Development of microsatellite markers for the bracken fern, Pteridium aquilinum

We isolated eight novel polymorphic microsatellite loci from Pteridium aquilinum. These loci were characterized in 30 individuals, one from Bolivia, two from Peru, one from the USA, one from Japan, and 25 from Northeast China to Southwest China. The number of alleles per locus ranged from two to seven. The observed heterozygosity (H(O) ) ranged from 0.000 to 0.600 with an average of 0.3051, and the expected heterozygosity (H(E) ) ranged from 0.0966 to 0.7780 with an average of 0.4267. One locus deviated from Hardy-Weinberg equilibrium and four pairs of loci were found to be in linkage disequilibrium. These polymorphic loci will be useful in the study of the population genetic structure of Pteridium.     

Estimates of gene flow, genetic substructure and population heterogeneity in bracken (Pteridium aquilinum)

Bracken [ Pteridium aquilinum (L.) Kuhn] is a cosmopolitan species and is a noxious weed in many areas. Because of its abundance, particularly in Britain, bracken affords an ideal system for investigating various aspects of population genetics and evolution. High mobility of dispersal units (spores) suggests that rates of gene flow among distant populations should be high. Gene flow is a major evolutionary force that influences the genetic structure of populations. To examine the effects of gene flow on population heterogeneity and population substructuring in bracken, starch gel electrophoresis of enzymes was used to provide the necessary genetic database. Allele frequency data at 21 loci were obtained for seven British populations, one Majorcan and one from the eastern United States. A model was employed to estimate the amount of gene flow ( Nm ) at several levels. Gene flow among British populations was extremely high ( Nm = 36.51), one of the highest estimates reported for plants. Among eight European populations gene flow was lower (but still considered high) at Nm = 2.47. Trans-Atlantic gene flow was low ( Nm = 0.0926).
F -statistics were used to assess population heterogeneity and substructuring. The data indicate that, compared with other species, there is very little genetic differentiation among British populations of bracken. Indeed, it appears that the whole island is behaving as a single randommating population. This result is consistent with high levels of gene flow. Only one population (on the Isle of Arran) showed statistically significant genetic substructuring. Habitat heterogeneity on the island and age structure are hypothesized as possible causes of this result.
The data reported here support previous studies demonstrating that bracken is genetically polymorphic and is an outcrossing species.     

The influence of position on genet growth: a simulation of a population of bracken (Pteridium aquilinum (L.) Kuhn) genets under grazing

In a spatially explicit simulation model of vegetation dynamics (VegeTate), I labelled the initial mass of Pteridium aquilinum in each of 225 cells as a single, unique genet or clone. The physical environment was homogeneous and all genets shared the same phenotype. The aim was to discover whether and how the success of each genet was affected by its initial position relative to other genets and a competing grass species. In a scenario in which grazing generated a grass-bracken mosaic with complex spatial dynamics, the amount of growth of each genet ranged widely, from frequent extinction to mass increase by over 300 times. The main factor in the impact of position on genet growth was shown to be a benefit from the initial presence or proximity of a large mass of P. aquilinum. This was because a high density of P. aquilinum reduced local grazing intensity, allowing plant mass to accumulate and shifting the balance of competition in favour of P. aquilinum. Thus variations between cells in initial mass of P. aquilinum were greatly amplified. The implications of this amplification of initial differences between sites for population genetics are briefly discussed. Qualitative features of the spatial distribution of genets at the end of simulations matched reported observations on patchily distributed field populations of P. aquilinum. These features included dominance of a large population by a small minority of genets, widespread mixtures of a dominant genet and one or more subordinate genets, and the presence of patches of P. aquilinum formed both by agglomeration from neighbouring foci and by spread of dominant genets. Under less intense grazing, which allowed little or no development of vegetation mosaics, genet growth varied relatively little and initial variations in relative mass between genets were little changed. Based on this study, I hypothesize that any processes that generate non-linear spatial dynamics will also generate complex genet dynamics.     

