Response of nitrogen-transforming microorganisms to arbuscular mycorrhizal fungi

 We studied fluctuations in the numbers of autotrophic ammonium oxidizers, ammonifying microorganisms and denitrifying microorganisms in pot cultures of mycorrhizal and non-mycorrhizal maize. The populations were enumerated after 0, 15, 30, 45 and 60 days of plant growth. Two arbuscular mycorrhizal (AM) fungi belonging to different Glomus species were investigated. Pot cultures with AM-infected maize had significant quantitative and qualitative changes in the root-associated population of N-transforming bacteria compared with the non-mycorrhizal controls. The occurrence of autotrophic ammonium oxidizers in pot cultures of the AM fungi Glomus mosseae and G. fasciculatum was significantly higher than in non-mycorrhizal cultures throughout maize growth. The occurrence of these bacteria was delayed by 15 days in non-mycorrhizal as opposed to Glomus-colonized soil. Ammonifying and denitrifying bacterial populations were significantly decreased in the pot cultures of AM plants compared with the control. The distribution patterns of the physiological groups of bacteria tested were similar for both AM treatments but different from that of the non-mycorrhizal controls. Activity measurements expressed on a per cell basis showed changes with respect to the form of N in the mycorrhizal soil. G. fasciculatum was more active than G. mosseae during the earlier stages of plant growth. Received: 8 July 1997     

Effect of salinity and arbuscular mycorrhizal fungi on growth and some physiological parameters of Citrus jambheri

Arbuscular mycorrhizal (AM) fungi alleviate the unfavorable effects of salinity stress on plant growth. A pot study was conducted to determine the effects of AM fungi and salt on growth and some physiological parameters of Citrus jambheri rootstock. Four levels of salinity (2, 4, 6, and 8 dS m^?1 as NaCl) and three mycorrhizal treatments (Glomus etunicatum, Glomus intraradices and non-mycorrhizal (NM) control) were used. As salinity increased, all measured characteristics of plants after 4.5-month growth except Na uptake, proline content, and electrolyte leakage decreased. Shoot dry weight and K uptake were significantly higher in G. intraradices-colonized seedlings than NM controls at all salinity levels. Root dry weight and shoot P uptake were significantly higher in G. etunicatum-colonized seedlings than NM controls at all salinity levels. G. intraradices-colonized seedlings had significantly higher proline content than NM controls and G. etunicatum-colonized seedlings at salinity levels of 4, 6 and 8 dS m^?1. The electrolyte leakage percentage was significantly lower in G. intraradices-colonized seedlings than NM controls at all salinity levels. The data demonstrated that mycorrhizal citrus seedlings exhibited greater tolerance to salt stress than NM seedlings and the enhanced proline content seems to be one of the mechanisms involved.     

Influence of arbuscular mycorrhizae on growth, leaf yield and phosphorus uptake in mulberry (Morus alba L.) under rainfed, lateritic soil conditions

 A field experiment was conducted to study the effect of arbuscular mycorrhizal association in mulberry saplings var. S₁, together with two levels of phosphate, on growth characters, leaf yield and phosphorus uptake. The experiment was conducted in the period 1995 to 1997 at the Technical Service Centre, Kashipur, Purulia (West Bengal) under rainfed, lateritic soil conditions. Pooled data analysis revealed that there was no significant variation among the treatments in most of the characters except height of plant and leaf moisture percentage. Plant height in both treatments with Glomus mosseae was significantly less than in the others, while there was significant reduction in moisture percentage in G. fasciculatum with 25 kg of phosphate and G. mosseae with 10 kg of phosphate ha^–1 year^–1 than in other treatments. There was no significant season–treatment effect on any of the parameters. Considering all the quantitative and qualitative characters, G. fasciculatum together with the most economic dose of phosphate showed more promising performance than G. etunicatum and G. mosseae with different doses of phosphate fertilizer. At 80% of the recommended dose of inorganic phosphate fertilizer, Rs 534.0 were earned as net profit. Received: 11 February 1998     

