubes. As a result of our experiments, It is concluded that the degeneration speed of the stigmatic cells may be a sign of incompatibility and the crossability in progamic phase may arrange as follows: P. lasiocarpa × P. nigra, P. lasiocarpa × P. simonii, P. lasiocarpa × P. adenopoda. The first one is compatible while the second and the third are incompatible.     

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     

Directional Guidance of Nicotiana alata Pollen Tubes in Vitro and on the Stigma

Pollen tubes navigate the route from stigma to ovule with great accuracy, but the cues that guide them along this route are not known. We reproduced the environment on the stigma of Nicotiana alata by immersing pollen in stigma exudate or oil close to an interface with an aqueous medium. The growth of pollen in this culture system mimicked growth on stigmas: pollen grains hydrated and germinated, and pollen tubes grew toward the aqueous medium. The rate-limiting step in pollen germination was the movement of water through the surrounding exudate or oil. By elimination of other potential guidance cues, we conclude that the directional supply of water probably determined the axis of polarity of pollen tubes and resulted in growth toward the interface. We propose that a gradient of water in exudate is a guidance cue for pollen tubes on the stigma and that the composition of the exudate must be such that it is permeable enough for pollen hydration to occur but not so permeable that the supply of water becomes nondirectional. Pollen tube penetration of the stigma may be the most frequently occurring hydrotropic response of higher plants.     

龙须草花粉活性及结实率与其无融合生殖的关系

以两种生态型无融合生殖龙须草为材料,经去雄套袋、去掉柱头及相互杂交等处理,比较几种方式下的结实率。结果表明：去雄套袋、截去柱头以破坏授粉和杂交授粉等处理与去雄不套袋作对照之间结实率的差异不显著。两种生态型龙须草的成熟花粉粒都具有活力,在蔗糖溶液中的离体萌发率分别为28％和34％;花粉在柱头上的附着和生长的荧光观察表明：花粉附着率较低,花粉在柱头上可以萌发,但不能进入柱头和花柱。证明龙须草花粉对其结实率没有显著影响;龙须草具有很高的无融合生殖度,是优良的无融合生殖基因供体。     

Pistil anatomy and pollen tube development in Polygala vayredae Costa (Polygalaceae)

Low seed ovule ratios have been observed in natural populations of Polygala vayredae Costa, a narrowly endemic species from the oriental pre-Pyrenees. To evaluate physical and nutritional constraints and pollen tube attrition in this endemic species, stigma and style anatomy, as well as pollen tube development along the pistil were investigated using light and fluorescence microscopy. The structural morphology of the stigmatic region was also examined with scanning electron microscopy. Pollen grains that reached the stigmatic papillae came into contact with a lipid-rich exudate and germinated easily. Although a large number of pollen grains reach the stigmatic papillae, few pollen tubes were able to grow into the style towards the ovary. The style was hollow, with the stylar channel beginning a few cells below the stigmatic papillae. Initially, the stylar channel area was small compared to other levels of the style, and was surrounded by lipid-rich, highly metabolic active cells. Furthermore, lipid-rich mucilage was detected inside the stylar channel. At subsequent style levels towards the ovary, no major reserves were detected histochemically. The reduced intercellular spaces below the stigmatic papillae and the reduced area of the stylar channel at its commencement are suggested to physically constrain the number of pollen tubes that can develop. In subsequent levels of the style, the stylar channel could physically support a larger number of pollen tubes, but the lack of nutritional reserves cannot be disregarded as a cause of pollen tube attrition. Finally, the number of pollen tubes entering the ovary was greater than the number of ovules, suggesting that interactions occurring at this level play a major role in the final reproductive outcome in this species.     

Role of phenolics and boron in reproductive success in seasonally transient sterile Tecoma stans L

Quantitative and qualitative analysis of phenolics and boron in stigma of transient sterile Tecoma stans L. during seedless (May-July), partially seedbearing (August-November, April) and seedbearing periods (December-March) was made. UV absorption profile of stigmatic exudates indicated the presence of simple phenolics. Total phenolics were higher in stigma during seedless period. Thin layer chromatographic analysis of stigmatic extracts exhibited only three principal spots. Mass spectrophotometry showed the presence of derivatives of cinnamic acid, namely, caffeic acid in these spots. Quantity of boron in stigma during seedless period was lowest but the difference with other periods was not significant. It was suggested that the accumulation of higher quantity of caffeic acid in the stigma during seedless period due to high temperature (40 degrees-45 degrees C) could lead to inhibition of pollen germination in vivo, thereby rendering the plants seedless. This was confirmed by inhibition of in vitro pollen germination in the basal medium containing higher quantity of caffeic acid.     

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.     

Stigma development and receptivity of two Kalanchoë blossfeldiana cultivars

Several members of the Kalanchoë genus are popular as ornamental plants. Cross-breeding and wide hybridisation are essential to continuously introduce novel traits into cultivated plant material. This study aimed to identify the major factors related to the stigma affecting cross-pollination in the Kalanchoë blossfeldiana. Pollen tube growth after pollination of K. blossfeldiana ‘Jackie’ and ‘Reese’ was examined at different stigma developmental stages. Five distinct developmental stages were identified based on changes in morphology and activity of stigmatic peroxidase. After reciprocal pollination at the five stigma developmental stages, fluorescence microscopy was used to estimate the number of pollen tubes in situ. Both cultivars had receptive stigmas from stage I to IV, which concurred with the continuous expansion of the stigma covered with exudates. No pollen tube growth was observed at stage V for both cultivars. The number of pollen tubes was significantly higher in carpels pollinated at stage III, characterized by loose arrangement of the papillae and maximal amount of exudates, compared to all other developmental stages. Stigmas showing drying exudates and absence of peroxidase exhibited a relatively decreased number of pollen tubes in situ. No pollen tubes germinated on wilting stigmas. The arrangement of the papillae, the presence of exudates and peroxidase activity affected the number of pollen tubes in cross-pollination of K. blossfeldiana cultivars ‘Jackie’ and ‘Reese’. These results will help breeders to better select the optimal time for effective pollination. The findings may be applicable for other cultivars of K. blossfeldiana and relevant for different species of Kalanchoë.     

