Pollen-limited seed set in pseudogamous blackberries (Rubus L. subgen. Rubus)

Summary Pseudogamous blackberry species are polyploid and usually exhibit meiotic irregularities causing severe reduction in pollen viability. When species means were compared, relative seed set was strongly correlated with the number of good pollen grains produced per flower divided by the number of ovules (good-pollen/ovule ratio).Partial correlation analysis for percentage viable pollen and relative seed set, controlling for good-pollen/ovule ratio, revealed an almost significant relationship, presumably because meiotic irregularities have a similar effect on both pollen viability and the viability of meiotically derived embryo sacs. Seed germination, on the other hand, showed no relationship with relative seed set or with pollen viability.The automatic selfing capacity exhibited by most of these blackberry species does not appear to be related to either pollen production or relative seed set.     

Fertility and meiotic chromosome behaviour in autotetraploid pearl millet

Summary Tetraploidy was induced in outbred pearl millet and selection for high and low seed set was started in the C₁ generation. Segregation in the C₃ generation was observed for fertility and also for meiotic features: per cent seed set in selfed earhead, chiasma frequency, chromosome association and chromosome distribution in pollen mother cells were all affected. However, variation in seed set was observed even between samples not differing in meiotic features. It is apparent that factors regulating seed set in autotetraploid pearl millet were genic as well as chromosomal.A high frequency of univalents and trivalents was the main cause of sterility; quadrivalent misdisjunction was not a significant factor. As univalency decreased with increased chiasma formation, the gain was in the form of quadrivalents. However, individuals not differing in chiasma frequency did differ in chromosome association frequencies, indicating that the dependence of chromosome pairing behaviour on chiasmata was subject to genotypic influence.     

Impaired Male Meiosis due to Irregular Synapsis Coupled with Cytomixis in a New Diploid Cytotype of Dianthus angulatus (Caryophyllaceae) from Indian Cold Deserts

Dianthus angulatus (Caryophyllaceae) is cytologically examined here for the first time for the area of India. The diploid chromosome count of 2n?=?30, ascertained here, represents a new cytotype, supplementing the earlier report of a hexaploid cytotype with 2n?=?90 from outside of India. We report here the occurrence of two plants showing impaired meiosis due to irregular synapsis and cytomixis collected from Kinnaur district, Himachal Pradesh (India). The other plants of this species collected in Lahaul-Spiti region showed normal male meiosis (n?=?15) with a high (95%?100%) pollen fertility and normal seed set, and they reproduce sexually. Irregular synapsis in two plants from the Kinnaur region is characterized by the complete absence of chromosome pairing and the presence of 30 univalents at diakinesis and meta-anaphase. In addition, other meiotic irregularities were found, such as unoriented chromosomes, laggards, precocious movements of univalents at anaphase-I and micronuclei at telophase. Microsporogenesis was also abnormal, resulting in the formation of monads, dyads, triads, polyads, and tetrads with micronuclei. The occurrence of intra- and intermicrosporal chromatin material transfer during microsporogenesis was observed, which is a rather rarely observed phenomenon. The synaptic irregularities coupled with chromatin transfer in these plants seem to be responsible for the high pollen sterility (38%?42%) and heterogeneously sized pollen grains. In these plants no seeds were set, and plants reproduced vegetatively through root suckers.     

Pollination intensity and potential seed set in Passiflora vitifolia

Summary Initial seed set and fruit set were pollen-limited in a Costa Rican population of Passiflora vitifolia, a self-incompatible species with 200–350 ovules per flower. Pollination intensity was measured by counting the number of allogamous pollen grains on stigmas of the large one-day flowers. Hand-pollinations demonstrated that 25–50 pollen grains are required for fruit set, and >450 are needed for maximum seed set, with a pollen:seed ratio of about 1.6:1.0. Hummingbirds (Phaethornis superciliosus) delivered sufficient allogamous pollen for maximum seed set to only 28% of the flowers examined. Naturally pollinated flowers yielded fewer fruits and fewer seeds per fruit than those pollinated by hand. Most pollen transferred by humming-birds was self-incompatible; emasculated flowers yielded higher seed set than flowers with intact anthers. Visitation rates did not provide a good index of effective pollination.There were significant differences in ovule number, maximum seed set, and maximum per cent seed set among individual vines. More than half of an individual's flowers failed to set fruit, whether pollinated by birds or by hand. In this population, maximum reproductive potential may be limited by maternal resources for fruit development, but seed set varies with pollination intensity. Pollen-limited seed set may be a disadvantage of self-incompatibility, especially in species with many-seeded fruits.     

