Reproductive systems in conifer seed orchards

Summary In order to quantify female and male fitness values of clones in a Pinus sylvestris L. seed orchard, multilocus-genotypes of parental clones were compared with those of open pollinated seeds in the bulked orchard crop. Female and male contributions to individual seeds were distinguished by observing enzyme gene loci active in both endosperm and embryo tissue. Seed probes from two successive flowering periods were surveyed. The female and male fitnesses of five parental clones measured relative to the population mean were derived. The contributions of four clones were found to be sexually asymmetric. One clone, for instance, made exclusively female contributions in one flowering period. Variations existed in fitness values between clones. Deviations in sex specificity occurred between flowering periods: one clone contributed asymmetrically in both periods, but in sexually reversed proportions. A method to comprehensively quantify and illustrate the observed phenomena is proposed.     

INDIVIDUAL FLOWERING TIME IN A GOLDENROD (SOLIDAGO CANADENSIS): FIELD EXPERIMENT SHOWS GENOTYPE MORE IMPORTANT THAN ENVIRONMENT

Individual plants of an old field population of the clonal perennial goldenrod, Solidago canadensis L. are consistent in rank order of flowering time over years. In order to analyze whether this consistency is due more to environmental variations among microsites of individual clones or due to genetic variation among clones, similar-sized sections of rhizomes were transplanted to an experimental garden in a randomized block design with five replicates per clone. Three years later we examined the transplants and the original plants for their flowering phenology. In the experimental garden there was an unexpected soil moisture gradient across blocks, with associated gradients in height of goldenrods and height of surrounding vegetation. No gradient was significantly correlated with the flowering time of the transplants. That is, the time of flowering of transplants was consistent among replicates of individual clones across the soil moisture gradient and regardless of the size of the transplants themselves, or the height of surrounding vegetation. Further, the flowering times of the transplants were significantly positively correlated with the flowering times of the original plants in the same year. We conclude that the differences in flowering time among clones within a goldenrod population is largely determined by genetic variation, rather than by environmental factors.     

Consequences of variation in flowering phenology for seed head herbivory and reproductive success in Erigeron glaucus (Compositae)

Summary We examined the relationship between flowering phenology, reproductive success (seed production only), and seed head herbivory for 20 similarly sized clones of Erigeron glaucus growing at Bodega Bay Reserve, northern California, USA. Although clones tended to reach peak flowering on the same date, they differed in the proportion of their total flowers produced around that date (flowering synchrony). Clones also differed in the number and density of flower heads presented at any one time to pollinators and herbivores (floral display). Both of these characteristics had consequences for herbivory and plant reproductive success. The proportion of flower heads damaged by insect herbivores was greater for clones that concentrated flowering activity during the main flowering period for the population as a whole (high synchrony) compared to clones that spread flowering out temporally. The primary reason for this result was that clones with low flowering synchrony produced a significant proportion of their flower heads during the fall and therefore, escaped attack by the tephritid fly, Tephritis ovatipennis. Clones with intermediate synchrony had lower seed success (total number of viable seeds produced over the year) than clones with either low or high synchrony. The proportion of flower heads damaged by insect herbivores and number of tephritid flies reared from flower heads were both negatively correlated to floral display while seed head mass and germination rates were positively related to display. Thus, clones which produced dense floral displays were favored both in terms of reduced herbivory and increased successful seed production.     

Prevention of the flowering of a tree,silver birch

Genetic modification of trees presents great advantages but it is hampered by the possible spread of introduced genes to native populations. However, the spread would be prevented if the modified trees would be sterile. We have previously shown that the induction of sterility by the prevention of flowering is possible in tobacco and Arabidopsis by introducing a gene construct composed of the ribonuclease gene BARNASE ligated to the flower-specific promoter of the birch gene BpMADS1. In the present study, we test this gene construct in silver birch (Betula pendula Roth). When this gene construct was introduced into very early-flowering birch clones, 81 kanamycin resistant lines were obtained. In 38 lines, the vegetative development was disturbed, e.g., the leaves were small and the plants were short and bushy or the growth of plants was weak. More importantly, in 7 other lines no male inflorescences formed or they aborted early. If male inflorescences were formed, they did not contain any stamens. The initial growth of these lines was similar to the non-transgenic control lines. Later, however, the growth of the non-flowering lines differed from that of the controls in showing some dichotomic branching and a reduced number of branches. Preliminary results showed that the gene construct can prevent the development of female inflorescences as well. The results show clearly that BpMADS1::BARNASE can prevent the flowering in a tree but the prevention of flowering may cause some side effects. Studies with ordinary birch clones will show whether the side effects are a property of the early flowering clones or all birches.     

