Induced cytomictic variations and syncyte formation during microsporogenesis in <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.

The intercellular translocation of chromatin material along with other cytoplasmic contents among the proximate meiocytes lying in close contact with each other commonly referred as cytomixis was reported during microsporogenesis in Phaseolus vulgaris L., a member of the family Fabaceae. The phenomenon of cytomixis was observed at three administered doses of gamma rays viz. 100, 200, and 300 Gy respectively in the diploid plants of Phaseolus vulgaris L. The gamma rays irradiated plants showed the characteristic feature of inter-meiocyte chromatin/chromosomes transmigration through various means such as channel formation, beak formation or by direct adhesion between the PMC’s (Pollen mother cells). The present study also reports the first instance of syncyte formation induced via cytomictic transmigration in Phaseolus vulgaris L. Though the frequency of syncyte formation was rather low yet these could play a significant role in plant evolution. It is speculated that syncyte enhances the ploidy level of plants by forming 2n gametes and may lead to the production of polyploid plants. The phenomenon of cytomixis shows a gradual inclination along with the increasing treatment doses of gamma rays. The preponderance of cytomixis was more frequent during meiosis I as compared to meiosis II. An interesting feature noticed during the present study was the channel formation among the microspores and fusion among the tetrads due to cell wall dissolution. The impact of this phenomenon is also visible on the development of post-meiotic products. The formation of heterosized pollen grains; a deviation from the normal pollen grains has also been reported. The production of gametes with unbalanced chromosomes is of utmost importance and should be given more attention in future studies as they possess the capability of inducing variations at the genomic level and can be further utilized in the improvement of germplasm.     

Flow Cytometric Determination of Relative Nuclear DNA Contents in Bicellulate and Tricellulate Pollen

Nuclear DNA content in mature pollen was measured with a flowcytometer Pollen of Lilium longiflorum, Dendranthema grandiflora(syn Chrysanthemum monfolium) and Zea mays was chopped and stainedwith the DNA fluorochrome DAPI DNA levels, expressed as arbitraryC values, were compared with those of nuclei isolated from leafor root material of the same plants In mature tricellulate pollen the generative cell is dividedafter second pollen mitosis into two sperm cells Tricellulatepollen from maize and chrysanthemum gave rise to one large 1Cpeak and, only in the case of chrysanthemum, a much smallerone at the 2C level These results suggest that the haploid nucleiof the vegetative as well as both sperm cells in tricellulatepollen are arrested in the G₁ stage of nuclear division Thesmall 2C peak in the case of chrysanthemum probably arose froma fraction of pollen with the sporophytic chromosome number(2n pollen) In contrast to this, mature bicellulate lily pollengave rise to two identical peaks at the 1C and the 2C levelFrom this result it was concluded that in bicellulate pollen,the 1C peak is caused by the signal of the haploid vegetativenucleus arrested in the G₁ stage of nuclear division, whereasthe 2C peak originates from the haploid generative nucleus whichhas already undergone DNA synthesis and is arrested in G₂ Lilium longiflorumThunb, lily, Dendranthema grandiflora Tzelev (syn Chrysanthemum morifolium Ramat ), chrysanthemum, Zea maysL, maize, male gametophytic cells, vegetative cells, generative cells, sperm cells, unreduced pollen, sporophytic cells, relative nuclear DNA contents, replication stage     

The involvement of ethylene in in vitro maturation of mid-binucleate pollen of Nicotiana tabacum

The role of ethylene during in vitro maturation of Nicotianatabacum pollen from the mld-binucleate (MB) stage was analysedby the addition of aminooxyacetic acid (AOA), aminoethoxyvinylglycine(AVG), CoCl₂ and AgNO₃ to the maturation medium (AMGLu). Anincrease in ethylene production was obtained in both isolatedpollen and pollen surrounded by sporophytic tissue during insitu maturation. in vitro maturation of pollen was inhibitedby AOA and AVG; ACC and ethrel were able to overcome this inhibitoryeffect. Cyclohexylamine (CHA) reverted the inhibition provokedby both Ag⁺ and Co²⁺ The results reported in this paper indicatethat ethylene is one of the factors implicated in in vitro maturationof MB pollen of Nicotiana tabacum. Key words: Nicotiana tabacum, maturation, germination, pollen, ethylene     

