Utilisation of organic nitrogen and phosphorus sources by mycorrhizal endophytes of Woollsia pungens (Cav.) F. Muell. (Epacridaceae)

 The ability of four ericoid mycorrhizal endophytes isolated from roots of Woollsia pungens (Cav.) F. Muell. (Epacridaceae) to utilise organic forms of nitrogen and phosphorus during growth in axenic culture was assessed. All isolates were able to utilise glutamine, arginine and bovine serum albumin (BSA), along with NH₄ ⁺ or NO₃ ^–, in most cases yielding at least as much biomass as the ericoid mycorrhizal endophyte Hymenoscyphus ericae (Read) Korf & Kernan. All isolated endophytes were able to utilise BSA, arginine and glutamine as sole sources of N and C. With the exception of a single isolate (C40), which showed little growth on glutamine, biomass yields on glutamine as the sole N and C source was significantly greater for all isolates than on either of the other two organic N sources. Two isolates from W. pungens (C40 and A43) utilised DNA and sodium inositol hexaphosphate as sole P sources, in each case yielding significantly more biomass than H. ericae. The results suggest that mycorrhizal endophytes from epacrid plant hosts and those from ericaceous hosts have similar abilities to utilise organic forms of N and P. Accepted: 4 September 1998     

Response of conifer seedlings to nitrate and ammonium sources of nitrogen

Summary Differences in growth responses of Douglas fir, western hemlock, Sitka spruce, and white spruce to nitrate and ammonium N sources were examined in sand culture and artificial soil culture. Effects of the two forms of N on growth, needle area, and N uptake of three Douglas fir halb-sib progenies were examined in a second sand culture. Response of Douglas fir to the two forms of N was followed over two years in nursery soil of different pH levels. In sand culture 1 mean seedling dry weight of all species, except hemlock, was greatest when ammonium N and nitrate N were provided in equal amounts. In all species, except Sitka spruce, ammonium alone resulted in greater growth than nitrate alone. Use of ammonium N resulted in greater growth of all species, than was obtained with nitrate N, at pH values in the region 5.4 and 7.5 in artificial soil culture. Only Douglas fir showed substantial differences due to N source below pH 5. Growth of all species was greater at pH 5.4 than at 7.5 in each N source treatment. Growth of Douglas fir seedlings was greatest with ammonium N and least with nitrate N in sand culture 2. Supply of nitrate and ammonium in equal proportions resulted in intermediate growth. Leaf area/plant weight ratio was unaffected by N source. Analysis of nutrient solutions showed appreciable nitrification of ammonium N during the 7 days between solution changes. In the three greenhouse experiments, with little exception, increase in proportion of ammonium in N supply resulted in increase of seedling tissue N concentration. This effect was more pronounced in roots than shoots. Total N uptake by ammonium fed seedlings was about double the N uptake of nitrate fed seedlings in sand culture 2. Nursery grown Douglas fir seedlings showed greater growth response to ammonium sulphate than to calcium nitrate, and this appeared due entirely to form of N supply in the first year. A similar response in the second year was partly due to greater soil acidification by ammonium sulphate. Compared with calcium nitrate, ammonium sulphate increased N concentration of one-year old shoots, but this difference was not detected by foliar analysis of two-year old seedlings.     

Response of Thlaspi caerulescens to nitrogen, phosphorus and sulfur fertilisation

The main limiting factor for cleaning-up contaminated soils with hyperaccumulator plants is the low production of aerial biomass and the number of successive crops needed to reach the objective of remediation. The aim of this study was to contribute to the determination of a fertilisation strategy to optimise soil metal phytoextraction by Thlaspi caerulescens. A pot experiment was conducted on an agricultural soil and on a contaminated soil from the vicinity of a former Pb/Zn smelter. The nitrogen (N) treatment consisted of 4 levels (0, 11, 21.5 and 31 mg N kg(-1) dry soil (DS)) added as NH4NO3. The highest N treatment was combined with 4 levels of phosphorus (P) (0, 20, 40 and 80 mg P kg(-1) DS as KH2PO4) and sulfur (S) additions (0, 10, 20 and 30 mg S kg(-1) DS as MgSO4). The highest N fertilisation contributed significantly to enhance biomass production of T. caerulescens and to decrease the concentration of Cd and Zn in the biomass. At constant N addition, P supply did not affect metal extraction by T. caerulescens but negatively affected plant health. Sulfur supply slightly increased phytoextraction of Cd. Our results show that N and S fertilisation might interact but further investigations on the effect of such interaction on Cd extraction efficiency are needed.     