Activity of the CaMV 35S promoter in various parts of transgenic early flowering birch clones

Early flowering together with small size would be useful for various biotechnical or genetic studies on trees. We report here the selection and micropropagation of early flowering birch (Betula pendula) clones (BPM1–12) obtained from seeds of birches bred elsewhere for early flowering. Under conditions that accelerate flowering (a high CO₂ level, strong and continuous illumination), the first male inflorescences emerged in 3–5 months, the trees then being 20–80 cm high. Transgenic lines (CaMV 35S-GUS INT) were produced through Agrobacterium-mediated gene transfer from BPM2, BPM5 and JR1/4 (a normally flowering birch). β-Glucuronidase (GUS) activities in the different lines, assayed 1–1.5 years after transformation, varied greatly. During further in vitro culture for 10 months, the activities decreased to 0.3–7% of the original values. GUS activities were detected in all organs studied, including the developing male inflorescences; the highest activity was in the roots. Received: 28 April 1997 / Revision received: 5 September 1997 / Accepted: 30 November 1997     

Sexual reproductive biology of the endangered Japanese red maple (<Emphasis Type="Italic">Acer pycnanthum</Emphasis>)

Japanese red maple, Acer pycnanthum, is an endangered wetland species having dioecious sex expression. Prior studies conclude that land development has caused severe loss of habitat, and seedling regeneration is rare. However, information is lacking on sexual reproduction, which is the first critical stage of regeneration. My research objective was to describe flowering and fruiting characteristics of Japanese red maple, focusing on tree sizes at the onset of flowering, factors affecting flowering regularity and fruit abundance, periodicity of fruit production, and sex ratio. In a study of a young population, Japanese red maple initiated flowering at a small size under 100% light intensity. Initiation of flowering occurred at relatively lower height in males than females, and males flowered more regularly than females. A survey of 1,106 clones from 46 natural populations revealed that male clones were significantly more abundant than female clones. In a 4-year study of mature populations, nearly all clones flowered regularly, and fruit abundance of dominant females was typically high. Fruit abundance at the population level also remained high, although fruit abundance of individual female clones varied year by year. Therefore, sexual reproductive ability is highly vigorous. Sex ratio exhibited a major deviation from a 1:1 ratio in populations with a small number of clones. In conservation planning, we should prioritize avoidance of an unbalanced sex ratio by increasing population sizes. In addition, populations having many clones >40 cm dbh with large, well-lighted crowns have a high potential of regular and abundant seed production.     

Distribution of Cashew Flower Sex-types between Clones and Sides of Tree Canopies in Tanzania

Production of male, hermaphrodite and abnormal flowers was studiedon the north, east, south and west sides of the cashew treecanopy from Jul. to Dec. 1992 at the Agricultural Research InstituteNaliendele, Southern Tanzania. Flowers were recorded daily onselected clones throughout the main flowering season and subsequentlyyield of each clone was also recorded. Results showed differencesin the number of flower types both between clones and betweensides; however, there were consistently more male than othertypes of flower. Significant yield differences between cloneswere not detected and so could not be used to provide unambiguousevidence of the relationship between yield and flower characteristicsbut did indicate that clonal differences were present in termsof earliness of yield production. Anacardium occidentale L.; cashew; flowering; sex-types; clones; Tanzania; sides of canopy; yield     

