Effects of tillage systems on soil characteristics, glomalin and mycorrhizal propagules in a Chilean Ultisol

Tillage affects the soil physical and chemical environment in which soil microorganisms live, thereby affecting their number, diversity and activity. However, soil disturbance generally has the greatest impact on biological properties, including both free and symbiotic fungal populations. Interest in more ecologically sustainable agricultural systems is rising with increasing recognition that agricultural intensification can adversely affect environmental quality. This paper discusses the effect of tillage system on some soil characteristics, such as pH, C, N and S levels, total and Olsen-P contents including some P forms associated with organic matter, glomalin contents and arbuscular mycorrhizae (AM) parameters, such as root colonization, spore number and total and active hyphal length. Measurements were in the sixth year of an on-going tillage-rotation experiment conducted on an Ultisol under no-till (NT), reduced tillage (RT) and conventional tillage with stubble mixed into the soil (CTS) or stubble burnt (CTB). Soil was sampled at two dates; after wheat (Triticum aestivum) harvest (autumn) and 6 months after subsequent grassland seeding (spring). Higher C, N, S, total P and fulvic acid-P concentrations and pH occurred under NT and RT than under CTS and CTB after wheat harvest. However, results at the second sampling were not consistent. AM spore number and active hyphal length were highest under NT having the greatest incidence on AM root colonization and P concentration in shoots of the pasture. Glomalin concentration was higher under NT and RT than under CTS and CTB but no differences in calculated glomalin to total C (ca. 5%) were found. It is concluded that a less disruptive effect of NT influences positively all soil characteristics and also increases P acquisition by the following crop in the rotation system.     

Characterization of glomalin as a hyphal wall component of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal fungi (AMF) produce a protein, glomalin, quantified operationally in soils as glomalin-related soil protein (GRSP). GRSP concentrations in soil can range as high as several mg g⁻¹ soil, and GRSP is highly positively correlated with aggregate water stability. Given that AMF are obligate biotrophs (i.e. depending on host cells for their C supply), it is difficult to explain why apparently large amounts of glomalin would be produced and secreted actively into the soil, since the carbon could not be directly recaptured by the mycelium (and benefits to the AMF via increased soil structure would be diffuse and indirect). This apparent contradiction could be resolved by learning more about the pathway of delivery of glomalin into soil; namely, does this occur via secretion, or is glomalin tightly bound in the fungal walls and only released after hyphae are being degraded by the soil microbial community? In order to address this question, we grew the AMF Glomus intraradices in in vitro cultures and studied the release of glomalin from the mycelium and the accumulation of glomalin in the culture medium. Numerous protein-solubilizing treatments to release glomalin from the fungal mycelium were unsuccessful (including detergents, acid, base, solvents, and chaotropic agents), and the degree of harshness required to release the compound (autoclaving, enzymatic digestion) is consistent with the hypothesis that glomalin is tightly bound in hyphal and spore walls. Further, about 80% of glomalin (by weight) produced by the fungus was contained in hyphae and spores compared to that released into the culture medium, strongly suggesting that glomalin arrives mainly in soil via release from hyphae, and not primarily through secretion. These results point research on functions of glomalin and GRSP in a new direction, focusing on the contributions this protein makes to the living mycelium, rather than its role once it is released into the soil.     