Microbial and chemical sources of phosphorus supply modulate the yield and chemical composition of essential oil of rose-scented geranium (Pelargonium species) in sodic soils

Rose-scented geranium (Pelargonium sp.) is a highly valued aromatic crop. Its growth is limited by soil salinity and sodicity stress. Arbuscular mycorrhizal (AM) fungus, phosphate-solubilizing bacteria (PSB), and P fertilizers may enhance the growth and secondary metabolism in geranium plants. In this context, a pot experiment was conducted to study the effects of PSB, AM fungi (Glomus intraradices), and P fertilizer on the yield, chemical composition of essential oil, and mineral element acquisition of geranium. The dry matter yield of shoot and essential oil yield, and mineral element (P, K, Ca, Mg, Na, Fe, Cu, and Zn) uptake in shoot tissues of geranium were significantly increased by the inoculation with AM fungi, co-inoculation with AM fungi and PSB, and P fertilization as compared to control. While the co-inoculation of geranium with AM fungi and PSB significantly enhanced the content of the monoterpenes such as citronellol, geraniol, geranial, and a sesquiterpene (10-epi-γ eudesmol), the P fertilization only enhanced the content of a sesquiterpene, 10-epi-γ eudesmol in the volatile oil. We conclude that the co-inoculation of PSB and AM fungi could be the best natural alternative to phosphate fertilizers to enhance the yield and quality of essential oil from geranium plants grown in sodic soils.     

Effectiveness of two arbuscular mycorrhizal fungi on concentrations of essential oil and artemisinin in three accessions of Artemisia annua L.

A field experiment was conducted to study and compare the effectiveness of two arbuscular mycorrhizal fungi (AMF), Glomus macrocarpum (GM) and Glomus fasciculatum (GF) on three accessions of Artemisia annua. The AM inoculation significantly increased the production of herbage, dry weight of shoot, nutrient status (P, Zn and Fe) of shoot, concentration of essential oil and artemisinin in leaves as compared to non-inoculated plants. The extent of growth, nutrient concentration and production of secondary plant metabolites varied with the fungus–plant accession combination. The mycorrhizal dependency of the three accessions was related to the shoot: root ratio. Comparing the two fungal inoculants in regard to increase in essential oil concentration in shoot, the effectiveness of GF was more than that of GM. While in two accessions, GM was more effective in enhancing artemisinin concentration than GF. Increase in concentration of essential oil was found to be positively correlated to P-status of the plant. Conversely, no correlation was found between shoot-P and artemisinin concentration.     

Interaction of arbuscular mycorrhizal fungi and rhizobia: Effects on flax yield in spoil‐bank clay

This study focused on the application of native strains of arbuscular mycorrhizal fungi (AMF) and Sinorhizobium in effective crop production during reclamation of coal‐mine spoil banks. Two greenhouse experiments were conducted in spoil‐bank clay with a low dose of organic amendment to determine whether the microbial inoculation improves growth and utility qualities of two cultivars of Linum usitatissimum L. (oil and fiber flax). Inoculation with two native AMF isolates (Glomus mosseae, G. intraradices, and their mixture) significantly increased growth and shoot phosphorus (P) concentration of both flax cultivars. Inoculated fiber flax plants produced fivefold more fibers than the uninoculated ones. In oil flax, mycorrhizal inoculation significantly but quantitatively to a minor degree decreased the concentration of nonsaturated fatty acids in the seed oil. A mixture of five native Sinorhizobium sp. strains supported growth and P uptake of oil flax only in the absence of AMF. However, these beneficial effects of the bacteria were significantly lower as compared to AMF. No synergic action of Sinorhizobium strains and AMF was observed, and their interactions were often even antagonistic. Inoculation with AMF significantly decreased population density of Sinorhizobium in the soil. These results suggest that a careful selection of suitable bacterial strains is necessary to provide effective AMF combinations and maximize flax‐growth support.     