Effects of nitrogen and phosphorus on blue grama growth and mycorrhizal infection

Summary The purpose of this study was to examine the growth response of Bouteloua gracilis, with and without the vescular-arbuscular mycorrhiza (VAM), Glomus fasciculatus, to varying levels of phosphorus and nitrogen (as NH ₊ ⁴ ) and to determine whether nitrogen and phosphorus levels influence VAM establishment. Bouteloua gracilis was grown in 225 g of soil in a factorial experiment combining four levels of ammonium nitrogen (4, 30, 60, and 126 g/g), four levels of phosphorus (3, 7, 12, and 22 g/g), and with VAM spores or no spores. Bouteloua gracilis showed enhanced growth with increased nutrients over the entire range of experimental amendments. Shoot nitrogen concentration for all plants ranged from an average of 0.73% at the low amendment level to 1.61% at the high level, whereas shoot total averages ranged from 2.43 mg at the low amendment to 16.4 mg at the high amendment. Mean shoot phosphorus concentrations ranged from 0.109% at the low amendment level to 0.150% at the high amendment, while totals averaged 5.29 mg at the low amendment and 11.8 mg at the high amendment level. Infected plants were consistently smaller than uninfected plants. This reduction was significant at high nitrogen-low phosphorus, where percent infection was highest (71%). At low nitrogen levels, moderate infection (17%) was established at all phosphorus levels. No infection occurred when both nitrogen and phosphorus levels were high. The lack of a positive nutrient or biomass response to VAM establishment is contrary to most published reports, but is similar to a lack of response shown with certain grasses and other plants. It is possible that the parasitic nature of the response to infection represents the early phase of infection.     

Nitrogen and phosphorus requirements for raising mycorrhizal seedlings of Leucaena leucocephala in containers

A greenhouse study was undertaken to determine the nitrogen and phosphorus fertilization requirements for raising mycorrhizal seedlings in soil in containers. Seedlings of Leucaena leucocephala were grown for 40 days in dibble tubes containing fumigated or nonfumigated soil uninoculated or inoculated with Glomus aggregatum. The soil was fertilized with NH₄NO₃ solution to obtain 25–200 mg N kg^-1 soil, and with a KH₂PO₄ solution to establish target soil solution P concentrations of 0.015–0.08 mg P l^-1. At the end of 40 days, seedlings were transplanted into pots containing 5-kg portions of fumigated soil. Posttransplant vesicular arbuscular mycorrhizal fungal (VAMF) effectiveness, measured as pinnule P content, plant height, shoot dry weight and tissue N and P concentrations, was significantly increased by pretransplant VAMF colonization in both soils. The best posttransplant mycorrhizal colonization and mycorrhizal growth responses were observed if the nonfumigated pretransplant soil was amended with 50 mg N kg^-1 soil and 0.04 mg P l^-1 or if the fumigated pretransplant soil was amended with 100 mg N kg^-1 soil and 0.04 mg P 1^-1. There was no relationship between NP ratios of nutrients added to the pretransplant soil medium and shoot NP ratios observed after transplanting. Shoot NP ratio was also not correlated with root colonization level.Contribution from the Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No. 4025     

构树(Broussonetia papyrifera)幼苗氮、磷吸收对接种AM真菌的响应

对石灰岩适生植物构树(Broussonetia papyrifera)进行菌根真菌摩西球囊霉(Glomus mosseea)、地表球囊霉(G.versiforme)、透光球囊霉(G.diaphanum)的单独接种、混合接种和不接种处理,幼苗生长3个月后测定其根茎叶N、P含量和土壤酶活性,结果表明:接种AM真菌后构树幼苗生物量显著增加,植株根茎叶N含量较对照组显著提高,其含量为根<茎<叶。各处理中除了透光球囊霉处理的茎含量与对照差异不显著外,其余各处理含量均呈显著或极显著差异。AM真菌对构树苗P的促进效应主要体现在根、茎的利用上,而叶片P含量除混合接种外则有所降低,但M (接种处理)与M-(非接种处理)间无显著差异。构树苗根茎叶等量干物质P含量依次为叶>根>茎。进一步研究表明,接种AM真菌显著提高了土壤酶活性,这种显著性主要体现在蛋白酶和碱性磷酸酶上,植株N含量与土壤蛋白酶和碱性磷酸酶活性有显著相关性,而P含量只是根部吸收与多酚氧化酶活性显著相关。结果表明,接种AM真菌促进了宿主植物构树生物量的积累,提高了根际土壤酶活性,增加了植株对N、P的吸收。     

