Carbon allocation during fruiting in Rubus chamaemorus

Background and Aims

Rubus chamaemorus (cloudberry) is a herbaceous clonal peatland plant that produces an extensive underground rhizome system with distant ramets. Most of these ramets are non-floral. The main objectives of this study were to determine: (a) if plant growth was source limited in cloudberry; (b) if the non-floral ramets translocated carbon (C) to the fruit; and (c) if there was competition between fruit, leaves and rhizomes for C during fruit development.

Methods

Floral and non-floral ramet activities were monitored during the period of flower and fruit development using three approaches: gas exchange measurements, ¹⁴CO₂ labelling and dry mass accumulation in the different organs. Source and sink activity were manipulated by eliminating leaves or flowers or by reducing rhizome length.

Key Results

Photosynthetic rates were lower in floral than in deflowered ramets. Autoradiographs and ¹⁴C labelling data clearly indicated that fruit is a very strong sink for the floral ramet, whereas non-floral ramets translocated C toward the rhizome but not toward floral ramets. Nevertheless, rhizomes received some C from the floral ramet throughout the fruiting period. Ramets with shorter rhizomes produced smaller leaves and smaller fruits, and defoliated ramets produced very small fruits.

Conclusions

Plant growth appears to be source-limited in cloudberry since a reduction in sink strength did not induce a reduction in photosynthetic activity. Non-floral ramets did not participate directly to fruit development. Developing leaves appear to compete with the developing fruit but the intensity of this competition could vary with the specific timing of the two organs. The rhizome appears to act both as a source but also potentially as a sink during fruit development. Further studies are needed to characterize better the complex role played by the rhizome in fruit C nutrition.Key words: Allocation pattern, ¹⁴C labelling, carbon translocation, carbon reserves, cloudberry, defoliation, fruit production, gas exchange, Rubus chamaemorus, source–sink relationship, flowering     

Between-year variation in flowering and fruit set in frost-prone and frost-sheltered populations of dioecious Rubus chamaemorus

Summary The flowering and fruiting patterns of the dioecious perennial herb Rubus chamaemorus L. were studied in frost-prone (open) and frost-sheltered (Shaded) habitats in northern Sweden over 6 years. The number of ramets with flower buds, the proportion of flower buds that opened, and fruit set varied markedly between years. In the frost-prone populations, the occurrence or absence of detrimental frosts during the development of flowers and fruit could explain much of the variation, both in the proportion of flower buds that developed into flowers, and in fruit set. In the frost-sheltered populations, most female flowers that did not develop into fruit aborted without any signs of physical damage and before any ovules had begun to enlarge. Flower mortality caused by herbivores feeding on reproductive parts was commonly low, but reached values higher than 10% in one of the shaded populations. Hand-pollination increased the proportion of ovules producing seeds in the mature fruits by about 20%, and in one year also increased fruit set significantly in one population. Fruit-producing female ramets had a higher mortality and a lower probability of flowering in the subsequent year than male ramets and non-fruiting female ramets. In R. chamaemorus, the conditions for fruit maturation are highly unpredictable at the time of flower initiation. It is suggested that the apparent over-initiation of flower buds is advantageous, as it allows the plant to attain a high reproductive success in years favourable for flowering and fruit development.     

In vitro Propagation of Cloudberry (Rubus chamaemorus)

The purpose of this study was to establish conditions for micropropagation of cloudberry (Rubus chamaemorus L.). Cultures were initiated from meristem cultures. When cultures were subcultured from clusters of 3–5 shoots, approximately 70 and 50 shoots were produced per cluster within 6 weeks at 8.9 μM BAP for the female cv. Fjellgull and the male cv. Apollen, respectively. Addition of 5.5 μM GA₃ reduced the number of shoots. Auxins (IBA, NAA) promoted root development in vitro, but inhibited formation of new shoots. However, as much as 85% of shoots rooted without auxin treatment when planted in a peat:sand (80:20 v/v) mixture. Some of the male plants regenerated from shoot tip cultures flowered in the greenhouse within a year after transfer to soil.     