AM真菌对桑树根围土壤团聚体的影响机制

为揭示丛枝菌根 (Arbuscular mycorrhizal,AM)真菌对植桑土壤的影响及机制,采用盆栽试验研究接种摩西管柄囊霉 (Funneliformis mosseae,Fm)和根内根生囊霉 (Rhizophagus intraradices,Ri)对土壤有机碳(Soil organic carbon, SOC)、球囊霉素相关土壤蛋白 (Glomalin related soil protein, GRSP)及团聚体组成与稳定性的影响。结果表明：⑴ 接种Ri显著增加土壤大团聚体百分比,并提高平均质量直径 (Mean weight diameter, MWD)和几何平均直径 (Geometric mean diameter, GMD)、显著降低团聚体破坏率 (Percentage of aggregate destruction, PAD)。⑵ 接种Fm和Ri均显著增加微团聚体SOC含量,接种Fm显著降低大团聚体总GRSP含量,而接种Ri却显著增加大团聚体和微团聚体总GRSP含量及易提取GRSP含量。⑶ 接种AM真菌对整体SOC的效应为负,土壤总GRSP对SOC占比在25.5%~76.5%之间,土壤易提取GRSP对SOC占比在4.87%~5.93%之间,且Ri的接种效应高于Fm。⑷ 总GRSP、易提取GRSP和SOC对团聚体组成表现均为正向显著影响,其中易提取GRSP是主要驱动因子,而总GRSP是土壤团聚体稳定性的主要影响因子。综上,AM真菌作用下桑树根围土壤团聚体得以改善并趋于稳定,Ri的接种效应明显大于Fm;土壤团聚体的形成主要依赖易提取GRSP,而其稳定性主要受总GRSP影响。     

Dynamics of arbuscular mycorrhizal fungi and glomalin in the rhizosphere of Artemisia ordosica Krasch. in Mu Us sandland, China

To understand the ecological significance of arbuscular mycorrhizal (AM) associations in semi-arid and arid lands, the temporal and spatial dynamics of AM fungi and glomalin were surveyed in Mu Us sandland, northwest China. Soil samples in the rhizosphere of Artemisia ordosica Krasch. were collected in May, July and October 2007, respectively. Arbuscular, hyphal and total root infection and spore density of AM fungi peaked in summer. The mean contents of total Bradford-reactive soil proteins (T-BRSPs, TG) and easily extractable Bradford-reactive soil proteins (EE-BRSPs, EEG) reached maximal values in spring. Spore density and two BRSPs fractions were the highest in the 0-10 cm soil layer, but the ratios of two BRSPs fractions to soil organic carbon (SOC) were the highest in the 30-50 cm soil layer. Hyphal infection was negatively correlated with soil enzymatic activity (soil urease and acid phosphatase) (P < 0.05). Arbuscular infection was negatively correlated with soil acid phosphatase (P < 0.01). Spore density was positively correlated with edaphic factors (soil available N, Olsen P, and SOC) and soil enzymatic activity (soil acid and alkaline phosphatase) (P < 0.01). Two BRSPs fractions were positively correlated with edaphic factors (soil available N and SOC) and soil enzymatic activity (soil urease, acid and alkaline phosphatase) (P < 0.01). TG was positively correlated with soil Olsen P (P < 0.05). We concluded that the dynamics of AM fungi and glomalin have highly temporal and depth patterns, and influenced by nutrient availability and enzymatic activity in Mu Us sandland, and suggest that glomalin are useful indicators for evaluating soil quality and function of desert ecosystem on the basis of its relationship to AM fungal community, soil nutrient dynamics and carbon cycle.     

Bradford-reactive soil proteins and aggregate stability under abandoned versus tilled olive groves in a semi-arid calcisol

Glomalin, a substance produced by arbuscular mycorrhizal fungi, is reported to play a role in soil aggregation, but this role has been questioned in soils rich in calcium carbonate. We studied the relationship between aggregation stability and glomalin in a Haplic Calcisol comparing abandoned and active cultivation of olive groves. Abandonment was associated with increases in soil organic carbon, the percentage of water stable aggregates (WSA_1-2mm), and easily extractable and total Bradford-reactive soil protein. WSA_1-2mm was strongly positively correlated with both easily extractable and total Bradford-reactive soil protein. While easily extractable Bradford-reactive soil protein measured in both stable and unstable aggregates did not show any significant differences, Bradford-reactive soil protein was twice as high in stable than in unstable aggregates under both tillage and abandonment. Our results suggest that Bradford-reactive soil protein influences aggregate stability, even in soils with low organic matter and high calcium carbonate contents. However, more research is needed to elucidate the role of easily extractable Bradford-reactive soil protein in soil aggregation.     