Flowering phenology in natural populations of Iris pumila

In the present study, I examined flowering phenology of dwarf bearded ins, Iris pumila , using naturally occurring clones (three sites, four microsites) and clones transplanted between two sites representing two habitats Naturally occurring clones in contrasting habitats and microsites differed significantly in phenology, with exposition of the site or microsite being the most important Genotypes from more exposed habitats flowered earlier in both habitats and these differences were statistically significant Patterns of between-habitat and between-population differences were stable over years even though years differed markedly in the flowering onset Within-population genetic variability for flowering phenology was also found to be significant Although clones with dark-colored flowers tended to flower earlier in all habitats and microsites I failed to detect statistically significant differences in flowering time among color morphs Flowering phenology in I pumila is highly susceptible to environmental variability, but this factor has not prevented population divergence in flowering time Between-habitat differences in flowering time turned out to be a result of both phenotypic plasticity of individuals and genetic differentiation of populations     

Temporal and intraclonal variation of flowering and pseudovivipary in Poa bulbosa

Background and Aims

Versatility in the reproductive development of pseudoviviparous grasses in response to growth conditions is an intriguing reproduction strategy. To better understand this strategy, this study examined variation in flowering and pseudovivipary among populations, co-occurring clones within populations, and among tillers in individual clones of Poa bulbosa, a summer-dormant geophytic grass that reproduces sexually by seed, and asexually by basal tiller bulbs and bulbils formed in proliferated panicles.

Methods

Clones were collected from 17 populations across a rainfall gradient. Patterns of reproduction were monitored for 11 years in a common garden experiment and related to interannual differences in climatic conditions. Intraclonal variation in flowering and pseudovivipary was studied in a phytotron, under daylengths marginal for flowering induction.

Key Results

Clones showed large temporal variability in their reproductive behaviour. They flowered in some years but not in others, produced normal or proliferated panicles in different years, or became dormant without flowering. Proliferating clones did not show a distinct time sequence of flowering and proliferation across years. Populations differed in incidence of flowering and proliferation. The proportion of flowering clones increased with decreasing rainfall at the site of population origin, but no consistent relationship was found between flowering and precipitation in the common garden experiment across years. In contrast, flowering decreased at higher temperatures during early growth stages after bulb sprouting. Pulses of soil fertilization greatly increased the proportion of flowering clones and panicle production. High intraclonal tiller heterogeneity was observed, as shown by the divergent developmental fates of daughter plants arising from bulbs from the same parent clone and grown under similar conditions. Panicle proliferation was enhanced by non-inductive 8 h short days, while marginally inductive 12 h days promoted normal panicles.

Conclusions

Interannual variation in flowering and proliferation in P. bulbosa clones was attributed to differences in the onset of the rainy season, resulting in different daylength and temperature conditions during the early stages of growth, during which induction of flowering and dormancy occurs.     

Flowering phenology,floral display and reproductive success in dioecious,Aralia nudicaulis L. (Araliaceae)

Summary Aralia nudicaulis L. is a dioecious, perennial, herbaceous plant that is commonly found in the understory vegetation throughout the boreal forest of North America. Female remets have fewer flowers per inflorescence, initiate flowering earlier, and reach peak flowering before male ramets. The consequences of the asynchrony in flowering between the sexes on pollination and seed set were examined during a two-year study. In both years there was significant variation in seed set associated with the flowering times of individual female ramets. In 1983, seed production was highest in the middle of the flowering season. In 1984, seed production was greatest in the later stages of flowering. Variation in seed set was not attributed to lack of pollination in 1983. In 1984, pollination limited seed set per flower during peak flowering. However, seed production never reached the potential five seeds per flower, suggesting that resource limitation was the most important factor affecting fecundity in both years. The asynchronous pattern of flowering is suggested to be the result of the different inflorescence sizes between the sexes.     