Self-pollen interference is absent in wild radish (Raphanus raphanistrum, Brassicaceae), a species with sporophytic self-incompatibility

Explaining the diversity of mating systems and floral forms in flowering plants is a long-standing concern of evolutionary biologists. One topic of interest is the conditions under which self-pollination can interfere with seed set for flowering plants with a self-incompatibility system. We investigated the effect of self-pollen interference for wild radish, Raphanus raphanistrum, which has sporophytic self-incompatibility. We performed pollinations and determined seed set for plants grown in the greenhouse, using pollen mixtures representing either self- with outcross-pollen or outcross-pollen alone. Stigmas were collected for a subset of pollinated flowers to determine the number of pollen grains applied. Average seed set for the self/cross (5.13 seeds/pollination) and cross treatments (5.09 seeds/pollination) did not differ significantly. Stigmatic pollen loads averaged around 700 grains, an amount close to observed natural pollen loads on R. raphanistrum. We concluded that for R. raphanistrum in natural populations, self-pollen is unlikely to interfere with outcross-pollen success. This study is the first to investigate effects of self-pollen interference on seed set for a homomorphic species with sporophytic self-incompatibility where rejection occurs at the stigmatic surface.     

Male sterility, protogyny, and pollen-pistil interference in Plantago maritima (Plantaginaceae), a wind-pollinated, self-incompatible perennial

Evolution and maintenance of male sterility in seed plants can be explained by the maternal inheritance of mitochondria, which encode the trait, and by adaptive functions that enhance female fecundity in male-sterile compared to hermaphrodite individuals. Protogyny and male sterility can independently decrease the negative effect of pollen-pistil interference in self-incompatible species. In Plantago maritima, which possesses both traits, protogyny increases seed set in hermaphrodite individuals. This is shown both by a significantly positive association between seed set and retarded dehiscence of the anthers and by a more than 50% reduction in seed set following self-pollination. Male sterility does not seem to increase seed set further, as female and hermaphrodite plants do not differ significantly in mean seed set per capsule. Bagging experiments demonstrate strong self-incompatibility in the study populations. Hence, in P. maritima male sterility seems neither to prevent selfing nor to reduce the effect of pollen-pistil interference. Females had significantly larger stigmas than hermaphrodites, but seed set varied negatively with stigma length among females, indicating that the evolution of unisexuality in P. maritima is not due to prefertilization sex allocation. I therefore conclude that the genetical system of nucleocytoplasmic determination of gender is the main cause for maintenance of male sterility in P. maritima.     

Specific features of meiosis in the Siberian Fir (<Emphasis Type="Italic">Abies sibirica</Emphasis>) in the forest Arboretum of the V. N. Sukachev Institute,Russia

Investigating the tolerance of plant reproductive systems to environmental changes has become a research priority under current climate change scenarios. Successful plant conservation requires knowledge of plant reproductive biology, particularly the meiotic characteristics of planted species. Meiosis, as part of microsporogenesis, is a critical plant developmental stage controlling future pollen quality. Meiosis in a Siberian fir (Abies sibirica) plantation, established in the Forest Arboretum of the Sukachev Institute, Russia, was studied from 2002 to 2004. The microsporogenesis pattern found for the Siberian fir appeared to be largely similar to that exhibited by other conifer species. Meiosis in the Siberian fir has the following characteristics: asynchrony, rapid progression of telophases I and II, and parallel and linear spindle arrangements at different meiosis II stages. General and specific meiosis irregularities were recorded at each stage. Some specific features of meiosis and the specific development of some irregularities were revealed. Pollen development analysis showed that irregular pollen grains made up less than 1% of all grains. The specific features of meiosis identified in fir trees growing in the Arboretum indicated low resistance of male reproductive structures to climatic changes and might account for high fir pollen sterility in this new environment.     