RNA and Protein Synthesis in Germinating Pine Pollen

The results of autoradiographic experiments demonstrate that,as with the pollen of most other species, both the generativeand vegetative nuclei of Loblolly Pine (Pinus taeda) activelyengage in RNA synthesis from the very early stages of pollengermination. Unlike most other species, however, this newlysynthesized RNA includes rRNA. Evidence is provided for theimportance of this newly synthesized RNA in the process of continuedpollen tube growth. One and two-dimensional gel electrophoretic analysis revealsa number of both qualitative and quantitative differences amongthe proteins synthesized during the early stages of germinationand the later stages of pollen tube growth. One of the mostnotable of these is a 36 kD protein, the synthesis of whichpredominates during the later stages of pollen germination.A similar pattern of 36 kD protein synthesis is observed whenmRNA extracted from pollen at each of these stages is translatedin vitro. Key words: Pinus, pollen tube growth     

Gynodioecy in Cirsium chikushiense Koidz. (Compositae)

A study of the morphology and function of flowers in Cirsiumchikushiense revealed that the species was distinctly gynodioecious.Self-incompatible hermaphrodite florets produced both seedsand pollen grains, while female ones produced seeds but no pollengrains at all. The degenerated stamens of females were not onlysmaller but also sometimes occurred at a lower position insidethe floral tube than in hermaphrodites. The stigmata of femalesoften developed more fully than those of hermaphrodites. Thefrequencies of female plants in natural populations varied from15·5 to 50%. Almost all the pollinators stayed on bothfemale and hermaphrodite heads only to collect nectar. The femaleplants of this species may be more specialized in their genderby saving the cost of not only pollen grains but also stamens,and may be maintained by sufficient pollinators in natural populations.This gynodioecy may provide an example of nuclear-cytoplasmicmale sterility.Copyright 1994, 1999 Academic Press Sex expression, gynodioecy, Cirsium chikushiense, Compositae, male sterility, degenerated stamen, female frequency     

Consistent mixing of near and distant resources in foraging bouts by the solitary mason bee Osmia lignaria

Female bees are usually confronted with a choice among severalflower species that differ in their location and abundance withinthe community, and in the efficiency with which their pollenand nectar can be harvested. We investigated the effects ofdistance and flower density of two flower species on pollencollection by providing nest locations for the mason bee Osmialignaria in natural settings. Distance weakly affected pollenuse; on average, bees nesting near a flower species tended tocollect more of its pollen than did bees nesting at a greaterdistance. Flower density did not predictably impact pollen use,and use did not track changes in density during the season.Bees consistently mixed pollen from more distant species, despitesubstantial added foraging costs, and also mixed when one specieswas an order of magnitude less abundant than the other. Beesrequire nectar as well as pollen to feed their offspring, andour preliminary data suggest that the efficiencies of pollenand nectar collection are inversely related between the twoflower species, which would favor visitation to both species.Bees appear to collect some pollen from the low-pollen, high-nectarplant while visiting it for nectar. Thus, a nectar-collectingconstraint may favor collecting pollen from mixtures of species.     

Range of Hevea brasiliensis Pollen Dispersal Estimated by Esterase Isozyme Markers

A study was carried out to estimate the distance Hevea brasiliensispollen could be dispersed under natural conditions. Seeds werecollected at varying distances from the boundary between twoadjacent fields that were each planted with a pure stand ofa genetic clone. Esterase isozyme markers were used to determineif the seeds had been derived from self- or cross-pollination.The incidence of cross-pollination was then examined in relationto the distance from the inter-clonal boundary. A logarithmicmodel gave the best fit (r²=0.864) and suggested that pollencould travel distances in the order of 0.3 to 1.1 km. Copyright1999 Annals of Botany Company Hevea brasiliensis (rubber tree), pollen dispersal, cross-pollination, self-pollination, esterase isozymes.     

