Plasticity of reproductive components at different stages of development in the annual plant Thlaspi arvense L.

Summary This study examines the effect of different densities and the removal of all neighbours at different stages of development on all components of reproduction in the inbreeding annual Thlaspi arvense L. A 64-fold increase in density significantly reduced all repooductive components. The number of flower buds per plant was decreased most strongly; the order of decreasing plasticity among the other components was number of capsules per flower, individual seed weight, ovule number per capsule, flowers per flower bud and seeds per ovule. Removing neighbours at all stages of development increased seed yield of plants in comparison to the control without density reduction, but patterns of plasticity depended strongly on time of treatment. The main effect of the removal of neighbours at the vegetative stage was to increase the number of flowers per plant, but the number of ovules per capsule and seed weight increased also, and abortion of capsules decreased. Removing neighbours at the onset of flowering initially failed to affect flower number per plant, instead it resulted in a strong reduction of capsule abortion and an increase in seed weight. However, several weeks after flowering had initially ceased, fresh lateral inflorescences were produced, resulting in a second flush of reproduction. Removing neighbours at the stage of fruit ripening resulted at first only in an increase in seed witht, but later a second reproductive phase occurred. Fresh lateral branches were produced, but the apical meristem was also reactivated. The overall pattern of plasticity among all reproductive components in response to a removal of neighbours was the same as in response to density. The position of a capsule along the inflorescence influenced its number of ovules, the rate of seed abortion and the mean weight of seeds, with the type of effect depending on the developmental stage at which neighbours were removed. Significant negative correlations were found between the mean weight of seeds and the number of seeds in a capsule under all treatments.     

Structural aspects of ovule and seed development and nonrandom abortion inMelilotus officinalis (Fabaceae)

Summary Only one ovule matures into a seed inMelilotus officinalis. Although eight ovules form within an ovary, only the basal ovule develops into a mature seed, whereas the other ovules degenerate. The investigation of ovule and seed structure at different developmental stages and a comparison of quantitative characters of differently fated ovules within an ovary were undertaken by light, phase contrast, and fluorescence microscopy. In this species, campylotropous ovules develop simultaneously on marginal placentae in an apocarpous unilocular gynoecium. Megasporo- and megagametogenesis proceed normally and are completed in bud. The maturation of the Polygonum type embryo sac takes place after the flower opens. Shortly before fertilization, synergids show signs of degeneration in all ovules. At this stage, neither the structure nor the sizes of ovules within one ovary differ significantly. In spite of this, only the basal ovule develops into a seed. Rarely, one of the upper-situated ovules or the basal and another ovule mature into seeds. Seed enlargement is insignificant until the stage when globular embryo and nuclear endosperm are formed. At the seed-filling stage, other ovules have collapsed and the seed gradually comes to occupy the total volume of the pod. The fruit-to-seed length ratio decreases considerably during seed ripening. At fertilization, ovary length is four times greater than ovule length. In the mature state, the fruit and seed lengths are approximately equal. Seed size and weight diminish with an increase in seed number within a pod, although pod size remains constant. It is assumed that nonrandom abortion of young seeds inM. officinalis is under maternal control and is not related to structural abnormalities in ovule development or with limitation in pollen. We suppose that evolution of this species may have proceeded in the direction of a decrease in seed number and an increase in its sizes, which may play an important role in seed dispersal and seedling establishment.     

Order of fertilization within the ovary in Phaseolus coccineus L. (Leguminosae)

Summary Phaseolus coccineus typically has six linearly arranged ovules per ovary. The three ovules near the stylar end of the fruit (positions one, two, and three) are more likely to produce mature seeds, to produce heavier seeds, and to produce more vigorous progeny than the ovules in positions near the peduncular/basal end of the fruit (ovule positions four, five, and six). We conducted a series of field experiments designed to supplement our understanding of the mechanisms determining these position effects. We found that approximately 98% of the ovules in 752 fruits were fertilized — about 0.6% of the stylar ovules were not fertilized, whereas 3.2% of the basal ovules were unfertilized. Moreover, we found that only about 49% of the ovules in these 752 fruits produced mature seeds. Over 60% of the stylar ovules produced mature seeds, whereas only 37% of the basal ovules produced mature seeds. Consequently, the proportion of fertilized ovules cannot explain the differences in seed maturation among the ovule positions. We found that after 6.5 h most of the fertilized ovules were located in the stylar ovule positions, and that there were no fertilized ovules in ovule positions five and six, indicating that the stylar ovules are fertilized first. When only the fastest growing pollen tubes were permitted to enter the ovary (due to exision of the style), only the ovules at the stylar end were fertilized, indicating that the ovule positions that are fertilized first are indeed fertilized by the fastest growing pollen tubes.On leave from the Escuela de Biologia, Universidad de Costa Rica, Cuidad Universitaria Rodrigo Facio, San Jose, Costa Rica, Central America     