Chlamydia trachomatis as a cause of abortion

The possible role of C. trachomatis as a causative agent of abortion in humans was investigated using the tissue culture method for the isolation of chlamydia. Two cases of spontaneous abortion and another seven of recurrent abortion due to C. trachomatis were positively identified. Among those with recurrent abortion were four patients with history of second trimester abortion.     

Seasonal changes in frost hardiness in cloudberry (Rubus chamaemorus) in relation to carbohydrate content with special reference to sucrose

Monthly determinations of frost hardiness of cloudberry ( Rubus chamaemorus L.) buds and rhizomes were done from October 1978 to October 1979. For buds LT₅₀ (lethal temperature for 50% of the plant material) was calculated from the percentage of bud breaking and for rhizomes from visual estimations of the degree of coloring by triphenyltetrazolium chloride. The frost hardiness varied from—11.5°C in November to—4°C in May to July for buds and from—16°C in January to—3°C in June— July for rhizomes. Dehardening started in February while the plants were still covered with snow. In connection with the determinations of frost hardiness, carbohydrate analyses were done. There was a good correlation between the degree of frost hardiness and the amount of soluble carbohydrates determined with anthrone. Sucrose, determined by gas chromatography, seemed to be the sugar contributing most in this correlation.     

Vegetative growth and frost hardiness of cloudberry (Rubus chamaemorus) as affected by temperature and photoperiod

The effect of photoperiod and temperature on growth and induction and development of frost hardiness in cloudberry ( Rubus chamaemorus L.) was examined in two experiments. The photoperiods were 8, 12 or 24 h and the temperatures were 18, 15, 12, 9, 4, 3, –3 or –4°C depending on the experiment. The level of hardiness was expressed as LT₆₆ or LT₅₀ (the lethal temperature for 66 or 50% of the plant material) for percentage of bud break and for the degree of coloring by triphenyltetrazolium chloride for rhizomes. The vegetative growth was clearly affected by daylength; petiole elongation, leaf growth, shoot dry weight and number of shoots per plant were all reduced under short days compared with long days. However, the photoperiod had no significant effect on hardening of buds or rhizomes. Hardening increased with successively decreasing temperatures. To get the maximum hardiness, plants had to be exposed to freezing temperatures.     

The Carbon Economy of Rubus chamaemorus L. II. Respiration

Respiratory activity and seasonal changes in carbohydrate contentof the storage organs of Rubus chamaemorus L. have been investigated.Leaf dark respiration rate increases in a non-linear mannerfrom 0·7 mg CO₂ evolved dm^–2 h^–1 at 0 °Cto 4·6 rng CO₂ evolved dm^–2 hh^–1 at 30 °C.Root and rhizome respiration rates increase from 1 µ1O₂ uptake g^–1 fresh weight h^–1 at 0.7 ° C to10 µ10, uptake g^–1 f. wt h^–1 at 20 °C.Rhizome carbohydrate reserves decline from a September peakof 33 per cent alcohol insoluble d. wt to 16 per cent in May. The circumpolar distribution of R. chamaemorus is discussedin relation to the evidence presented here and in the precedingpaper of the series.     

The Carbon Economy of Rubus chamaemorus L. I. Photosynthesis

Studies on the photosynthetic activity of Rubus chamaemorusL. in controlled environment conditions are reported. Theseshow that material collected from Moor House National NatureReserve, England has a photosynthetic light saturation pointof 100 J m^–2s^–1 (380–720 nm) and a temperatureoptimum for photosynthesis between 10 and 15 °C. A markeddecline in net CO₂ uptake is evident at temperatures in excessof 18 ° C; this persists for some time after return to anoptimum temperature regime. Leaves show rapid responses to changesin both light intensity and temperature, and show no evidenceof an endogenous rhythm in photosynthetic rate.     