蕨配子体发育及卵发生的显微结构观察

运用显微观察技术对蕨(Pteridium aquilinum var. latiusculum)配子体发育和卵发生进行了研究。结果表明：(1)蕨孢子黄褐色,四面体形,具三裂缝,接种后3～7 d萌发,经丝状体和片状体阶段发育成原叶体,成熟原叶体雌雄异株或同株。(2)蕨颈卵器产生于生长点下方的表面细胞(颈卵器原始细胞),该细胞经2次分裂形成3层细胞,其上层和下层细胞发育为颈卵器壁细胞,中间细胞为初生细胞,它经2次不等分裂产生3个细胞,分别为卵细胞、腹沟细胞和颈沟细胞;刚产生时,此3个细胞紧贴颈卵器壁,细胞质内液泡较多,随着发育,卵细胞和腹沟细胞之间产生了分离腔,但二者通过孔区相连,在卵细胞上表面可观察到卵膜;此后,颈沟细胞和腹沟细胞逐渐退化,颈卵器壁细胞内具有黑色颗粒物质。连续切片观察发现,成熟卵细胞上表面中央具有受精孔。卵发生的细节尚需超微结构的研究。     

蕨菜多酚氧化酶的酶学性质

研究了蕨菜[Pteridium aquilinum (L．) Kuhn var．latiusculum (Desv．) Underw．]多酚氧化酶的动力学性质,结果表明：以邻苯二酚为底物,该酶最适pH为7．4,最适温度为25℃,60℃以上使酶迅速失活,动力学方程v=619．08[S]/(0．031 [S]),Vc、异Vc钠、NaHSO3、L-Cys可完全抑制酶活性,饱和NaCl能显著抑制酶活性,蔗糖、SDS对酶有激活作用。该酶能催化邻苯二酚、焦性没食子酸氧化,但对焦性没食子酸亲和力更强。     

The effects of control on the biomass, carbohydrate content and bud reserves of bracken (Pteridium aquilinum (L.) Kuhn), and an evaluation of a bracken growth model

This paper examines the initial effects of bracken control on frond numbers and biomass, and the biomass, carbohydrate reserves and bud densities of bracken stands cut once per year, twice per year, subject to a single application of asulam or left untreated. The seasonal dynamics of these parameters are displayed; carbohydrate and biomass are both removed from the rhizome system to produce frond tissue, and are replenished at the end of the growing season. Asulam application reduced densities of both active and dormant buds, and both frond biomass and density. It did not significantly reduce rhizome biomass or carbohydrate reserves in the two years after treatment. Cutting, either once or twice per year reduced both rhizome biomass and rhizome carbohydrate reserves, as well as bud densities, though the latter were reduced in proportion to biomass. Cutting twice a year reduced the production of fronds, both in numbers and biomass. The collected data were used to evaluate a model of bracken growth, and subsequently to improve estimates of some of the model parameters. The model simulations of control treatments were compared to field data. The effects of cutting once per year and spraying with asulam were predicted accurately, but the bracken stand was more resilient to cutting twice per year than would be expected from model predictions. The combination of cutting and spraying is discussed as a potential tool in land management and the deficiencies of the model are discussed in relation to the need for future research into the biology of bracken.     

A ten year study of the incidence of spores of bracken, (Pteridium aquilinum (L.) Kuhn.) in an urban rooftop airstream in south east Scotland

Field studies of bracken during sporing haveindicated very high numbers of sporesreleased per cubic metre of air. Thecarcinogenic nature of bracken spores poses ahealth hazard to workers and visitors in areasof sporing strands.A retrospective study of bracken spores trappedon slides exposed during Augustand September over the ten year period, 1988–1997, on a rooftop site in Edinburgh,has added an urban dimension to the aerobiologyof bracken.Results have revealed, apart from 1989,consistently low counts. The spore samplerwas situated downwind approximately 1.6 km fromthe nearest sporing stand.A spore calendar, based on the ten years ofcumulative data, has been constructed.     

Long-term recovery of bracken (Pteridium aquilinum (L.) Kuhn) after asulam spraying

This paper investigates the effects of a single asulam application, sprayed from the air, on the rhizome biomass, bud density, fronds and carbohydrate reserves of bracken (Pteridium aquilinum (L.) Kuhn) using a time sequence approach. Regression models were used to investigate how these characteristics varied with time after spraying, and were used, where appropriate, to calculate the time taken for full recovery after treatment. Frond density and biomass recovered in approximately eight years, bud numbers in seven, but rhizome biomass and total carbohydrate reserves required 10 to 12 years to recover. The consequences of these results are compared with predictions from a computer model and discussed in relation to the best timing of re-treatment and the management needed for long term control.     