Seed germination and dormancy in the medicinal woodland herbs Collinsonia canadensis L. (Lamiaceae) and Dioscorea villosa L. (Dioscoreaceae)

We used a double germination phenology or “move-along” experiment (sensu Baskin and Baskin, 2003) to characterize seed dormancy in two medicinal woodland herbs, Collinsonia canadensis L. (Lamiaceae) and Dioscorea villosa L. (Dioscoreaceae). Imbibed seeds of both species were moved through the following two sequences of simulated thermoperiods: (a) 30/15 °C→20/10 °C→15/6 °C→5 °C→15/6 °C→20/10 °C→30/15 °C, and (b) 5 °C→15/6 °C→20/10 °C→30/15 °C→20/10 °C→15/6 °C→5 °C. In each sequence, seeds of both species germinated to high rates (>85%) at cool temperatures (15/6 and 20/10 °C) only if seeds were previously exposed to cold temperatures (5 °C). Seeds kept at four control thermoperiods (5, 15/6, 20/10, 30/15 °C) for 30 d showed little or no germination. Seeds of both species, therefore, have physiological dormancy that is broken by 12 weeks of cold (5 °C) stratification. Morphological studies indicated that embryos of C. canadensis have “investing” embryos at maturity (morphological dormancy absent), whereas embryos of D. villosa are undeveloped at maturity (morphological dormancy present). Because warm temperatures are required for embryo growth and cold stratification breaks physiological dormancy, D. villosa seeds have non-deep simple morphophysiological dormancy (MPD). Neither species afterripened in a 6-month dry storage treatment. Cold stratification treatments of 4 and 8 weeks alleviated dormancy in both species but C. canadensis seeds germinated at slower speeds and lower rates compared to seeds given 12 weeks of cold stratification. In their natural habitat, both species disperse seeds in mid- to late autumn and germinate in the spring after cold winter temperatures alleviate endogenous dormancy.     

Chromosomal evolution inEpilobium sect.Epilobium (Onagraceae), III.

A cytological analysis of 22 experimental interspecific hybrids withinEpilobium sect.Epilobium, involving 15 taxa, has indicated that the generalized western North AmericaE. obcordatum has the BB chromosome arrangement. This strengthens greatly our earlier contention that BB is the original chromosome arrangement in the section. The very distinctive northwestern North AmericanE. luteum, only species in the section with cream-colored petals, has the CC arrangement, like the members ofHaussknecht'sAlpinae. This chromosome arrangement apparently originated in the North Pacific region in plants that were of normal stature. The North AmericanE. leptocarpum, which closely resembles the Japanese endemicE. fauriei and, like it, forms gemmae in the axils of the upper leaves, also resembles it in having the CC chromosome arrangement. The AsianE. platystigmatosum has a novel chromosome arrangement derived from BB, which we here designate the EE arrangement.     

Plant regeneration through shoot formation from callus of Areca catechu L.

In order to establish and optimize an in vitro micropropagation protocol of Venus fly trap (Dionaea muscipula Ellis), a carnivorous plant, the effects of medium type, MS medium concentration, pH, and cytokinin and auxin types on shoot proliferation and root formation were investigated using 3-month-old shoots. The shoot proliferation was most effective in 2.3 M kinetin-supplemented 1/3MS medium at pH 5.5. The best conditions for rooting were 1/3MS medium supplemented with 0.5 M IBA. All subcultured shoots produced extensive root systems after 5–6 weeks culture. When plantlets after rooting were planted in plastic pots filled with 1:1 peat moss and sand, the survival rate of plantlets was almost 100%, exhibiting normal development. With subculture every 8 weeks, hundreds of the plants were propagated from a single plant within a year.     