Vibratory Collection of Actinidia deliciosa (Kiwifruit) Pollen

Dry, powdery pollen grains were expected from 'buzz pollinated'flower species. However, vibration of Actinidia deliciosa (kiwifruit)anthers (a buzz pollinated species) by a mechanical shaker,at similar vibrations to Bombus terrestris caused clumps ofpollen joined by small droplets of tapetal fluid to be ejected.Pollen that was largely covered with tapetal fluid could notbe removed by vibration, whereas dehydrated pollen was easilyremoved, even without vibration. The late desiccation of A.deliciosa anthers after anthesis meant that pollen removed byvibration depended on the anther maturity. The presence of thedroplets gives insects which vibrate while foraging advantagesover insects which do not. A vibration attachment on a commercialpollen harvester increased the weight of pollen collected by57% over the whole day; 91% in the afternoon.Copyright 1994,1999 Academic Press Actinidia deliciosa, Bombus spp., bumblebees, buzz pollinate, kiwifruit, pollen collection, tapetal fluid, vibration     

Exploring the 'Most Effective Pollinator Principle' with Complex Flowers: Bumblebees and Ipomopsis aggregata

The ‘most effective pollinator principle’ impliesthat floral characteristics often reflect adaptation to thepollinator that transfers the most pollen, through a combinationof high rate of visitation to flowers and effective depositionof pollen during each visit. We looked for the expected positivecorrelation between quantity and quality of visits in Ipomopsisaggregata, whose red, tubular flowers are considered to be adaptedto hummingbirds. Hummingbirds were indeed the most common floralvisitors in 5 years of observation. However, long-tongued bumblebeesdeposited on average three-times as much outcross pollen pervisit to virgin flowers, and elicited four-times as much seedproduction, as did hummingbirds. Hence visitors that are relativelyinfrequent, and unexpected given the ‘pollination syndrome’of the plant, can be surprisingly good pollinators. One interpretationof this observation is that natural selection favours a specializedfloral morphology that excludes all but a single type of visitor,but that there are constraints on achieving this outcome. Analternative is that selection favours some degree of floralgeneralization, but that flowers can retain features that adaptthem to a particular type of pollinator in spite of this generalization.Copyright 2001 Annals of Botany Company Bumblebees, floral evolution, hummingbirds, Ipomopsis aggregata, pollination syndromes, pollinator effectiveness, scarlet gilia, seed set, specialization, temporal variation     

Souroubea guianensis Aubl.: Quest for its Legitimate Pollinator and the First Record of Tapetal Oil in the Marcgraviaceae

The floral biology of Souroubea guianensis Aubl. was studiedat Dois Irmãos forest, a remnant of Atlantic forest inRecife, Pernambuco state, northeastern Brazil. The flowers ofS. guianensis have attributes which are not easily related toa particular pollination syndrome. They also have some uncommoncharacteristics, among them the presentation of pollen immersedin an oily fluid. The occurrence of this kind of pollen presentationin species of Marcgraviaceae is described here for the firsttime. The term tapetal oil is proposed for this unusual fluid.The flowers are protandrous, self-incompatible, last for 6 to7 d, and fruit-set under natural conditions is low. Sugar concentrationsin the nectar vary from 27.6 to 58.8%. After 105 h of observationfor pollinators in 3 different years, no efficient pollinatorwas found. Fragmentation of natural habitats could be one ofthe causes for the local loss of the legitimate pollinator ofSouroubeaguianensis . Copyright 2000 Annals of Botany Company Marcgraviaceae, Souroubea, pollination, tapetal oil, pollen presentation, protandry, psychophily, ornithophily, Atlantic forest, fragmentation, northeastern Brazil     

