Use of specific phospholipid fatty acids for identifying and quantifying the external hyphae of the arbuscular mycorrhizal fungus Gigaspora rosea

The external hypha of arbuscular mycorrhizal (AM) fungi, extending from roots out into soil, is an important structure in the uptake of phosphate from the depletion zone around each root. In this paper, we analysed some phospholipid fatty acids (PLFAs) derived from external hyphae of four AM fungi (Glomus etunicatum, Glomus clarum, Gigaspora margarita and Gigaspora rosea) to find fatty acids which may be useful as specific markers for identifying and quantify the external hyphae of Gigaspora species. Leek (Allium porrum L.) seedlings inoculated with each AM fungus were grown in river sand. Sand samples were collected and four PLFAs (16:1ω5, 18:1ω9, 20:1ω9 and 20:4) in the sand were analysed. In addition, the hyphal biomass in the sand was determined by the direct microscopic method. PLFAs 18:1ω9 and 20:4 were found in all the AM-inoculated and non-inoculated sand samples. PLFA 16:1ω5 was detected in the sand inoculated with G. etunicatum, G. clarum and Gi. rosea. PLFA 20:1ω9 was detected only in the sand inoculated with Gi. rosea. PLFAs 16:1ω5 and 20:1ω9 were not found in the sand inoculated with Gi. margarita. The amount of PLFA 20:1ω9 was closely correlated with the amount of biomass of external hyphae of Gi. rosea (r=0.937, P<0.001), whereas no correlation was observed for PLFA 16:1ω5. The 20:1ω9 content of Gi. rosea was approximately 6.56 nmol mg⁻¹ hyphal biomass. We suggest that PLFA 20:1ω9 can be used as a specific marker for identifying and quantifying the external hyphae of Gi. rosea, at least in controlled experimental systems.     

Monitoring extra-matrical hyphae of a vesicular-arbuscular mycorrhizal fungus with an immunofluorescence assay and the soil aggregation technique

An immunofluorescence assay (IFA) was used to compare the amount of extra-matrical hyphae of the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus versiforme (Karst) Berch colonizing seedlings of White Cedar, Chamaecyparis lawsoniana (A. Murr.) Parl., with different root densities in three soil mixes. Restriction (binding) of the root mass with nylon screening decreased root density and also decreased the density of VAM hyphae detected per root length or per soil volume. Top growth of transplanted seedlings was not affected by root restriction. More VAM fungal external hyphae, determined by IFA and the sand aggregation method, formed in river sand than in silt loam with similar percentages of intraradical VAM colonization. More soil aggregates formed in silt loam than in sand regardless of the presence of VAM fungal hyphae. Depending on experimental design, the aggregation method for estimating VAM external hyphae may be limited to use in sandy soil. IFA reacted specifically with G. versiforme hyphae in mineral soil or sand, but was ineffective in peat soil due to nonspecific staining and autofluorescence of organic matter.     

Selective enrichment of a pyrene degrader population and enhanced pyrene degradation in Bermuda grass rhizosphere

Rhizosphere soil has a more diverse and active microbial community compared to nonvegetated soil. Consequently, the rhizosphere pyrene degrader population (PDP) and pyrene degradation may be enhanced compared to nonvegetated bulk soil (NVB). The objectives of this growth chamber study were to compare (1) Bermuda grass (Cynodon dactylon cv. Guymon) growth in pyrene-contaminated and noncontaminated soils and (2) pyrene degradation and PDP among NVB, Bermuda grass bulk (BB), and Bermuda grass rhizosphere soil (BR). Soils were amended with pyrene at 0 and 500 mg kg^–1, seeded with Bermuda grass, and thinned to two plants per pot 14 days after planting (DAP). Pyrene degradation was evaluated over 63 days. The PDP was enumerated via a most probable number (MPN) procedure at 63 DAP. Bermuda grass root growth was more sensitive to pyrene contamination than shoot growth. Pyrene degradation followed first-order kinetics. Pyrene degradation was significantly greater in BR compared to BB and NVB with rate constants of 0.082, 0.050, and 0.052 day^–1, respectively. The PDPs were 8.01, 7.30, and 6.83 log₁₀ MPN g^–1 dry soil for BR, BB, and NVB, respectively. The largest PDP was in soil with the most rapid pyrene degradation. These results indicate that Bermuda grass can grow in pyrene-contaminated soil and enhance pyrene degradation through a rhizosphere effect.     