Differences in response to defoliation between males and females of Silene dioica

Summary The response by male and female plants to herbivory was studied by experimental defoliation of the dioecious perennial herb Silene dioica in a green-house. Male and female plants were defoliated prior to and during the early flowering phase at two intensities (50% and 100% of leaf-area removed) in two consecutive years. Defoliation resulted in a decrease in the number of flowers initiated in both sexes, while a larger delay of peak flowering and a higher mortality was observed in males compared to females. In female plants, severe defoliation resulted in a reduction in seed number per capsule and in seed size compared to control. Females showed a negative correlation between the production of flowers in the first and second season in all treatments, while flowering in males the first season was not correlated with flowering in the second season. Females also showed a lower frequency of flowering than males during the two seasons studied. However, during the flowering period, males allocated significantly more biomass to flowers than did females. This outcome supports the idea that females may have a higher total reproductive expenditure than males, but males have a higher reproductive effort during flowering. Male rosette leaves were significantly preferred by the generalist herbivore Arianta arbustorum in experiments. This preference was most pronounced in trials with leaves from fertilized plants compared to nonfertilized plants. A greater storage of resources in aboveground leaves during winter by males compared to females may explain the higher preference for male leaves and the higher male mortality following early defoliation. Furthermore, males are smaller than females and may have a lower ability than females to replace lost resources needed for reproduction when defoliated early in the season.     

Specific features of development of the male generative organs in clones of the Siberian stone pine (Pinus sibirica De Tour) of different origins in the Krasnoiarsk forest-steppe

The development of male generative organs of the Siberian stone pine clones of different origin was analyzed in the Krasnoyarsk forest-steppe for many years. Geographical and individual variability was described as concerns the number of microstrobils and pollen variability. The beginning of "flowering", abundance of microstrobils, and their regular formation were shown to markedly vary and, all other conditions being equal, be determined by the presence of clones, their hereditary features, and environmental conditions, as well by the influence of stock.     

Wind pollination and insect pollination of two temperate willow species,Salix miyabeana and Salix sachalinensis

Pollination systems of Salix miyabeana and Salix sachalinensis were studied at a riverside in northern Japan in order to measure the balance of wind pollination and insect pollination. In 1996, 19 clones of each species were selected, and seed-set success by a net-bagging (to exclude insect visitation) and an artificial pollination (to remove pollen limitation) were compared to by natural pollination. For S. miyabeana, the same experiment was repeated in two populations in 1997. Proportion of seed set through wind pollination dominated in both species in this study. Pollen limitation was common under natural conditions, and S. sachalinensis relied more on insect pollination for seed production than S. miyabeana. Meteorological factors such as precipitation and hours of sunshine during the flowering season influenced the potential reproductive activity of the willow between years. In the wet and cloudy spring of 1996, clones which obtained high seed set depended more on insect pollination for both species, whereas in the dry and sunny spring of 1997, such clones depended more on wind pollination for S. miyabeana. Because the efficiency of wind pollination seemed to be more sensitive to fluctuating weather conditions than insect pollination, insect pollination was considered to play an assurance role for seed production in these willows.     

The causes of selection on flowering time through male fitness in a hermaphroditic annual plant

Flowering is a key life‐history event whose timing almost certainly affects both male and female fitness, but tests of selection on flowering time through male fitness are few. Such selection may arise from direct effects of flowering time, and indirect effects through covariance between flowering time and the environment experienced during reproduction. To isolate these intrinsically correlated associations, we staggered planting dates of Brassica rapa families with known flowering times, creating populations in which age at flowering (i.e., flowering time genotype) and Julian date of flowering (i.e., flowering time environment) were positively, negatively, or uncorrelated. Genetic paternity analysis revealed that male fitness was not strongly influenced by seasonal environmental changes. Instead, when age and date were uncorrelated, selection through male fitness strongly favored young age at flowering. Strategic sampling offspring for paternity analysis rejected covariance between sire age at flowering and dam quality as the cause of this selection. Results instead suggest a negative association between age at flowering and pollen competitive ability. The manipulation also revealed that, at least in B. rapa, the often‐observed correlation between flowering time and flowering duration is environmental, not genetic, in origin.     