有花植物触敏柱头的闭合行为及其适应意义

有花植物触敏柱头的闭合行为一直是传粉生态学和进化生物学研究的热点问题之一。植物花的多裂片柱头在传粉者接触后迅速闭合的现象主要存在于玄参目的一些类群中。早在上个世纪初,植物学家就对花的柱头闭合现象进行过初步的研究,认为柱头闭合主要是由于花粉在柱头上萌发时吸收水分导致柱头细胞膨压降低而引起的;并对柱头闭合的适应性提出了一些假说。近年来,许多学者研究证实柱头闭合能促进花粉萌发及花粉管伸长,或减少花粉散出与柱头接受花粉间的相互干扰,提高传粉者访问过程中花粉的散播量,并且柱头的状态会直接影响传粉者的取食行为。到目前为止,对柱头闭合的机制及其避免自交的假说还存在许多争议。关于触敏柱头的闭合行为,尤其是其适应机制及其生态学意义还有待更加系统和深入的研究。本文概述了触敏柱头的研究进展,并对研究中尚存在的一些问题进行了讨论。     

Flowering and pollination patterns of Magnolia denudata with emphasis on anatomical changes in ovule and seed development

Magnolia denudata is an excellent ornamental and ecologically important tree that exhibits low fecundity because of seed abortion. To better understand the cause of this low sexual reproduction, pistil receptivity, and pollen quality were investigated using culture medium and histochemical staining, with emphasis on pollen viability at different flowering stages. Cross-pollination, geitonogamy, and self-pollination were undertaken on flowers of sample trees. We found that flowers produce a large number of pollen grains available for effective pollination. Pollen viability varied at different stages, but was highest during stage 3, in which the stamens detach from the axis, the anthers dehisce, and the gynoecium stigma starts to fade. The duration of pistil receptivity was approximately 5 h. These results indicate that poor fertilisation may occur if the optimum period for pollination is missed or the stigma receives poor-quality pollen grains from stages other than stage 3. Although dichogamy avoids selfing, it has led to considerable geitonogamy. Our anatomical analysis of ovule and seed development further revealed that fertilisation occurred in samples with geitonogamy, but that the embryo degenerated in the torpedo stage, demonstrating that aborted seeds were produced by fertilisation, rather than by pseudogamy. Our results provide new insights into the mechanism of reduced seed set under natural conditions.     

On factors influencing fruit-set and sterility in arecanut (Areca Catechu Linn.)

Summary (1) Histological studies on the stigmas of female flowers of areca indicate the possibility of the stigmatic surfaces being impacted upon by air spora. The presence of spores or hyphae together with pollen grains has been shown in stigmatic surface.(2) Air spora of arecanut plantation chiefly consist ofAspergillus, Penicillium, Actinomycetes, Mucor, Alternaria, Cladosporium andBacteria.(3) The types of fungi, bacteria and actinomycetes adhering to the stigma are isolated.(4) The metabolites of the fungi, bacteria and actinomycetes isolated from the stigmas of arecanut flowers inhibit,in vitro, considerably the germination of pollen grains and rate of growth of pollen tubes. In certain instances there is no germination of the pollen grains in the metabolites of fungi, bacteria and actinomycetes. In view of the results the significance of the air spora in affecting the normal course of pollination and fertilization is discussed.     

The functional significance of a stigma color polymorphism in Acer pictum subsp. mono (Aceraceae)

Aims Exploring the characteristics and function of a flower color polymorphism contributes to our understanding of floral evolution in angiosperms. However, little information is available on stigma color polymorphisms in flowering plants despite their important functional role in plant reproduction.Methods We studied a stigma color polymorphism at the individual level in Acer pictum subsp. mono (aka A. mono) by investigating stigma-color morph proportion and comparing stigma performance and components of female fitness (pollen adhesion, pollen germination, fruit set, seed set and fruit weight) between the two morphs (red and greenish-yellow stigma) within natural populations at Dongling Mountain in the north of China. In the flowering period, we conducted hand pollination in natural populations and then made microscopic observations using the aniline blue fluorescence method.Important findings Individuals in the studied populations did not change their stigma color between years, and flowers produced by a single tree were uniform in their stigma color. This strongly suggests that stigma color variation is genetically controlled. The percentage of the red stigma flowers with germinated pollen grains was significantly higher than that of the greenish-yellow stigma flowers when hand pollination was conducted in the early flowering period, but not so in the late flowering period. There was no significant difference in the percentage of flowers with pollen adhesion to the stigma between the two morphs. Fruit set of the red morph was significantly higher than that of the greenish-yellow morph. Our findings suggest that the red morph may be more advantageous for pollen germination and fruit set than the greenish-yellow morph, which may provide a functional explanation for the high incidence of the red stigma morph in the studied populations. Alternative explanations for the stigma color polymorphism are discussed to stimulate further work.     

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

Background and Aims

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

Methods

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

Key Results

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

Conclusions

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