Genetic analysis of morphological variability in three Plantago species with different mating systems

Summary Morphological variability in three Plantago species possessing different mating systems — P. Major, P. coronopus, and P. lanceolata — was analyzed. From each species several populations were analyzed. In comparing the species, it appeared that both the inbreeding P. major and the outbreeding P. lanceolata showed a high degree of morphological differentiation between populations. It was hypothesized that the absence of population differentiation for the species with a mixed mating system, P. coronopus, was caused by high adaptive plasticity. Plasticity is probably the predominant mechanism by which this species copes with different environments. There were considerable differences between populations within each species for their levels of variability and their heritabilities. It appeared that besides the influence of the mating system, selection may diminish morphological variability in the case of strong directional selection, especially in P. major and P. lanceolata.Grassland Species Research Group Publication No. 135     

Analysis of allozyme variability in three Plantago species and a comparison to morphological variability

Summary The level of electrophoretic variability in three Plantago species, P. major, P. coronopus, and P. lanceolata, was analyzed in relation to their breeding systems and compared with their morphological variability. From each species several populations were analyzed. The outcrossing P. lanceolata had the highest level of electrophoretic variability and the lowest population differentiation. The inbreeding P. major showed the opposite: a low level of electrophoretic variability and a high population differentiation. P. coronopus, with an intermediate breeding system, had an intermediate level of variability and differentiation. In comparing the species, it appeared that P. coronopus and P. major showed good concordance in the distribution of both kinds of variability, each having only a slightly higher morphological than electrophoretic differentiation between populations. P. lanceolata showed a higher morphological than electrophoretic differentiation between populations. A comparison of populations, within species, revealed good concordance of electrophoretic and morphological variability only within P. coronopus, while some populations of the other two species had relatively lower morphological variability compared with electrophoretic variability.Grassland Species Research Group Publication No. 182     

Nitrate production in the rhizosphere of Plantago species

Summary The production of nitrate in an old established dune grassland soil and its uptake by plants was studied by comparing amounts of mineral nitrogen and numbers of nitrifying bacteria in the rhizosphere on the one hand, and on the other accumulated nitrate and levels of nitrate reductase (NaR) of individual plants of three Plantago species,i. e., P. major, P. lanceolata andP. coronopus. For these three Plantago species andP. media basal levels of NaR in the absence of nitrate were determined in plants grown in culture solutions. The basal NaR levels ofP. major andP. media (species occurring on nutrient-rich soils) were significantly higher than those ofP. lanceolata andP. coronopus (species found on nutrient-poor soils). NaR activity increased in the presence of nitrate and was suppressed by ammonium.From the numbers of nitrifying bacteria in the rhizosphere and NaR activity in the leaves it was concluded that nitrate was produced in the root environments of the three Plantago species and that the compound was taken up by the plants. NaR activities and numbers of nitrifying bacteria were higher for individuals ofP. major than for those ofP. lanceolata andP. coronopus. No correlation was found between the ammonium levels and the numbers of nitrifying bacteria in the soil, and no indications of inhibition of nitrifying bacteria in the rhizosphere were obtained. For individuals ofP. lanceolata a correlation was found between the numbers of nitrifying bacteria in the soil and NaR activity in the leaves. The results are discussed in relation to the ecological habitats of the three species.Grassland Species Research Group Publication No.38.     

The response ofPlantago lanceolata L. andP. major L. spp.major to nitrate depletion

