Yield response of various wheat genotypes to inoculation withAzospirillum brasilense

Summary Twenty different wheat genotypes representing a wide genetic variability, were tested for their yield and yield components response to inoculation withAzospirillum brasilense, at two levels of N fertilization. Only two cultivars responded by a significant yield increase of 7.4 and 8.0 per cent — both at the higher N level. The response reflected an increase in the number of grains per plant added as additional spikes. The importance of the host plant genotype for a successful wheat-bacteria association is discussed.     

Symbiotic N2 fixation in pea and field bean estimated by15N fertilizer dilution in field experiments with barley as a reference crop

Summary The total amount of nitrogen derived from symbiotic nitrogen fixation in two pea and one field bean cultivar, supplied with 50 kg N ha⁻¹ at sowing (‘starter’-N), was estimated to 165, 136, and 186 kg N ha⁻¹, respectively (three-year means). However, estimates varied considerably between the three years. At the full bloom/flat pod growth stage from 30 to 59 per cent of total N₂ fixation had taken place. The proportion of total N derived from N₂ fixation at maturity was higher in seeds than in vegetative plant parts and amounted to 59.5, 51.3 and 66.3 per cent of total above-ground plant N in the two pea cultivars and field bean, respectively (three-year means). The recovery of fertilizer N was 62.2, 70.2, 52.1, and 69.5 per cent in the two pea cultivars, field bean and barley, respectively. Growth analysis indicated that barley did not meet the claims for an ideal reference crop in the¹⁵N fertilizer dilution technique for estimating N₂ fixation in pea and field bean. ‘Starter’-N neither increased the seed yield nor the N content of the grain legumes.     

Yield response of a common spring wheat cultivar to inoculation withAzospirillum brasilense at various levels of nitrogen fertilization

Summary The yield response of a common spring wheat cultivar,Triticum aestivum, to inoculation withAzospirillum brasilense was studied at four levels of N fertilization. Plant yield increased due to the inoculation treatment only at medium and high levels of N fertilization, with a maximum yield increase of about 8.0 per cent at the highest level (approximately 1.0 g of pure N per plant). Yield increase was mostly due to an increase in the number of grains per spike, and at the highest level of fertilization, also due to a higher number of spikes per plant. At all N levels, the inoculation caused an increase of 0.5–1.4 per cent in the number of fertile spikelets per main spike.Grain protein percentage was unaffected by the inoculation, though significantly increase due to the fertilization treatments.The occurrence of maximum yield response at the highest N level, the response by early-determined yield components, i.e. spikelet number, and the unaffected grain protein content are in accord with the suggestion that the contribution ofAzospirillum brasilense to wheat yield is not through N₂-fixation.     

Correlation of greenhouse bioassay with field response to fertilizer by paper birch

Summary Bulk soil samples were collected from the top 15 cm of untreated areas adjacent to field fertilizer trials at 2 locations. Amounts of N, P, K, and lime equivalent to the field treatments were mixed with the soil in 15-cm diameter pots. Paper birch (Betula papyrifera March.) trees were grown from seed for a greenhouse bioassay. Height and dry weight of the bioassay seedlings were significantly correlated with 3-year volume growth of 10-year-old paper birch seedlings in the field. Correlation coefficients were 0.88 for height growth and 0.91 for dry weight growth on one site, and 0.72 and 0.63 on the other. With further refinements and observations on a larger number of sites, this bioassay technique should be a valuable tool for estimating potential response to fertilizer by young paper birch in the field, and for ranking the relative productivity of different soils.     