Crop–weed hybrids are more frequent for the grain amaranth ‘Plainsman’ than for ‘D136-1’

White-seeded cultivated amaranth [Amaranthus caudatus L., A. cruentus L., and A. hypochondriacus L.] grain is less valuable if contaminated with brown off-type seeds from the hybrid weedy progeny of spontaneous crop–weed crossing. Crop–weed crossing frequency was estimated by using two grain amaranth cultivars. Both cultivars were planted at the same nine locations in the Midwest United States and exposed to pollen from local weedy Amaranthus species. Harvested seeds were grown in a greenhouse, and the frequency of crop–weed hybrids was determined by observing dominant weed traits in the progeny. The cultivar ‘Plainsman’ was approximately tenfold more likely to hybridize with weedy amaranths than was ‘D136-1’. These are the first amaranth cultivars to be evaluated in this way. The reduced hybridization potential of ‘D136-1’ or similar material can be exploited to reduce the occurrence of off-type seed contamination in grain amaranths.     

The maize root system in situ: Evaluation of structure and capability of utilization of phytate and inorganic soil phosphates

• 1 The dependence of the morphology of the maize (Zea mays L.) seminal root system on physical, chemical and biotic parameters was investigated with pot cultures in quartz sand and in a natural loamy sand soil. Low O₂-supply to the soil resulted in a substantially smaller root biomass despite a relative increase in total root length. Reduced N-supply also stimulated root length growth, but also enhanced the formation of laterals. The presence of soil microorganisms, in comparison to sterile cultures, resulted in a reduced length of the main roots, and the production of slender laterals with a decreased root hair density. Generally, the structural variability of laterals in response to different growth conditions was much more pronounced than that of the main roots.
• 2 A major part of the work reported here was dedicated to a detailed study of phosphate (P) acquisition by the maize root system under field conditions. Radioactive labelling of the roots and radioautography of soil cores revealed the in situ distribution pattern of the maize root system. Controlled labelling of the soil with radioactive phosphate allowed the documentation of the development and replenishment of the phosphate depletion zone around roots. Finally, the longevity and phosphate uptake activity of the different parts and tissues of the primary root system of maize was examined by electron microscopy and tracer studies including pulse chase experiments. From these studies the phosphate-acquiring strategy of the maize root system appears as follows: The capability of P uptake decreases in the order: root hairs, 1^st order laterals, 2^nd order laterals, main root. The life-spans of the components of the maize root system increase by the sequence: root hairs, laterals, main root. Inorganic P uptake, therefore, mainly occurs during the first weeks of root development. Dying back of the root occurs in an ordered manner resulting in a relocation of stored P predominantly into the main root cortex. Furthermore, it could be shown that competition for P between roots of the same or of adjacent maize and/or lupin plants virtually does not occur in situ.
• 3 The utilization of phytate-P was studied with ¹⁴C/³²P-labelled Camyo-inositol-hexaphosphate supplied to maize plants grown in sterile quartz sand or in hydroponic cultures. The ratio of P- and C-uptake as well as the incidence of phytate hydrolysis products in the rooting medium indicated the capability of maize roots to acquire P from phytate by enzymatic hydrolysis. This was confirmed by enzyme studies of the root tissues. A specific hydrolyzing enzyme (phytase; molecular weight 51 kD) could be detected in the cell wall of the root, especially in the root tip, which initiates phytate dephosphorylation. Further breakdown is presumably accomplished by monophosphoric phosphohydrolases.

     

Transfer of rubidium‐86 (potassium) from deeply‐rooted alfalfa (Medicago sativa L.) to associated,shallow‐rooted maize (Zea mays L.) or grain sorghum (Sorghum vulgare pers.).

Circumstantial evidence exists for non‐N‐mineral element transfer in legume‐grass associations. Three experiments were conducted in an effort to directly demonstrate transfer of a non‐N‐mineral element in alfalfa (Medicago sativa L.)‐maize (Zea mays L.) and alfalfa‐grain sorghum (Sorghum vulgare Pers.) associations in two rooting media. Associations were established in double‐tube apparatus so that a single alfalfa plant was rooted in media of top‐ and bottom‐tubes, while an associated grass plant was rooted exclusively in the top‐tube (Intact treatment). Severed treatments (the control) were identical to the Intact treatments except the alfalfa roots in an air gap between the top‐and bottom‐tubes were excised.