To study aspects of the ecology of grassland species, in a comparative experiment, plants ofP. lanceolata andP. major were grown in pots in a greenhouse, and subjected to a gradual nitrate depletion for several weeks. Control plants were weekly supplied with nitrate. Growth, leaf appearance and disappearance, concentrations of cations and inorganic anions, soluble and insoluble reduced nitrogen concentrations,in vivo nitrate reductase activity (NRA) and the concentration of non-structural carbohydrates in several parts of the plants were followed. Depletion of nitrate caused a reduction of shoot growth, both in biomass and number of leaves. Withering of leaves increased. Accumulation of root dry matter was little (P. lanceolata), or not (P. major) affected. The concentration of reduced nitrogen in all tissues also decreased, both that of the soluble and that of the insoluble fraction. As a result, nitrogen use efficiency (NUE, g dry matter produced per mmol N incorporated) increased by nitrate depletion. NRA was higher in the roots than in the leaves, and decreased with increasing nitrate depletion. In control plants, nitrate became also limiting. This resulted in decreasing nitrate concentrations in leaves and roots. In the leaves, the decrease in nitrate concentration was preceded by a decrease in NRA. The decrease of the nitrate concentration was parallelled by an increase in the concentration of soluble sugar. No major differences in the response towards nitrate depletion were observed between the two species. Grassland Species Research Group, publication no. 129     

Population and species variation of minisatellite DNA in Plantago

Three Plantago species were surveyed for within- and between-population variation at DNA sequences detected with the M13 minisatellite probe. The levels and patterns of variation detected by this probe correspond to those expected from the mating systems of the species. The highly-selfing species P. major has a relatively low variability of minisatellite sequences within populations and considerable differentiation between populations. The outcrossing species P. lanceolata exhibits higher minisatellite variability within populations and moderate differentiation between populations. P. coronopus, with a mixed mating system, has levels of variation intermediate between P. major and P. lanceolata. The levels of variation within and between populations corresponds, in general, to the levels of allozyme variation determined in an earlier study. Mating system and population structure appear to have a major influence on M13-detected fragment variation.     

Polymorphic microsatellite loci in Plantago lanceolata

The genus Plantago is particularly interesting for evolutionary studies because of its wide range of mating systems. We have developed primers for five highly polymorphic microsatellite loci isolated from P. lanceolata. All five loci amplified and were polymorphic in the two populations examined, Lowsteads Beach in the United Kingdom and Duke in the United States. These new markers will allow a comparison of population structure between the outcrossing species P. lanceolata, and the highly selfing species P. major.     

Intraspecific variation in the response to CO2 enrichment in seeds and seedlings of Plantago lanceolata L.

Summary Four genotypes of P. lanceolata were grown to maturity at combinations of two levels of atmospheric CO₂ concentrations and two temperature conditions. Seed weight was determined, and seed germination and seedling growth were measured for the progeny of each genotype under the same environmental conditions. Overall, high CO₂ levels decreased seed weight, increased germination percentage and rate, and increased seedling size. Families differed in their response to CO₂ enrichment, and to combinations of CO₂ and temperature levels for several characters. These results suggest the existence of genetic variability in P. lanceolata in response to CO₂ enrichment.     

Growth reduction in Plantago lanceolata in relation to biotic factors in the soil environment

Summary To investigate the influence of soil-borne pathogens on the population biology of a grassland species in natural vegetation, soil samples were collected, partly disinfected or sterilized and then planted with Plantago lanceolata. Those on some of the untreated soils showed growth reduction due to a biotic factor in the soil. This growth reduction was not associated with the density of P. lanceolata plants at the sampling sites. It was only visible when differences in abiotic factors — especially nutrient levels —were eliminated. In the natural situation the nutritional status of the soil has a far greater impact than biotic soil factors. Micro-organisms harmful to Plantago roots are considered to be of minor importance in the distribution of the individuals in the P. lanceolata population studied.Study performed by Grassland Species Research Group No 113     

Neighbor species differentially alter resistance phenotypes in Plantago

In this study, we investigated how neighbors (i.e., competitors) altered resistance phenotypes, namely plant size and levels of secondary compounds (iridoid glycosides), of individual plants and specifically tested whether neighbor identity mattered. We conducted a greenhouse experiment with Plantago lanceolata and Plantago major (Plantaginaceae) in which each species served as focal plants as well as neighbors in a factorial design. In addition, we harvested plants six and nine weeks after transplantation to test whether effects changed as plants grew. In both species, competition reduced plant size, and this effect increased over time. Plantago lanceolata neighbors suppressed growth of both focal plant species more than P. major neighbors. Effects of competition on levels of secondary compounds were more complex. Concentrations of iridoid glycosides were increased by competition in both species at harvest one. By the second harvest, an effect of competition on iridoid glycosides was found only in P. major. Neighbor identity influenced levels of iridoid glycosides in P. lanceolata at harvest one; concentrations were higher in plants grown with P. lanceolata neighbors than in plants grown with P. major neighbors. We also tested whether there was a trade-off between growth (biomass) and defense (levels of iridoid glycosides). Biomass and iridoid glycoside content were significantly correlated only in plants grown with competition and harvested at nine weeks, and this relationship was positive in both species, indicating that there was no trade-off between growth and defense. This study suggests that neighbor identity could play an important role in interspecific interactions, including the interactions of plants with other trophic levels.     