Field response of wheat to arbuscular mycorrhizal fungi and drought stress

Mycorrhizal plants often have greater tolerance to drought than nonmycorrhizal plants. This study was conducted to determine the effects of arbuscular mycorrhizal (AM) fungi inoculation on growth, grain yield and mineral acquisition of two winter wheat (Triticum aestivum L.) cultivars grown in the field under well-watered and water-stressed conditions. Wheat seeds were planted in furrows after treatment with or without the AM fungi Glomus mosseae or G. etunicatum. Roots were sampled at four growth stages (leaf, tillering, heading and grain-filling) to quantify AM fungi. There was negligible AM fungi colonization during winter months following seeding (leaf sampling in February), when soil temperature was low. During the spring, AM fungi colonization increased gradually. Mycorrhizal colonization was higher in well-watered plants colonized with AM fungi isolates than water-stressed plants. Plants inoculated with G. etunicatum generally had higher colonization than plants colonized with G. mosseae under both soil moisture conditions. Biomass and grain yields were higher in mycorrhizal than nonmycorrhizal plots irrespective of soil moisture, and G. etunicatum inoculated plants generally had higher biomass and grain yields than those colonized by G. mosseae under either soil moisture condition. The mycorrhizal plants had higher shoot P and Fe concentrations than nonmycorrhizal plants at all samplings regardless of soil moisture conditions. The improved growth, yield and nutrient uptake in wheat plants reported here demonstrate the potential of mycorrhizal inoculation to reduce the effects of drought stress on wheat grown under field conditions in semiarid areas of the world.     

The effects of placing small amounts of phosphate fertilizer close to the seed on growth and nutrient concentrations of lettuce

Summary Two glasshouse experiments are described in which the effects of applying starter phosphate fertilizer, 1 cm beneath the seeds, on early growth and nutrient concentrations of lettuce (Lactuca sativa L. cv. Avondefiance) in well fertilized soil were determined. In Experiment 1 various rates of starter P in the form of NH₄H₂PO₄ were applied to soil containing a range of rates of incorporated triple superphosphate. Although there was little response of lettuce dry weight to the incorporated triple superphosphate there was a large response (about 65% increase after 36 days) to the starter. N and P concentrations within the plants were increased by the starter treatments whereas K concentration was reduced. The per cent P in the plants at 36 days from sowing could account for 60% of the variation in plant dry weight. In Experiment 2 the starter P was added as either the Ca, Na or K salt, with or without added (NH₄)₂SO₄. Adding the starter P without ammonum increased the P concentration of the plants by an average of 12% and the dry weight by an average of 39% at 30 days from sowing. The addition of ammonium ions increased plant concentrations of P, Mg and N but decreased plant K concentration. The effect of the ammonium ions on growth depended on the form of phosphate supplied as the starter. This variation in effect of ammonium ions was attributed to the effects of other starter ions on the relative concentration of ammonium in the soil solution.     

In-vitro interaction between chloroplasts and peroxisomes as controlled by inorganic phosphate

Peroxisomes, whole chloroplasts, mitochondria, and broken chloroplasts of spinach (Spinacia oleracea L.), each form 1 band at its typical density, when isolated in sucrose gradients by isopycnic centrifugation in glycylglycine buffer. In potassium-phosphate buffer peroxisomes form a 2nd band at the density of whole chloroplasts. The phosphate effect is half-saturated at a concentration of 10–20 mM. If whole chloroplasts are removed by differential centrifugation before isopycnic centrifugation no second band is formed. Arsenate can be substituted for potassium phosphate while KCl, NaCl, KNO₃ and glycolate cannot, showing inorganic phosphate to be the active ion. Evidence is presented showing that during isopycnic centrifugation more slowly sedimenting peroxisomes have to move through faster sedimented bands of whole and broken chloroplasts. In the presence of inorganic phosphate this leads to a specific interaction between whole chloroplasts and peroxisomes visible as a second peroxisomal band at the density of the whole chloroplasts. The relationship of the interaction in vitro to the known association of the two organelles in vivo is considered.     