⁸⁶Rb was dispensed onto the medium of bottom tubes with movement of the radioisotope determined by analyzing the legume and grass tissues over time. ⁸⁶Rb was detected in: i) soil‐grown maize associated with alfalfa within a 40‐day treatment period; ii) sand‐grown maize associated with alfalfa within 20 days after treatment and iii) sand‐grown sorghum associated with alfalfa within 10 days. Detection of ⁸⁶Rb in grass plants associated with alfalfa demonstrated that transfer of this potassium analog can occur via the root systems of legume‐grass associations.     

Polysaccharides and monosaccharides in the hyphosphere of the arbuscular mycorrhizal fungi Glomus E3 and Glomus tenue

 Plants colonised with the arbuscular mycorrhizal fungi (AMF) Glomus E3 and Glomus tenue were grown in microcosms that permitted separation into root:hyphae and hyphae compartments. Hydrolysed polysaccharides from the hyphae and water-soluble sugars released into the hyphosphere were assayed using chromatography. Total sugars and most monosaccharides were elevated in the hyphosphere of Glomus E3 but not in the hyphosphere of G. tenue. Differences in the levels of sugars did not depend on hyphal surface area. It is suggested the diversity in sugars produced in the hyphosphere of AMF may drive some of the spatial and temporal variation in microbial diversity and function in soils.     

The contribution from hyphae, roots and organic carbon constituents to the aggregation of a sandy loam under long-term clover-based and grass pastures

Water-stable macro-aggregate size fractions (>2.0 mm, 1.0–2.0 mm, 0.5–1.0 mm and 0.25–0.5 mm) and non-aggregated soil from a sandy loam under long-term clover-based pasture and from grass pasture were analysed to determine the role of acid- and water-extractable carbohydrate C, total hyphal length, microbial biomass, organic C and total and mycorrhizal root length in stabilization of the aggregates. Aggregates were examined by scanning electron microscopy (SEM) and the particle-size distribution of the size fractions was also determined. Macro-aggregation increased under grass, relative to clover-based pasture; however, the properties of the aggregate fractions measured did not reflect this difference. Microbial-biomass C, extractable-carbohydrate C, hyphal length, total and mycorrhizal root length and organic C content of the soils were poorly correlated with macro-aggregation. Within the aggregates, the proportion of 250–1000-km sand was smaller and clay, silt and fine sand (20–250 μm) were greater relative to non-aggregated soil, suggesting that the >250-μm sand in the non-aggregated soil limited the stabilization of macro-aggregates. Under SEM, no enmeshment of aggregates by hyphae and roots was apparent. Although 50–160 m hyphae g^?1 soil was found within the aggregates, calculations showed that on average only 5 to 13 lengths of hyphae were associated with each 250-μm cube of soil within the aggregates, and suggested little potential to stabilize the aggregates by enmeshing. On average, all >2.0-mm aggregates contained less than 3.6 mm of roots and less than 50% by weight of <2.0-mm aggregates contained a single length of root. The findings cast doubt about the role of hyphae and fine roots in the stabilization of macro-aggregates through an enmeshing mechanism in sandy soils.     

Root morphological traits related to phosphorus‐uptake efficiency of soybean,sunflower, and maize

Many of the plant acquisition strategies for immobile nutrients, such as phosphorus (P), are related to the maximization of soil exploration at minimum metabolic cost. Previous studies have suggested that soybean (Glycine max L.), sunflower (Helianthus annuus L.), and maize (Zea mays L.) differ in their P uptake efficiency. In this investigation we employed these three species to evaluate: (1) the effect of suboptimal P conditions on root morphological traits related to root porosity and fineness and (2) how these traits are related to P‐uptake efficiency. Opaque 25‐L plastic containers were used to grow plants hydroponically. The three species were compared under two P availability levels (low P and high P). Most of the observed responses were in the direction to favor P uptake under low‐P conditions. Compared to P‐sufficient plants, P‐stressed plants of the three species showed higher root‐to‐shoot ratio, specific root length, root porosity and root aerenchyma, and a lower root density. For example, P‐stress increased root porosity by a factor of 2.0, 1.4, and 1.4 in soybean, sunflower, and maize, respectively. Soybean and sunflower were the species with the highest P‐uptake efficiency, expressed as P uptake either per unit root biomass or length. The results demonstrate the central role of aerenchyma development in modifying root length per unit root biomass and, thus, reducing the root's foraging costs. Consequently, aerenchyma is suggested to be a possible mechanism for better P‐uptake efficiency.     