Plantago lanceolata L. and Rumex acetosella L. differ in their utilisation of soil phosphorus fractions

To establish relationships between soil phosphorus (P) fractions and leaf P, a mycorrhizal species (Plantago lanceolata L.) was compared with a typically non-mycorrhizal species (Rumex acetosella L.) in a glasshouse experiment. The plants were grown in 40 soils from non-fertilised, abandoned pastures or abandoned arable fields and leaf P concentration were found to be related to various soil P fractions after six weeks of growth. The differences in the P fractions in soil can account for a large share of the variation in leaf P concentration in both species, but the two species differed in their utilisation of P fractions. Leaf P concentration of R. acetosella was more related to extractable soil P than that of P. lanceolata. Rumex acetosella showed a higher maximum P concentration. The P fractions accounting for the largest share of the variation in leaf P concentration was the Bray 1 extractable and the weak oxalate (1 mM) extractable P, and for P. lanceolata also the Na₂SO₄+NaF extractable P fraction. P extracted with these methods accounted for up to 80% of the variation in P concentration in leaves of R. acetosella and 65% of the variation in leaves of P. lanceolata. More P extractable with weak oxalate, Na₂SO₄+NaF and strong oxalate (50 mM) was released from the soil than was taken up by the plants during the experimental period. The Bray 1 extractable P fraction, however, decreased in both unplanted and planted soils. Phosphatase release was not induced in any of the plants during the experimental period, indicating that they were not mobilising soil organic P. However, some of the methods extracted a large share of the organic P and still explained much of the variation in leaf P concentration. Mycorrhizal colonisation of P. lanceolata was inversely related to the extractable soil P. The consistently fast P uptake of R. acetosella indicates that this species have a high demand for P. The differences in P utilisation between R. acetosella and P. lanceolata could be caused by their different mycorrhizal status.     

Chloroplast DNA variation within and among five Plantago species

Chloroplast DNA variation was studied within and among five Plantago species. We determined polymorphism by surveying 86% of the chloroplast genome using 9 or 11 restriction enzymes for intra or interspecific variation, respectively. All five species were polymorphic for both site and length mutations. The outcrossing P. lanceolata has six chloroplast haplotypes of which some were found in the Netherlands as well as in the United States. The highly selfing P. major had five haplotypes and Dutch and United States populations were differentiated from each other in cpDNA. Plantago species are highly differentiated from each other. Of 252 restriction-sites, 52% were variable among species. Most of this variation was localized in part of the large single copy region. We derived a molecular phylogeny of the five species from restriction-site variability using PAUP 3.0. P. coronopus and P. maritima form a group as do P. major and P. media. P. lanceolata is more distantly related to the other four species. In a consensus tree both P. major haplotypes and both P. media haplotypes were connected to one node.     

The effect of nitrate concentration in a flowing solution system on growth and nitrate uptake of twoPlantago species

Summary Juvenile plants ofPlantago lanceolata andP. major ssp.major were grown in a flowing solution system at 7.5 mM or 9.5 M NO₃. The parameters investigated were: RGR, shoot weight percentage, leaf length, length of main root axis, shoot concentrations of major ions and organic N, and the specific uptake rate for NO₃. At 9.5 M NO₃ growth ofP. major was not hampered, whereas shoot growth and leaf length ofP. lanceolata were reduced. The NO₃ concentration ofP. lanceolata decreased more than that ofP. major. The different performances of the species at 9.5 M NO₃ were associated with different specific uptake rates. In both treatments the root system ofP. major was shorter than that ofP. lanceolata. P. lanceolata accumulated more NO₃ in the leaves. The performance of thePlantago species is discussed in relation to the availability of nutrients in their habitats.Grassland Species Research Group. Publication no. 37.     

