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.     

Stylar trimorphism in four functionally andromonoeciousProsopis species (Mimosaceae)

The reproductive system ofProsopis chilensis, P. pugionata, P. flexuosa, andP. torquata is described. Observations were made of floral morphology, stigmatic receptivity, pollen grain viability, and the nature of pollen reserves. Scanning electrone microscopy and statistical analysis were carried out. The results allow us to conclude that: 1) The studied species have three distinct floral morphs characterized by significantly different style lengths and, to a lesser extent, different stamen lengths; 2) style precocity found in all species does not indicate protogyny; 3) flowers with shortest styles are not receptive; 4) the pollen grains of all floral morphs showed high viability; 5) peroxidase is present in both the pollen and pistil; 6) viable pollen grains invariably have both starch and pollenkitt; 7) the studied species are andromonoecious.     

Chromosome numbers of European blackberries (Rubus subg.Rubus,Rosaceae)

Chromosome numbers are presented for 32 collections of 29 European blackberry species (Rubus subg.Rubus) from Germany. One species is triploid (2n = 21), 27 species are tetraploid, (2n = 28), and one species is pentaploid (2n = 35). Chromosome numbers are reported for the first time ofR. adspersus, R. amisiensis, R. calvus, R. conothyrsoides, R. contractipes, R. demissus, R. elegantispinosus, R. ferocior, R. foliosus, R. hypomalacus, R. leucandrus, R. nemorosus, R. platyacanthus, R. praecox, R. rhombifolius, andR. rhytidophyllus. Chromosome numbers forR. dasyphyllus, R. gelertii, R. glandithyrsos, R. lamprocaulos, R. lindebergii, R. macrophyllus, R. montanus, R. muenteri, R. pedemontanus, R. polyanthemus, R. senticosus, R. silvaticus, andR. vigorosus are confirmed.     

DNA fingerprint variation in some blackberry species (Rubus subg.Rubus,Rosaceae)

We have analysed samples from Sweden, Denmark, and Germany of six facultatively apomictic blackberry species to investigate the accordance between a taxonomy based on morphological characters on the one hand, and distribution of genetic variation estimated by DNA fingerprinting on the other hand. DNA fingerprint variation was found to be quite restricted in all species investigated. The first taxonomic group included three species related toRubus nessensis, two being characterized by one very widespread DNA fingerprint in all three countries and a few rare ones, whereas the third species differed between Sweden and Germany. The second taxonomic group included species related toR. gracilis. Two of these species exhibited very similar DNA fingerprints, whereas the third species deviated clearly. The utilization of DNA fingerprinting as a tool in taxonomy is discussed; most likely this method could become a useful complement to morphology, especially in plant groups with reduced levels of genetic recombination.     

Crossing experiments within theCampanula dichotoma group (Campanulaceae)

Four supposedly closely related taxa were crossed artificially to test for hybridization barriers. Intra- as well as inter-specific crossings were made. The intra-specific crossings resulted in a high portion of viable hybrids, whereas hybridization barriers were found in all inter-specific combinations except one. The hybrids were non-viable at different levels, viz., (1) no seeds germinated, (2) seeds formed lethal chlorotic seedings, or (3) seeds formed chlorophyllous seedlings which survived the cotyledonary stage, but developed into intermediate forms with low pollen production and/or a strongly reduced pollen viability. Crossings betweenCampanula afra and any of the other three taxa resulted in a non-viable progeny. The crossing capacity amongC. occidentalis, C. kremeri, andC. dichotoma varied. Combinations with large-flowered maternal taxa and small-flowered paternal ones usually failed, whereas a high number of the reciprocal crossings produced hybrids which survived the cotyledonary stage. The hybrid plants varied in pollen viability and pollen production, however. It is concluded that theCampanula dichotoma group should be treated as comprising four distinct species.     

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.     