Sex Ratios and Resource Allocation among Sexually Reproducing Plants of Rubus chamaemorus

Sex ratios and patterns of size variation and resource allocationwere investigated in the dioecious species Rubus chamaemorus.Sex ratios among flowering ramets varied from 6% to 40% of females.Female ramets were slightly, although not significantly, tallerthan males. It appeared that population effects (including bothgenetic population and environmental site effects) on plantsize and allocation patterns at flowering are considerably greaterthan sex effects. If both flowering and fruit production areconsidered, then female allocation to reproduction clearly exceedsmale allocation. In females, no significant relationship wasdetected between the mass of reproductive and vegetative tissues,while males did exhibit such a relationship. Reproductive effortwas less for tall males than for small males. Despite the occurrence of sexual reproduction, the main modeof reproduction in R. chamaemorus is vegetative propagation,which is the best strategy for reproduction in the unpredictableclimate of high latitudes but which leads to skewed sex ratios.As a consequence of vigorous vegetative reproduction, individualclones can grow to be large. The results of electrophoreticstudies show that the numbers of clones per population are low.Copyright1994, 1999 Academic Press Rubus chamaemorus, cloudberry, sex ratios, resource allocation, clonal structure, electrophoresis     

Clonal structure of Rubus chamaemorus populations: comparison of different molecular methods

The clonal structure of Rubus chamaemorus populations was investigated using DNA fingerprinting. The PCR-based methods included the use of 10-base RAPD primers and 16-base simple sequence repeat primers. In the hybridization method variation was studied using hypervariable multilocus probes, one derived from the M13 bacteriophage and the other a synthetic (AC)/(TG) polynucleotide. Although R. chamaemorus expresses clear variation in morphology, the level of genetic differentiation appears to be fairly low. The observed numbers of clones in the three populations examined in Finland varied from 2 to 4. The total number of genotypes across populations was 5, of which one was unique. The results obtained using the two fingerprinting methods were comparable but lead to a slightly different grouping of clones.     

Where did the northern peatland species survive the dry glacials: cloudberry (Rubus chamaemorus) as an example

Aim During the last ice age large parts of the north boreal and subarctic zones were covered by ice, while the climate in ice‐free regions of northern Asia was extremely cold and dry. The extensive peatlands of these zones with their characteristic vegetation developed at the beginning of the Holocene. We combine a phylogeographical approach with maps of pollen records to identify regions where Rubus chamaemorus, a plant of moist, peaty soils, was likely to grow during this period. Location Circumarctic/circumboreal. Methods Samples were collected from 45 locations throughout much of the range of R. chamaemorus and 398 plants were analysed with amplified fragment length polymorphism (AFLP). Estimates of diversity and differentiation, principal coordinates analysis and Bayesian clustering methods were used for the analysis of genetic data. Dated pollen records were retrieved from the European and the Global Pollen Databases. Results The plants from Sakhalin are highly divergent from the rest of the material and represent the previously described var. pseudochamaemorus. The main genetic division in R. chamaemorus sensu stricto is found in the Taymyr region in central Eurasia. Genetic diversity and the relative number of rare markers are highest in central Siberia and eastern Asia and decrease towards Europe and to a lesser extent eastwards through North America. Pollen dating back to the last ice age is found in central and eastern Siberia and Alaska. The maximum observed clone size is about 250 m, and more than one clone is found in nearly every local population. Main conclusions The genetic data are consistent with the pollen records and indicate that R. chamaemorus was growing in several areas of northern central Siberia and Beringia during the last glaciation. This finding suggests that sufficient humidity for this and other species of peaty soils was present locally in different parts of the generally dry ice‐free areas of northern Asia, as had been previously documented for Beringia. The AFLP data show that var. pseudochamaemorus, which is also morphologically quite divergent, clearly represents a distinct genetic entity.     

Intersexual differences in phenology and damage by herbivores and pathogens in dioecious Rubus chamaemorus L.

Summary Shoot development and damage by herbivores and pathogens to male, female and non-floral ramets of the dioecious, perennial herb Rubus chamaemorus were studied in the field during three consecutive years. Leaves on male ramets were usually consumed more by herbivores and attacked more by fungi than were leaves on female ramets. Male ramets unfolded their leaves later than did female ramets. In 1983, when the level of herbivory was comparatively high, ramets that were fully developed in early June were more damaged by herbivores than were ramets that unfolded their leaves later. Non-floral ramets usually showed intermediate levels of damage compared to male and female flowering ramets.Defoliation caused a greater increase in ramet mortality among females than among males in a field experiment. It is suggested that the different reproductive roles of males and females differently constrain the evolution of defense against herbivores and pathogens in dioecious plants. This may result in intersexual differences both in palatability to herbivores and susceptibility to pathogens.     