Pollen Dimorphism--Origin and Significance in Pollen Plant Formation by Anther Culture

Pollen dimorphism during the ripening of Nicotiana tabacum antherstakes the form of differentiation at the binucleate pollen stageinto normal (N) grains, characterized by their high frequency,larger size, densely–staining cytoplasm and high starchcontent and into smaller (S) grains characterized by their variableand low frequency and weakly–staining cytoplasm. Mostof the S grains show distinctive vegetative and generative nuclei(A grains); a small number have two vegetative–type nuclei(B grains). Evidence is presented that when excised anthersare cultured, pollen plants arise only from S grains. It issuggested that the differentiation into N and S grains arisesby an abnormal second meiotic division in the pollen mothercells. Nicotiana tabacum, tobacco, pollen dimorphism, anther culture     

Studies on Microspore Development in Liliaceous Plants III. Pollen Tube Development in Lily Pollens Cultured from the Uninucleate Microspore Stage

Uninucleate microspores of Lilium longiflorum isolated at theGi phase of the cell cycle were cultured on a solid nutrientmedium for 12 days. The typical pollens produced in the culturegerminated on a germination medium and developed a structureresembling a pollen tube. The elongating pollen tubes occasionallycontained dividing generative nuclei or two sperm nuclei. (Received September 29, 1980; Accepted November 25, 1980)     

Involvement of Polypyrimidine Tract-Binding Protein (PTB)-Related Proteins in Pollen Germination in Arabidopsis

The pollen grains of most angiosperms contain stores of RNAsand their translation products required for pollen germinationand subsequent early elongation of pollen tubes. Polypyrimidinetract-binding protein (PTB), which is involved in the regulationof pre-mRNA alternative splicing, internal ribosomal entry site(IRES)-mediated translation and mRNA localization/sorting, isknown to act as a bridging molecule between RNAs and a varietyof cellular factors to fulfill cellular functions in both thenucleus and cytoplasm. Moreover, it has been reported that PTBplays roles in the differentiation and development of animalcells and tissues. In the Arabidopsis genome, there are twoPTB-related genes, tentatively termed AtPTB1 and AtPTB2. Inthe present study, the physiological functions of AtPTBs wereinvestigated using genetic and cytological approaches. The AtPTBpromoter was highly active in vegetative cells of mature pollengrains, and AtPTB was localized in the nucleus and cytoplasmof these vegetative cells. Mutations in the AtPTB genes resultedin decreased germination efficiency, and this effect was rescuedby introduction of the AtPTB2 promoter::AtPTB2–GFP. Takentogether, these findings suggest that AtPTB is involved in pollengermination through possible RNA metabolism processes in late-maturingand mature pollen grains.     

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     

The Reproductive Biology of Totara (Podocarpus totara) (Podocarpaceae)

A reproductive cycle of totara (Podocarpus totara) in New Zealandis complete within 2 years. After strobilus initiation in September,there is a 9 month period of winter dormancy until emergenceduring the growth flush in July–August of the followingyear. Female strobili bear only one or two ovules which arepollinated mid-October to mid-November at the megaspore tetradstage. Pollen germination and fertilization occur rapidly duringDecember. The pollen tube carries the body cell, sterile andtube nuclei with at least three prothallial nuclei, and branchesout after reaching the archegonia. The four–six archegoniacontain egg nuclei with no distinct perinuclear zone and a largechalazal vacuole. Fertilization by the larger of the two unequalmale gametes is accompanied by a degradation of egg cell cytoplasm.The three-tier proembryo contains a binucleate embryonal tierof only one cell. Soon after embryo penetration into the megagametophyte,the binucleate embryonal-tier cell undergoes mitosis and cellwall formation resulting in four uninucleate cells. Some ovulescontained secondary embryos thought to be the product of suspensorcleavage. Embryo maturation occurs by February.Copyright 1999Annals of Botany Company Podocarpus totara, totara, conifer, podocarp, reproductive biology, embryology.     