毛竹各器官和根际土浸提液对杉木种子萌发的化感作用

随着毛竹杉木混交林面积的不断扩大,它们之间既会产生促进作用,又会产生抑制作用。本研究对比分析了不同浓度毛竹各器官(鲜叶、干叶、枝条、竹杆、竹鞭和鞭根)的浸提液及枯落物、根际土浸提液对杉木种子发芽率的影响,探讨了毛竹各器官浸提液及枯落物、根际土浸提液对杉木种子萌发的化感作用。结果表明:(1)处理前期,毛竹各器官浸提液对杉木种子萌发具有抑制作用,且当浸提液浓度为1∶25和1∶50的高浓度时,相对于低浓度浸提液对杉木种子萌发的抑制作用更为持久;(2)处理后期,低浓度的毛竹各器官浸提液对杉木种子萌发促进作用较快,而高浓度的浸提液仍呈现抑制作用,促进作用出现较为缓慢;(3)枯落叶浸提液总体上对杉木种子萌发具有抑制作用,在整个实验过程中低于对照组的发芽率,仅在1∶50浓度时略高对照组实验;而根际土浸提液对杉木种子萌发自始至终都具有促进作用,其中低浓度的促进作用十分显著。故在实际的毛竹杉木混交林生产营林过程中可选用毛竹根际土制备浸提液,促进杉木种子萌发,为毛竹杉木混交林的生产经营提供参考。     

Efficiency of Rumex dentatus L. Leaves Extract for Enhancing Phytoremediation of Plantago major L. in Soil Contaminated by Carbosulfan

Carbosulfan and carbofuran induce water pollution and health issues. Such issues may be solved by phytoremediation. Here we tested the potential of adding Rumex dentatus L. leaves extract to Plantago major L. for enhancing phytoremediation of carbosulfan-contaminated soil. The phytoremediation efficiency of P. major L. amended with R. dentatus L. leaves extract was significantly greater than that of P. major only and R. dentatus with respect to the removal of carbosulfan from contaminated soil. The contribution of R. dentatus leaves extract to the degradation of carbosulfan in the soil were 20.95%, 18.85%, 5.2% and 1.85% after 2, 4, 8 and 16 days of treatments, respectively. The presence of P. major amended with R. dentatus leaves extract showed significant effect on uptake of carbosulfan into roots at 2 and 4 days, the uptake ratio was about 2.39 and 1.31 times higher compared to P. major alone, respectively. Carbofuran appeared in the P. major roots amended with R. dentatus leaves extract within 2 days of treatment (5.49 mg/kg) and reached the maximum over 8 days (10.19 mg/kg), while carbofuran appeared in the P. major over 4 days of treatment (4.09 mg/kg) and reached the maximum (6.92 mg/kg) over 8 days of exposure. Carbosulfan taken up into P. major leaves reached 4.36 mg/kg over 4 days, while it reached 2.75 mg/kg in P. major leaves amended with R. dentatus leaves extract over the same time. Carbofuran translocated into the P. major leaves amended with R. dentatus leaves extract and reached the maximum over 16 days of exposure (10.43 mg/kg), followed by P. major (9.47 mg/kg) and R. dentatus leaves (9.5 mg/kg), respectively. This study indicates that R. dentatus leaves extract can improve the efficiency of phytoremediation of carbosulfan.     