Pollination ecology of the high latitude, dioecious cloudberry (Rubus chamaemorus; Rosaceae)

In a 3-yr study, we examined the pollinator guild and intersexual floral characteristics of the dioecious, perennial cloudberry (Rubus chamaemorus), which flowers in early spring. The findings contribute to our general understanding of pollination ecology at high latitudes and provide important information for the commercialization of cloudberry. Female flowers were smaller than males but provided more nectar, although this resource was low in both sexes. Insects from 43 families visited cloudberry flowers, yet four families (Apidae, Halictidae, Muscidae, Syrphidae) represented ca. 87% of all visitors observed. Introduction experiments revealed that apids and muscids are significantly poorer pollinators (based on fruit production) than halictids and syrphids, but when fruit mass or seed set was considered, there were no significant differences between families. Pollinator importance, a product of flower visitation frequency and seed set effectiveness, revealed that the dipterans were of paramount importance to the pollination of cloudberry. Furthermore, they are limited to cloudberry because their lapping mouthparts exclude them from accessing the nutritional rewards of competing Ericaceae flowers. While the total number of pollinator families observed suggest a generalist pollination system, if one considers the dominant pollinators (flies) as a functional group, then this insect-flower relationship could be considered a specialized one.     

Genetic differentiation of Rubus chamaemorus populations in the Czech Republic and Norway after the last glacial period

The population structure of cloudberry (Rubus chamaemorus L.), collected from Krkonose Mountains (the Czech Republic), continental Norway and Spitsbergen, was examined using microsatellite analyses (SSR). Among 184 individuals, 162 different genotypes were identified. The overall unbiased gene diversity was high (). A high level of genetic differentiation among populations (F_ST = 0.45; p < .01) indicated restricted gene flow between populations. Using a Bayesian approach, six clusters were found which represented the genetic structure of the studied cloudberry populations. The value of correlation index between genetic and geographical distances (r = .44) indicates that gene flow, even over a long distance, could exist. An exact test of population differentiation showed that Rubus chamaemorus populations from regions (Krkonose Mountains, continental Norway and Spitsbergen) are differentiated although some individuals within populations share common alleles even among regions. These results were confirmed by AMOVA, where the highest level of diversity was found within populations (70.8%). There was no difference between 87 pairs of populations (18.7%) mostly within cloudberry populations from continental Norway and from Spitsbergen. Based on obtained results, it is possible to conclude that Czech and Norwegian cloudberry populations are undergoing differentiation, which preserves unique allele compositions most likely from original populations during the last glaciation period. This knowledge will be important for the creation and continuation of in situ and ex situ conservation of cloudberry populations within these areas.     

Pollination by deceit,floral sex ratios and seed set in dioecious Rubus chamaemorus L.

Summary Male and female flowers of the dioecious perennial herb Rubus chamaemorus L. are similar in general appearance. However, female flowers are somewhat smaller, do not produce any pollen, and contain very small amounts of nectar. Syrphids and bumblebees, which are important pollinators of R. chamaemorus, showed a strong preference for male flowers. Male flowers were also less often rejected by flower visitors than were female flowers, and two different groups of syrphid species stayed longer in male than in female flowers. These observations suggest that female flowers of R. chamaemorus attract pollinators by deceit.Hand-pollination experiments indicated that pollen availability limited seed production of R. chamaemorus in female dominated habitats but not in areas with an equal floral sex ratio. We suggest that the relative importance of factors limiting female reproductive success is not constant, but is influenced by the floral sex ratio of the population. This should apply also to other dioecious species that show variable sex ratios on either a local or regional scale.     