Growth and iron nutrition of broccoli (Brassica oleracea L. var. italica Plenck), grown in a Typic Ustochrept, as influenced by vesicular-arbuscular mycorrhizal fungi in the presence of pyrite and farmyard manure

Greenhouse experiments were conducted using potted soil (Fe-deficient Typic Ustochrept) to study the influence of the vesicular-arbuscular mycorrhizal fungi (VAM), Glomus macrocarpum and G. fasciculatum, on the mobilisation of Fe in broccoli (Brassica oleracea L. var. italica Plenck) in the presence of pyrite and farmyard manure (FYM). Individual applications of either VAM or pyrite with NPK fertiliser significantly enhanced both the Fe²⁺ content in leaf tissue and total uptake of Fe and resulted in increased curd and straw yields of broccoli compared to those observed with NPK alone. Though the application of FYM decreased the Fe²⁺ content in leaf tissue relative to plants supplied NPK alone, this result was not statistically significant. The available Fe content in soil, after harvest of broccoli, was found to be lower in the presence of VAM than in the control. Received: 18 June 1997     

砷污染土壤中接种丛枝菌根真菌和蚯蚓对土壤线虫群落和植物砷吸收的影响

The influences of arbuscular mycorrhizal fungi (MF, Acaulospora spp. and Glomus spp.), rice straw and earthworms (RE, Eisenia foetida) on nematode communities and arsenic (As) uptake by maize (Zea mays L.) in As-contaminated soils were examined in a field experiment conducted in Wujiang, Jiangsu Province, China. The experiment was designed as a 2 × 2 factorial with the factors of MF (inoculated or uninoculated) and RE (added or not added). The results demonstrated that MF inoculation led to significantly higher root colonization of MF and root dry weight. Plants inoculated with both MF and RE had the highest As concentrations in root. The number of total nematodes increased with MF inoculation when RE was absent, and decreased with RE addition when MF was inoculated. The improved abundance of nematodes with the MF treatment implied that the tested MF acted as food sources for fungivores. The abundances of omnivores-predators and plant parasites were reduced by earthworm activity. Twenty-seven genera of nematodes were identified, with Filenchus dominant in all treatments. Trophic diversity (TD), Shannon-Weaver diversity (H′), Simpson dominance index (λ), and species richness (SR) indicated higher species diversity, more proportionate species composition, evenly distributed species, and more food sources in the MF, RE, and their interaction treatments. Maturity index (MI) showed a moderately disturbed environment due to As pollution. Besides enhancing plant uptake of contaminants, MF and RE amendments could also improve soil health by restoring the structure of soil communities, as reflected by more stable nematode community structure.     

Behaviour of arbuscular-mycorrhizal fungi on Vigna luteola growth and its effect on the exchangeable (32P) phosphorus of soil

 The effect of the inoculation of Glomus mosseae and Scutellospora fulgida, singly or in mixed inocula, was tested on the growth and mycorrhizal characteristics of Vigna luteola. Soil depletion by mycorrhizal inoculation was determined as exchangeable ³²PO₄. Five treatments were performed: non-inoculated control (C); inoculated with a suspension of microorganisms free of mycorrhizae (M); inoculated with S. fulgida plus microorganisms (Sf+M); inoculated with G. mosseae plus microorganisms (Gm+M) and a treatment inoculated with both S. fulgida and G. mosseae plus microorganisms (Sf+Gm+M). G. mosseae was the most efficient fungus in promoting growth of V. luteola. This fungus produced higher shoot dry weight, P uptake in shoots (P_shoot) and P_shoot/arbuscules ratio than the S. fulgida inoculum, even though percent arbuscular-mycorrhizal (AM) colonization was similar for the two single AM-inoculated treatments. The highest value of isotopically exchangeable P was recorded for treatment M (P<0.05). In comparison with M, Gm+M treatment reduced the different P pools more than Sf+M treatment. C_p values were reduced to half by Gm+M and Sf+M treatments and were minimum for the combined treatment. Pools A (exchangeable phosphate between 1 min and 1 day) and B (exchangeable phosphate between 1 day and 3 months) were reduced by Gm+M treatment in contrast to Sf+M, which left them unchanged. The depression of isotopically exchangeable P for all pools obtained for the Sf+Gm+M treatment suggested that specific characteristics of fungi caused differences in P absorption, which in addition was altered by the possible interactions among them, when multiple inocula were used in a soil low in native P. Received: 29 January 1999     

