Biological nitrogen fixation in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) by <Superscript>15</Superscript>N isotope-dilution and identification of associated culturable diazotrophs

The nitrogen-fixing capacity of a range of commercial cultivars of maize (Zea mays L.) was evaluated by the ¹⁵N isotope-dilution method. Biological nitrogen fixation (BNF) expressed as percent nitrogen derived from air (Ndfa) ranged from 12 to 33 regardless of nitrogen fertilization. BNF was not affected by mineral nitrogen fertilization except on cultivar Topacio and PAU-871 cultivars. Subsequently, culturable bacterial diazotrophs were isolated from endophytic tissue of maize: seed, root, stem, and leaf. All isolates were able to grow on N-free semisolid medium. Eleven bacteria isolates showed nitrogen-fixing capacity by the reduction of acetylene to ethylene and confirmed by PCR the presence of nifH gene in their genome. Identification of the 11 isolates was performed by bacteriological methods, 16S rRNA gene sequences, and phylogenetic analysis, which indicated that the bacteria isolated were closely related to Pantoea, Pseudomonas, Rhanella, Herbaspirillum, Azospirillum, Rhizobium (Agrobacterium), and Brevundimonas. This study demonstrated that maize cultivars obtain significant nitrogen from BNF, varying by maize cultivar and nitrogen fertilization level. The endophytic diazotrophic bacteria isolated from root, stem, and leaf tissues of maize cultivars may contribute to BNF in these plants.     

Isolation and partial characterization of endophytic diazotrophs associated with Japanese sugarcane cultivar

Endophytic bacteria were isolated from the juice of a 6 month-old sugarcane cv. NiF-8, which was collected from Miyako, Okinawa and planted in a field lysimeter in Tsukuba, Ibaraki, Japan. The most probable number of N₂-fixing endophytes was 4.5 × 10⁵ cells per gram of fresh weight sample and 21 isolates have a positive reaction for ARA in an N₂-free semi-solid medium with 10% crystallized sugarcane sugar and 0.5% sugarcane juice. Analyses of some of the biochemical properties of the N₂-fixing isolates indicated that 13 isolates were putative strains of Acetobacter diazotrophicus, 4 isolates showed similar characteristics to those of Herbaspirillum seropedicae, and 4 isolates consisted of Herbaspirillum rubribalbicans-like strains. This study confirmed the existence of N₂-fixing endophytic bacteria in the Japanese sugarcane cv. NiF-8.     

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.     

Pre-inoculation with arbuscular mycorrhizal fungi suppresses root knot nematode (<Emphasis Type="Italic">Meloidogyne incognita</Emphasis>) on cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>)

A pot experiment was conducted to evaluate the influence of pre-inoculation of cucumber plants with each of the three arbuscular mycorrhizal (AM) fungi Glomus intraradices, Glomus mosseae, and Glomus versiforme on reproduction of the root knot nematode Meloidogyne incognita. All three AM fungi tested significantly reduced the root galling index, which is the percentage of total roots forming galls. Numbers of galls per root system were significantly reduced only in the G. intraradices + M. incognita treatment. The number of eggs per root system was significantly decreased by AM fungus inoculation, no significant difference among the three AM fungal isolates. AM inoculation substantially decreased the number of females, the number of eggs g⁻¹ root and of the number of eggs per egg mass. The number of egg masses g⁻¹ root was greatly reduced by inoculation with G. mosseae or G. versiforme. By considering plant growth, nutrient uptake, and the suppression of M. incognita together, G. mosseae and G. versiforme were more effective than G. intraradices.     

Isolation of an endophytic diazotroph,Klebsiella oxytoca,from sweet potato stems in Japan

