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1.
Extraction of arbuscular mycorrhizal fungal (AMF) spores from soil is widely used to assess AMF community structure and abundance. The most widely used protocol relies on a water-sucrose gradient flotation technique. Na-hexametaphosphate has also been used to deflocculate soil aggregates prior to spore extraction in order to optimize recovery, but its effect on spore viability remains unknown. Here, we report that Na-hexametaphosphate increases average spore yield in a high clay soil by about 15%, but decreases average spore viability by about 20%. Na-hexametaphosphate should therefore be used cautiously where the extracted spores are destined to be used as inoculum for subsequent studies.  相似文献   

2.
The effects of carbon (C) and nitrogen (N) sources on N utilization and biosynthesis of amino acids were examined in the germinating spores of the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith after exposure to various N substrates,CO2,glucose,and/or root exudates.The N uptake and de novo biosynthesis of amino acids were analyzed using stable isotopic labeling with mass spectrometric detection.High-performance liquid chromatography-based analysis was used to measure amino acid levels.In the absence of exogenous N sources and in the presence of 25 mL L-1 CO2,the germinating AM fungal spores utilized internal N storage as well as C skeletons derived from the degradation of storage lipids to biosynthesize the free amino acids,in which serine and glycine were produced predominantly.The concentrations of internal amino acids increased gradually as the germination time increased from 0 to 1 or 2 weeks.However,asparagine and glutamine declined to the low levels;both degraded to provide the biosynthesis of other amino acids with C and N donors.The availability of exogenous inorganic N (ammonium and nitrate) and organic N (urea,arginine,and glutamine) to the AM fungal spores using only CO2 for germination generated more than 5 times more internal free amino acids than those in the absence of exogenous N.A supply of exogenous nitrate to the AM fungal spores with only CO2 gave rise to more than 10 times more asparagine than that without exogenous N.In contrast,the extra supply of exogenous glucose to the AM fungal spores generated a significant enhancement in the uptake of exogenous N sources,with more than 3 times more free amino acids being produced than those supplied with only exogenous CO2.Meanwhile,arginine was the most abundant free amino acid produced and it was incorporated into the proteins of AM fungal spores to serve as an N storage compound.  相似文献   

3.
The effects of bacteria associated with arbuscular mycorrhizal fungal (AMF) spores on spore germination, growth in vitro and on the pea-AMF symbiosis were evaluated. Bacterial colonies were recovered from untreated Glomus clarum NT4 spores and NT4 spores decontaminated with 5% chloramine-T for 30, 45 and 60 min on five different media. Both G+ and G− bacteria were recovered from untreated NT4 spores, whereas only G+ bacteria were isolated from decontaminated spores. An in vitro assessment of the effect of spore-associated bacteria on clean, decontaminated NT4 spores revealed that (i) most of the bacteria isolated from untreated spores generally did not significantly alter spore function, (ii) some bacteria isolated from clean, decontaminated spores inhibited or stimulated NT4 spore germination, (iii) stimulation of spore germination occurred only when bacteria were in contact with spores, and (iv) inhibition of spore germination was the result of volatile bacterial metabolites. A stimulatory bacterial isolate, Bacillus pabuli LA3, significantly (P<0.05) enhanced the shoot growth, AMF-colonization, shoot N content and P use efficiency of NT4-inoculated 6 week-old pea plants over that of plants co-inoculated with an inhibitory bacterial isolate, Bacillus chitinosporus LA6a and NT4.  相似文献   

4.
Spontaneous yeast mutants isolated in continuous culture as resistant to toxic amino acid analogues, able to increase up to 40 times their free amino acid pool of Thr, up to 160 times their pool of Met, or up to 20 times their pool of Lys, were characterized with regard to properties of industrial interest. Growth rate, mu (h(-1)), and biomass yield, Y (g/L), of the amino acid overproducing mutants (AA(S)) were in many cases similar to those of the wild type, whereas their free amino acid content was substantially increased in laboratory and industrial media (molasses). Doughs fermented with 3% baker's yeast and 0.5% AA(S) mutants produced bakery products that displayed texture similar to those fermented with 3.5% baker's yeast, but the former had a considerable improvement of their taste and aroma. On the other hand, bread content of the essential amino acids Lys, Met, and Thr provided by yeast was also increased.  相似文献   