Tree seed rain and seed removal,but not the seed bank,impede forest recovery in bracken (Pteridium aquilinum (L.) Kuhn)‐dominated clearings in the African highlands

Considerable areas dominated by bracken Pteridium aquilinum (L.) Kuhn occur worldwide and are associated with arrested forest recovery. How forest recovery is impeded in these areas remains poorly understood, especially in the African highlands. The component processes that can lead to recruitment limitation—including low seed arrival, availability and persistence—are important determinants of plant communities and offer a potential explanation for bracken persistence. We investigated key processes that can contribute to recruitment limitation in bracken‐dominated clearings in the Bwindi Impenetrable National Park, Uganda. We examined if differences in seed rain (dispersal limitation), soil seed bank, or seed removal (seed viability and persistence) can, individually or in combination, explain the differences in tree regeneration found between bracken‐dominated areas and the neighboring forest. These processes were assessed along ten 50‐m transects crossing the forest–bracken boundary. When compared to the neighboring forest, bracken clearings had fewer seedlings (bracken 11,557 ± 5482 vs. forest 34,515 ± 6066 seedlings/ha), lower seed rain (949 ± 582 vs. 1605 ± 335 tree seeds m^?2 year^?1), comparable but sparse soil seed bank (304 ± 236 vs. 264 ± 99 viable tree seeds/m²), higher seed removal (70.1% ± 2.4% vs. 40.6% ± 2.4% over a 3‐day interval), and markedly higher rodent densities (25.7 ± 5.4 vs. 5.0 ± 1.6 rodents per 100 trapping sessions). Camera traps revealed that rodents were the dominant animals visiting the seeds in our seed removal study. Synthesis: Recruitment limitation contributes to both the slow recovery of forest in bracken‐dominated areas, and to the composition of the tree species that occur. Low seed arrival and low persistence of unburied seeds can both explain the reduced density of seedlings found in bracken versus neighboring forest. Seed removal, likely due to rodents, in particular appears sufficient to constrain forest recovery and impacts some species more severely than others.     

Herbivorous arthropods on bracken (Pteridium aquilinum (L.) Khun) in Australia compared with elsewhere

Abstract The herbivorous arthropod fauna of bracken fern (Pteridium aquilinum (L.) Khun), at a site near Sydney, Australia, is described and compared with previously reported bracken faunas in other geographic regions. Monthly sampling over 18 months found 17 species of herbivorous arthropods (15 insect and two mite species) from five orders. At the ordinal level, the mixture differed substantially from the bracken faunas of sites in Britain and Papua New Guinea. Notable was the presence of Thysanoptera and Acari, and the absence of Coleoptera and Hymenoptera. The mixtures of orders/families represented in the site bracken faunas in Britain, and less so in Australia, resembled those in the pool of herbivorous arthropods in those regions. Further, the mixture of orders on bracken was more similar to the mixture of orders on other ferns than to the mixture of orders among herbivorous insects on all plants; such similarity was not evident at family level. Compared with sites in other regions, the Sydney site had an abundance of pinna-sucking species and a dearth of mining species. Differences between regions in feeding niches most occupied tended to correspond with the differences in orders represented. Not all features of the fauna of bracken near Sydney reflected differences in the general herbivorous arthropod fauna of Australia compared with other regions, or differences between the herbivore faunas of ferns and seed plants. Its composition must be attributed in part to stochastic aspects of the speciation of herbivorous arthropods onto host plants.     

The effects of cutting pre-treatments on the performance of asulam on bracken (Pteridium aquilinum)

The performance of asulam was assessed from a field experiment in which bracken (Pteridium aquilinum) was subjected to a variety of cutting pre-treatments prior to spraying with the herbicide. Both cutting bracken twice in the year prior to asulam application, and spraying frond regrowth 10 wk after a single mid-June cut, improved the performance of asulam. Summer cutting removes nutrients and dry matter from the system and reduces frond height, but increases frond density and the number of active buds on the rhizome system. The enhanced efficiency of asulam on pre-cut plots may be due to any one of these factors, or a complex interaction between some (or all) of them. In areas of tall bracken a cutting pre-treatment has the important practical advantage of reducing the height of the canopy, making the subsequent use of ground spraying equipment easier.     