AM真菌接种对甘薯产量和品质的影响

通过田间试验方法研究了接种AM真菌对甘薯产量和品质的影响。结果表明,种植6周时接种能够提高甘薯的菌根侵染率、生长和吸P量,收获时可提高甘薯的产量和品质。从接种效果看,本地分离的菌株接种效果好于异地分离菌株,混合菌株好于单一菌株。     

Influence of arbuscular mycorrhizal fungi and salinity on growth,biomass, and mineral nutrition of<Emphasis Type="Italic"> Acacia auriculiformis</Emphasis>

The effect of salinity on the efficacy of two arbuscular mycorrhizal fungi, Glomus fasciculatum and G. macrocarpum, alone and in combination was investigated on growth, development and nutrition of Acacia auriculiformis. Plants were grown under different salinity levels imposed by 0.3, 0.5 and 1.0 S m^-1 solutions of 1 M NaCl. Both mycorrhizal fungi protected the host plant against the detrimental effect of salinity. The extent of AM response on growth as well as root colonization varied with fungal species, and with the level of salinity. Maximum root colonization and spore production was observed with combined inoculation, which resulted in greater plant growth at all salinity levels. AM fungal inoculated plants showed significantly higher root and shoot weights. Greater nutrient acquisition, changes in root morphology, and electrical conductivity of soil in response to AM colonization was observed, and may be possible mechanisms to protect plants from salt stress.     

Synergistic effects of vesicular-arbuscular mycorrhizal fungi and diazotrophic bacteria on nutrition and growth of sweet potato (Ipomoea batatas)

Summary Sweet potatoes were micropropagated and then transplanted from axnic conditions to fumigated soil in pots in the greenhouse. Spores of Glomus clarum were obtained from Brachiaria decumbens or from sweet potatoes grown in soil infected with this fungus and with an enrichment culture of Acetobacter diazotrophicus. Three experiments were carried out to measure the beneficial effects of vesicular-arbuscular mycorrhizal (VAM) fungi-diazotroph interactions on growth, nutrition, and infection of sweet potato by A. diazotrophicus and other diazotrophs obtained from sweet potato roots. In two of these experiments the soils had been mixed with ¹⁵N-containing organic matter. The greatest effects of mycorrhizal inoculation were observed with co-inoculation of A. diazotrophicus and/or mixed cultures of diazotrophs containing A. diazotrophicus and Klebsiella sp. The tuber production was dependent on mycorrhization, and total N and P accumulation were increased when diazotrophs and G. clarum were applied together with VAM fungal spores. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM fungi or when present within G. clarum spores. More pronounced effects on root colonization and intraradical sporulation of G. clarum were observed when A. diazotrophicus was co-inoculated. In non-fumigated soil, dual inoculation effects, however, were of lower magnitude. ¹⁵N analysis of the aerial parts and roots and tubers at the early growth stage (70 days) showed no statistical differences between treatments except for the VAM+Klebsiella sp. treatment. This indicates that the effects of A. diazotrophicus and other diazotrophs on sweet potato growth were caused by enhanced mycorrhization and, consequently, a more efficient assimilation of nutrients from the soil than by N₂ fixation. The possible interactions between these effects are discussed.     

