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Nitrogen fixation by Azospirillium brasilense in soil and the rhizosphere under controlled environmental conditions (1991)

Christiansen-Weniger, C. ; Veen, J. A.

Springer

Biology and fertility of soils 12 (1991), S. 100-106

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ISSN: 1432-0789

Keywords: Azospirillum ; 15N-isotope dilution ; Nitrogen fixation ; Acetylene reduction activity ; ARA ; Rhizosphere ; Mineral nitrogen ; Oxygen tension

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition

Notes: Summary Acetylene reduction activity by Azospirillum brasilense, either free-living in soils or associated with wheat roots, was determined in a sterilised root environment at controlled levels of O2 tension and with different concentrations of mineral N. In an unplanted, inoculated soil nitrogenase activity remained low, at approximately 40 nmol C2H4 h-1 per 2kg fresh soil, increasing to 300 nmol C2H4 h-1 when malic acid was added as a C source via a dialyse tubing system. The N2 fixation by A. brasilense in the rhizosphere of an actively growing plant was much less sensitive to the repressing influence of free O2 than the free-living bacteria were. An optimum nitrogenase activity was observed at 10 kPa O2, with a relatively high level of activity remaining even at an O2 concentration of 20 kPa. Both NO inf3 sup- and NH inf4 sup+ repressed nitrogenase activity, which was less pronounced in the presence than in the absence of plants. The highest survival rates of inoculated A. brasilense and the highest rates of acetylene reduction were found in plants treated with azospirilli immediately after seedling emergence. Plants inoculated at a later stage of growth showed a lower bacterial density in the rhizosphere and, as a consequence, a lower N2-fixing potential. Subsequent inoculations with A. brasilense during plant development did not increase root colonisation and did not stimulate the associated acetylene reduction. By using the 15N dilution method, the affect of inoculation with A. brasilense in terms of plant N was calculated as 0.067 mg N2 fixed per plant, i.e., 3.3% of the N in the root and 1.6% in the plant shoot were of atmospheric origin. This 15N dilution was comparable to that seen in plants inoculated with non-N2-fixing Psudomonas fluorescens.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00341483

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N2-fixation by ammonium-excreting Azospirillum brasilense in auxine-induced root tumours of wheat (Triticum aestivum L.) (1992)

Christiansen-Weniger, C.

Springer

Biology and fertility of soils 13 (1992), S. 165-172

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ISSN: 1432-0789

Keywords: Azospirillum brasilense ; 15N-isotope enrichment ; Nitrogen fixation ; Auxine ; 2,4-dichlorphenoxy acetic acid ; 3,5-dichlor-phenoxy acetic acid ; Acetylene reduction assay ; Wheat

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition

Notes: Summary Wheat seedlings, treated with the auxine 2,4-dichlor-phenoxy acetic acid (2,4-D) during germination developed only a residual root system. Root elongation was extremely restricted and root tips were deformed to thick club-shaped tumours. When 2,4-D was added in a later stage of plant growth the plants developed additional nodule-like knots along primary roots. Root and shoot dry-matter production was slightly repressed in all 2,4-D treatments and N translocation from roots to shoots was repressed as well. When transferred to an auxine-free growth medium, the 2,4-D-affected roots were not capable of complete recovery. In plants inoculated gnotobiotically with Azospirillum brasilense, either with the wild type or with the NH 4 + -excreting mutant strain C3, a 2,4-D addition increased rhizosphere acetylene-reduction activity at pO2 1.5 kPa. The O2 sensitivity of root-associated nitrogenase activity tended to be reduced. The number of root-colonizing bacteria, at approximately 108 colony-forming units (cfu) per g dry root, was similar in the 2,4-D treatments and untreated controls. Plant treatment with high concentrations of the chemical isomer 3,5-dichlor-phenoxy acetic acid (3,5-D) did not have comparable effects, either on plant development or on rhizosphere-associated nitrogenase activity. Root-tumour tissue inhabited by A. brasilense showed purple staining when subjected to a tetrazolium chloride solution, which may indicate intensive local nitrogenase activity in this tissue. Exposed to an 15N2-enriched atmosphere, plants treated with 2,4-D and with A. brasilense incorporated significantly higher amounts of 15N than untreated controls. In all cases the highest values of 15N enrichment were found following inoculation with the NH 4 + -excreting mutant strain C3.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00336274