Response of cassava to VA mycorrhizal inoculation and phosphorus application in greenhouse and field experiments

Cassava (Manihot esculenta Crantz) was grown in the greenhouse and in the field at different levels of phosphorus applied, with or without inoculation with VA mycorrhiza in sterilized or unsterilized soil. When grown in a sterilized soil to which eight levels of P had been applied the non-inoculated plants required the application of 3200 kg P ha⁻¹ to reach near-maximum yield of plant dry matter (DM) at 3 months. Inoculated plants, however, showed only a minor response to applied P. Mycorrhizal inoculation in the P check increased top growth over 80 fold and total P uptake over 100 fold. Relating dry matter produced to the available P concentration in the soil (Bray II), a critical level of 15 ppm P was obtained for mycorrhizal and 190 ppm P for non-mycorrhizal plants. This indicates that the determination of critical levels of P in the soil is highly dependent on the degree of mycorrhizal infection of the root system. In a second greenhouse trial with two sterilized and non-sterilized soils it was found that in both sterilized soils, inoculation was most effective at intermediate levels of applied P resulting in a 15–30 fold increase in DM at 100 kg P ha⁻¹. In the unsterilized soil inoculation had no significant effect in the quilichao soil, but increased DM over 3 fold in the Carimagua soil, indicating that the latter had a native mycorrhizal population less effective than the former. When cassava was grown in the field in plots with 11 levels of P applied, uninoculated plants grown in sterilized soil remained extremely P deficient for 4–5 months after which they recuperated through mycorrhizal infection from unsterilized borders or subsoil. Still, after 11 months inoculation had increased root yields by 40%. In the non-sterilized soil inoculation had no significant effect as the introduced strain was equally as effective as the native mycorrhizal population. These trials indicate that cassava is extremely dependent on an effective mycorrhizal association for normal growth in low-P soils, but that in most natural soils this association is rapidly established and inoculation of cassava in the field can only be effective in soils with a low quantity and quality of native mycorrhiza. In that case, plants should be inoculated with highly effective strains.     

Nodulation and nitrogen accretion response of Cercocarpus betuloides seedlings to phosphorus supplementation and water availability

Greenhouse experiments were conducted to assess the effect of phosphorus (P) supplementation and water availability on incidence of nodulation and nitrogen (N) fixation by Cercocarpus betuloides seedlings. Phosphorus supplementation resulted in a two-fold increase in shoot and total dry matter of seedlings grown in a P-deficient soil. Seedlings grown in P-supplemented soil supported a larger number (4.8 vs. 0.3) and fresh mass (0.179 vs. 0.009 g) of nodules than did controls. Nitrogenase activity (acetylene reduction), N concentration and N accretion were greater for seedlings grown in P-amended soil than for controls.Seedlings were grown at four soil water potentials –0.1, –0.2, –0.4, and –0.7 MPa for 214 days. Biomass production, nodulation, acetylene reduction activity and N accretion were greatest at –0.2 MPa and declined at higher and lower soil water potentials. Independently-measured dependent variables were highly correlated, suggesting that water availability was influencing a primary plant process, most likely photosynthesis, on which other plant processes are dependent.     

Conidia of Trichoderma virens as a phosphorus source for mycorrhizal Pinus sylvestris seedlings

In this study, the mobilization and further translocation of phosphorus from conidia of saprotrophic fungus Trichoderma virens into Pinus sylvestris seedlings by nondestructive measuring of ³²P was assessed. The radioactive phosphorus flux from the conidia to the Scots pine seedlings forming mycorrhiza with Laccaria laccata and Suillus bovinus amounted up to 27.82% and 7.42%, respectively, on the 28th day of the experiment, while at the same time in nonmycorrhizal pine seedlings, the detected radioactivity reached only 0.56%. Our studies revealed that both ectomycorrhizal fungi: L. laccata and S. bovinus, mobilized the phosphorus from radioactive conidia of T. virens. On this basis, we conclude that activities of the mycosymbionts may facilitate absorption and further translocation of phosphorus from organic matter into the host plants.     