Production of somatic hybrids between frost-tolerant Solanum commersonii and S. tuberosum: characterization of hybrid plants

Somatic fusion of mesophyll protoplasts was used to produce hybrids between the frost-tolerant species Solanum commersonii (2n=2x=24) and dihaploid S. tuberosum (2n=2x=24). This is a sexually incompatible combination due to the difference in EBN (Endosperm Balance Number, Johnston et al. 1980). Species with different EBNs as a rule are sexually incompatible. Fifty-seven hybrids were analysed for variation in chromosome number, morphological traits, fertility and frost tolerance. About 70% of the hybrids were tetraploid, and 30% hexaploid. Chloroplast counts in stomatal guard cells revealed a low frequency of cytochimeras. The frequency of aneuploids was relatively higher at the hexaploid level (hypohexaploids) than at the tetraploid level (hypotetraploids). The somatic hybrids were much more vigorous than the parents, and showed an intermediate phenotype for several morphological traits and moderate to profuse flowering. Hexaploid hybrid clones were less vigorous and had a lower degree of flowering than the tetraploid hybrid clones. All of the hybrids were female fertile but male sterile except for one, which was fully fertile and self-compatible. Many seeds were produced on the latter clone by selfing and on the male-sterile clones by crossing. The somatic hybrid plants showed an introgression of genes for frost tolerance and an adaptability to cold from S. commersonii. Therefore, the use of these somatic hybrids in breeding for and in genetic esearch on frost tolerance and cold-hardening is suggested.     

浙江低山地区多用途植物无患子的开花物候特征

无患子(Sapindus mukorossi Gaertn.)是我国长江以南地区传统的重要绿化树种,其果皮富含皂苷,种仁富含油脂,是国家林业局审定的新型木本油料树种之一。为研究多用途树种无患子在浙江低山地区的开花特征,2012年和2013年连续两年对位于浙江省天台县9年生无患子人工林在群体、个体、花序和单花水平进行开花物候观测和比较,并运用开花日期、相对开花强度和同步性等指数研究了无患子开花物候特征。观察结果显示:无患子花呈浅黄白色,花的类型有雄花和两性花,没有发现雌花。雄花较两性花大(花径分别为5.09 mm和3.72 mm),雄蕊多为8枚,个别7或9、10枚;雌蕊退化仅留下浅绿色凸起。两性花花萼稍抱拢,花药藏于花被片下,雄蕊大多8枚,极少数7枚或9枚,柱头高于花药并伸出花苞,子房一般具3室,极少数4室或仅2室。无患子2012年的开花进程略早于2013年,其花期集中在在5月中旬至6月上旬,单花从花蕾膨大到花朵凋谢一般为8—9 d。在2012年和2013年,无患子在群体、个体和花序水平的花期约为30 d、20 d、11 d和28 d、19 d、13 d。个体水平的开花振幅均呈单峰曲线,年际间相似性较高;开花同步性在个体水平同步性较高(同步指数为0.868),表现出一种大量、集中的开花式样;相对开花强度在单株间分布范围相对宽泛,但主要分布在30%—40%,在年际间和年际内均呈现极显著差异。花期同步指数在两年的变异范围分别是0.81—0.97和0.70—0.98,不同单株开花同步性在年际内差异极显著,但在年际间差异性则不显著。由此可见,无患子的生殖资源分配存在明显的时空差异,较长的花期可以减少非法花粉的干扰、保持其种群基因多样性,遗传因子是决定无患子开花物候的主要因素,生态环境对无患子开花物候的影响还需进一步研究,本研究以期为探索无患子开花的主要限制因子奠定基础。     

Reproduction in Polemonium: factors influencing outbreeding potential

Summary Three measures of potential outbreeding efficiency were quantified throughout two flowering seasons in a population of the mass-flowering Polemonium foliosissimum: 1) the distances flown between plants by bumblebee pollinators; 2) the proximity of visited plants; and 3) the diversity of individuals visited. Results were consistent between years. Individual plants did not function differentially with respect to female (i.e., pollen receipt) and male (i.e., pollen donation) function. Both female and male components of outbreeding, as estimated by all three measures, were positively correlated with the number of flowers per plant. Significantly more outbreeding could occur during the final phase of flowering when most individuals had relatively few blossoms. Although the potential for outbreeding was the same at any time through an individual's blooming regime, individuals for which the peak of blooming occurred after the peak for the population as a whole have significantly more outbreeding potential via both female and male function. These factors may combine such that any pressure to increase outbreeding in this self-incompatible species manifests itself in directional selection for delayed flowering time.     