Comparative spatial distribution, size, biomass and growth rate of two varieties of bracken fern (Pteridium aquilinum L. Kuhn) in a neotropical montane habitat

A comparative field study of caudatum and arachnoideum, the two Pteridium aquilinum varieties of the caudatum subspecies known to grow in the neotropics, was performed in a montane savanna habitat of the Venezuelan Andes that was affected by wildfire. Frond size, ramet density and spatial distribution, blade and rhizome biomass, and frond elongation and expansion rates were measured in separate, isolated stands each containing only one bracken variety and covering approximately the same area (540 m²). In addition to clearly discernible morphological differences, caudatum and arachnoideum were found to possess distinct features: caudatum tends to develop open stands of relatively shorter blades of 76.6±0.89 cm (±SE) of rachis length and lower ramet density (1.6 fronds m^-2, max.=7 fronds m^-2) whereas arachnoideum grows into longer, more expanded fronds 124±1.6 cm tall and significantly higher ramet density (5.1 fronds/m², max.=14.6 fronds m^-2). The sum of aerial and underground biomass was found to be notably larger for arachnoideum (8522±614 Kg/ha) than for caudatum (1929±131 Kg ha^-1) in stands growing under the same habitat conditions. Therefore the spatial distribution of arachnoideum appeared considerably more compact than that of caudatum. Blade growth rates and development time were also very different. Newly emerged caudatum croziers developed into mature blades within 42 to 48 days following an inverse exponential curve whereas arachnoideum blades required 70 to 75 days to reach maturity following a linear development. All the above dissimilarities are interpreted as the hitherto unreported indication of diverging growth strategies of two cohabitant bracken varieties following fire.     

SEM studies on vessels in ferns. 2. Pteridium

Xylem from roots and rhizomes of two infraspecific taxa of Pteridium aquilinum was studied by means of scanning electron microscopy (SEM). All tracheary elements proved to be vessels. End wall perforation plates were all scalariform, lacked pit membrane remnants in at least the central part of the perforation plate, and varied with respect to width of bars, from wide to tenuous, and with respect to presence of pit membrane remnants. In addition, porose pit membranes on walls that are likely all lateral vessel-to-vessel walls must be considered to be perforations also, although different from those on end walls. Lateral wall perforation plates, hypothesized by one worker on the basis of tylosis presence but denied by another on the basis of light microscopy, were confirmed by demonstration of pores with SEM. In addition, lateral walls of Pteridium vessels bear some grooves interconnecting pit apertures; this feature is newly figured by SEM for ferns. Lateral wall pitting that is not porose may either have striate thickenings of the primary wall or be smooth. Vessel presence and degree of specialization in Pteridium vessels may bear a relationship to the wide ecological tolerances of the genus.     

蕨菜黄酮苷元的组成及含量测定研究

以80%乙醇浸提经乙醚脱脂预处理的干蕨菜粉,减压回收乙醇得浓缩液.浓缩液以8%硫酸沸水浴水解3 h后,加入20%NaOH中和至弱酸性,过滤,滤渣以三氯甲烷索氏提取获总蕨菜黄酮苷元.TLC法检测总黄酮苷元的组成,HPLC法测定其主要苷元的含量.结果表明,通过酸水解获得总蕨菜黄酮苷元,其主要成分为山奈酚,含量约占蕨菜干重的1.9%.     

蕨菜黄酮分离鉴定

采用75%的乙醇提取经乙醚脱脂的蕨菜粉末,提取液回收乙醇后以石油醚、氯仿、乙酸乙酯、正丁醇依次萃取粗分段,硅胶柱层析、乙酸乙酯-甲醇系统对乙酸乙酯部位梯度洗脱,对其乙酸乙酯洗脱部分反复凝胶柱层析获得2个黄酮单体,分别鉴定为紫云英苷(1)、芦丁(2),其中化合物1为首次从蕨菜中分离得到。