土地利用方式对球囊霉素土层分布的影响

球囊霉素对维持土壤有机碳平衡和土壤团聚体稳定性具有明显作用,但不同土地利用方式下土壤球囊霉素的土层分布模式及其影响因素尚不清楚.本研究选取农田、人工草地、果园和撂荒地4种不同土地利用方式,分别采集0～10 cm、10～20 cm、20～30 cm和30～40 cm 4个土层土样,通过测定土壤球囊霉素.pH、速效磷、有机碳和蛋白酶活性,研究不同土地利用方式下土壤球囊霉素土层分布的模式及影响因素.结果表明,土壤球囊霉素平均含量为1.41～3.18 mg·g-1,占土壤有机碳的6.98%～31.34%,是土壤中的一个重要碳库.土壤球囊霉素在不同土地利用方式和土层剖面之间表现出显著差异(P<0.01),具有明显的垂直分布特征,除撂荒地外,其随土层深度的增加表现出降低趋势.土壤球囊霉素分别与土壤速效磷、蛋白酶呈显著正相关(P<0.01).土壤速效磷在很大程度上决定土壤球囊霉素的含量和分布.土壤球囊霉素含量和土壤蛋白酶活性之间为间接相关关系,该关系有待于进一步研究.建议把球囊霉素作为研究丛枝菌根真菌生长状况和土壤生态系统波动的一个重要指标.     

Glomalin-related soil protein responses to elevated CO2 and nitrogen addition in a subtropical forest: Potential consequences for soil carbon accumulation

According to the economy theory, plants should preferentially allocate photosynthate to acquire below-ground resources under elevated atmospheric carbon dioxide (eCO₂) but decrease below-ground C allocation when nitrogen (N) is sufficient for plant growth. Arbuscular mycorrhizae (AM) represent a critical mechanism of below-ground nutrient acquisition for plants. The dynamics of arbuscular mycorrhizal fungi (AMF) could therefore reflect the response of plant C allocation under eCO₂ and N addition. We examined the responses of glomalin-related soil protein (GRSP) to eCO₂ (approximately 700 μmol mol⁻¹ CO₂) and/or N addition (100 kg N ha⁻¹ yr⁻¹ as NH₄NO₃) in a modeled subtropical forest to better understand its potential influence on soil C storage. We hypothesized that GRSP would increase under eCO₂ and decrease under N addition. Furthermore, the positive effects of eCO₂ on GRSP would be offset by extra N addition, and GRSP would remain unchanged under combined eCO₂ and N addition. Our results showed that the mean concentrations of easily extractable GRSP (EE-GRSP) and total GRSP (T-GRSP) were 0.35 ± 0.05 and 0.72 ± 0.13 mg C cm⁻³, respectively, which accounted for 2.76 ± 0.53% and 5.67 ± 0.92% of soil organic carbon (SOC) in the 0–10 cm soil layer. Elevated CO₂ significantly increased T-GRSP by 35.02% but decreased EE-GRSP by 5.09% in the top 10 cm soil layer. The opposite responses of T-GRSP and EE-GRSP to eCO₂ might result from an unchanged photosynthate investment to AMF with possible changes in their decomposition rates. The effect of N on GRSP was contrary to our hypothesis, i.e., there was a 1.72%–48.49% increase in T-GRSP and a slightly increase in EE-GRSP. Both EE-GRSP and T-GRSP concentrations increased under the combination of eCO₂ and N addition, which was inconsistent with our hypothesis. The significant increase of EE-GRSP under the combination of eCO₂ and N addition was partly caused by more rapid plant growth and reduced microbial diversity, and the marginal increase of T-GRSP indicated that the interaction between eCO₂ and N addition offset their independent effects. In addition, the relatively higher accumulation ratios of GRSP (22.6 ± 13.6%) compared with SOC (15.9 ± 9.4%) indicated that more rapid GRSP deposition in the soil might accelerate SOC accumulation under eCO₂ and N addition. Our results will improve the understanding of the functioning of GRSP in soil C sequestration under global environmental change scenarios.     