Arbuscular Mycorrhizal Association in Plants Growing on Metal-Contaminated and Noncontaminated Soils Adjoining Kanpur Tanneries, Uttar Pradesh, India

A pot experiment was conducted to study the effect of 7 intercrops on Cd uptake by maize. The intercrops included cowpea (V. unguiculata (L.) Walp.), purple haricot (L. purpureus (L.) Sweet.), chickpea (C. arietinum L.), alfalfa (M. sativa L.), teosinte (E. mexicana Schrad.), amaranth (A. paniculatus L.) and rape (B. napus L.). The results showed that most legumes substantially increased Cd uptake by maize during vegetative growth. Leaf tissue of maize grown with legumes averaged 5.05 mg kg^?1 higher Cd than that grown with nonlegumes, or 2.42 mg kg^?1 higher than the control. However, the effect of intercrops on Cd uptake by maize became small during reproductive growth. Since chickpea resulted in a relatively large maize bioconcentration factor of 2.0 and large transfer factor of 0.55, it is regarded as the most valuable intercrop for enhancing Cd extraction from soil by maize. The results suggest that intercropping might be a feasible practice in facilitating phytoremediation.     

Land use in agricultural landscapes with chernozems contaminated after Chernobyl accident: Can we be confident in radioecological safety of plant foodstuff?

Agricultural land use in the area of the post-Chernobyl Plavsk radioactive hotspot (Tula region, Central Russia) has raised a problem of radioecological safety of obtained plant foodstuff. Verification of ¹³⁷Cs activities and inventories in components of “soil-plant” systems of the territory has been conducted in 2014–2017 in 10 agrosystems and 2 semi-natural meadows. It was revealed that density of ¹³⁷Cs contamination of arable chernozems and alluvial calcareous soils nowadays varies in a range 140–220 kBq/m² and exceeds radiation safety standard by ˜ 3.5–6 times. Deep plowing of the arable soils up to 30-cm in 1986–1987 resulted in decreasing of ¹³⁷Cs inventories in rooting zone by ≈ 70% for crops cultivated with shallow disk plowing (wheat, barley), and by ≈ 35% for crops cultivated with middle plowing (buckwheat, amaranth, white mustard). The investigated plants and their compartments can be grouped on the basis of transfer factor values as follows: maize (stems and leaves) > amaranth > bromegrass > vegetation of dry meadow, galega, sunflower (seeds), vegetation of wet meadow > maize (grain), soybean (pods), barley (grain), buckwheat (grain), potatoes (tubers) > white mustard (seeds), wheat (grain). It is noticeable that generative plant compartments are characterized by less ¹³⁷Cs activities in comparison with stems and leaves; and that ¹³⁷Cs root uptake is not coincide with total flux of mineral nutrients in “soil-plant” systems. In sum, ¹³⁷Cs soil-to-plant transfer in the area of the Plavsk radioactive hotspot is characterized by considerable discrimination, so ¹³⁷Cs activities in plants are completely in accordance with national standards.     

Field survival of the phytostimulator Azospirillum lipoferum CRT1 and functional impact on maize crop, biodegradation of crop residues, and soil faunal indicators in a context of decreasing nitrogen fertilisation