The evaluation of pollen quality,and a further appraisal of the fluorochromatic (FCR) test procedure

Summary Methods currently available for evaluating pollen quality in vitro include, (a) tests of germinability; (b) tests of the stainability of the vegetative cell contents; (c) tests for enzyme activity, and (d) the fluorochromatic procedure (FCR), which tests principally the integrity of the plasmalemma of the vegetative cell. Using germinability in vitro as a standard, a comparison has been made between histochemical methods of classes (b), (c) and (d) in application to various pollens, immature, mature, and treated in ways known to affect viability and membrane state. Predictably, the lowest correlation was obtained with tests of stainability. The highest was given by the FCR, which generally provided an excellent guide to potential germinability. The FCR procedure is subject to various limitations, however, (a) A high correlation between FCR and germinability can only be expected when mature, ripe pollen is used; with immature pollen, the FCR will predict excessively high potential germinability. (b) The FCR may also predict a higher potential level of pollen function than in vitro germinability when the germination medium is sub-optimal. In this situation, however, it will generally give a better guide to fertilising capacity, (c) The FCR is not a test of pollen viability. Like germinability in vitro, it can yield a negative score with pollen which is nevertheless capable of functioning. For example, false negatives will be obtained with some species if the pollen is not properly pre-conditioned by rehydration before testing, an important point in monitoring stored pollen. The paper includes a brief discussion of the rationale of pollen testing.     

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.     

Germination-associated events and the desiccation sensitivity of recalcitrant seeds — a study on three unrelated species

The storage behaviour of recalcitrant seeds was assessed using three diverse species: a gymnosperm, Araucaria angustifolia (Bert.) O. Kuntze; a herbaceous monocotyledon, Scadoxus membranaceus (Bak.) Friis Nordal; and a woody dicotyledon, Landolphia kirkii Dyer. Seeds were stored under conditions of high relative humidities that maintained seed moisture content and under low relative humidities that caused drying. At regular intervals moisture content was determined, germinability assessed and the ultrastructure of radicle meristem cells examined. Under storage at high relative humidity, seed moisture content was maintained at the original level and subcellular germination events were initiated in the short-term. Such seeds showed enhanced rates of germination when removed from storage and planted. Long-term storage under these conditions resulted in the initiation of subcellular damage which intensified with time and ultimately resulted in the loss of viability. The rate at which germination events proceeded varied among the three species, and could be directly correlated with the period of viability retention under humid storage conditions. Storage under desiccating conditions resulted in subcellular damage and rapid loss of viability. The rate at which the seeds dried varied among the three species. The proportion of water loss tolerated by the different species before loss of viability, correlated with the rate of drying. The storage behaviour of the seeds of these three species is discussed in terms of a previously described model.     

Isolation of protoplasts from edible seaweeds

Protoplasts were isolated enzymatically from three species of Chlorophyta (Enteromorpha linza, Monostroma zostericola andUlva pertusa) with high yield and viability. An enzyme solution appropriate for protoplast isolation from the marine green algae was the following: 2% Cellulase Onozuka R-10, 1.0.M mannitol, pH 6.0. Protoplasts could not be obtained from members of Phaeophyta or Rhodophyta.     

Breeding system and crossability in JapaneseEpimedium (Berberidaceae)

A series of experimental pollinations involving six species ofEpimedium provided strong evidence for an outbreeding system and no internal (postmating, postpollination) barrier to hybridization in the taxa concerned. Self-pollinations of four species,E. diphyllum, E. trifoliatobinatum, E. grandiflorum andE. sempervirens indicated high self-incompatibility (at most 5.4%, usually 0% in capsule-set). Field experiments in a population revealed the occurrence of cross-pollination. On the other hand, interspecific cross-pollinations showed high crossability at any combination of species (33.3%–100% in capsule-set). Furthermore, the interspecific F₁s obtained germinated at a high rate (usually more than 20%) and three of them, which bloomed, are highly fertile (more than 68.6% in pollen viability). The results were discussed in connection with the isolating mechanisms between the species ofEpimedium.     