Abstract

The population density of endophytic bacteria in the stem of field-grown sweet potato cultivars (Beniotome [BO], Koganesengan [KS], and Shiroyutaka [SYD in Miyakonojo, Miyazaki, Japan, ranged from 10² to 10⁴ cells g^?1 fresh weight sample using a semi-solid nitrogen-free medium. Eleven strains were isolated from the stems and two isolates, BO-1 and BO-5, showed a positive reaction in the acetylene reduction activity (ARA) test. BO-1 and BO-5 were isolated from cv. Beniotome in September 1999 and 2000, respectively. Morpho-physiological characterization of these isolates revealed that BO-1 and BO-5 showed a similar colony color in potato sucrose agar slants, produced bubbles in a modified semi-solid medium, acidified the medium, and displayed similar characteristics using the API 20NE rapid diagnostic kit. Partial sequence analysis of 16S rRNA from BO-1 revealed a 100% similarity (491 bp) to that of Klebsiella oxytoca. The other 9 isolates showed a negative reaction in the ARA test, slightly acidified or did not acidify the medium. Partial sequence analysis of the 16S rRNA revealed that the isolate SY-2 corresponded to Methylobacterium sp. (99.3% similarity for 1,241 bp), BO-3 to Pantoea agglomerans (99.1% similarity for 469 bp), and BO-8 to Sphingomonas sanguinis (98.8% similarity for 419 bp).     

Nitrogen-fixing stem nodules on Aeschynomene afraspera

Summary Aeschynomene afraspera is a wild annual legume growing in periodically waterlogged soils in western Africa. This legume is characterized by a profuse stem nodulation. Nodules are formed on the stem at the emergence of lateral root primordia, called nodulation sites. These sites are irregularly distributed on vertical rows all along the stem and branches. Stem nodules are hemispherically shaped. Their outside is dark green and they contain a red-pigmented central zone. Stem nodules exhibit a high nitrogen-fixing potential. Acetylene reduction assays result in stem nodule activity of 309 mol C₂H₄ g^–1 dry nodule h^–1. Field-grown stem nodulated Aeschynomene accumulated more N (51 g N m^–2 in 10 weeks) than the root nodulated one. Because of this nitrogenfixing potential and its ability to grow in waterlogged conditions, A. afraspera could probably be introduced into tropical rice cropping systems.     

Biodiversity of diazotrophs associated to the plant cover of north sinai deserts: Biodiversität diazotropher assoziiert mit der pflanendecke der wüsten nordsinais

Efforts are made to record biodiversity of microflora and diazotrophs associated with the plant cover of the major agricultural development areas in north Sinai, around the El-Salam canal, a newly-constructed canal that brings Nile water westward across the Suez canal. Natural plant communities were collected from three major areas. Ectorhizosphere, endorhizosphere and phyllosphere samples were examined for total microbial population and diazotrophs. The vegetation of South Qantara (area I) is characterized by the dominance of Stipagrostis scoparia followed by Nitraria retusa, Convolvulus lanatus, Cornulaca monacantha and Filago desertorum. Rabaa-Bir El Abd (area II) is dominated by Artemisia monosperma, Panicum turgidum and Zygophyllum album. Euphorbia terracina, Oligomeris linifolia, Astragalus kahiricus, Hyoscyamus muticus and Thymelea hirsuta represent the major plants of El Ser and Al Quarir (area III). Microorganisms colonized root surfaces of all tested plants ranging from >?10⁵ to 10¹⁰ cfu g^???1. Diazotrophs were common residents (10¹⁰ cfu g^???1), invaded the root tissue and established endophytically (10²?–?10⁶ cfu g^???1). Fifty-one N₂-fixing isolates were obtained. Among the 32 bacilli isolates, Bacillus polymyxa and Bacillus circulans were more common compared to Bacillus macerans. BNF Gram-negative isolates belonged to Enterobacter agglomerans, Enterobacter gergoviae, Enterobacter amnigenus, Klebsiella pneumoniae, Pseudomonas luteola, Pseudomonas cepacia, Agrobacterium radiobacter and Azospirillum spp.     

Influence of N fertilisation on the isolation of Acetobacter diazotrophicus and Herbaspirillum spp. from Indian sugarcane varieties

 Observations of newer diazotrophs like Acetobacter diazotrophicus and Herbaspirillum spp. in Indian sugarcane varieties is reported here for the first time. A. diazotrophicus was isolated from sugarcane plants where N fertilisation was nil, and numbers in the Indian cultivars were estimated to be up to 10⁶–10⁷ colony forming units g¹ fresh weight. N fertilisation was the limiting factor for the isolation of A. diazotrophicus, as evidenced by selective growth patterns in different concentrations of N sources, but Herbaspirillum spp. were found to occur both in N fertilised and unfertilised samples. The fact that N is not the limiting factor for Herbaspirillum spp. was evidenced in a similar study of its growth pattern in different N sources. Received: 14 May 1998     