Seed and fruit abortion as a process of self organization among developing sinks

Fruit and seed abortion in plants have been traditionally attributed to limiting factors such as resources or to chemically mediated dominance hierarchy among the developing sinks. These hypotheses however do not satisfactorily explain the observed patterns of seed and fruit abortion and have also not withstood the test of critical experimentation. In this paper, we propose an alternative model based on the process of self organization used in physics to explain the flow of liquids etc. As proposed here, the model assumes one simple rule, that the probability of any given sink getting a resource molecule is a function of (a) the sink-drawing ability of each unit of the tissue and (b) the amount of resource molecules already moved to that sink. Accordingly, any resource molecule moving to a sink, autocatalytically increases the probability of the latter getting further resources.
We show that starting from a set of identical sinks, all with equal sink-drawing ability, the differential development of sinks and thence their abortion might occur purely as a consequence of the autocatalytic or feedback movement of the resource molecules to the developing sinks. We also show that, such a process can lead to abortion even under resource-abundant conditions, though resource limitation aggravates the extent of abortion. Further, the extent of abortion is shown to be highly dependent on the sink-drawing ability of the species. We offer the test of this prediction in two ways: (a) in Derris indica the extent of abortion increases with the sink drawing ability of the fertilized ovules and (b) treatments involving growth regulators that increase the sink-drawing ability result in increased abortion of seeds and fruits. Based on these, we argue that self organization could be the basic process regulating resource flow to sinks and factors such as resource limitation and chemical inhibition might be additionally aggravating the asymmetry among the developing sinks.     

Mediterranean fruit fly as a potential vector of bacterial pathogens

The Mediterranean fruit fly (Ceratitis capitata) is a cosmopolitan pest of hundreds of species of commercial and wild fruits. It is considered a major economic pest of commercial fruits in the world. Adult Mediterranean fruit flies feed on all sorts of protein sources, including animal excreta, in order to develop eggs. After reaching sexual maturity and copulating, female flies lay eggs in fruit by puncturing the skin with their ovipositors and injecting batches of eggs into the wounds. In view of the increase in food-borne illnesses associated with consumption of fresh produce and unpasteurized fruit juices, we investigated the potential of Mediterranean fruit fly to serve as a vector for transmission of human pathogens to fruits. Addition of green fluorescent protein (GFP)-tagged Escherichia coli to a Mediterranean fruit fly feeding solution resulted in a dose-dependent increase in the fly's bacterial load. Flies exposed to fecal material enriched with GFP-tagged E. coli were similarly contaminated and were capable of transmitting E. coli to intact apples in a cage model system. Washing contaminated apples with tap water did not eliminate the E. coli. Flies inoculated with E. coli harbored the bacteria for up to 7 days following contamination. Fluorescence microscopy demonstrated that the majority of fluorescent bacteria were confined along the pseudotrachea in the labelum edge of the fly proboscis. Wild flies captured at various geographic locations were found to carry coliforms, and in some cases presumptive identification of E. coli was made. These findings support the hypothesis that the common Mediterranean fruit fly is a potential vector of human pathogens to fruits.     

An assessment of tree availability as a possible cause of population declines in scavenging raptors

Lack of suitable nesting trees is an increasingly common issue for avian conservation given rampant habitat and tree destruction around the world. In the African savannah, habitat loss and particularly tree damage caused by elephants have been suggested as possible factors in the decline of large bird species. Given the recent declines of vultures and other scavenging raptors, it is critical to understand if nest availability is a limiting factor for these threatened populations. Loss of woodland, partially due to elephant populations, has been reported for the Mara‐Serengeti ecosystem. Data on characteristics of trees used for nesting were collected for white‐backed, lappet‐faced, white‐headed vulture, and tawny eagle nests in Masai Mara National Reserve, Kenya. Nest tree characteristics were compared with the distribution of a random subsample of trees to assess nest preferences and determine suitability of available trees. Nearest neighbor distances were estimated as well as availability of preferred nesting trees to determine if tree availability is a limiting factor for tree‐nesting vultures. Tree availability was found to greatly exceed nesting needs for African vultures and tawny eagles. We thus conclude that on a landscape scale, tree availability is not a limiting factor for any of the species considered here (white‐backed, lappet‐faced, white‐headed vultures and tawny eagles).