Variation in life-cycle between three rare and endangered floodplain violets in two regions: implications for population viability and conservation

We studied the demography of Viola elatior, V. pumila, and V. stagnina, three rare and endangered Central European floodplain species, to (i) analyse variation in life-cycles among congeners and between regions (Dyje-Morava floodplains, Czech Republic; Upper Rhine, Germany), (ii) to define sensitive stages in the life-cycles, and (iii) to identify possible threats for population viability and species conservation. Matrix models were based on the fate of marked individuals from a total of 27 populations over two years. We analysed population growth rate (λ), stage distribution, net reproductive rate (R ₀), generation time, age at first reproduction, and elasticity and calculated a life table response experiment (LTRE). Most populations were declining and λ did not differ between species or regions during the observed interval. Despite higher probabilities for survival and flowering in the Dyje populations, R ₀ was higher in the Rhine populations. Also other demographic traits showed consistent differences between regions and/or species. Complex life-cycles and large variation in λ precluded unequivocal identification of sensitive stages or vital rates for conservation. Variation between regions may be a consequence of differences in habitat quality. Our results suggest that deterministic processes such as reduced management, succession, habitat destruction, and lack of disturbance through reduced or eliminated flooding present the strongest threat for the viability and persistence of populations of the three floodplain violets as compared with stochastic processes. However, the persistent seed bank of the species may buffer populations against environmental variation and represents a reservoir for recovery after resumption of suitable land-use management.     

Genetic analysis of natural hybrids between endemic and alien Rubus (Rosaceae) species in Hawai`i

A population of putative hybrids between theendemic Rubus hawaiensis and naturalizedR. rosifolius was discovered inKpahulu Valley, on the island of Maui inthe Hawaiian archipelago. The goal of thisstudy was to molecularly characterize thisnatural hybridization event, investigate themode of hybridization, and determine the malefertility of the hybrid individuals. Bothmorphological and RAPD marker data indicatethat the putative hybrid individuals are theprogeny of R. rosifolius and R.hawaiensis. All 39 hybrid individuals sampledhad the chloroplast DNA haplotype of R.rosifolius. Thus hybridization appears to beasymmetric, with R. rosifolius acting asthe maternal parent. All hybrid individualsassessed for pollen stainability were sterile,and there was no evidence of backcrossing toeither parent. This result suggests thathybrids are of the first filial generation andthat variation among hybrids reflectsdifferences within the parental populations.Sympatric populations of R. hawaiensisand R. rosifolius occur on four islandsand six additional alien species of Rubusare naturalized and sympatric with R. hawaiensisin Hawai`i. Further investigationis merited to assess whether hybridization maypose a threat to the long term viability ofR. hawaiensis. This study highlights theincreasing frequency and negative consequencesof native-alien hybridization and theimportance of maintaining active alien speciescontrol programs in the Hawaiian Islands.     

3种雌雄异熟樟科植物开花动态的比较研究

为了解樟科植物雌雄异熟的繁育系统特点,对3种樟科植物阴香(Cinnamomum burmannii)、紫楠(Phoebe sheareri)和浙江楠(Phoebe chekiangensis)雌雄异熟花的开花动态进行了比较研究.结果表明,3种植物雌性功能期的开始时间、雌性功能期和雄性功能期的时间分配有差异.3种植物的主...     