Heavy nitrogen fertilisation is often implemented in maize cropping systems, but it can have negative environmental effects. Nitrogen-fixing, phytohormone-producing Azospirillum plant growth-promoting rhizobacteria (PGPR) have been proposed as crop inoculants to maintain high yield when decreasing nitrogen fertilisation. In this context, agronomic and ecological effects of the inoculation of maize seeds with the PGPR Azospirillum lipoferum CRT1 were studied in two consecutive years. The inoculant was recovered from maize at 10⁵ CFU g⁻¹ root or higher. Inoculation enhanced root growth and development based on results of root biomass, rooting depth and/or parameters describing root system architecture, and a transient positive effect on shoot height was observed in the first year. Inoculation did not increase yield, but reducing mineral nitrogen fertilisation had only a minor effect on yield. This suggests that the lack of positive effect of the PGPR on yield was due to the fact that the whole field was heavily fertilised in years prior to the start of the experiment. Soil nitrogen levels decreased during the 2 years of the study, and the inoculant had no effect on residual soil nitrogen levels at harvest. Inoculation had no impact on Fusarium symptoms and concentration of the mycotoxin deoxynivalenol in maize kernels, but both were influenced by the interaction between inoculation and nitrogen fertilisation level. Inoculation did not influence meso/macrofaunal soil populations, but had a small but significant effect (smaller than the effect of added nitrogen) on decomposition, nitrogen mineralisation and mesofaunal colonisation of maize leaves (in litter bags). Overall, the ecological impact of seed inoculation with the PGPR A. lipoferum CRT1 was small, and its magnitude was smaller than that of chemical nitrogen fertilisation.     

Partitioning root and microbial contributions to soil respiration in Leymus chinensis populations

Based on the enclosed chamber method, soil respiration measurements of Leymus chinensis populations with four planting densities (30, 60, 90 and 120 plants/0.25 m²) and blank control were made from July 31 to November 24, 2003. In terms of soil respiration rates of L. chinensis populations with four planting densities and their corresponding root biomass, linear regressive equations between soil respiration rates and dry root weights were obtained at different observation times. Thus, soil respiration rates attributed to soil microbial activity could be estimated by extrapolating the regressive equations to zero root biomass. The soil microbial respiration rates of L. chinensis populations during the growing season ranged from 52.08 to 256.35 mg CO₂ m⁻² h⁻¹. Soil microbial respiration rates in blank control plots were also observed directly, ranging from 65.00 to 267.40 mg CO₂ m⁻² h⁻¹. The difference of soil microbial respiration rates between the inferred and the observed methods ranged from −26.09 to 9.35 mg CO₂ m⁻² h⁻¹. Some assumptions associated with these two approaches were not completely valid, which might result in this discrepancy. However, these two methods' application could provide new insights into separating root respiration from soil microbial respiration. The root respiration rates of L. chinensis populations with four planting densities could be estimated based on measured soil respiration rates, soil microbial respiration rates and corresponding mean dry root weight, and the highest values appeared at the early stage, then dropped off rapidly and tended to be constant after September 10. The mean proportions of soil respiration rates of L. chinensis populations attributable to the inferred and the observed root respiration rates were 36.8% (ranging from 9.7 to 52.9%) and 30.0% (ranging from 5.8 to 41.2%), respectively. Although root respiration rates of L. chinensis populations declined rapidly, the proportion of root respiration to soil respiration still increased gradually with the increase of root biomass.     

Impact of 12-year field treatments with organic and inorganic fertilizers on crop productivity and mycorrhizal community structure

The effect of manure and mineral fertilization on the arbuscular mycorrhizal (AM) fungal community structure of sunflower (Helianthus annuus L.) plants was studied. Soils were collected from a field experiment treated for 12 years with equivalent nitrogen (N) doses of inorganic N, dairy manure slurry, or without N fertilization. Fresh roots of tall fescue (Festuca arundinacea Schreb.) grass collected from the field plots without N fertilization and unfumigated field soils were used as native microbial inoculum sources. Sunflower plants were sown in pots containing these soils, and three different means of manipulating the microbial community were set: unfumigated soil with fresh grass roots, fumigated soil with fresh grass roots, or fumigated soil with sterilized grass roots. Assessing the implications with respect to plant productivity and mycorrhizal community structure was investigated. Twelve AM fungal OTUs were identified from root or soil samples as different taxa of Acaulospora, Claroideoglomus, Funneliformis, Rhizophagus, and uncultured Glomus, using PCR-DGGE and sequencing of an 18S rRNA gene fragment. Sunflower plants grown in manure-fertilized soils had a distinct AMF community structure from plants either fertilized with mineral N or unfertilized, with an abundance of Rhizophagus intraradices-like (B2). The results also showed that AM inoculation increased P and N contents in inorganic N-fertilized or unfertilized plants, but not in manure-fertilized plants.     