Apomixis versus sexuality in blackberries (Rubus subgen.Rubus,Rosaceae)

InRubus L. a connection seems to exist between the degree of meiotic disturbances on the one hand, and the production of unreduced embryo sacs, pollen fertility and relative seed set on the other hand. Severe meiotic disturbances commonly encountered in apomictic taxa decrease pollen fertility and thereby seed set since pollen is necessary for endosperm development. By contrast interspecific hybrids between apomictic taxa appear to be sexual and exhibit high pollen fertilities, probably due to an improved meiosis. Thus, apomixis leads to a decreased fertility inRubus, not the opposite, as often discussed.     

Cytotypes within the family Umbelliferae with special reference to the genera Seseli L. and Oenanthe L. (subtribe Seselinae)

Cytotypes within the genera Seseli L. and Oenanthe L. show variation in chromosome number in pollen mother cells. A correlation of low chiasma frequency with meiotic irregularities and pollen sterility was noted. In addition to the meiotic irregularities in the two genera occurrence of polyspory and triple pollen formation during meiosis have been observed in Seseli diffusum (Roxb.) Sant. Wagh. The presence of different chromosome numbers in different populations of the same species and karyological investigation in 4 species of Oenanthe L. suggests that structural changes of chromosomes associated with numerical differences have possibly played a role in their evolution. Diminution in chromosome size and complete absence of metacentric chromosomes in Oenanthe lachenalli Gmel. suggest that probably this species is more evolved than 3 species of Oenanthe (i.e., O. benghalensis Benth., O. pimpinelloides L. and O. thomsoni Clarke). The interrelationships of the two genera are discussed.     

Cyto-morphological studies op some species and hybrids in the Eu-Sorghums

Summary Important morphological features such as plant height, leaf size and number of leaves, shape of the panicle and sessile spikelets, staminate condition of the pedicellate florets, nature of lemma, colour of the stigmatic surface and seeds etc., were studied in 8 Sorghum species and 10 F₁ hybrids between them. Based on the data, interrelationship amongst the species are discussed.Pachytene pairing was complete and apparently normal, followed by regular meiosis at later stages resulting in high pollen stainability and good seed setting in all the parental species except the male sterile Kafir. Studies on the pairing properties of the differentially stained regions showed that synapsis starts from the proximal to the distal end and separation of the split chromosomes starts from the distal to the proximal.The interspecific hybrids studied are classified into four types based on pachytene pairing and pollen sterility. 1. normal pairing accompanied by high pollen fertility, 2. normal pairing accompanied by partial pollen sterility. 3. irregularities in the pachytene pairing followed by partial pollen sterility and 4. irregularities in the pachytene pairing accompanied by normal pollen fertility. Suitable explanations are advanced to explain the meiotic aberrations noted in some of the hybrids under study.Cytogenetical mechanisms underlying species differentiation in the Eu-Sorghum species are discussed.     

Male meiosis,morphometric analysis and natural propagation in the 2× and 3× cytotypes of Tordyliopsis brunonis (Apiaceae) from northwest Himalayas (India)

Tordyliopsis brunonis (Apiaceae) is cytologically investigated here for the first time from India. The chromosome count of 2n = 33, ascertained here, represents a new intraspecific triploid cytotype in the species, supplementing the earlier report of a diploid cytotype with 2n = 22 from Nepal Himalayas. The diploid chromosome count (n = 11) has also been found in some of the presently investigated individuals which showed perfectly normal meiosis with 100 % pollen fertility and normal seed set. However, the individuals with triploid chromosome count showed irregular meiotic behaviour and abnormal microsporogenesis resulting in high pollen sterility (56.26 %) and no seed set. The irregular meiotic behaviour in the triploid individuals is attributed to the occurrence of variable number of univalents (1–7) at diakinesis and metaphase-I. In the subsequent meiotic stages, these univalents lagged at anaphases and constituted micronuclei in sporads. The triploid plants were also observed for natural propagation and it was noticed that no seeds were set. These plants were noticed to propagate vegetatively by rootstocks. Chromosomal pairing in triploid cytotype is typical of an allopolyploid. Based on the characterization of chromosomal pairing during meiosis, we assumed that the triploid individuals are probably alloploid in nature. Hypotheses concerning the possible origin of allotriploid in T. brunonis are also discussed.     