Infection and colonization of aseptically micropropagated sugarcane seedlings by nitrogen-fixing endophytic bacterium, Herbaspirillum sp. B501gfp1

In this study, we used Herbaspirillum sp. B501gfp1 (B501gfp1), an isolate from wild rice, to investigate the interaction between a non-host nitrogen-fixing endophytic bacterium and micropropagated sugarcane plants under aseptic condition. Two Japanese sugarcane plants (Saccharum sp.) cultivars (cvs) NiF8 and Ni15 were inoculated using B501gfp1 in two inoculum doses of 10⁸ and 10² bacterial-cells-per-milliliter suspension. The results showed that bacterial cells colonized both the root and stem tissues, and colonization was apparent in the intercellular spaces. Higher bacterial numbers were detected in plant tissues inoculated with the higher inoculum concentration treatment. Bacterial numbers also varied between the two cultivars, with the higher values determined in cv Ni15. This study provides evidence that Herbaspirillum sp. B501gfp1, a rice isolate, could colonize sugarcane tissues, suggesting non-specificity of host plant among endophytes.     

Diversity and plant growth promoting evaluation abilities of bacteria isolated from sugarcane cultivated in the South of Brazil

Plant growth promoting (PGP) bacteria associated with sugarcane are a promising alternative for the expansion of this crop in Southern Brazil. In this study bacterial strains from different sugarcane fields were isolated to estimate their diversity, to evaluate some of their PGP activities and to use them as inoculant strains in field experiment. Samples of rhizospheric soil, roots, and stems of sugarcane were collected in six Rio Grande do Sul localities. The isolation of bacteria was made in three different N-free media. DNA from each isolate was subjected to nifH or 16S rDNA PCR-RFLP, and to the 16S rDNA partial sequencing. Five hundred and sixteen strains were isolated and several PGP characteristics were analyzed. Shannon index was used to evaluate the bacterial diversity. Indexes varying from 0.94 to 2.46 were obtained. Soil pH and clay were the characteristics most closely related to bacterial diversity. Achromobacter, Agrobacterium, Burkholderia, Gluconacetobacter, and Stenotrophomonas were the most abundant genera. Concerning the PGP activities, indolic compounds production was detected in 368 isolates; 138 isolates were able to solubilize phosphate; and 390 were siderophores producers. The inoculation of sugarcane with Gluconacetobacter diazotrophicus VI27 strain showed a significant increase in the number of sets germinated, in the amount of soluble solids, and in the yield of sugarcane juice compared with the control. As a conclusion, a diverse population of PGP bacteria was found in the sugarcane samples. These bacteria, especially G. diazotrophicus strain VI27, could be used as biofertilizers of sugarcane as well as other cereal crops under controlled conditions to avoid or reduce the use of standard N fertilizers.     

Characterization of cultivable methanotrophs from paddy soils and rice roots

Abstract

Characterization of methanotrophs isolated from paddy soils and rice (Oryza sativa) roots was investigated in the present study. The number of methanotrophs in root homogenates of the rice cultivar Mutsuhomare was 4.9 × 10⁷ most-probable-number (MPN) g^?1 dry roots, in Yumeakari it was 2.0 × 10⁸ MPN g^?1 dry roots and in Kirara it was 4.6 × 10⁷ MPN g^?1 dry roots. Although bacterial cells were observed infrequently on the surface and in the interiors of roots before incubation, a large number of colonies, measuring 0.5–5 mm in diameter, were observed on the sterilized roots after incubation on nitrate mineral agar plates under methane in air. In particular, a large number of colonies were observed at the emergence sites of lateral roots and root hairs. Strains MD5-1 and M1 were isolated from the roots of Mutsuhomare and strain R62 was isolated from the root homogenate of Yumeakari. All isolates were catalase-positive and oxidase-positive, Gram-negative, straight-rod-shaped and curved-rod-shaped bacteria, and formed exospores. The isolates were able to fix nitrogen and grew in the absence of copper. In addition, all were found to be positive for naphthalene-oxidizing activities (corresponding to soluble methane mono-oxygenase activities). Strains MD5-1, M1 and R62 were closely related to Methylosinus sporium. Methanotrophic strains W3-6, SD3-5 and 2-19, isolated previously from paddy field soils, were classified into Methylosinus (W3-6) and Methylocystis (SD3-5 and 2-19) type II methanotrophs. Isolates from the rice roots (MD5-1, R62 and M1) grew logarithmically when casamino acid was used as the nitrogen source; however, the growth of these strains was reduced on the nitrate medium. These strains preferred amino acids over inorganic nitrogen as a nitrogen source for growth.     

