Effect of the air pollution component ammonium sulphate on the VAM infection rate of three heathland species

Three heathand species, Antennaria dioica, Arnica montana and Hieracium pilosella, were artificially rained with ammonium sulphate solutions at increasing concentrations in a greenhouse experiment. The same species were also artificially rained with increasing ammonium sulphate solutions under field conditions. Dry weights of the plants in the field experiments did not change with increasing ammonium sulphate applications. Nor did the dry weights of plants in the greenhouse experiments change with increasing ammonium sulphate concentrations, except for Arnica montana, which showed an increase in dry weight. VAM infection percentage of Antennaria dioica increased in both the greenhouse and the field experiment. The results of the field experiment show that VAM infection rates are reduced after two years of artificial rain in the plant species Arnica montana, which grows naturally under nutrient poor conditions and is presently declining in its natural habitat in the Netherlands. In the greenhouse experiment, VAM infection of Arnica montana did not change with increasing ammonium sulphate concentrations. VAM infection rates of Hieracium pilosella, which presently is not declining, did not change with increasing ammonium sulphate concentrations.     

Acidification effects on vesicular-arbuscular mycorrhizal (VAM) infection, growth and nutrient uptake of established heathland herb species

A greenhouse experiment was carried out to determine whether the decline of Arnica montana L. in heathland vegetation in the Netherlands could be caused by a detrimental effect of soil acidification on vesicular-arbuscular mycorrhiza of this species. Arnica montana and two non-declining species from the same habitat, Hieracium pilosella L. and Deschampsia flexuosa (L.) Trin., were grown with and without the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus fasciculatum (Thaxter sensu Gerdemann) Gerd. and Trappe in pots with an extremely nutrient-poor, sandy soil. They were percolated weekly with nutrient solution with different pH values, viz. 5.5, 4.5, 3.5 and 2.5. At intervals of three weeks and up to 12 weeks, measurements were made on growth, nutrient uptake and VAM infection.In the most acid treatments growth and nutrient uptake were reduced in all species. VAM infection decreased only slightly with decreasing pH of the treatments. Without VAM, Arnica montana died and Hieracium pilosella hardly grew at the most acid treatments. Therefore it is concluded that VAM decreased the stress caused by the most acid treatments. Leachate from the most acid treatment had a pH of approximately 4, and contained considerable amounts of aluminium, dissolved from the solid phase of the soil. This might have played a role in the detrimental effects on the plants in the case of the most acid treatment. No evidence was found in this experiment that the decline of Arnica montana was due to detrimental effects of soil acidification on VAM of this species.     

Effects of inoculation with arbuscular mycorrhizal fungi on photosynthetic physiology in females and males of Populus deltoides exposed to cadmium pollution

Aims Populus deltoides is an important plantation tree species in the middle and lower reaches of the Yangtze River and in Huai River Plain. The extensively cultivated varieties are consisted of both females and males of P. deltoides. The objective of this paper was to characterize the difference in cadmium (Cd) tolerance between the sexes and the effects of symbiosis with Rhizophagus intraradices on their Cd tolerance.
Methods The experiment was carried out under semi-controlled conditions in a natural light greenhouse, protected from rain. Rhizophagus intraradices was inoculated on roots of both females and males when transplanting to a sand culture substrate. After one month, half of cuttings were exposed to Cd pollution (10 mg·kg^-1). All cuttings were irrigated with sterile water to avoid infection by other microorganism. About three months later, the gas exchange rate, net photosynthesis rate (P_n)-intercellular CO₂ concentration (C_i) curve, chlorophyll fluorescence, osmotic adjustment and phytohormone content of both females and males of P. deltoides were measured.
Important findings Our main results are as follows: (1) when compared to the controls, decreases in P_n, stomatal conductance, C_i, transpiration rate, maximum photochemical efficiency of photosystem II (PSII), effective quantum yield of PSII, photo-chemical quenching coefficient, maximum rate of carboxylation of Rubisco, photosynthetically active radiation-saturated rate of electron transport and rate of triose phosphate utilization to a different degree in both sexes of P. deltoides under Cd pollution were found, and females exhibited a greater decrease in such parameters than males. Rhizophagus intraradices inoculation mitigated the toxic effect of Cd on such parameters to a different degree in females, not in males. (2) Under Cd pollution, there was an increase in proline content in both sexes when compared to the controls. A further increase in proline content occurred in females, not in males, when inoculated with R. intraradices. (3) When compared to the controls, there was a decrease in indoleacetic acid, but an increase in abscisic acid in leaves of both sexes when exposed to Cd pollution. The amplitude changed in both phytohormones in females was greater than that in males. Rhizophagus intraradices inoculation was helpful for recovery of phytohormone balance in females, which was not observed in males. Therefore, our results indicated that (1) there were a greater negative effect exerted by Cd pollution on gas exchange rate, carbon fixation capacity and phytohormone balance and a more impairment of photosynthetic apparatus in females when compared to males, showing a less tolerance to stress conditions in females; (2) Rhizophagus intraradices inoculation could enhance the osmotic adjustment capacity in females, thus mitigate the negative effect of Cd stress on ability of carbon fixation and phytohormone balance in females. However, such positive effects derived from R. intraradices symbiosis were not observed in males.     