5.
Plants can take up intact amino acids, even in competition with soil microbes, yet we lack detailed information on which amino acids dominate the soil and whether amino acid composition varies seasonally. This study tested the hypotheses that 1) the pool of amino acid N is generally larger than inorganic N; 2) temporal changes in the concentration of amino acid N is related to changes in the size of the microbial N pool; and 3) amino acid N is dominated by simple, neutral amino acids during warm months, whereas during cold months the amino acid N is dominated by more complex aromatic and basic amino acids. Approximately every month for two years we collected soil from a temperate, sub-alpine grassland in the Snowy Mountains of Australia. We quantified exchangeable pools of amino acids, nitrate and ammonium in 1 M KCl extracts. Microbial N was quantified by chloroform fumigation. Averaged across the 21 monthly samples, nitrate was 13% of the quantified pool of soluble non-protein N, ammonium was 34% and amino acid N was 53%. These data are consistent with our hypothesis that the pool of amino acid N is larger than inorganic N. There was substantial variation between months in concentrations of amino acids and inorganic N, but no clear temporal pattern. Microbial N did not vary between months, and thus changes in amino acid N were unrelated to microbial N. Principal components analysis indicated multivariate groupings of the different pools of N that were broadly indicative of function and/or biosynthetic relationships. Thus PCA identified a grouping of aromatic amino acids (Phe and Try) with amino acids derived from oxaloacetate (Asp, Ala, Val, Leu, Ile), and a second group comprising microbial N, nitrate and glycine. The pool of exchangeable amino acid N was dominated by Arg (26% of amino N) Val (20%) Gln (18%), Try (8%) and Asn (8%). Contrary to our hypothesis, the composition of the amino acid pool did not vary in a consistent way between months, and there was no evidence simple amino acids were relatively more abundant in warm months and complex amino acids in cool months.  相似文献   

6.
Regulation of amino acid biodegradation in soil as affected by depth   总被引：1，自引：0，他引：1  
Dissolved organic nitrogen (DON) and in particular free amino acids represent a key pool in the terrestrial soil C and N cycle. The factors controlling the rate of turnover of this pool in soil, however, remain poorly understood. We investigated the factors regulating the rate of amino acid turnover at different depths (up to 1.2 m) in five low-input, acid soil profiles. Within the root zone (0–60 cm), amino acids constituted 8% of the DON and represented only a small fraction of plant available N. In all the soil profiles, the rate of amino acid mineralisation decreased progressively with depth. The average half-life of the exogenously added amino acids in the soil was 5.8 h in topsoils (0–10 cm), falling to 20 h at a depth of 50 cm and to 33 h at 100 cm. Generally, the rate of amino acid mineralisation correlated positively with total soil C and N, soil microbial activity (basal soil respiration rate) and soil content. The relatively rapid rates of microbial amino acid assimilation in subsoils below the root zone (>60 cm) indicate that long-term transport of amino acids (e.g. from soil to freshwaters) will be low. Based upon the C-to-N ratio of the amino acid substrate and the microbial C assimilation efficiency, we estimate that approximately 40–60% of the amino acid-N will be excreted as . In conclusion, the rapid rate of free amino acid turnover and their low concentration in soil solution indicate that the formation of inorganic N ( and ) in soil is limited primarily by the rate of free amino acid appearance in soil and not by the rate of amino acid mineralisation.  相似文献   

7.
New data on the content and composition of the amino acids in a typical chernozem of Moldova were obtained. Eighteen amino acids were identified. Among the usually found amino acids, tryptophan was not identified. It was found that the total content of free and bonded amino acids in the studied chernozem varies from 197 to 317 mg of N/kg of soil, which constitutes 8–11% of the total pool of nitrogen. The predomination of alanine (13.88–17.26%), glycine (10.66–14.51%), aspartic (9.15–13.29%), glutamine (7.26–11.30%), and arginine (8.12–11.99%) is typical of the chernozem. The size and composition of the amino acid pool in this chernozem correlated well with the soil’s organic matter content.  相似文献   

8.
Lentil flours were extracted with 80% ethanol at 25 and 50 degrees C for 1, 2, or 3 h. The various nitrogen fractions, soluble carbohydrates, three amino acids (Lys, His, and Tyr), available lysine, protein digestibility, and vitamins B(1) and B(2) were analyzed to evaluate the effect of extraction. Extraction resulted in an increase in the total nitrogen content of the extracted flours, with extraction temperature affecting the nature of the nitrogen (protein or nonprotein) content. There was also a large reduction in the oligosaccharides of the raffinose family, although the effect of temperature was appreciable only in the case of stachyose. There was hardly any effect on the concentrations of the amino acids analyzed or on protein digestibility; however, a positive correlation between protein digestibility and the available lysine was recorded in the samples. The vitamin B(1) and B(2) contents underwent variable decreases depending on extraction temperature.  相似文献   