Effects of pollen reward removal on fecundity in a self-incompatible hermaphrodite plant

The pollen of hermaphrodite plants is often utilised by flower-visiting animals. While pollen production has obvious benefits for plant male fitness, its consequences for plant female fitness, especially in self-incompatible hermaphrodite species, are less certain. Pollen production could either enhance seed production though increased pollinator attraction, or reduce it if ovules are discounted by deposition of self pollen, as can occur in species with late-acting self-incompatibility. To test the effects of pollen reward provision on female fitness, we artificially emasculated flowers in two populations of the succulent Aloe maculata (Asphodelaceae), which has a late-acting self-incompatibility system, over the course of its flowering period. Flowers of this species are visited by sunbirds (for nectar) and native bees (for pollen and nectar). We measured floral visitation rates, floral rejection rates, pollen deposition on stigmas and fruit and seed set in both emasculated and non-emasculated plants. We found that flowers of emasculated plants suffered reduced visitation and increased rejection (arrival without visitation) by bees, but not by sunbirds; had fewer pollen grains deposited on stigmas and showed an overall decrease in fruit set and seed set. Rates of seed abortion were, however, greatly reduced in emasculated flowers. This study shows that pollen rewards can be important for seed set, even in self-incompatible plants, which have been assumed to rely on nectar rewards for pollinator attraction. Seed abortion was, however, increased by pollen production, a result that highlights the complexity of selection on pollen production in hermaphrodite flowers.     

Persistent occurrence of meiotic abnormalities in a new hexaploid cytotype of <Emphasis Type="Italic">Thalictrum foetidum</Emphasis> from Indian cold deserts

Thalictrum foetidum L. (Ranunculaceae), a morphologically variable and widely distributed species of temperate and alpine Himalayas is worked out cytologically for the first time from India. Earlier studies from outside India were restricted to chromosome counts and karyotypic analysis. We studied the male meiosis, microsporogenesis and pollen viability in the wild accessions from the cold deserts of Lahaul-Spiti, Kinnaur and Pangi Valley of Himachal Pradesh. Present cytomorphological surveys in the species record the existence of two distinct morphotypes involving plant size; colour and size of leaf/leaflet; dentation of leaflet lobes; and degree of leaf pubescence. All the accessions in the two morphovariants share the same meiotic chromosome number (n = 21) and adds a new intraspecific hexaploid cytotype. The accessions show the phenomenon of cytomixis involving transfer of chromatin material among proximate pollen mother cells (PMCs) and associated meiotic abnormalities like, out of plate bivalents, interchromosomal connections, and laggards, bridges and micronuclei at anaphases/telophases. Microsporogenesis results into abnormal sporads (tetrads with micronuclei, dyads, triads and polyads). The products of such sporads resulted into some pollen sterility and pollen grains of heterogeneous sizes. The persistent occurrence of phenomenon of cytomixis and associated meiotic abnormalities and consequently pollen sterility and pollen grains of heterogeneous sizes in the hexaploid cytotype of T. foetidum seems to be under some genetic factors associated with the genome.     