Response of early and late semiarid seral species to nitrogen and phosphorus gradients

Above- and below-ground biomass production, nitrogen (N) and phosphorus (P) tissue concentrations, and root: shoot ratios were examined for five species that are characteristic of a semiarid successional sequence under controlled greenhouse conditions. In two simultaneous experiments, seedlings of one forb, two grass, and two shrub species important in a sagebrush successional sere, were subjected to seven levels of N and P. Results of the experiments suggest distinct differences in nutrient response patterns between early and late seral species. Early seral species produced more biomass but had lower tissue nutrient concentrations than late seral species. As N and P availabilities decreased, late seral species displayed characteristics indicative of increasing competitive advantage over those of early seral species. Root: shoot ratios of the five species primarily reflected patterns related to lifeform, but with some early and late seral characteristics. Results from this study 1) confirm that nutrient use pattern, nutrient availability, and seral position relationships characteristic of mesic ecosystems hold equally true for semiarid systems, and 2) suggest that nutrients are important organizing factors in semiarid ecosystems.     

Response of irrigated dry-seeded rice to nitrogen and phosphorus in a semi-arid environment

Summary Under semi-arid conditions, three field experiments were conducted at Gezira Research Station to determine response of irrigated dry-seeded rice (Oryza sativa L. var IR 2053-206-1-3-6) to addition of nitrogen and phosphorus fertilizers. The experimental treatments included the factorial combinations of seven levels of nitrogen applied as urea and four levels of phosphorus applied as super phosphate. Plant growth and grain yield were significantly and progressively increased with the rise in the levels of added nitrogen and phosphorus. However, response to added phosphorus was restricted by the applied level of nitrogen. The responses of grain yield to nitrogen and phosphorus levels are given by quadratic regression equations. Without addition of nitrogen or phosphorus grain yield averaged 1.52 t/ha compared to 6.07 t/ha with addition of the optimal levels (160 kg N plus 35 kg P/ha). The high potential for rice production in semi-arid environment is evidently restricted by addition of relatively high rates of nitrogen and phosphorus.     

Response of Douglas fir seedlings to phosphorus fertilization and influence of temperature on this response

Summary Effects of P fertilizers on growth of Douglas fir (Pseudotsuga menziesii var.menziesii (Mirb.) Franco.) seedlings were examined in pots and nursery beds. In pot experiments levels of P equivalent to 300 kg/ha were adequate for maximum growth over 14–18 weeks and resulted in available soil P levels of 80 ppm after 15 weeks' growth. Maximum growth in pots was obtained with shoot P concentrations of 0.18%–0.20%, with higher values at lower temperatures, but the optimum concentration for one-year old (1-0) nursery seedlings was 0.16% P. Growth of seedlings was greatly restricted at a soil temperature of 5°C and an air temperature of 12°C. At a soil temperature of 10°C and an air temperature of 14°C seedling P requirement was greater than at soil and air temperatures of 20°C.Comparison showed that monammonium phosphate was more effective than calcium superphosphate in stimulating growth in pots and nursery. Triple superphosphate was also effective in the nursery. Diammonium phosphate, potassium dihydrogen phosphate and phosphoric acid had no advantages as P sources in the nursery. Available P levels of 100–130 ppm, in the loamy sand and sandy loam nurseries studied, and needle P concentrations of 0.18%, when sampled in October, were associated with maximum growth of two-year old (2-0) seedlings.P fertilization decreased root/shoot ratio, but did not alter the allometric relationship of shoot to root. Improving P status from a low level increased root growth capacity in 2-0 seedlings and P fertilization of potted seedlings increased dry weight/height ratio. Uptakes per seed bed ha of 236 kg N, 31 kg P, 81 kg K and 73 kg Ca by 2-0 seedlings were comparable with, or greater than, uptake rates of agricultural crops. Recoveries of 6–11% of P from fertilizer were recorded in the nursery.     