9.
For the purpose of explaining the forming process of soil humic acids, the author determined the absorption spectra of various humic acids. From soils and peats which were pretreated with 5% HCl at 70°C for 30 minutes or from those which were not, humic acids were extracted by treating with 0.5% NaOH at boiling temperature for 30 minutes. In these humic acids, the one which is extracted after acid pretreatment is provisionally designated as SrL humic acid and the other as L humic acid. The supernatant alkaline solutions obtained by centrifuging the above mentioned extracts were acidified with hydrochloric acid, and precipitated humic acids were filtered and washed with water until Cl' free. Humic acids were dissolved in 0.1% NaOH and ultrafiltered using collodion membrane. The filtrates were acidified with hydrochloric acid and humic acids were collected by centrifuging, transferred on the filter paper, washed with dilute hydrochloric acid and water successively, then air-dried and pulverized.  相似文献   

10.
Two garlic subspecies (n = 11), Allium sativum L. var. opioscorodon (hardneck) and Allium sativum L. var. sativum (softneck), were evaluated for their free amino acid composition. The free amino acid content of garlic samples analyzed ranged from 1121.7 to 3106.1 mg/100 g of fresh weight (mean = 2130.7 +/- 681.5 mg/100 g). Hardneck garlic had greater methiin, alliin, and total free amino acids contents compared to softneck garlic. The major free amino acid present in all but one subspecies was glutamine (cv. Mother of Pearl had aspartic acid as the major free amino acid). Cv. Music Pink garlic (a rocambole hardneck variety) contained the most methiin, alliin, and total free amino acids. The solid-phase extraction, alkylchloroformate derivatization, GC-FID, and GC-MS methods used in this study were simple and rapid, allowing 18 free amino acids in garlic to be separated within 10 min.  相似文献   

11.
New developments in the estimation of free spores of Pasteuria penetrans, a hyperparasite of plant-parasitic nematodes, including Meloidogyne spp., have been tested in contrasted textured soils. They were dedicated to improving the recovery of spores. Different methods of increasing energy of aggregate dispersion were compared in their efficiency in recovering spores inoculated in a sandy clay soil and a clay soil. The dispersion of the soils by the less energetic method (method A) allowed only 50% and 20% of the spore inoculum to be recovered from the sandy clay soil and clay soil, respectively. For these soils, 76% and 81% of the particle-size fraction (0-20 µm) isolated by this method were still aggregated in coarser structures. With increasing energy (methods B and C), these coarse aggregates disappeared entirely in both soils. At the same time, the recovery of spores increased sharply, representing about 87% and 75% of the inoculum of the sandy clay soil and clay soil, respectively. Therefore, at most 25% of the pool of spores remained undetectable. The formation of artificial aggregates during the enumeration procedure could not be advocated to explain this result, since the dispersion of the fraction collected for the enumeration did not improve spore recovery.  相似文献   

12.
The microfungal community of an andosol, under an acid beech forest located in a zone of the Province of Navarra (Spain) characterized by a temperate climate and high rainfall was studied. Mycelial length, number of spores and fungal biomass were eslimated by the agar-film technique. The number of fungal propagules were estimated by plate counting. There was a pronounced decrease of fungal propagules and mycelial biomass with soil depth; the highest values were obtained in the spring and the autumn. Seasonal variations were attenuated with increasing depth. In autumn the leaves of the A00 horizon contained a great number of fungal spores but there was little mycelial growth.  相似文献   

13.
Selenium-enriched broccoli florets, harvested from plants grown on soil fertilized with four levels of sodium selenate, were evaluated for their free amino acid composition using alkylchlorformate derivatization, solid-phase extraction, and GC-FID or GC-MS. The selenium-enriched florets contained 0.4 (control), 5.7 (treatment A), 98.6 (treatment B), and 879.2 (treatment C) microg/g Se (dry weight). Twenty-one free amino acids were identified in the control and all three treatments. The total free amino acid content of the broccoli florets ranged from 178 mmol/kg (dry weight), for the control, to 479 mmol/kg (dry weight), for treatment C. Broccoli from treatment C contained the highest level of Se, had the most total free amino acids, and had an extremely high level of glutamine (Gln) when compared to the control and the other two treatments. In general, the smallest addition of Se to the soil (treatment A) induced increased levels of all detectable amino acids when compared to the control, whereas increased additions of Se (treatments B and C) produced mixed responses. Florets from treatment A contained the highest essential amino acid content.  相似文献   