MEGASPOROGENESIS AND GAMETOPHYTE SELECTION IN PAEONIA CALIFORNICA

Walters , James L. (U. California, Goleta.) Megasporogenesis and gametophyte selection in Paeonia californica. Amer. Jour. Bot. 49(7): 787–794. Illus. 1962.—In the ovules of Paeonia californica, a massive archesporium produces numerous (estimated at 30–40) megasporocytes, many of which complete meiosis. Several continue into gametophyte development, which is of the Polygonum type, and at the time of fertilization there are from 1 to 4 gametophytes per ovule. Rarely does more than 1 seedling per seed appear in germination. This species is characterized by extensive translocation heterozygosity, and other meiotic irregularities, in its natural populations. It shows a complete range from plants forming only pairs to those with all their chromosomes in rings at meiosis. The latter types have as high as 90% bad pollen. The course of events in the ovules is compared with the “Renner-effect” found in Oenothera. The multiple megasporocytes and subsequent events are seen as a mechanism which insures each ovule a high probability of containing a viable egg in spite of meiotic behavior which can produce 90% sterility, and thus insures high seed set in the translocation heterozygotes.     

Cytological investigation of male sterility in sorghum caused by a dominant mutation (<Emphasis Type="Italic">Ms</Emphasis><Subscript><Emphasis Type="Italic">tc</Emphasis></Subscript>) derived from tissue culture

Male sterility mutations are an important tool in the investigation of anther and pollen development and for obtaining hybrid seeds in plant breeding. Cytological analysis of microsporo- and microgameto-genesis in sorghum plants with dominant mutation of male sterility (Ms_tc) derived from tissue culture has been carried out. Using substitution backcrosses, this mutation was introduced first into the nuclear background of the fertile sorghum line SK-723 and from this line into Volzhskoe-4w (V-4w). The mechanism of Ms_tc action on anther and pollen development differed in different nuclear backgrounds. In SK-723, phenotypic expression of Ms_tc began before the beginning of meiosis, which resulted in degeneration of sporogenous tissue in some anthers and in significant disturbances of anther morphology. In microsporocytes that did not degenerate, the frequency of non-specific meiotic abnormalities characteristic of the fertile line SK-723 significantly increased. In addition, in the mutant plants, a number of specific meiotic abnormalities--almost complete desynapsis, and formation of syncytial structures--were observed, apparently the consequence of Ms_tc action. In mono- or bi-nucleate microspores, degenerative processes resulting in formation of empty or anomalously coloured pollen grains led to almost complete male sterility. In the V-4w nuclear background, changes in anther structure and meiotic disturbances were infrequent. The degenerative processes began at the uni- or binucleate microspore stage and resulted in formation of empty or abnormally coloured pollen grains, and in partial pollen sterility. Thus, the same nuclear male sterility-inducing mutation in different nuclear backgrounds affects different stages of pollen development.     

Impact of cytomixis on meiosis, pollen viability and pollen size in wild populations of Himalayan poppy (Meconopsis aculeata Royle)

We report the occurrence of cytomixis in wild populations of Himalayan poppy (Meconopsis aculeata Royle), which is considered to be an important and threatened medicinal plant growing in the high hills of the Himalayas. The impact of cytomixis on meiotic behaviour, reduced pollen viability and heterogeneous-sized pollen grains was also studied. Cytological studies in the seven wild populations from the high hills of Himachal Pradesh revealed that all the Himalayan populations exist uniformly at the tetraploid level (2n=56) on x=14. The phenomenon of chromatin transfer among the proximate pollen mother cells (PMCs) in six populations caused various meiotic abnormalities. Chromatin transfer also resulted in the formation of coenocytes, aneuploid, polyploid and anucleated PMCs. Among individuals that showed chromatin transfer, chromosome stickiness and interbivalent connections were frequently observed in some PMCs. The phenomenon of cytomixis in the species seems to be directly under genetic control; it affects the meiotic course considerably and results in reduced pollen viability.     

Microsporogenesis in the endangered species Cupressus dupreziana A. Camus: evidence for meiotic defects yielding unreduced and abortive pollen

To understand the reproductive biology of Cupressus dupreziana A. Camus (Cupressaceae), a highly endangered Mediterranean conifer, the processes of microsporogenesis and pollen differentiation were investigated cytologically. Pre-meiotic development proved to be similar to the coniferous pattern: the microsporangia differentiated sporogenous tissue in which microsporocytes separated and underwent meiosis. As the meiotic steps proceeded, unexpected irregularities were observed concerning chromosomal and nuclear behaviour. This mainly included: abnormal chromosome segregation and cytokinesis, and nuclear fusion of the meiotic products. The result was the formation, in the same microsporangium, of heterogeneous microspore populations arranged in monads, dyads, triads, tetrads, and polyads, and cytoplasts giving rise to pollen grains of different sizes. This indicates that in C. dupreziana both abortive and unreduced pollen grains are generated. The significance of the finding is discussed in relation to reproductive biology and vulnerability to extinction.     