Sexual differentiation in Asparagus officinalis L.

Summary Sexual dimorphism in the dioecious plant Asparagus officinalis L. was examined by two-dimensional (2-D) electrophoresis of both total proteins and newly synthesized proteins from cladophylls (leaves), whole mature flowers and homologous sex organs (i.e. true female ovaries and small sterile ovaries from male flowers). Polypeptides isolated from cladophylls of male and female plants were practically indistinguishable; the flowers, however, showed a distinct set of specific proteins, some of which differed between the two sexes. While the total protein profiles of isolated ovaries from male and female plants were very similar, the patterns were strikingly different after the tissues were pulsed with ³⁵S-methionine: mature male ovaries showed a number of newly synthesized proteins, while in female ovaries only a few molecular species were actively synthesized.     

Intra-fruit seed abortion in a wind dispersed tree,Dalbergia sissoo Roxb: Proximate mechanisms

Dalbergia sissoo, a tropical tree with wind-dispersed pods, exhibits a highly positively skewed distribution of seeds per pod with predominantly only one of the four or five ovules maturing into seed. The abortion cannot be attributed to lack of pollen or resources. This study examines the hypothesis that the abortion is due to an intense rivalry among the developing sibs to gain dispersal advantage. Aqueous extract of the dominant embryos at the stigmatic end that generally develop to maturity significantly inhibited the uptake of labelled sucrose by the young developing (subject) embryos in an in vitro assay system. Extracts of tissues such as subordinate (peduncular embryos), unfertilized ovules and pod coat did not cause such inhibition. Aqueous diffusate of dominant embryos also inhibited the uptake of labelled sucrose by subject embryos. The chemical substance responsible for the inhibition appears to be heat-stable and non-proteinaceous. HPLC analysis indicated the presence of two retention time peaks, different from that of standard indole acetic acid, but with considerable overlap. We hypothesize that the compound could be an indole derivative. We propose that the stigmatic embryos have a head start due to earlier fertilization and produce a chemical that either directly (by metabolically killing) or indirectly (by preventing the uptake of assimilates) kills the proximally placed peduncular embryos.     

Unique tetrads of <Emphasis Type="Italic">Epilobium luteum</Emphasis> (Onagraceae: Onagreae) pollen from Alaska

Aperture morphology of tetrad pollen of Epilobium luteum (Onagraceae: Epilobieae) from three Alaskan collections is highly variable. The first collection appears to lack apertures altogether and is presumed to consist of immature pollen gains in a genus known to achieve mature size before the apertures become distinctly protruding. A second collection has tetrads with 3- and 4-apertured grains, the apertures in the latter are often irregularly spaced and not in apposition with the apertures of neighboring members. The third collection consists of the more typical 3-apertured members that characterize the majority of Epilobium pollen grains. In all of these collections individual pollen grains (monads) are interspersed among the tetrads. The variations in the number of apertures emphasize the importance of having a comprehensive understanding of the stage of development of the pollen (taxon) examined when describing pollen collections. In the first collection this would mean the recognition that in Onagraceae apertures occur in the later stages of microspore ontogeny. In the latter two collections a thorough background of the literature of the pollen morphology on this largest Onagraceae taxon is useful for the understanding of the significance of a range of aperture numbers on Epilobium pollen grains.     

Silencing of chaperonin 21, that was differentially expressed in inflorescence of seedless and seeded grapes,promoted seed abortion in tobacco and tomato fruits

Vitis vinifera L. cv. ‘Thompson Seedless’ presents a type of stenospermocarpy in grape where fertilization occurs but seeds abort and fail to develop. To unravel the molecular basis for stenospermocarpy in grapes, subtractive hybridization was carried out in order to isolate differentially regulated genes that participate in the seedlessness machinery. Two ‘Thompson’ lines, a seeded and a seedless, were screened during different flower developmental stages. One of the genes, that was differentially expressed between the seeded and seedless lines, was the chloroplast chaperonin 21 (ch-Cpn21). ch-Cpn21 is a 21-kDa co-chaperonin polypeptide formed by two GroES-like domains fused together in tandem. Silencing of ch-Cpn21 in Nicotiana benthamiana plants resulted in leaf stunting, chlorosis, as well as ovary necrogenesis leading to seed abortion. Moreover, organ-specific silencing of ch-Cpn21 only in Lycopersicum esculentum fruits resulted in the development of seedless tomatoes. These results suggest that ch-Cpn21 may play a role in seed abortion in stenospermocarpic grapes.     