14.
Soil amino acids are important sources of organic nitrogen for plant nutrition, yet few studies have examined which amino acids are most prevalent in the soil. In this study, we examined the composition, concentration, and seasonal patterns of soil amino acids across a primary successional sequence encompassing a natural gradient of plant productivity and soil physicochemical characteristics. Soil was collected from five stages (willow, alder, balsam poplar, white spruce, and black spruce) of the floodplain successional sequence on the Tanana River in interior Alaska. Water-extractable amino acid composition and concentration were determined by HPLC. Irrespective of successional stage, the amino acid pool was dominated by glutamic acid, glutamine, aspartic acid, asparagine, alanine, and histidine. These six amino acids accounted for approximately 80% of the total amino acid pool. Amino acid concentrations were an order of magnitude higher in coniferous-dominated late successional stages than in early deciduous-dominated stages. The composition and concentration of amino acids were generally constant throughout the growing season. The similar amino acid composition across the successional sequence suggests that amino acids originate from a common source or through similar biochemical processes. These results demonstrate that amino acids are important components of the biogeochemical diversity of nitrogen forms in boreal forests.  相似文献   

15.
We describe a simple technique for the germination of arbuscular mycorrhizal (AM)–fungal spores and their multiplication in pots. Glomus fasciculatum, G. mosseae, and Gigaspora margarita were used. A single wheat seedling was tied to a glass slide, previously covered with filter paper with the help of thread. One single surface‐sterilized AM‐fungal spore was placed on the middle portion of the root of the wheat seedling using a sterilized syringe. The slide was placed vertically in a 100 mL glass beaker filled with 25 mL of root exudates–water (1:4, v/v) solution, which was collected by growing twenty wheat seedlings in a 150 mL beaker filled with 100 mL sterilized distilled water for 7 d. The slide was observed daily using a compound microscope to follow the time course of germination. In this technique, the spore is directly in contact with the host root, and a visualization of spore germination, hyphal development, and appressorium formation is possible without disrupting fungal growth or the establishment of the symbiosis. The method allows to document the germination events and to assess hyphal‐elongation rates by photographing the same spore on consecutive days. The inoculated seedling was used to initiate single‐spore multiplication in a sterilized (autoclave on 3 alternate days at 120°C for 120 min at 1.05 kg cm–2 pressure) potted sandy soil (150 mL volume) into which the slide with the inoculated seedling was inserted carefully through a previously made slit. The wheat seedlings in all pots (4 treatments and 15 replications) became colonized by mycorrhiza, confirming that the establishment of the AM‐fungal symbiosis is highly reproducible. Our technique permits the relatively undisturbed growth of the symbiotic partners, the visualization of germinating AM‐fungal spores, and their multiplication in pots. This simple and low‐cost method facilitates the production of pure lines of AM fungi from single spores, allowing for the study of intraspecific variation and potentiality for cytological, biochemical, physiological, and taxonomical studies.  相似文献   

16.
A survey on the endorhizal status of 39 fruit crops of 25 families, indicated that 22 fruit crops had arbuscular mycorrhizal (AM)–, four had dark septate endophyte (DSE)–fungal association, and 13 had dual colonization of AM and DSE fungi. Fruit crops were capable of forming Arum‐, Paris‐, or intermediate‐types of AM morphologies of which intermediate‐type was common. To our knowledge, we report for the first time AM in 10 fruit crops and DSE‐fungal association in 17 fruit crops. The extent of AM‐ and DSE‐fungal colonization ranged from 41% to 98% and < 1% to 89.9%, respectively, in different fruit crops. Arbuscular mycorrhizal–fungal spore numbers in the rhizosphere ranged from 6 to 61 spores per 25 g of soil. Arbuscular mycorrhizal–fungal spores belonging to Acaulospora, Glomus, and Scutellospora were isolated from the rhizosphere soil.  相似文献   