Losses of glomalin-related soil protein under prolonged arable cropping: A chronosequence study in sandy soils of the South African Highveld

Residues of arbuscular mycorrhizal fungi (AMF) may be important for agroecosystem functioning due to their ability to promote soil aggregation, especially in coarse textured soils with little biomass input and low capacity to conserve soil organic matter (SOM). Our aim was to assess the fate of AMF residues with prolonged arable cropping in coarse textured soils in a subtropical savannah assuming that glomalin-related soil protein (GRSP), especially the MAb32B11-immunoreactive fraction, mainly constitutes material of AMF origin. In three agroecosystems on the South African Highveld, surface soils were sampled. The former grassland soils had a history of up to 98 yr of cropping. We measured four GRSP fractions: Bradford-reactive soil protein (BRSP) and immunoreactive soil protein (IRSP), and easily extractable fractions of both. The primary grassland sites exhibited generally low contents of SOM and low GRSP contents. Prolonged arable land use of former grassland soils reduced the content of GRSP further. The decline could be described with a mono-exponential function with rate constants ranging from 0.04 to 0.41 yr⁻¹. Depending on the GRSP fraction, steady-state conditions were reached after 11-92 yr on a level of 39-69% of the initial contents. We conclude that even though GRSP fractions had the same hypothesized origin, they comprised pools with different stability or replacement rate. Easily extractable IRSP was lost most rapidly. In contrast to carbon, nitrogen and microbial residue dynamics, GRSP contents were not reduced below a certain steady-state level, despite potentially negative management effects on AMF, such as tillage, inclusion of fallows into crop rotation and fertilization with inorganic phosphorus. The steady-state GRSP contents coincided with low, but steady agroecosystem yields under the given cropping management.     

Carbon and nitrogen in operationally defined soil organic matter pools

Humic substances [humic acid (HA), fulvic acid (FA), and insoluble humin], particulate organic matter (POM), and glomalin comprise the majority (ca 75%) of operationally defined extractable soil organic matter (SOM). The purpose of this work was to compare amounts of carbon (C) and nitrogen (N) in HA, FA, POM, and glomalin pools in six undisturbed soils. POM, glomalin, HA, and FA in POM, and glomalin, HA, and FA in POM-free soil were extracted in the following sequence: (1) POM fraction separation from the soil, (2) glomalin extraction from the POM fraction and POM-free soil, and (3) co-extraction of HA and FA from the POM fraction and POM-free soil. Only trace amounts of HA and FA were present in the POM fraction, while POM-associated glomalin (POM-glomalin) and POM alone contributed 2 and 12%, respectively, of the total C in the soil. Mean combined weights for chemically extracted pools from POM and from POM-free soil were 9.92 g glomalin, 1.12 g HA, and 0.88 g FA kg⁻¹ soil. Total protein and C, N, and H concentrations showed that glomalin and HA were, for the most part, separate pools, although protein was detected in HA extracts. Even though percentage carbon was higher in HA than in glomalin, glomalin was a larger (almost nine times) operationally defined pool of soil organic C. Glomalin was also the largest pool of soil N of all the pools isolated, but all pools combined only contained 31% of the total N in the soil.     

Polyphenolic compounds interfere with quantification of protein in soil extracts using the Bradford method

 The Bradford protein quantification assay is based on an absorbance shift in Coomassie brilliant blue G-250 (CBB). Samples extracted for glomalin, a protein produced by arbuscular mycorrhizal (AM) fungi, are quantified using the Bradford assay. CBB is known to react with polyphenolic substances, and co-extraction of glomalin and humic substances is known to occur. The effects of increasing concentrations polyphenolic compounds were measured. The addition of any amount of polyphenolic compounds increased the Bradford reactive fraction (BRF) of soil extract. Caution is required when interpreting BRF data, as comparison of BRF data from different studies or different field sites is problematic. The BRF may represent recalcitrant organic material in soil, though its relationship to AM fungi remains unclear.