Generative Cell Division and Sperm Cell Association in the Pollen Grain of Sambucus nigra

Generative cell division in tricellular pollen grains of Sambucusnigra L. (Caprifoliaceae) has been examined with light and electronmicroscopy. During division the generative cell is located inthe centre of the pollen grain, near to the nucleus of a surroundingvegetative cell. Conventional mitosis of the generative cellis followed by cytokinesis through centrifugal cell plate formation.Two sister sperm cells remain in spatial contact with each otherand are surrounded, as formerly their progenitor cell was, bythe vegetative cell. From the changes of shape of the generativecell during division and of the sperm cells it may be assumedthat the space between these cells and the vegetative one containsa labile, non-rigid, wall material. No plastids have been observedin the generative cell and its mitochondria appear to be unequallydistributed between the two future sperm cells during division. Sambucus nigra L., generative cell division, pollen, sperm cell association     

Anther Culture of Lolium temulentum, Festuca pratensis and Lolium x Festuca Hybrids. II. Anther and Pollen Development In Vivo and In Vitro

The various pathways of pollen development were investigatedin cultured anthers of Lolium temulentum, Festuca pratensisand the L. multiflorum x F. pratensis hybrid ‘Elmet’.In all three, development from the vegetative cell was the predominantpathway of pollen callus development. However, there were characteristicdifferences in the behaviour of the generative cell. In L. temulentumit remained attached to the pollen wall and degenerated, whereasin F. pratensis it divided. In ‘Elmet’ it detachedfrom the pollen wall and remained undivided. Both polarizedand unpolarized partitioned calluses were observed. Developmentof the fusion product of the vegetative and generative nucleiwere also observed in anthers of L. temulentum. Anomalous grainswere not found to be major source of pollen calluses. Sections of anthers of L. temulentum were used to investigatethe origin of S pollen grains, the small pale-staining grainswhich denote pollen dimorphism. Such grains form out of contactwith the tapetum and are therefore determined before or duringmeiosis (i.e. before harvest of anthers for culture). Sectionswere also used to demonstrate the influence of the durationof pretreatment on the development of the middle layer of theanther wall. Festuca pratensis, Lolium temulentum, Lolium x Festuca, anther culture, haploid, microspore, pollen     

Anatomy of Watermelon Embryo Sacs Following Pollination, Non-pollination or Parthenocarpic Induction of Fruit Development

There is little information on the fate of embryo sacs in plantovules if pollination is prevented. In this study embryo sacsfrom watermelon were observed over a 13 day period followingflowering with (a) normal pollination, (b) non-pollination and(c) induction of parthenocarpic fruit development with naphthaleneacetic acid. Following pollination, and prior to fertilizationapproximately 2 days later, the embryo sacs completed developmentand consisted of two synergids with prominent filiform apparatus,an egg cell, a central cell with two polar nuclei and threeantipodal cells. Sperm nuclei were observed within the embryosac at 2 days and by 4 days the endosperm was proliferating.In the non-pollination treatment the embryo sac was still intactafter 4 days although the antipodal nuclei were becoming hardto distinguish. By 7 days only the two synergids and the eggcell were still well defined, the polar nuclei appeared in somepreparations to be fused, and the antipodals had degenerated.By 10 days the embryo sac was a structure-less watery mass.In parthenocarpic fruit the fate of the embryo sac was similarto that in non-pollinated fruit except that final breakdownwas delayed past 10 days. Maturity of the majority of embryo sacs in an ovary appearedto be contemporaneous with penetration of the pollen tube, andon the basis of the anatomical results it seems possible thatembryo sacs could be fertilized up to 2 days beyond the normaltime. Citrullus lanatus, watermelon, embryo sac, anatomy, pollination, parthenocarpy