Functional diversity of external hyphae of AM fungi: Ability to colonise new hosts is influenced by fungal species,distance and soil conditions

The aim of this paper was to investigate the effects of soil conditions and distance from a host plant on the ability of hyphae of arbuscular mycorrhizal (AM) fungi to grow and colonise a new host. Two glasshouse experiments were conducted using compartmented pots. The first investigated the effects of distance between a colonised and uncolonised host plant (Trifolium subterraneum L.) and average pore size of the growth substrate (100 μm, 38 μm) on the ability of two AM fungi, G. intraradices and G. mosseae, to colonise a new host plant. The second experiment determined if the pore size of the substrate (100 μm, 38 μm) affected the growth of AM fungi in the absence of a new host. In Experiment 1, both G. mosseae and G. intraradices grew successfully through the two sand substrates and colonised new host plants. Both fungi reached and colonised new hosts fastest when hosts were separated by the shortest distance (2.5 cm), with largest pore size substrate (100 μm). G. mosseae produced more external hyphae per unit of colonised root and colonised new host plants more rapidly than G. intraradices. However, receiver plants colonised by G. mosseae exhibited a negative mycorrhizal growth response following colonisation. Experiment 2 showed that G. mosseae grew further from its host than G. intraradices. The results support the theory that some AM fungal species may produce large amounts of external hyphae primarily to increase the probability of locating and colonising a new host plant.     

伊犁河谷不同草地类型坡面水土保持效应的模拟降雨试验

[目的]研究一定降雨条件下,伊犁河谷地区3个不同草地类型径流小区的减流减沙效果,以期为该区的水土保持工作提供借鉴。[方法]采用野外人工模拟降雨的方法。[结果](1)几种草地类型初始产流时间的长短依次为:角果藜伊犁蒿狗牙根裸地;(2)径流速率由大到小依次为:裸地伊犁蒿狗牙根角果藜;(3)泥沙含量由大到小依次为:裸地伊犁蒿狗牙根角果藜;(4)狗牙根减流因子的数值处于0.21~0.58之间,伊犁蒿的减流因子处于0.14~0.87之间,角果藜的减流因子则在0.02~0.11之间,狗牙根减沙因子的数值在0.16~0.76之间,角果藜的减沙因子处于0~0.42之间,伊犁蒿的减沙因子则在0.42~0.88之间。[结论]水土保持效益最好的是角果藜,其次为狗牙根与伊犁蒿,裸地效果最差。     

Carbon sequestration in soils with annual inputs of maize biomass and maize-derived animal manure: Evidence from C abundance

The abundance of ¹³C was determined over a period of nine years in two soils (LUN, coarse sand; ASK, sandy loam) following their conversion from C3-crops and to the C4-crop silage maize (Zea mays L.). The soils were exposed to identical management and climatic conditions, and were sampled every second year. The aboveground maize biomass was either removed (stubbles and roots left), chopped and added to the soil, or fed to sheep and the faeces then added to the soil. Annual inputs of maize biomass and sheep faeces were similar (0.8 kg DM m⁻²). The study included soils maintained under C3-crops (beet roots, Beta vulgaris L.). After nine years of maize cropping, soil C from stubbles and roots accounted for 12 and 16% of the total-C in the LUN and ASK soil, respectively. Without additional organic amendment the content of total-C in the ASK soil remained constant and similar to that of soil retained under C3-crops whereas total-C tended to decrease in the LUN soil. When maize biomass and sheep faeces were added, soil total-C increased and C from these C4-sources averaged 14% and 21% of the soil total-C, respectively. Following nine annual additions, retention of C added in aboveground maize biomass averaged 19% while the retention of C added in maize-derived faeces was 30%. Our study infers that that ruminant manure C contributes about 50% more to soil C sequestration than C applied in crop residues.     

Savanna-derived organic matter remaining in arable soils of the South African Highveld long-term mixed cropping: Evidence from C and N natural abundance