Effects of heavy metals on nitrogen uptake by Paxillus involutus and mycorrhizal birch seedlings

The effects of the heavy metals Cu, Cd, Ni, Pb and Zn on [(14)C]methylamine and [(14)C]aminoisobutyric acid uptake were studied in the free-living fungus Paxillus involutus and in mycorrhizal and non-mycorrhizal birch roots. The uptake of both N sources by P. involutus was inhibited by the five metals tested. However, Cu(2+) and Pb(2+) had a greater inhibitory effect. Non-competitive inhibitions were determined between heavy metals and [(14)C]methylamine uptake. [(14)C]Methylamine uptake was reduced by one third by 2 μM Cd(2+) and Cu(2+) in non-mycorrhizal roots, whereas that of mycorrhizal roots was not affected. However, it was reduced by 30 to 80% by 200 μM Cd(2+) and Cu(2+) irrespective of the mycorrhizal status. [(14)C]Aminoisobutyric acid uptake in mycorrhizal roots was not significantly affected by Cd(2+) and Cu(2+), whereas that of non-mycorrhizal roots was decreased by 77% at 200 μM Cu(2+). [(14)C]Aminoisobutyric acid uptake was 4.5 to 6 fold higher in mycorrhizal roots, compared with non-mycorrhizal roots, even under metal exposure. The high efficiency of N acquisition by mycorrhizal birch seedlings under metal exposure might be regarded as a mechanism of stress avoidance.     

Culture of mycorrhizal tree seedlings under controlled conditions: Effects of nitrogen and aluminium

A sand culture system was developed for growth of mycorrhizal seedlings under monoxenic conditions, with frequently renewed nutrient solution The composition of the nutrient solution resembled that of a forest soil solution, based on long-term measurements from forest sites at Soiling, northern Germany. Seedlings of Picea abies (L.) Karst. inoculated with Lactarius rufus (Scop.) Fr. were grown in this culture system. Plants developed rapidly, having almost totally mycorrhizal root systems. Nitrate at 2.7 mM in the nutrient solution and applied over a 13 week period had no negative effect on mycorrhizal development. Ammonium at 2.7 mM reduced the degree of mycorrhizal infection slightly, in such a way that the degree of mycorrhizal infection was reduced to a much less extent than the total number of root lips. Hence, the impact of NH⁺₄ may be primarily on root development and not on mycorrhizal fungal colonization. When the concentrations of NO-₃ and NH⁺₄ used in the present study are compared to those found in forest soil solutions, NO-₃ and NH₄⁺ would not appear to influence mycorrhizal development negatively under natural conditions. Aluminium at 0.8 mM and applied over a 13 week period reduced Mg uptake into roots and needles by 52 and 64%, respectively, resulting in needle chlorosis and strongly reduced photosynthetic activity. From a comparison of this study with others, no major difference in physiological response to aluminium exposure between non-mycorrhizal seedlings and seedlings colonized with Lactarius rufus was found.     

Response of strawberry to inoculation with arbuscular mycorrhizal fungi under very high soil phosphorus conditions

A field study was done to assess the potential benefit of arbuscular mycorrhizal (AM) inoculation of elite strawberry plants on plant multiplication, under typical strawberry nursery conditions and, in particular, high soil P fertility (Mehlich-3 extractible P=498 mg kg⁻¹). Commercially in vitro propagated elite plants of five cultivars (‘Chambly,’ ‘Glooscap,’ ‘Joliette,’ ‘Kent,’ and ‘Sweet Charlie’) were transplanted in noninoculated growth substrate or in substrate inoculated with Glomus intraradices or with a mixture of species (G. intraradices, Glomus mosseae, and Glomus etunicatum) at the acclimation stage and were grown for 6 weeks before transplantation in the field. We found that AM fungi can impact on plant productivity in a soil classified as excessively rich in P. Inoculated mother plants produced about 25% fewer daughter plants than the control in Chambly (P=0.03), and Glooscap produced about 50% more (P=0.008) daughter plants when inoculated with G. intraradices, while the productivity of other cultivars was not significantly decreased. Daughter plant shoot mass was not affected by treatments, but their roots had lower, higher, or similar mass, depending on the cultivar–inoculum combination. Root mass was unrelated to plant number. The average level of AM colonization of daughter plants produced by noninoculated mother plants did not exceed 2%, whereas plants produced from inoculated mothers had over 10% of their root length colonized 7 weeks after transplantation of mother plants and ∼6% after 14 weeks (harvest), suggesting that the AM fungi brought into the field by inoculated mother plants had established and spread up to the daughter plants. The host or nonhost nature of the crop species preceding strawberry plant production (barley or buckwheat) had no effect on soil mycorrhizal potential, on mother plant productivity, or on daughter plant mycorrhizal development. Thus, in soil excessively rich in P, inoculation may be the only option for management of the symbiosis.