Genetic differentiation in Plantago major L. in growth and P uptake under conditions of P limitation

To study possible adaptive mechanisms inbred lines from three populations of Plantago major from sites that were found to differ in P availability were compared. In a pot experiment the growth and P uptake either in the presence or absence of Glomus fasciculatum was determined. Under these P-limited conditions it was shown by partitioning the relative growth rate (RGR, in mg g^-1 day^-1) in the components root weight ratio (RWR, in g_roots g_plant ^-1), specific P uptake rate (SPUR, in mol P g_roots ^-1 day^-1), and P-efficiency (PEFF, in mg mol P^-1), that the increase in RGR of mycorrhizal infected plants was related to an increase in SPUR, and a decrease in RWR and PEFF. P. major ssp. major had a lower RGR (related to a lower PEFF and SPUR) and a higher RWR than P. major ssp. pleiosperma. In a second experiment three inbred lines were compared upon P depletion in a nutrient solution. The P. major ssp. major line had a lower RGR and higher RWR, and a higher accumulation of P in the roots than the P. major ssp. pleiosperma lines under optimal growing conditions. There were no differences among the inbred lines in the relative contribution of inorganic P to the total P concentration in the shoot. The results are discussed in relation to the characteristics of the habitats of the investigated P. major populations.     

Ammonium and nitrate nutrition inPlantago lanceolata L. andPlantago major L. ssp.major

With the aims (1) to test whether the different natural occurrence of twoPlantago species in grasslands is explained by a different preference of the species for nitrate or ammonium; (2) to test whether the different occurrence is explained by differences in the flexibility of the species towards changes in the nitrogen form; (3) to find suitable parameters as a tool to study ammonium and nitrate utilization of these species at the natural sites in grasslands, plants ofPlantago lanceolata andP. major ssp.major were grown with an abundant supply of nitrate, ammonium or nitrate+ammonium as the nitrogen source (0.5 mM). The combination of ammonium and nitrate gave a slightly higher final plant weight than nitrate or ammonium alone. Ammonium lowered the shoot to root ratio inP. major. Uptake of nitrate per g root was faster than that of ammonium, but from the mixed source ammonium and nitrate were taken up at the same rate. In vivo nitrate reductase activity (NRA) was present in both shoot and roots of plants receiving nitrate. When ammonium was applied in addition to nitrate, NRA of the shoot was not affected, but in the root the activity decreased. Thus, a larger proportion of total NRA was present in the shoot than with nitrate alone. In vitro glutamate dehydrogenase activity (GDHA) was enhanced by ammonium, both in the shoot and in the roots.In vitro glutamine synthetase activity (GSA) was highest in roots of plants receiving ammonium. Both GDHA and GSA were higher inP. lanceolata than inP. major. The concentration of ammonium in the roots increased with ammonium, but it did not accumulate in the shoot. The concentration of amino acids in the roots was also enhanced by ammonium. Protein concentration was not affected by the form of nitrogen. Nitrate accumulated in both the shoot and the roots of nitrate grown plants. When nitrate in the solution was replaced by ammonium, the nitrate concentration in the roots decreased rapidly. It also decreased in the shoot, but slowly. It is concluded that the nitrogen metabolism of the twoPlantago species shows a similar response to a change in the form of the nitrogen source, and that differences in natural occurrence of these species are not related to a differential adaptation of nitrogen metabolism towards the nitrogen form. Suitable parameters for establishing the nitrogen source in the field are thein vivo NRA, nitrate concentrations in tissues and xylem exudate, and the fraction of total reduced nitrogen in the roots that is in the soluble form, and to some extent thein vitro GDHA and GSA of the roots. Grassland Species Research Group. Publ. no 118.     

Comparison of the Respiratory Metabolism of Plantago lanceolata L. and Plantago major L.

Bryce, J. H. and ap Rees, T. 1985. Comparison of the respiratorymetabolism of Plantago lanceolata L. and Plantago major L.—J.exp. Bot. 36 1559–1565. The aim of this work was to discover if the respiratory metabolismof the roots of Plantago lanceolata L. differed from that ofthe roots of Plantago major L. Measurements of oxygen uptakeand dry weight of excised root systems during growth of seedlingsprovided evidence that the two species differed in the amountof respiration needed to support a given increase in dry weight.Excised root systems were given a 6-h pulse in [U-¹⁴C]sucrosefollowed by a 16.5-h chase in sucrose. The detailed distributionof ¹⁴C amongst the major components of the roots at the endof the pulse and the chase revealed no significant differencebetween the two species. Patterns of ¹⁴CO₂ production from [1-¹⁴C],[2-¹⁴C], [3,4-¹⁴C], and [6-¹⁴C]glucose of excised root systemsfrom plants of three ages were similar for the two species.It is suggested that there is no conclusive evidence for anysignificant inherent difference in the respiratory metabolismof the roots of the two species. Key words: ¹⁴C sugar metabolism, respiration, roots, Plantago     