Similarities and relationships among populations of the bulb onion as estimated by nuclear RFLPs

Random nuclear restriction fragment length polymorphisms (RFLPs) were used to assess similarities and relationships among open-pollinated (OP) populations of the cultivated bulb onion (Allium cepa). Seventeen OP populations and 2 inbreds of contrasting daylength response [termed by convention as long (LD) and short (SD) day], 1 shallot (A. cepa var. ascalonicum), and one cultivar of bunching onion (Allium fistulosum) were examined with 104 cDNA clones and two to four restriction enzymes. Sixty (58%) clones detected at least 1 polymorphic fragment scorable among the OP populations and were used for analyses. The average number of polymorphic fragments per polymorphic probe-enzyme combination was 1.9, reflecting that numerous monomorphic fragments were usually present. Similarities were estimated as the proportion of polymorphic fragments shared by 2 populations. Average similarity values among LD, among SD, and between LD and SD OP populations were 0.79, 0.67, and 0.68, respectively. Relationships among the OP populations were estimated by parsimony, cluster analysis of similarities using the unweighted-pair-group method (UPGMA), and multivariate analysis using principle components. Parsimony analysis generated a strict consensus tree that grouped all but 1 LD onion with unresolved relationships to the SD OP populations. The UPGMA analysis placed together the LD storage OP populations. Principal component analysis grouped all but 2 LD onions; the other OP populations were dispersed. The results suggest that LD and SD onions do not represent distinct germ plasm, but that LD storage onions represent a derived group selected for production at higher latitudes. If it is assumed that the sampled populations are representative of all onion OP populations, the lower similarities among SD OP populations indicate that their collection and maintenance in germ plasm collections is important for the preservation of genetic diversity.     

Native soil and fresh fertilizer phosphorus uptake as affected by rate of application and P fertilizers

³²P labelled fertilizers were used to measure native soil and fresh fertilizer phosphorus uptake byLolium perenne L. in greenhouse experiments. The P source evaluation was carried out for multiple rates of application for a standard P fertilizer (DAP) on low and medium soil P levels and for North Carolina rock phosphate (RP) at the medium soil P level only. At the low soil P level, the native P uptake increase was independent of P-DAP applied, and represented 19% of the nil P uptake. At the medium soil P level, the variability of the native soil as a nutrient P source depended on the phosphate fertilizer applied, and the rate of application. Consequently the amount of total P uptake could conceal differences in P fertilizer evaluations as the nutrient P source. Fresh P uptake increased linearly with the rates of P applied as standard or tested P fertilizer. The comparison of various P sources by means of fresh P uptake ratio (i.e. fresh P uptake from tested phosphate divided by fresh P uptake from standard phosphate) was independent of the rate of application. It was therefore suggested that various phosphorus sources be evaluated by measuring the fresh P uptake for a single rate of application.     

The role of root-associatedKlebsiella pneumoniae in the nitrogen nutrition ofPoa pratensis andTriticum aestivum as estimated by the method of15N isotope dilution

Summary The technique of¹⁵N isotope dilution was used to verify that nitrogen was fixed and transferred to the plant byKlebsiella pneumoniae strain Pp in association withPoa pratensis orTriticum aestivum. Surface sterilized, sprouting seeds were inoculated withK. pneumoniae and grown in sand in modified Leonard jars. Potassium nitrate enriched with¹⁵N was used to provide N concentrations ranging from 10–40 mg Nl^–1 nutrient solution. After 10–18 weeks the shoots and roots were analyzed separately for dry matter, N content, total N, and atom %¹⁵N excess. The acetylene reduction technique was used to test for the presence of N₂-fixing organisms on the roots. The data from¹⁵N isotope dilution demonstrated that up to 33.8% of N in the shoots ofP. pratensis and 15.9% in those ofT. aestivum were derived from associative N₂ fixation byK. pneumoniae. In most experiments the dry matter yield, N content, and total N yield of the shoots ofP. pratensis were increased byK. pneumoniae inoculation, whereas inoculation had no significant effect on the dry matter yield, N content or total N of the shoots ofT. aestivum.     