MES [2-(N-morpholine)-ethane sulphonic acid] buffer promotes the growth of external hyphae of the arbuscular mycorrhizal fungus Glomus intraradices in an alkaline sand

In this work we investigated the hypothesis, suggested by unpublished experiments carried out previously, that application of MES [2-(N-morpholino)-ethane sulphonic acid] buffer to the substrate of plants hosting the arbuscular mycorrhizal fungus Glomus intraradices might favour the growth of external hyphae. The substrate of Trifolium pratense plants, with or without G. intraradices inoculum, was treated twice weekly, between weeks 5 and 11 of growth, plant with distilled water, P-supplemented 10% Hewitt nutrient solution (H+P), 51mM in H+P or 1mM TRIS-HCl in H+P. MES buffer increased the length of external G. intraradices hyphae and the density of propagules in the substrate, but plant growth was not affected. Although both TRIS-HCl and MES caused substrate pH to fall (from 8.51 to 7.35 and 7.17, respectively), only MES induced a significant increase in external hypha length. We conclude that the effect of MES on external hypha length was not due to plant growth or to substrate pH per se. Received: 10 March 1996     

枯草芽孢杆菌SO113分泌蛋白的抑菌作用及抗菌蛋白的分离纯化

枯草芽孢杆菌（Bacillus subtilis)SO113对水稻白叶枯病菌（Xanthomonas oryzaepv.oryzae）有强烈的抑菌作用（表现出一透明抑菌圈）。培养液上清中加入60％饱和度的硫酸铵沉淀所得的蛋白粗提液对热稳定，对链霉蛋白酶E不敏感，对蛋白酶K部分敏感，对中国各稻区白叶枯病菌的7种致病型都有强烈的抑杀作用，蛋白粗提液100℃加热15min后脱盐，经DEAE0－Sepharose Fast Flow柱层析得3个未分开的抗菌活性峰，示吸附部分经CM－Sepharose Fast Flow柱层析，得到1个主活性峰，由于B.subtilis SO113的分泌蛋白对水稻白叶枯病菌表现出良好的广谱抗性以及丰富的抗菌活性峰，因此对其进一步的研究和设法隆编码抗菌蛋白的基因都具有重要意义。     

Rhodotorulic acid enhances root colonization of tomato plants by arbuscular mycorrhizal (AM) fungi due to its stimulatory effect on the pre-symbiotic stages of the AM fungi

Exudates of Rhodotorula mucilaginosa, a yeast commonly found in the rhizosphere, increased hyphal length of the arbuscular mycorrhizal (AM) fungi Gigaspora rosea and Gigaspora margarita. Rhodotorulic acid (RA), a siderophore compound obtained from R. mucilaginosa exudates, increased hyphal length and branching. Thus, the increase in the number of entry points and the higher AM root colonization of tomato plants in the presence of RA can at least partially be explained by the positive effect of RA on the pre-symbiotic stages of the AM fungi.     

Impacts of earthworms and arbuscular mycorrhizal fungi (Glomus intraradices) on plant performance are not interrelated

Earthworms and arbuscular mycorrhizal fungi (AMF) might interactively impact plant productivity; however, previous studies reported inconsistent results. We set up a three-factorial greenhouse experiment to study the effects of earthworms (Aporrectodea caliginosa Savigny and Lumbricus terrestris L.) and AMF (Glomus intraradices N.C. Schenck & G.S. Sm.) on the performance (productivity and shoot nutrient content) of plant species (Lolium perenne L., Trifolium pratense L. and Plantago lanceolata L.) belonging to the three functional groups grasses, legumes and herbs, respectively. Further, we investigated earthworm performance and plant root mycorrhization as affected by the treatments. Our results accentuate the importance of root derived resources for earthworm performance since earthworm weight (A. caliginosa and L. terrestris) and survival (L. terrestris) were significantly lower in microcosms containing P. lanceolata than in those containing T. pratense. However, earthworm performance was not affected by AMF, and plant root mycorrhization was not modified by earthworms. Although AMF effectively competed with T. pratense for soil N (as indicated by δ¹⁵N analysis), AMF enhanced the productivity of T. pratense considerably by improving P availability. Remarkably, we found no evidence for interactive effects of earthworms and AMF on the performance of the plant species studied. This suggests that interactions between earthworms and AMF likely are of minor importance.     