Phytometric estimation of fertility of waterlogged rich-fen peats usingEpilobium hirsutum L.

Summary The above-ground net production of herbaceous vegetation in 16 stands of waterlogged rich fen in Broadland, Norfolk UK, has been compared with the growth ofEpilobium hirsutum seedlings (in controlled environment) on replicate peat samples from each site and with the chemical composition of extracts from sub-samples of each peat (Ca, Mg, Na, K, Fe, Mn, inorganic P, inorganic N). There were no consistent or significant relationships between measured concentrations of major nutrients (K, N, P) in peat samples and netin situ production of the vegetation or with growth ofE. hirsutum seedlings on each peat. There was a strong, significant (p<0.001) relationship between above ground productionin situ and growth ofE. hirsutum seedlings. Seedling growth thus provided a simple, sensitive phytometric measure of fertility of the waterlogged peats (pH range 6.0–7.0). One possible limitation is the apparent sensitivity ofE. hirsutum seedlings to high manganese concentrations.     

Induction of adventitious buds in cultured petal explants ofChelidonium majus L.

Petal explants ofChelidonium majus L. (Papaveraceae) formed noteworthy adventitious buds without any intermediate callus when cultured under appropriate conditions. Bud formation was favored by combinations of 1–2 mg/l indoleacetic acid (IAA) and/or 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1–0.5 mg/l kinetin (K). In the present study, neither bud formation nor callus formation occurred in cultures of excised leaves. A histological study revealed that adventitious bud formation occurred only in single epidermal layers of petals, while several subepidermal parenchyma layers did not join in its formation. Activation zones arising from the epidermis underwent intense cell divisions to initiate buds on the epidermal surface. These buds later turned green in color, developing into shoots which eventually grew into plantlets after root formation.     

Aggregated seed dispersal by wreathed hornbills at a roost site in a moist evergreen forest of Thailand

Hornbills (Bucerotidae) are widely regarded as important seed dispersers in tropical forests in Africa and Asia. We investigated how the roosting behavior of wreathed hornbills (Aceros undulatus) influences seed deposition and seedling survival at a roost site in a moist evergreen forest of Khao Yai National Park, Thailand. Fallen fruits and seeds were collected in traps that were placed around a roosting site for 14 months, and seedlings were monitored in adjacent quadrats for 3 years. Seedfall and seedlings of species represented in the hornbill diet occurred at significantly higher densities in the traps and quadrats located beneath the crown of the roosting tree than in those located beyond the crown. With the exception of Cinnamomum subavenium, the seeds and seedlings of most diet species rarely survived beyond the first year. The quality of hornbill dispersal to this roosting site may be poor due to the highly concentrated seedfall, which results in high seed and seedling mortality. However, the number of seeds deposited by each hornbill each day at roosting sites is relatively low. Wreathed hornbills are primarily scatter dispersers during the day and probably serve as agents of seed dispersal in the moist evergreen forest of Khao Yai.     

Hydroponic culture of Catharanthus roseus (L.) G. Don and studies on seed production

Under climatic conditions of Poland, Catharanthus roseus may be grown in the open field only as annual crop, using transplants raised in a greenhouse or in a plastic tunnel. An effective method of hydroponic culture has been elaborated and tested. Herbal material obtained in hydroponics showed considerable concentration of alkaloids in different plant parts. Seeds of C. roseus may also be produced under these conditions. Characteristics of seeds as well as the effect of different methods of their storage are presented. Seeds stored in paper bags at room temperature may retain germinability for 7 years and those stored in a refrigerator, at 5°C, up to 15 years. Storage in a freezer, at -10°C, resulted in a quick and sharp reduction of germination rate.     