Sustainable agriculture requires the formation of new humus from the crops. We utilized ¹³C and ¹⁵N signatures of soil organic matter to assess how rapidly wheat/maize cropping contributed to the humus formation in coarse-textured savanna soils of the South African Highveld. Composite samples were taken from the top 20 cm of soils (Plinthustalfs) cropped for lengths of time varying from 0 to 98 years, after conversion from native grassland savanna (C4). We performed natural ¹³C and ¹⁵N abundance measurements on bulk and particle-size fractions. The bulk soil δ¹³C values steadily decreased from −14.6 in (C4 dominated) grassland to −16.5‰ after 90 years of arable cropping. This δ¹³C shift was attributable to increasing replacement of savanna-derived C by wheat crop (C3) C which dominated over maize (C4) inputs. After calculating the annual C input from the crop yields and the output from literature data, by using a stepwise C replacement model, we were able to correct the soil δ¹³C data for the irregular maize inputs for a period of about one century. Within 90 years of cropping 41-89% of the remaining soil organic matter was crop-derived in the three studied agroecosystems. The surface soil C stocks after 90 years of the wheat/maize crop rotation could accurately be described with the Rothamsted Carbon Model, but modelled C inputs to the soil were very low. The coarse sand fraction reflected temporal fluctuations in ¹³C of the last C₃ or C₄ cropping and the silt fraction evidenced selective erosion loss of old savanna-derived C. Bulk soil ¹⁵N did not change with increasing cropping length. Decreasing δ¹⁵N values caused by fertilizer N inputs with prolonged arable cropping were only detected for the coarse sand fraction. This indicated that the present N fertilization was not retained in stable soil C pool. Clearly, conventional cropping practices on the South African highlands neither contribute to the preservation of old savanna C and N, nor the effective humus reformation by the crops.     

Untapped amaranth (<Emphasis Type="Italic">Amaranthus </Emphasis>spp.) genetic diversity with potential for nutritional enhancement

Significant genetic diversity was observed in 218 out of a total of 1309 accessions of amaranth (Amaranthus hypochondriacus L.) and its seven wild relatives, A. spinosus L., A. dubius Mart. ex Thell., A. hybridus L., A. tricolor L., A. cruentus L., A. caudatus L., A. retroflexus L. for 24 nutritional parameters including total oil content, fatty acid profile, total protein content and amino acid profile. Diversity for total oil content (6.42–12.53%), linoleic acid (25.68–54.34%), oleic acid (21.97–42.01%) of the total fatty acids, total protein content (7.84–18.01%), among important essential amino acids; lysine content (0.66–11.12 g/16 g N), methionine (0.35–4.80 g/16 g N) and half cystine and (0.12–8.32 g/16 g N) was reported. The un-weighted pair-group method using arithmetic average cluster analysis based on pair wise Euclidean genetic distance grouped the accessions into seven major clusters. Histidine, half cystine, tyrosine, essential amino acids, total oil content, linoleic acid and oleic acid content were the major parameters contributing significantly to genetic diversity. Present findings indicate that significant diversity exists for nutritional parameters in amaranth germplasm. The promising accessions with higher multiple nutritive traits; protein content (>16%), oil content (>11%), lysine content (>7.5 g/16 g N) and EAA higher than the FAO reported values, were identified. This is the first report on detailed nutritional analysis of diversity collected worldwide. These could be used as potential breeding material for nutritional enhancement through genetic improvement. This will help in overcoming the “triple burden” of malnourishment, hidden hunger, and obesity.     

Effects of epigeic earthworm (Eisenia fetida) and arbuscular mycorrhizal fungus (Glomus intraradices) on enzyme activities of a sterilized soil–sand mixture and nutrient uptake by maize

A pot experiment was conducted to investigate the effect of epigeic earthworm (Eisenia fetida) and arbuscular mycorrhizal (AM) fungi (Glomus intraradices) on soil enzyme activities and nutrient uptake by maize, which was grown on a mixture of sterilized soil and sand. Maize plants were grown in pots inoculated or not inoculated with AMF, treated or not treated with earthworms. Wheat straw was added as a feed source for earthworms. Mycorrhizal colonization of maize was markedly increased in AM fungi inoculated pots and further increased by addition of epigeic earthworms. AM fungi and epigeic earthworms increased maize shoot and root biomass, respectively. Soil acid phosphatase activity was increased by both earthworms and mycorrhiza, while urease and cellulase activities were only affected by earthworms. Inoculation with AM fungi significantly (p?<?0.001) increased the activity of soil acid phosphatase but decreased soil available phosphorus (P) and potassium (K) concentrations at harvest. Addition of earthworms alone significantly (p?<?0.05) increased soil ammonium-N content, but decreased soil available P and K contents. AM fungi increased maize shoot weight and root P content, while earthworms improved N, P, and K contents in shoots. AM fungi and earthworm interactively increased maize shoot and root biomass through their regulation of soil enzyme activities and on the content of available soil N, P, and K.