Ectomycorrhizal and arbuscular mycorrhizal colonization of two species of floodplain willows

To understand the relationships between the distribution of Chosenia arbutifolia and Salix sachalinensis and their mycorrhizal colonization, changes in the quality and types of ectomycorrhizas and arbuscular mycorrhizas of the seedlings of two species were studied at five different sites with different soil conditions in the floodplain of the Satsunai River, Hokkaido. High ectomycorrhizal and low arbuscular mycorrhizal colonization were found in roots of both plants. Ectomycorrhizal colonization of S. sachalinensis in wet sandy or muddy soil conditions was at the same level as that in dry gravelly sites. In contrast, ectomycorrhizal colonization of C. arbutifolia seedlings was lower from wet sandy sites than that from dry gravelly sites. In all study sites, the same three morphological types of ectomycorrhizas were dominant.     

Effects of ectomycorrhizal and vesicular-arbuscular mycorrhizal fungi on drought tolerance of four leguminous woody seedlings

Seedlings ofAcacia auriculiformis A. Cunn. ex. Benth.,Albizia lebbeck (L.) Benth.,Gliricidia sepium (Jac.) Walp andLeucaena leucocephala (Lam.) de Wit. were inoculated with an ectomycorrhizal (Boletus suillus (l. ex. Fr.) or indigenous vesicular-arbuscular mycorrhizal (VAM) fungi in a low P soil. The plants were subjected to unstressed (well-watered) and drought-stressed (water-stressed) conditions. InGliricidia andLeucaena, both mycorrhizal inoculations stimulated greater plant growth, P and N uptake compared to their non-mycorrhizal (NM) plants under both watering regimes. However, inAcacia andAlbizia, these parameters were only stimulated by either ectomycorrhiza (Acacia) or VA mycorrhiza (Albizia). Growth reduction occurred as a result of inoculation with the other type of mycorrhiza. This was attributed to competition for carbon betweenAcacia and VA mycorrhizas and parasitic association betweenAlbizia and ectomycorrhiza. Drought-stressed mycorrhizal and NMLeucaena, and drought-stressed mycorrhizalAcacia tolerated lower xylem pressure potentials and larger water losses than the drought-stressed mycorrhizal and NMAlbizia andGliricidia. These latter plants avoided drought by maintaining higher xylem pressure potentials and leaf relative water content (RWC). All the four leguminous plants were mycorrhizal dependent. The higher the mycorrhizal dependency (MD), the lower the drought tolerance expressed in terms of drought response index (DRI). The DRI may be a useful determinant of MD, as they are inversely related.     

Differential effects of ammonium and nitrate on three heathland species

Arnica montana and Cirsium dissectum, typical of species-rich heathlands and acidic grasslands, have declined rapidly in The Netherlands in recent years. Field surveys suggest that the decline is caused by soil acidification as a result of enhanced atmospheric N and S deposition. Therefore, the survival, growth and development of these species were studied in a water culture experiment, using nutrient solutions which differed both in mineral nitrogen form and in ammonium concentration. For comparison, the performance of a third, acid tolerant species, Calluna vulgaris, was studied. The results showed that both Arnica and Cirsium performed better using nitrate than when using ammonium as a sole nitrogen source, whereas ammonium toxicity became apparent when ammonium concentrations were raised above 100 µM. Ammonium toxicity was expressed by an increase in mortality of Arnica plants with increasing ammonium concentrations and by a reduction of biomass in Arnica and Cirsium. Furthermore, cation concentrations in both roots and shoots decreased when ammonium was supplied as a nitrogen source. In contrast, Calluna showed optimal development when using ammonium as a sole nitrogen source. In this species, only root biomass was negatively affected by high ammonium concentrations. The ecological implications of these preferences are discussed in relation to soil acidification.     