Microevolution of S-allele frequencies in wild cherry populations: respective impacts of negative frequency dependent selection and genetic drift

Negative frequency dependent selection (NFDS) is supposed to be the main force controlling allele evolution at the gametophytic self-incompatibility locus (S-locus) in strictly outcrossing species. Genetic drift also influences S-allele evolution. In perennial sessile organisms, evolution of allelic frequencies over two generations is mainly shaped by individual fecundities and spatial processes. Using wild cherry populations between two successive generations, we tested whether S-alleles evolved following NFDS qualitative and quantitative predictions. We showed that allelic variation was negatively correlated with parental allelic frequency as expected under NFDS. However, NFDS predictions in finite population failed to predict more than half S-allele quantitative evolution. We developed a spatially explicit mating model that included the S-locus. We studied the effects of self-incompatibility and local drift within populations due to pollen dispersal in spatially distributed individuals, and variation in male fecundity on male mating success and allelic frequency evolution. Male mating success was negatively related to male allelic frequency as expected under NFDS. Spatial genetic structure combined with self-incompatibility resulted in higher effective pollen dispersal. Limited pollen dispersal in structured distributions of individuals and genotypes and unequal pollen production significantly contributed to S-allele frequency evolution by creating local drift effects strong enough to counteract the NFDS effect on some alleles.     

Density-dependent seed set in the Haleakala silversword: evidence for an Allee effect

Plant species may be subject to Allee effects if individuals experience a reduction in pollination services when populations are small or sparse. I examined temporal variation in reproductive success of the monocarpic Haleakala silversword (Argyroxiphium sandwicense subsp. macrocephalum) over five years, to determine if plants flowering out of synchrony with most of the population (i.e., in low flowering years) exhibited lower percent seed set than synchronously-flowering plants (i.e., those flowering in high flowering years). Through two pollination experiments conducted over multiple years, I also measured pollen limitation and self-incompatibility in this species. The number of flowering plants varied greatly among years, as did reproductive success. Percent seed set was significantly correlated with the number of plants flowering annually, such that plants flowering in high flowering years (1997 and 2001) exhibited significantly higher percent seed set than did plants flowering in low flowering years (1998-2000). In the 3-year pollen limitation study, plants flowering asynchronously were pollen-limited, whereas plants flowering synchronously were not. This species is strongly self-incompatible. Results of this study demonstrate that the Haleakala silversword experiences reduced reproductive success in low flowering years, and suggest that this Allee effect is pollinator-mediated. Allee effects in plants are an understudied yet potentially important force with implications for the population dynamics and conservation of rare species.     

A histological comparison of the development of pollen and female gametophytes in fertile and sterile Cryptomeria japonica

To determine a possible mechanism causing male and female sterility in Cryptomeria japonica male and female cones were collected from a C. japonica, tree, ShinDai2, that lacks pollen release and fertile seeds and specimens were processed to examine the development of pollen and female gametophytes using light microscopy and field emission scanning electron microscopy. Pre-meiotic development proceeded normally, but the formation of aberrant meiotic products was observed in cones of both sexes. In sterile microsporangia, heterogeneous microspore populations ranging from monads to polyads gave rise to mature pollen grains of non-uniform size. These pollen grains were covered with an amorphous layer and adhered to each other. In addition, they remained in the microsporangia and were not released even after the onset of pollen dissemination from fertile trees. In the ovules of sterile female cones, megaspores with abnormal shapes, numbers, and sizes formed, and the development of female gametophytes was arrested at the free nuclear or archegonium formation stages. These gametophytes collapsed, and no fertile embryo was generated. Results indicate that meiotic defects are important in the sterility mechanism.