Phosphorus (P) efficiencies and mycorrhizal responsiveness of old and modern wheat cultivars

A pot experiment was carried out in a growth chamber to investigate P efficiencies and mycorrhizal responsiveness of modern (Krichauff and Excalibur) and old (Khapstein, Bobin, Comeback and Purple Straw) wheat cultivars (Triticum aestivum). The arbuscular mycorrhizal fungus (AMF) used in this study was Glomus intraradices. The growth medium was a soil/sand mixture with NaHCO₃-extractable P of 9.4 mg P kg^–1 and no extra P was added. Plant P efficiencies (uptake, utilisation and agronomic) were found to differ significantly between cultivars, but no general trends of changes with the year of release of the cultivar were found. AMF colonisation was found to decrease plant growth under our experimental conditions with low light intensity. Mycorrhizal responsiveness (MR) was measured in terms of the improvement in plant P nutrition (shoot P concentrations). MR was found to be generally lower in modern cultivars than in old cultivars, indicating that modern breeding programs may have reduced the responsiveness of modern wheat cultivars to arbuscular mycorrhizal fungi. MR was also found to decrease in general with increased plant P utilisation efficiency.     

The response of rice to the growth and nitrogen fixation inAzolla caroliniana andAzolla pinnata at varying rates of phosphate fertilization

The response of rice toAzolla caroliniana, newly introduced in India, was compared with the reponse to the local isolate ofAzolla pinnata at varying rates of phosphate fertilizer (4.4–8.8 kg P ha^–1) during a wet and a dry season.Fresh weight, dry weight and fixed N were more for both species 21 DAI (days after inoculation) than 14 DAI, but acetylene reduction activity (ARA) was higher 14 DAI than 21 DAI. Dry weight of Azolla and fixed N were less 14 DAI forA. caroliniana than forA. pinnata during the wet season. Twenty-one DAI, fresh weight ofA. caroliniana was 62.1 and 27.6% higher than that ofA. pinnata during the wet and dry season, respectively. However, dry weight and fixed N were more 21 DAI inA. caroliniana than inA. pinnata during only the wet season. The ARA was higher inA. caroliniana both 14 and 21 DAI, irrespective of season. The presence of either species in the rice field increased grain yield, straw yield, number of panicles m^–2, number of grains per panicle and reduced percentage sterility during both the wet and the dry season. Phosphate application significantly increased fresh weight, dry weight, ARA and fixed N for both species as well as grain and straw yields of rice. The responses to phosphate fertilizer were similar for both Azolla species and for rice grown with either one of the Azolla species.     

Availability of phosphorus in a calcareous soil treated with rock phosphate or superphosphate as affected by phosphate-dissolving fungi

A study was undertaken to isolate some fungi exhibiting phosphate-dissolution ability, and to test whether these fungi are capable of increasing the amount of available P in a calcareous soil treated with rock phosphate (RP) or with triple superphosphate (TSP) and its subsequent uptake by sorghum (Sorghum bicolor L. Moench).Penicillium sp. and twoAspergillus foetidus (Naka) isolates significantly increased the availability of P in soil treated with RP or TSP during the growing season.Penicillium sp. isolate was more effective in increasing available P in the soil treated with RP or TSP than were Aspergillus isolates. However, the dry matter and P uptake responses to inoculation with these fungi were better in the soil treated with RP than in soil treated with TSP. In the TSP treated soil, the fungi achieved their maximum P releasing capacity two weeks earlier than in soil treated with RP. Positive and significant correlation coefficients among available P, P uptake and dry matter production at different periods of the growing season were observed following inoculation. However, none of these variables were found to be significantly correlated with the fungal populations.     

Differences in the heat-shock response between thermotolerant and thermosusceptible cultivars of hexaploid wheat

Summary Heat-shock protein (HSP) gene expression in two wheat lines cv Mustang (heat-tolerant) and cv Sturdy (heat-susceptible) were analyzed to determine if wheat genotypes differing in heat tolerance also differ in in-vitro HSP synthesis (translatable HSP mRNAs) and steady-state levels of HSP mRNA. Several sets of mRNA were isolated from seedling leaf tissues which had been heat-stressed at 37 °C for various time intervals. These mRNAs were hybridized with HSP cDNA or genomic DNA probes (HSP17, 26, 70, 98, and ubiquitin). Protein profiles were compared using in-vitro translation and 2-D gels. The Northern slot-blot data from the heat-stress treatment provide evidence that the heat-tolerant cv Mustang synthesized low molecular weight (LMW) HSP mRNA earlier during exposure to heat shock and at a higher level than did the heat-susceptible cv Sturdy. This was especially true for the chloroplast-localized HSP. The protein profiles shown by 2-D gel analysis revealed that there were not only quantitative differences of individual HSPs between the two wheat lines, but also some unique HSPs which were only found in the Mustang HSP profiles. The high level of RFLP between the two wheat lines was revealed by Southern blot hybridization utilizing a HSP17 probe. These data provide a molecular basis for further genetic analysis of the role of HSP genes in thermal tolerance in wheat.     