Differences in the location of subcotyledonary buds among Epilobium angustifolium L., E. dodonaei Vill. and E. fleischeri Hochst. (Onagraceae) and effects on architecture and population structure

STÖCKLIN, J., 1992. Differences in the location of subcotyledonary buds among Epilobium angustifolium L., E. dodonaei Vill. and E. fleischeri Hochst. (Onagraceae) and effects on architecture and population structure . Morphological and architectural features in the two closely related pioneer plant species Epilobium dodonaei and E. fleischeri are examined in cultivation and in the field and compared with E. angustifolium. In E. angustifolium , the aerial shoot system is renewed every year from buds on horizontal roots and results in a horizontal spread and a clonal growth form. In E. dodonaei , bud formation is restricted to the hypocotyl and in larger plants to the transitional region between root and shoot. Consequently this species shows no vegetative mobility and develops a shrub-like habit. The alpine E. fleischeri combines the habit of E. angustifolium and E. dodonaei and may either develop successive generations of shoots from the transitional region between root and shoot and/or exploit new areas by horizontal roots and the formation of shoots from root buds. The simple difference in the location of renewal buds is accentuated by cumulative growth. The study shows that E. dodonaei and E. fleischeri , which often are considered as subspecies, are separated by fundamental differences in their architectural models. The ecological and demographic implications of these differences are discussed.     

Immunoelectrophoretic analysis of seed proteins from Pisum sativum L.

Summary Soluble proteins of pea seed were investigated by quantitative immunological methods. Vicilin, legumin, pea seed lectin (PEA), 26 albumins and a globulin (B₁) were detected and observed during seed development, germination and under different extraction and fractionation procedures. Vicilin and legumin were found to be immunologically distinctly different. Legumin was found to be comprised of two similar proteins, Legumin species I and II. Vicilin, but no legumin, was detected in the embryonic axis.Three albumins, B₁ and PEA were found to be synthesized after the onset of legumin synthesis.Among the pea lines investigated, one line exhibited distinct differences with respect to the albumins and PEA.Some observations indicate that PEA might interact with other seed proteins of pea.     

Evidence for spontaneous diploid androgenesis in Brassica napus L.

Summary Seeds of androgenetic origin were obtained among the F₁ progenies of two crosses between resynthesized and cultivated forms of Brassica napus. The high-erucic, white-flowered, resynthesized line No7076 acted as the female, and the zero-erucic, yellow-flowered, cultivars Topas and Puma, as males. No androgenetic seeds were obtained in the reciprocal crosses. Resynthesized rape could thus be of potential use for the production of androgenetic plants. Of special interest is the high frequency (21%) of spontaneous androgenesis observed in one of the two crosses. One plant, determined from erucic acid content and flower colour analysis as androgenetic, had a diploid chromosome number. Further knowledge about the genetic control of spontaneous androgenesis in the present material and the origin of the cytoplasm in androgenetic plants are required in order to exploit this phenomenon in practical plant breeding.     

Effects of herbivore identity on plant fecundity

Herbivores are pervasive, yet their effects on plant fecundity are often variable. One potential source of variation in herbivore impacts results from differing feeding modes of herbivores. We examined the relative importance of inflorescence-feeding insects versus large ungulates in affecting the fecundity of Balsamorhiza sagittata (Asteraceae), a dominant native perennial forb in western Montana, USA. We quantified these effects across four sites that varied in elevation to determine how consistent herbivore impacts were across these divergent sites. Insect herbivores were present in flower heads at all sites but they significantly depressed plant fecundity at two of the four sites. At the two sites where herbivore suppression had significant effects, this treatment let to 1.6 and 3.4-fold increases in seed production, respectively. In contrast, across all sites ungulates had minimal impacts on balsamroot flower and seed production. Seed addition experiments revealed that at some sites and in some years B. sagittata is seed-limited, as there was a positive relationship between seed input and seedling recruitment. Thus, reductions in seed production from heavy insect herbivory may limit balsamroot recruitment in some locations. Overall, results show that inconspicuous insects have stronger effects on balsamroot fecundity than do mammalian herbivores, but the magnitude of negative impacts at both the individual and population-level vary by site.