Distinct life histories of peracarid crustaceans in a Ria Formosa salt marsh (S. Portugal)

Pitfall traps were exposed between February 1993 andSeptember 1994 every 4th to 6th week near the highwater mark in a Ria Formosa salt marsh (Portugal).With only a few exceptions, peracarid crustaceansaccounted for more than 95% of the individualscaught. Three amphipod species (Orchestiagammarellus(Pallas), Orchestia mediterraneaCosta, Talorchestia deshayesii(Audouin)) and3 isopod species (Tylos ponticus(Arcangeli),Halophiloscia couchi(Kinahan) and Porcellio lamellatus(Uljanin) Budde-Lund) wereidentified. In general, these species were most activefrom spring to autumn.Data collected included information on growth rate,life expectancy and timing of reproduction. Theamphipods displayed more opportunistic life-historypatterns with high growth rates, reproductive activityduring most of the year, early sexual maturity andrelatively short life expectancies. The isopods weremore persistent with slower growth rates, morerestricted reproductive periods and longer lifeexpectancies.There is no indication in the literature that most ofthese species are particularly common in salt marshes.In addition, peracarid crustaceans are not mentionedas dominant taxa in most studies of salt marsh fauna.We suggest that the peracarids play an important rolein the degradation of organic matter in salt marshesand hypothesize that the high numbers of peracaridsfound in Ria Formosa is due to the high contributionof leaf shedding plants.     

Reproductive systems and crossing potential in three species ofBulbophyllum (Orchidaceae) occurring in Brazilian ‘campo rupestre’ vegetation

Experimental self-, cross- and interspecific pollinations were carried out inBulbophyllum weddellii, B. involutum andB. ipanemense (Orchidaceae). The last two species are closely related, butB. weddellii is somewhat more distant. The three species are self-compatible, with pollination by a vector necessary for fruit formation. In all crossing types, high rates of fruit abortion and formation of seeds without embryos were observed, particularly in interspecific crosses that involvedB. weddellii. Crosses between the more closely relatedB. involutum andB. ipanemense produced fruit formation rates and seed viability similar to those obtained in intraspecific crosses. These results agree with current hypotheses that suggest that interspecific crossing rates reflect phylogenetic proximity. Examination of pollen tube growth and aborted fruits suggests that a series of factors may be involved in the high abortion rate for fruits and large proportion of seeds without embryos. The low interspecific fertility noted forB. weddellii andB. involutum is important in the maintenance of isolation between these species that are sympatric, flower in the same period and share the same pollinators.     

Suitability of different media for in vitro cultivation of the ruminal protozoa species Entodinium caudatum, Eudiplodinium maggii, and Epidinium ecaudatum

Three protozoal cultivation media were tested to determine the medium which best facilitated growth and viability of key B-type ciliates isolated from the sheep rumen. Entodinium caudatum and Eudiplodinium maggii were grown anaerobically in 50-ml flasks for 32 days in Caudatum-type (C), Kisidayova (K) or Dehority (M) medium. On day 32, in media K and M, E. caudatum cell counts were high with 5.6 × 10³ and 7.8 × 10³ mL⁻¹, respectively, and the proportion of dead cells was low with 0.6 and 1.4%, respectively. E. maggii concentrations when grown in medium M and C were 2.7 × 10³ and 2.4 × 10³ mL⁻¹, respectively, with 3.9 and 14.1% dead cells. Medium M, which favoured growth of both protozoa species, was tested again and Epidinium ecaudatum was included. Protozoa were grown for a 4-month period and samples were taken in the last two months on days 1, 7, 35 and 57. Average cell concentrations were 10.0, 0.8 and 0.5 × 10³ mL⁻¹ for E. caudatum, E. maggii, and E. ecaudatum, respectively. In conclusion, medium M would appear to be the best choice for cultivating these three species in one medium.     