Ontogenetic patterns in the mechanisms of tolerance to herbivory in Plantago

Background and Aims

Herbivory and plant defence differ markedly among seedlings and juvenile and mature plants in most species. While ontogenetic patterns of chemical resistance have been the focus of much research, comparatively little is known about how tolerance to damage changes across ontogeny. Due to dramatic shifts in plant size, resource acquisition, stored reserves and growth, it was predicted that tolerance and related underlying mechanisms would differ among ontogenetic stages.

Methods

Ontogenetic patterns in the mechanisms of tolerance were investigated in Plantago lanceolata and P. major (Plantaginaceae) using the genetic sib-ship approach. Pot-grown plants were subjected to 50 % defoliation at the seedling, juvenile and mature stages and either harvested in the short-term to look at plasticity in growth and photosynthesis in response to damage or allowed to grow through seed maturation to measure phenology, shoot compensation and reproductive fitness.

Key Results

Tolerance to defoliation was high in P. lanceolata, but low in P. major, and did not vary among ontogenetic stages in either species. Mechanisms underlying tolerance did vary across ontogeny. In P. lanceolata, tolerance was significantly related to flowering (juveniles) and pre-damage shoot biomass (mature plants). In P. major, tolerance was significantly related to pre-damage root biomass (seedlings) and induction of non-photochemical quenching, a photosynthetic parameter (juveniles).

Conclusions

Biomass partitioning was very plastic in response to damage and showed associations with tolerance in both species, indicating a strong role in plant defence. In contrast, photosynthesis and phenology showed weaker responses to damage and were related to tolerance only in certain ontogenetic stages. This study highlights the pivotal role of ontogeny in plant defence and herbivory. Additional studies in more species are needed to determine how seedlings tolerate herbivory in general and whether mechanisms vary across ontogeny in consistent patterns.     

Variations in pollen proportions of Plantago lanceolata and P. major/media at a Neolithic lake dwelling,Lake Chalain,France

Detailed pollen analytical investigations at a Neolithic lake dwelling site on Lake Chalain, Jura, France, show very characteristic variation in the Plantago lanceolata and P. major/media pollen curves in the period during which settlements are recorded (3030–2630 cal. B.C.). At first, P. lanceolata is the more important taxon but P. major/media representation gradually rises, to become the more important taxon in the uppermost settlement phase. After considering the present day ecology and phytosociology of the Plantago species in question, i.e. P. lanceolata, P. major and P. media, and the available archaeozoological and archaeological information, it is suggested that the changes in the representation of these two pollen taxa are the result of a change in the farming economy, at ca. 2800 cal. B.C., which involved a substantial rise in the numbers of domesticated grazing animals and more intensive land use.     

Genetic variability in Plantago species in relation to their ecology

Summary Populations of the three Plantago species P. major, P. lanceolata and P. coronopus were scored for allozyme variability. They showed normal variability levels compared with other, similar plant species. Differentiation among populations appeared to be rather low in comparison with other species, probably due to a considerable amount of long distance seed transport. In order to be able to make an analysis of small-scale gene flow, all (sub)populations were critically checked for the existence of population structure equilibrium. The allozyme variation was tested for neutrality by testing homogeneity of F values among loci: between populations (Lewontin-Krakauer test) and within populations. No systematic deviations from the prediction of the neutral theory could be established. From the population structure analyses, little gene flow in the species P. major (with high selfing levels) and P. coronopus could be concluded, whereas P. lanceolata showed relatively high levels of gene flow. The degree of homozygosity in the latter species was too high to explain with the available data. In P. coronopus, on the other hand, an unusually high number of heterozygotes were observed.Grassland Species Research Group Publication No. 73