Reproductive isolation of two sympatric subspecies ofJuniperus phoenicea (Cupressaceae) in southern Spain

The phenology, reproductive cycle and cone crop of two sympatric subspecies ofJuniperus phoenicea (Cupressaceae) were studied during three consecutive years in southern Spain. The flowering pattern of each subspecies was constant during this period: inJ. phoenicea subsp.turbinata flowering was always in October–November, whileJ. phoenicea subsp.phoenicea flowered in February–March. There was no overlap between the flowering periods of the two taxa. The reproductive cycle of the two subspecies was similar, though there were some important differences because of their flowering times. Male cone development in both subspecies occurred in autumn, but the male cones ofJ. phoenicea subsp.phoenicea did not shed pollen until the end of February. InJ. phoenicea subsp.turbinata, the gap between pollination and fertilisation was seven months while inJ. phoenicea subsp.phoenicea it was only three months, fertilisation taking place at the same time in the two taxa. Post-fertilisation events were similar in the two subspecies, leading to a cycle of two years inJ. phoenicea subsp.turbinata and a shorter one of about 20 months inJ. phoenicea subsp.phoenicea. Alternating good and slight seed cone crops were found in both taxa during the three years studied, the good crops of one subspecies coinciding with the slight crops of the other one. Although the proximity of the two subspecies and method of pollination could make hybridisation between the taxa possible, the different flowering seasons prevent hybridisation isolating them reproductively.     

Effect of growth substances on flowering and sex expression in isolated apical buds of Spinacia oleracea

Apical buds of Spinacia oleracea L. cv. Matador were isolated from 7-day-old vegetative seedlings and grown in sterile culture under inductive long, or non-inductive short photoperiods. Flowering of isolated apices was inducible in long days in approximately 75% of the plants, and in short days by gibberellin treatment (about 40%) or by raising the temperature to 30–32°C (13%). In long days the percentage of flowering was further increased by gibberellin treatment (up to 90%), while it was unaffected by abscisic acid and was strongly decreased by Amo 1618 (55%). In long days the ratio of male to female plants was near 1. The percentage of female plants in long days was increased by gibberellins, and the percentage of male ones decreased by kinetin; as a consequence, the ratio of male to female plants was lowered to about 0.50 in both cases. Abscisic acid and Amo 1618 had the opposite effect, probably because they decreased the flowering in female plants, so that the sex ratio was shifted to 2.6 and 6.8, respectively. Simultaneous treatment with GA₃ reversed the effect of abscisic acid on the sex ratio, but the reversal of the shift to maleness induced by Amo 1618 was only partial. In short days, gibberellins also stimulated the flowering in female plants more than in male. However, when the flowering was induced by higher temperature, most flowering plants were male and kinetin increased their percentage further. The above results have been discussed in terms of different requirements for flowering in male and female plants.     

Clonal genetic structure and diversity in populations of an aquatic plant with combined vs. separate sexes

Clonality is often implicated in models of the evolution of dioecy, but few studies have explicitly compared clonal structure between plant sexual systems, or between the sexes in dioecious populations. Here, we exploit the occurrence of monoecy and dioecy in clonal Sagittaria latifola (Alismataceae) to evaluate two main hypotheses: (i) clone sizes are smaller in monoecious than dioecious populations, because of constraints imposed on clone size by costs associated with geitonogamy; (ii) in dioecious populations, male clones are larger and flower more often than female clones because of sex‐differential reproductive costs. Differences in clone size and flowering could result in discordance between ramet‐ and genet‐based sex ratios. We used spatially explicit sampling to address these hypotheses in 10 monoecious and 11 dioecious populations of S. latifolia at the northern range limit in Eastern North America. In contrast to our predictions, monoecious clones were significantly larger than dioecious clones, probably due to their higher rates of vegetative growth and corm production, and in dioecious populations, there was no difference in clone size between females and males; ramet‐ and genet‐based sex ratios were therefore highly correlated. Genotypic diversity declined with latitude for both sexual systems, but monoecious populations exhibited lower genotypic richness. Differences in life history between the sexual systems of S. latifolia appear to be the most important determinants of clonal structure and diversity.