On the control of HAB species using low biosurfactant concentrations

Biosurfactants have been suggested as a method to control harmful algal blooms (HABs), but warrant further and more in-depth investigation. Here we have investigated the algicidal effect of a biosurfactant produced by the bacterium Pseudomonas aeruginosa on five diverse marine and freshwater HAB species that have not been tested previously. These include Alexandrium minutum (Dinophycaee), Karenia brevis (Dinophyceae), Pseudonitzschia sp. (Bacillariophyceae), in marine ecosystems, and Gonyostomum semen (Raphidophyceae) and Microcystis aeruginosa (Cyanophyecae) in freshwater. We examined not only lethal but also sub-lethal effects of the biosurfactant. In addition, the effect of the biosurfactant on Daphnia was tested. Our conclusions were that very low biosurfactant concentrations (5 μg mL⁻¹) decreased both the photosynthesis efficiency and the cell viability and that higher concentrations (50 μg mL⁻¹) had lethal effects in four of the five HAB species tested. The low concentrations employed in this study and the diversity of HAB genera tested suggest that biosurfactants may be used to either control initial algal blooms without causing negative side effect to the ecosystem, or to provoke lethal effects when necessary.     

三种厚朴叶绿体基因组的比较研究

为了深入发掘日本厚朴、厚朴、凹叶厚朴叶绿体基因组差异,筛选厚朴优良性状候选基因,开展三种厚朴的分子遗传研究,该文利用Illumina HiSeq高通量测序平台首次对日本厚朴叶绿体进行测序、组装,并与已有的厚朴、凹叶厚朴叶绿体基因组共同注释,获得三个物种叶绿体基因图谱,筛选出三个基因组中的差异基因,又与同科中11个亲缘物种进行叶绿体基因组比对,构建NJ遗传树。结果表明：(1)日本厚朴叶绿体基因组的Clean Reads为19 791 019,Q30为91.33%,组装后基因组全长160 051 bp, GC含量为39.2%,含tRNA 37个,rRNA 8个。(2)比对分析发现三种厚朴具有相似的IR、LSC和SSC结构,以及GC含量和tRNA数量,但编码基因种类和数量、内含子和外显子的数量和结构等存在差异。(3)日本厚朴的功能基因数目较厚朴、凹叶厚朴分别多6个和4个,主要分布于LSC区和IR区,涉及核糖体大亚基、核糖体小亚基和未知功能基因类群。(4)系统发育分析结果进一步显示日本厚朴与凹叶厚朴亲缘关系较近,其次是厚朴。该研究表明日本厚朴具有更丰富的叶绿体基因组结构、组成和变异特征,是其适...     

Thermotolerance of Photosystem 2 of Three Alpine Plant Species Under Field Conditions

The species specific response of photosystem 2 (PS2) efficiency and its thermotolerance to diurnal and seasonal alterations in leaf temperature, irradiance, and water relations were investigated under alpine field conditions (1 950 m) and in response to an in situ long-term heat treatment (+3 K). Three plant species were compared using the naturally occurring microstratification of alpine environments, i.e. under contrasting leaf temperatures but under similar macroclimatic conditions. Thermotolerance of PS2 showed a high variability in all three species of up to 9.6 K. Diumal changes (increases or even decreases) in PS2 thermotolerance occurred frequently with a maximum increase of +4.8 K in Loiseleuria procumbens. Increasing leaf temperatures and photosynthetic photon flux density influenced thermotolerance adjustments. Under long-term heating (+3 K) of L. procumbens canopies with infra-red lamps, the maxima of the critical (T_c) and the lethal (T_p) temperature of PS2 increased by at least 1 K. Thermotolerance of the leaf tissue (LT₅₀) increased significantly by +0.6 K. The effects of slight water stress on thermotolerance of PS2 were species specific. High temperature thresholds for photoinhibition were significantly different between species and increased by 9 K from the species in the coldest microhabitat to the species in the warmest. Experimental heating of L. procumbens canopies by +3 K caused a significant (p>0.01) upward shift of the high temperature threshold for photoinhibition by +3 K. Each species appeared to be very well adapted to the thermal conditions of its microhabitat as under the most frequently experienced daytime leaf temperatures no photoinhibition occurred. The observed fine scale thermal adjustment of PS2 in response to increased leaf temperatures shows the potential to optimise photosynthesis under varying environmental conditions as long as the upper thermal limits are not exceeded.