The influence of ammonium nitrate on the root growth and ericoid mycorrhizal colonization of Calluna vulgaris (L.) Hull from a Danish heathland

Conversion of European heathlands to grassland has been reported as a response to increased nutrient availability, especially of nitrogen; a direct effect upon mycorrhizal colonization has been proposed as an likely explanation.This hypothesis was tested in a random block experiment with four blocks and four replicates on a Danish inland heath, Hjelm Hede. Ammonium nitrate was applied (0, 35, 50 and 70 kg N ha^–1 year^–1) to a stand of Calluna vulgaris (L.) Hull four times annually for 2 years. Calluna roots were sampled on four occasions in the 2nd year of the nitrogen treatment. The extent of ericoid mycorrhizal colonization was determined by direct observation of the roots using a line-intersection method. The nitrogen content of the current-year shoots of Calluna increased when they were treated with nitrogen. Nitrogen fertilization had no significant effects on ericoid mycorrhizal colonization of Calluna nor on root biomass. The seasonal variation in mycorrhizal colonization of the Calluna roots was highly significant. The spatial variability of mycorrhizal colonization, both in replicated plots and in the two contrasted soil horizons – the mor layer and the bleached sand – within the plots, were considerable. I conclude that heather decline under enhanced nitrogen input is unlikely to be caused by a direct impact on the ericoid mycorrhizae of Calluna. Received: 1 December 1998 / Accepted: 7 December 1999     

Effects of nutrients and arbuscular mycorrhizal colonization on the growth of Salix gracilistyla seedlings in a nutrient-poor fluvial bar

A field survey and a pot culture experiment were conducted to examine the effects of nutrients (N and P) and arbuscular mycorrhizal (AM) fungi on the growth of Salix gracilistyla, a pioneer plant in riparian habitats. The plants growing in the field were colonized by AM and/or ectomycorrhizal fungi. However, the direct effect of AM colonization on seedling growth was not detected in the pot culture experiment. In contrast, N application significantly promoted plant growth, suggesting that the growth of S. gracilistyla seedlings is largely limited by the availability of N in the field.     

Metabolic activity of Glomus intraradices in Arum- and Paris-type arbuscular mycorrhizal colonization

Colonization of two plant species by Glomus intraradices was studied to investigate the two morphological types (Arum and Paris), their symbiotic interfaces and metabolic activities. Root pieces and sections were stained to observe the colonization and metabolic activity of all mycorrhizal structures. There were no growth responses observed in the plants caused by mycorrhizal symbiosis. The two morphological types had a similar percentage of root colonized, but the Arum-type had higher metabolic activity. Most of the mycorrhizal structures (88%) showed succinate dehydrogenase activity; about half showed acid phosphatase activity; and a small percentage showed alkaline phosphatase activity. Phosphatase activity was highest in arbuscules and low in intercellular hyphae in the Arum-type colonization. In the Paris-type, hyphal coils and arbusculate coils showed a similar intermediate percentage of phosphatase activity. We conclude that acid phosphatase is more important than alkaline phosphatase in both colonization types. We discuss the possibility that, whereas arbuscules in Arum-type are the main site for phosphorus release to the host plant, both the hyphal and arbusculate coils may be involved in the Paris-type.     