Isolation and characterization of endophytic bacteria from wild and cultivated soybean varieties

Endophytic bacteria were isolated from surface sterilized stems, root, and nodules of wild and cultivated soybean varieties. Various phenotypic traits that are expected to be involved in the persistence and functions of the bacteria were analyzed. Most of the isolates from soybean were motile and indoleacetic acid producers, and 70 and 33% of isolates excreted cellulase and pectinase, respectively. These traits may be involved in endophytic characteristics in soybean. Some isolates were resistant to Str₁₀₀, formed capsules, and produced fluorescent pigments. Molecular characterization of selected 35 endophytic bacteria by 16S rDNA–polymerase chain reaction–restriction fragment length polymorphism showed two main clusters at 48 and 43% similarity coefficients in which most of the endophytes belonged. The genetic variation was more among endophytes isolated from Glycine max tissues than from G. soja.     

Sesbania rostrata as a green manure for lowland rice: Growth,N2 fixation,Azorhizobium sp. inoculation,and effects on succeeding crop yields and nitrogen balance

Summary Root and stem nodulation, nitrogen fixation (acetylene-reducing activity), growth and N accumulation bySesbania rostrata as affected by season and inoculation were studied in a pot experiment. The effects ofS. rostrata as a green manure on succeeding wet-season and dry-season rice yields and total N balance were also studied.S. rostrata grown during the wet season showed better growth, nodulation, and greater acetylene-reducing activity than that grown during the dry season. Inoculation withAzorhizobium caulinodans ORS 571 StrSpc® (resistant to streptomycin and spectinomycin) on the stem alone or on both root and stem significantly increased N₂ fixation by the plants. Soil and seed inoculation yielded active root nodules under flooded conditions. Plants that were not inoculated on the stem did not develop stem nodules. The nitrogenase activity of the root nodules was greater than that of the stem nodules in about 50-day-oldS. rostrata. S. rostrata incorporation, irrespective of inoculation, significantly increased the grain yield and N uptake of the succeeding wet season and dry season rice crops. The inoculated treatments produced a significantly greater N gain (873 mg N pot^–1) than the noinoculation (712 mg N pot^–1) treatment. About 80% of the N gained was transferred to the succeeding rice crops and about 20% remained in the soil. The soil N in the flooded fallow-rice treatment significantly declined (–140 mg N pot^–1) but significantly increased in bothS. rostrata-rice treatments (159 and 151 mg N pot^–1 in uninoculated and inoculated treatments respectively). The N-balance data gave extrapolated values of N₂ fixed per hectare at about 303 kg N ha^–1 per two crops forS. rostrata (uninoculated)-rice and 383 forS. rostrata (inoculated)-rice.     

Effect of inoculation with different strains of Azospirillum brasilense on cotton (Gossipium barbadense)

Summary The response of the cotton plant to inoculation with six strains of Azospirillum brasilense was investigated under subtropical conditions in Egypt. Azospirilla populations and activities were increased as a result of root inoculation with liquid inoculum of Azospirillum sp. Highest C₂H₂ — reduction activities on roots were obtained with strains S631 and Sp Br 14 (means of 216.85 and 209.50 nmol C₂H₄g^–1 root h^–1 respectively) while strain M4 gave the lowest activity (mean of 100.8 nmol C₂H₄g^–1 root h^–1). Statistical analysis showed that Azospirillum strains 5631, Sp Br 14, E15 and SC22 significantly increased the plant dry weight and nitrogen uptake while inoculation with strains M4 and SE had no significant effect in that respect.     