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Ammonium-excreting Azospirillum sp. become intracellularly established in maize (Zea mays) para-nodules (1994)

Christiansen-Weniger, C. ; Vanderleyden, J.

Springer

Biology and fertility of soils 17 (1994), S. 1-8

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ISSN: 1432-0789

Keywords: Ammonium excretion ; Azospirillum brasilense ; Auxine ; 2,4-Dichlor-phenoxy-acetic acid ; Nitrogen fixation ; Paranodulation ; Maize ; Zea mays ; Electron microscopy

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition

Notes: Abstract Maize seedlings develop nodule-like tumour knots (para-nodules) along primary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Inoculated NH 4 + -excreting Azospirillum brasilense cells were shown to colonize these tumours, mostly intracellularly, promoting a high level of N2 fixation when microaerophilic conditions were imposed. The nitrogenase activity inside the para-nodules was less sensitive to free O2 than in non-para-nodulating roots. Both light and electron microscopy showed a dense bacterial population inside intact tumour cells, with the major part of the cell infection along a central tumour tissue. The bacteria colonized the cytoplasm with a close attachment to inner cell membranes. In an auxin-free growth medium, young 2,4-D-induced para-nodules grew further to become mature differentiated root organs in which introduced bacteria survived with a stable population. These results provide evidence that gramineous plants are potentially able to create a symbiosis with diazotrophic bacteria in which the NH 4 + -excreting symbiont will colonize para-nodule tissue intracellularly, thus becoming well protected.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00418663

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4

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Associative N2 fixation and root exudation of organic acids from wheat cultivars of different aluminium tolerance (1992)

Christiansen-Weniger, C. ; Groneman, A. F. ; Veen, J. A.

Springer

Plant and soil 139 (1992), S. 167-174

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ISSN: 1573-5036

Keywords: aluminium tolerance ; Azospirillum brasilense ; N2 fixation ; rhizosphere ; root exudation ; wheat

Source: Springer Online Journal Archives 1860-2000

Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition

Notes: Abstract Three wheat cultivars with different tolerances against free aluminium were grown monoxenically in association with Azospirillum brasilense. In situ nitrogen fixation, measured with the acetylene reduction assay, was higher by the aluminium-tolerant cultivars than by the sensitive cultivar. The transfer of fixed nitrogen to the host plant, determined by the 15N dilution technique, was also significantly higher in the aluminium-resistant wheat plants. The total accumulation of fixed nitrogen in the host plants due to an A. brasilense inoculation varied from approximately 13% to 17% of the total nitrogen in the root and 2.9% to 3.9% of the nitrogen in the shoot. The quantity and quality of exudates released in liquid nutrient solution were analysed separately for two of the wheat cultivars, one aluminium-tolerant and one aluminium-sensitive. After 29 days of growth the aluminium-tolerant plants exudated significantly higher total amounts of carbon than aluminium-sensitive plants. No differences between the two cultivars existed in the carbon exudation rate per gram dry root. Much higher concentrations of low molecular dicarboxylic acids i.e. succinic, malic and oxalic acid, were found in the exudates of aluminium-tolerant plants. Dicarboxylic acids are potential chelating compounds for positively charged metals such as aluminium and they may play an important role in protecting the plant against aluminium incorporation. They are also very suitable substrates for Azospirillum spp. It is therefore suggested that these factors may be causing the higher associative nitrogen fixation rates which was found in the aluminium-tolerant wheat cultivars.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00009307

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