Effect of arbuscular mycorrhizal colonization and two levels of compost supply on nutrient uptake and flowering of pelargonium plants

Two challenges frequently encountered in the production of ornamental plants in organic horticulture are: (1) the rate of mineralization of phosphorus (P) and nitrogen (N) from organic fertilizers can be too slow to meet the high nutrient demand of young plants, and (2) the exclusive use of peat as a substrate for pot-based plant culture is discouraged in organic production systems. In this situation, the use of beneficial soil microorganisms in combination with high quality compost substrates can contribute to adequate plant growth and flower development. In this study, we examined possible alternatives to highly soluble fertilizers and pure peat substrates using pelargonium (Pelargonium peltatum L’Her.) as a test plant. Plants were grown on a peat-based substrate with two rates of compost addition and with and without arbuscular mycorrhizal (AM) fungi. Inoculation with three different commercial AM inocula resulted in colonization rates of up to 36% of the total root length, whereas non-inoculated plants remained free of root colonization. Increasing the rate of compost addition increased shoot dry weight and shoot nutrient concentrations, but the supply of compost did not always completely meet plant nutrient demand. Mycorrhizal colonization increased the number of buds and flowers, as well as shoot P and potassium (K) concentrations, but did not significantly affect shoot dry matter or shoot N concentration. We conclude that addition of compost in combination with mycorrhizal inoculation can improve nutrient status and flower development of plants grown on peat-based substrates.     

Influence of three species of arbuscular mycorrhizal fungi on the persistence of aromatic hydrocarbons in contaminated substrates

Aromatic hydrocarbons are pollutants which have mutagenic and carcinogenic properties as well as relatively high hydrosolubility. Their presence in soils makes techniques such as bioremediation an important topic for research. In this work, the effect of arbuscular mycorrhiza (AM) on the persistence of benzene, toluene, ethylbenzene and xylene (BTEX) in artificially contaminated substrates was evaluated. Leek plants were grown with three AM fungal species using a specially designed mesocosm system, in which internal air and substrate samples were analyzed by gas chromatography for BTEX content. Strong reductions in the BTEX concentration in substrates were generally observed in the presence of mycorrhizal plants. Residual BTEX content ranged between nearly total disappearance (<2%) and 40% of the original concentration, whereas there was a high persistence of hydrocarbons in the samples of substrate alone or with non-mycorrhizal plants. These results provide first evidence for an influence of AM activity in reducing pollution of substrates by aromatic hydrocarbons.Andrea Volante and Guido Lingua contributed equally to this paper     

Effect of arbuscular mycorrhizal (AM) colonization on terpene emission and content of Artemisia annua L

Plant roots interact with a wide variety of rhizospheric microorganisms, including bacteria and the symbiontic arbuscular mycorrhizal (AM) fungi. The mycorrhizal symbiosis represents a series of complex feedbacks between plant and fungus regulated by their physiology and nutrition. Despite the widespread distribution and ecological significance of AM symbiosis, little is known about the potential of AM fungi to affect plant VOC metabolism. The purpose of this study was to investigate whether colonization of plant roots by AM fungi and associated soil microorganisms affects VOC emission and content of Artemisia annua L. plants (Asteraceae). Two inoculum types were evaluated: one consisted of only an arbuscular mycorrhizal (AM) fungus species (Glomus spp.), and the other was a mixture of different Glomus species and associated soil bacteria. Inoculated plants were compared with non-inoculated plants and with plants supplemented with extra phosphorus (P) to obtain plants of the same size as mycorrhizal plants, thus excluding potentially-confounding mycorrhizal effects on shoot growth. VOC emissions of Artemisia annua plants were analyzed by leaf cuvette sampling followed by off-line measurements with pre-concentration and gas chromatography mass spectrometry (GC-MS). Measurements of CO(2) and H(2)O exchanges were conducted simultaneously. Several volatile monoterpenes were identified and characterized from leaf emissions of Artemisia annua L. by GC-MS analysis. The main components identified belong to different monoterpene structures: alpha-pinene, beta-pinene, camphor, 1,8-cineole, limonene, and artemisia ketone. A good correlation between monoterpene leaf concentration and leaf emission was found. Leaf extracts included also several sesquiterpenes. Total terpene content and emission was not affected by AM inoculation with or without bacteria, while emission of limonene and artemisia ketone was stimulated by this treatment. No differences were found among treatments for single monoterpene content, while accumulation of specific sesquiterpenes in leaves was altered in mycorrhizal plants compared to control plants. Growth conditions seemed to have mainly contributed to the outcome of the symbiosis and influenced the magnitude of the plant response. These results highlight the importance of considering the below-ground interaction between plant and soil for estimating VOC emission rates and their ecological role at multitrophic levels.