Competition between inoculated and indigenous Rhizobium/Bradyrhizobium spp. strains for nodulation of grain and fodder legumes in Pakistan

Summary The competitive ability of inoculated and indigenous Rhizobium/Bradyrhizobium spp. to nodulate and fix N₂ in grain legumes (Glycine max, Vigna unguiculata, Phaseolus vulgaris) and fodder legumes (Vicia sativa, Medicago sativa, and Trifolium subterraneum) was studied in pots with two local soils collected from two different fields on the basis of cropping history. The native population was estimated by a most-probable-number plant infectivity test in growth pouches and culture tubes. The indigenous rhizobial/bradyrhizobial population ranged from 3 to 2×10⁴ and 0 to 4.4×10³ cells g^-1 in the two soils (the first with, the second without a history of legume cropping). Inoculated G. max, P. vulgaris, and T. subterraneum plants had significantly more nodules with a greater nodule mass than uninoculated plants, but N₂ fixation was increased only in G. max and P. vulgaris. A significant response to inoculation was observed in the grain legume P. vulgaris in the soil not previously used to grow legumes, even in the presence of higher indigenous population (>10³ cells g^-1 soil of Rhizobium leguminosarum bv phaseoli). No difference in yield was observed with the fodder legumes in response to inoculation, even with the indigenous Rhizobium sp. as low as <14 cells g^-1 soil and although the number and weight of nodules were significantly increased by the inoculation in T. subterraneum. Overall recovery of the inoculated strains was 38–100%, as determined by a fluorescent antibody technique. In general, the inoculation increased N₂ fixation only in 3 out of 12 legume species-soil combinations in the presence of an indigenous population of rhizobial/bradyrhizobial strains.     

Nitrogen fixation of Sesbania rostrata: Contribution of stem nodules to nitrogen acquisition

Abstract

Nitrogen contents, nodule numbers, and nodule dry weights of 6-week-oId Sesbania rostrata plants grown in sand culture with only root nodules, only stem nodules or with both were compared and the root nodules were found to contribute to nitrogen acquisition more significantly than the stem nodules. Similar findings were obtained in ¹⁵N₂-fixing experiments. An 8-week-old plant with both stem and root nodules fixed 1.50 mg nitrogen in a 12 h light period, while the fixation decreased to 1.15 mg nitrogen after the removal of the stem nodules, suggesting that root nodules played major role in nitrogen fixation. However, acetylene-reducing activities per nodule dry weight were higher in the stem nodules. Under flooding conditions, the aerenchyma tissues contributed to about 40% of N₂ transport to root nodules, and 60% was supplied through stem.     

Interactions of Glomus clarum with Acetobacter diazotrophicus in infection of sweet potato (Ipomoea batatas), sugarcane (Saccharum spp.), and sweet sorghum (Sorghum vulgare)

Summary Spores of the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus clarum obtained from sweet potatoes grown in soil inoculated with this fungus and with an enrichment culture of Acetobacter diazotrophicus contained A. diazotrophicus and several other bacteria, including a diazotrophic Klebsiella sp. Inoculation of micropropagated sweet potatoes with G. clarum and A. diazotrophicus enhanced spore formation in soil compared to VAM inoculation alone. Plants inoculated with VAM spores containing the bacteria showed additional increases in the number of spores formed within roots. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM or when present within VAM spores. Micropropagated sugarcane seedlings inoculated with the same VAM spores containing the diazotrophs also contained much higher numbers of A. diazotrophicus in aerial parts than seedlings inoculated in vitro with the bacteria alone. When grown in non-sterile soil, the sugarcane seedlings again showed the greatest infection of aerial parts after inoculation with VAM spores containing the diazotrophs. This treatment also increased VAM colonization and the numbers of spores formed within roots. Similar effects were observed in sweet sorghum except that the aerial plant parts were not infected by A. diazotrophicus.     