Vermicompost from biodegraded distillation waste improves soil properties and essential oil yield of Pogostemon cablin (patchouli) Benth

Pre-treatment of patchouli lignocellullosic distillation waste with a consortium of microbes viz. Trichoderma harzianum – ATCC PTA-3701, Pseudomonas monteilii – HQ995498, Bacillus megaterium – ATCC 14581 and Azotobacter chroococcum – MTCC 446 significantly enhanced the bio-degradation of cellulose, hemicelluloses and lignin by 58.44%, 29.44% and 65.23%, respectively and improved the yield of vermicompost by 15%. Application of bioinoculant-enriched vermicompost (BEVC) in patchouli reduced the percent disease index (PDI) of Rhizoctonia root-rot by 36.36% and improved the essential oil yields by 59.74%, 37.70% and 37.96 as compared to plots receiving untreated (not enriched) vermicompost (VC), bioinoculant-enriched compost (BEC) and chemical fertilizers (CF), respectively. Furthermore, BEVC application resulted in a marked improvement in physical (bulk density and water holding capacity) and chemical properties (pH, percent organic carbon, available N, P and K) of the soil. In conclusion, BEVC from distillation waste can be safely utilized as a bio-organic input unambiguously particularly in situations where the use of chemical fertilizers and pesticides is restricted like organic agriculture.     

Suppressive effects of seed-applied fungicides on arbuscular mycorrhizal fungi (AMF) differ with fungicide mode of action and AMF species

Seed-applied fungicides are commonly used to prevent or suppress fungal disease organisms in pulse crop production. However, non-target beneficial fungi, such as arbuscular mycorrhizal fungi (AMF), also may be affected. Seed-applied fungicides Agrox^® FL (active ingredient: captan), Allegiance™ FL (metalaxyl), Apron Maxx^® RTA^® (fludioxonil and metalaxyl), Thiram 75WP (thiram), Vitaflo^® 280 (carbathiin and thiram), Crown^® (carbathiin and thiabendazole), and Trilex^® AL (trifloxystrobin and metalaxyl) were assessed in a greenhouse study for their effects on colonization and development of AMF in pea and chickpea, and the consequent impact on plant growth. In the absence of disease pressure, systemic fungicides Allegiance™ FL, Apron Maxx^® RTA^®, Vitaflo^® 280, Crown^® and Trilex^® AL restricted mycorrhizal colonization, host growth and P uptake to different levels. In contrast, contact fungicides Agrox^® FL and Thiram 75WP had minimal effects on mycorrhizal colonization, host growth and P uptake. Although consequent sporulation and glomalin-related protein production were not significantly affected by fungicides at an early host growth stage, the compositional structure of the AMF community in host roots was significantly altered in response to Agrox^® FL, Allegiance™ FL, Apron Maxx^® RTA^®, and Trilex^® AL as revealed by pyrosequencing-based analysis of fungal 18S rRNA. These results indicate that the suppressive effects of seed-applied fungicides on AMF development depend on specific fungicide-AMF interactions.     

Influence of different dates of planting on growth,herb, oil yield and quality of essential oil of menthol mint (Mentha arvensis) in the North Indian Plain

A field experiment was conducted for two years (2007–2008 and 2008–2009) to study the effect of different dates of planting for three menthol mint (Mentha arvensis) cultivars (Saksham, Kushal and Kosi; developed by Central Institute of Medicinal and Aromatic Plants, Lucknow, India) on herb yield, oil yield and oil quality. All the cultivars produced higher herb and oil yields when planted on 18 February, compared with cultivars planted on other dates in both years. Average herb yield for Saksham increased by 12.89, 30.66, 54.01 and 85.71%, average herb yield for Kushal increased by 15.09, 30.57, 71.69 and 104.91%, and average herb yield for Kosi increased by 32.48, 53.28, 87.23 and 109.49% when planted on 4 January, 19 January, 3 February and 18 February, respectively, over cultivars planted on 20 December. Among the cultivars, Kosi produced a higher herb yield than Saksham and Kushal planted on 18 February. Saksham, Kushal and Kosi produced ～89.39, 152.04 and 170.22% higher oil yield, when planted on 18 February over their respective planting date of 20 December. Maximum menthol content was recorded in cultivar Saksham when planted on 18 February and in Kushal and Kosi when planted on 4 March.