17.
蚕豆枯萎病是土传病害,其发生与蚕豆根系分泌物有密切关系。本文以3个枯萎病不同抗性蚕豆品种——‘89-147’(高抗)、‘8363’(中抗)和‘云豆324’(感病)为材料,通过水培试验收集根系分泌物,测定根系分泌物对镰刀菌孢子萌发和菌丝生长的影响,分析对枯萎病表现出不同抗性的蚕豆品种根系分泌物中糖、氨基酸和有机酸的含量,分离鉴定了根系分泌物中氨基酸和有机酸的组分。结果表明,抗病品种的根系分泌物抑制了尖孢镰刀菌的孢子萌发和菌丝生长,在加入5 mL中抗品种根系分泌物时,显著促进尖孢镰刀菌孢子萌发,但对菌丝生长无显著影响;而在加入1 mL感病品种根系分泌物时,就能显著促进尖孢镰刀菌孢子萌发和菌丝生长。不同抗性蚕豆品种根系分泌物中氨基酸总量和总糖含量随抗性的降低而升高,有机酸分泌总量则随蚕豆品种对枯萎病的抗性增加而升高。感病品种和中抗品种中检出15种氨基酸,而高抗品种中检出14种,组氨酸只存在于中抗品种中,脯氨酸仅在感病品种中检出,3个蚕豆品种根系分泌物中均未检出精氨酸。蚕豆根系分泌物中天门冬氨酸、谷氨酸、苯丙氨酸、酪氨酸和亮氨酸含量高,可能会促进枯萎病的发生,而蛋氨酸、赖氨酸和丝氨酸含量高可能抑制枯萎病发生。酒石酸仅在抗病品种中存在,根系分泌物中有机酸种类丰富,有助于提高蚕豆对枯萎病的抗性。蚕豆对枯萎病的抗性不同,根分泌物对镰刀菌孢子萌发和菌丝生长的影响也不同,而这种抗病性差异与蚕豆根系分泌物中糖、氨基酸、有机酸的含量和组分密切相关。  相似文献   

18.
Extraction of a copper contaminated soil material by the percolation of an amino acid containing residue hydrolysate. 2. Time course of amino acid elution and input/output balance of amino acids During 16 days an amino acid containing blood meal hydrolysate (amino acid concentration: 188 mMol·L?1) was percolated through a column packed soil material (soil content per column: 4.1 kg dry weight, four parallels). The copper contaminated material (soil type: Typic Udifluvent, soil texture: sandy loam, loamy sand) was sampled from an area formerly used for cultivation of hop (Humulus lupus). Besides the investigation of the copper liberation the experiments aimed to determine the elution dynamic and input/output balance of amino acids (time span for amino acids balance 14 days). In total 11.7 L of hydrolysate, containing 2.2 Mol of amino acids, were introduced into each column. The mean amino acid output with the column effluent was 1.13 Mol. This corresponds to an elution degree of 51.2%, related to the sum of applicated amino acids, and to a mean substance specific elution degree of 48.4% reflecting the elution of 15 compounds. The substance specific elution ranged from 9.6% (serine) to 75.5% (valine). The highest concentrations of serine and threonine were determined in the effluents after two days, whereas the histidine concentration was highest at the last sampling. The differences in the percolation properties of the amino acids are discussed in terms of important retention and elimination processes (biodegradation, ad-/desorption, intercalation).  相似文献   

19.
Mineralization kinetics of corn and clover residues in quartz sand, loam, sand + 15% bentonite, and sand + 30% kaolinite have been studied. A scheme has been proposed for the transformation of plant residues in mineral substrates. Kinetic parameters of mineralization have been calculated with the use of a first-order two-term exponential polynomial. It has been shown that the share of labile organic carbon pool in the clover biomass is higher (57–63%) than in the corn biomass (47–49%), which is related to the biochemical composition of plant residues. The mineralization constants of clover residues generally significantly exceed those of corn because of the stronger stabilization of the decomposition products of corn residues. The turnover time of the labile clover pool (4–9 days) in all substrates and that of the labile corn pool (8–10 days) in sands and substrates containing kaolinites and bentonite are typical for organic acids, amino acids, and simple sugars. In the loamy substrate, the turnover time of labile corn pool is about 46 days due to the stronger stabilization of components of the labile pool containing large amounts of organic acids. The turnover time of the stable clover pool (0.95 years) is significantly lower than that of the stable corn pool (1.60 years) and largely corresponds to the turnover time of plant biomass.  相似文献   

20.
Vesicular-arbuscular mycorrhizae in tropical sedges of southern India   总被引：6，自引：0，他引：6  
Twenty-four species of sedges (representing six genera) from different vegetation types in Western Ghats, South India, were examined for vesicular-arbuscular mycorrhizal (VAM) associations. All the sedges had VAM fungal infections with hyphae and vesicles, but arbuscules were observed only in 42% of the total species. The VAM fungal colonization varied considerably between species, ranging from 9 to 62%. Root colonization was positively and negatively correlated with root diameter and root hair length, respectively. The number of VAM fungal spores in the rhizosphere varied from 5 to 86 g-1 soil. No significant relationship was found between spore numbers or root colonization and either soil pH or moisture. Four Glomus spp., one Acaulospora sp., one Sclerocystis spp., and one Gigaspora sp. were identified among the VAM fungal spores. The results reflect a high incidence of VAM in sedges occurring in Western Ghats and emphasize the need to assess the VAM status of plant species from different ecosystems to understand their mycorrhizal status.  相似文献   

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