不同连作年限马铃薯根系分泌物的成分鉴定及其生物效应

为探讨马铃薯连作障碍的可能机理,在大田条件下,收集连作1～5 a(CP1-CP5)马铃薯植株的根系分泌物,采用GC-MS对根系分泌物的主要成分进行了鉴定,并通过生物检测验证了根系分泌物的生物效应。结果表明:CP1-CP5鉴定出的物质主要有酸类、糖类、胺类、醇类、酯类和嘧啶类,但各类物质的数量和含量不同。CP1-CP5中均鉴定出棕榈酸,相对含量分别为0.55%、0.87%、1.24%、1.05%和0.95%,浓度分别为7.12 mg×L~(-1)、7.39 mg×L~(-1)、9.46 mg×L~(-1)、8.38 mg×L~(-1)和8.02 mg×L~(-1)。马铃薯根系分泌物显著抑制了马铃薯的生长,抑制作用随连作年限延长而增强。棕榈酸对马铃薯生长的抑制表现出明显的浓度效应,随浓度升高而增强。马铃薯根系分泌物明显促进了立枯丝核菌的生长,菌落直径和菌丝鲜质量表现为CP3最高,CP1最低,CP2、CP4和CP5之间没有显著差异。棕榈酸明显促进了立枯丝核菌的生长,随着棕榈酸浓度的增加,菌落直径和菌丝鲜质量先增加再减小, 10 mg×L~(-1)棕榈酸的菌落直径和菌丝鲜质量最大。由此说明,随马铃薯连作年限延长,根系分泌物的毒性越强;马铃薯根系分泌物对立枯丝核菌的促进作用加剧了马铃薯的连作障碍,棕榈酸是马铃薯根系分泌的化感自毒物质。     

Phosphate solubilization by <Emphasis Type="Italic">Penicillium</Emphasis> spp. closely associated with wheat roots

In this study we found that Penicillium spp. exhibiting P-solubilizing activity are common both on and in the roots of wheat plants grown in southern Australian agricultural soils. From 2,500 segments of washed and surface-disinfested root pieces, 608 and 223 fungi were isolated on a selective medium, respectively. All isolates were screened for P solubilization on solid medium containing hydroxyapatite (HA); 47 isolates (5.7%) solubilized HA and were identified as isolates of Penicillium or its teleomorphs. These isolates were evaluated for solubilization of Idaho rock phosphate (RP) in liquid culture. Penicillium bilaiae strain RS7B-SD1 was the most effective, mobilizing 101.7 mg P l^–1 after 7 days. Other effective isolates included Penicillium simplicissimum (58.8 mg P l^–1), five strains of Penicillium griseofulvum (56.1–47.6 mg P l^–1), Talaromyces flavus (48.6 mg P l^–1) and two unidentified Penicillium spp. (50.7 and 50 mg P l^–1). A newly isolated strain of Penicillium radicum (KC1-SD1) mobilized 43.3 mg P l^–1. RP solubilization, biomass production and solution pH for P. bilaiae RS7B-SD1, P. radicum FRR4718 or Penicillium sp. 1 KC6-W2 was determined over time. P. bilaiae RS7B-SD1 solubilized the greatest amount of RP (112.7 mg P l^–1) and had the highest RP-solubilizing activity per unit of biomass produced (up to 603.2 g P l^–1 mg biomass^–1 at 7 days growth). This study has identified new isolates of Penicillium fungi with high mineral phosphate solubilizing activity. These fungi are being investigated for the ability to increase crop production on strong P-retaining soils in Australia.     

Nitrogenase (C2H2) Activity in Roots of Non-Cultivated and Cereal Plants: Influence of Nitrogen Fertilizer on Populations and Activity of Nitrogen-Fixing Bacteria

Root samples of 11 non-cultivated monocotyledonous and 7 dicotyledonous species taken during a wet summer had low mean nitrogenase activities of 10.2 and 7.1 nmol C₂H₄·g^?1 DW·h^?1 after preincubation at pO₂ 0.02, respectively. Maxima of 139–169 nmol·g^?1·h^?1 were observed with Agrostis vulgaris and Agropyron repens on a sandy soil poor in C_org. Three of 6 early, but none of 4 late fodder maize cultivars had a very low activity up to 0.5 nmol·g^?1h^?1. Oat, rye and wheat roots from plots with organic or mineral N fertilizers had activities between 1.3 and 7.3 nmol·g^?1h^?1 at flowering, which were not correlated with their Azospirillum populations (10²-10⁷·g^?1 after preincubation). Winter wheat and barley roots given 0, 40, 80 and 120 kg. ha^?1 NH₄NO₃-N in 0–3 applications had mean activities of 0.08, 4.06, 0.09 and 0.08 nmol or 1.77, 2.67, 0.36 and 0.23 nmol C₂H₄g^?1·h^?1 after flowering, respectively. An appreciable part of this activity could be removed by root washing. In preincubated rhizosphere soil of wheat and barley populations of N₂-fixing, facultative anaerobic Klebsiella and Enterobacter spp. were 10–100 times higher than those of Azospirillum sp., both being higher in O N than in 80 kg N·ha^?1 trials.