Photoacclimation and photoinhibition inUlva rotundata as influenced by nitrogen availability

Clonal tissue of the marine chlorophyte macroalga, Ulva rotundata Blid., was transferred from 100 to 1700 mol photons · m^–2 · s^–1 under limiting (1.5 M NH ₄ ⁺ maximum, N/P=2) and sufficient (15 M NH ₄ ⁺ maximum, N/P=20) nitrogen supply at 18° C and 11 h light-13 h darkness daily. Photoinhibition was assayed by light-response curves (photosynthetic O₂ exchange), and chlorophyll fluorescence at 77 K and room temperature. Daily surface-area growth rate (_SA) in N-sufficient plants increased sixfold over 3 d and was sustained at that level. During this period, respiration (R _d) doubled and light-saturated net photosynthesis capacity (P _m) increased by nearly 50%, indicating acclimation to high light. Quantum yield () decreased by 25% on the first day, but recovered completely within one week. The ratio of variable to maximum fluorescence (F _v/F _m) also decreased markedly on the first day, because of an increase in initial fluorescence (F _o) and a decrease in F _m, and partially recovered over several days. Under the added stress of N deficiency, _SA accelerated fivefold over 4 d, despite chronic photoinhibition, then declined along with tissue-N. Respiration doubled, but P _m decreased by 50% over one week, indicating inability to acclimate to high light. Both and F _v/F _m decreased markedly on the first day and did not significantly recover. Changes in F _o, F _m and xanthophyll-cycle components indicate concurrent photodamage to photosystem II (PSII) and photoprotection by thermal deexcitation in the antenna pigments. Increasing _SA coincided with photoinhibition of PSII. Insufficient diel-carbon balance because of elevated R _d and declining P _m and tissue-N, rather than photochemical damage per se, was the apparent proximate cause of decelerating growth rate and subsequent tissue degeneration under N deficiency in U. rotundata.Abbreviations and Symbols Chl chlorophyll - F_o, F_m, F_v in the sequence given, initial, maximum and variable chlorophyll fluorescence - PFD photon flux density - P_m photosynthesis rate at light saturation - PSI, PSII photosystems I, II, respectively - R_d dark-respiration rate - RT room temperature - quantum yield of photosynthetic O₂ evolution - _SA daily surface-area specific growth rate We wish to thank Jud Kenworthy, National Marine Fisheries Service, Beaufort, N.C., USA for performing the C/N analysis, and Olle Björkman and Susan Thayer, Carnegie Institution of Washington, Stanford, Cal., USA for the analysis of xanthophylls. This research was supported by National Science Foundation (NSF) grant OCE-8812157 to C.B.O. and J.R. Support for G.L. was provided by a NSF-CNRS (Centre National de la Recherche Scientifique) exchange fellowship.     

Nitrogenase activity of pea bacteroids as affected by carbohydrates and ammonium chloride

Summary Regulation and efficiency of the nitrogen-fixing system of the rhizobium-pea symbiosis were investigated. Acetylene reduction of detached root nodules was measured with various substrates added. Succinate, fumarate and malate were most effective in stimulating nitrogenase activity; glucose, pyruvate and citrate were also active. Acetylene reducing activity of detached nodules was inhibited by the addition of NH₄Cl, irrespective of the substrate present. Nitrogenase activity of isolated bacteroids was not influenced by NH₄Cl.Respiration of detached nodules was not significantly stimulated by the addition of substrates. Ammonium chloride did not influence respiration. With detached nodules and isolated bacteroids a consumption of about 16 g of carbohydrate per g of nitrogen fixed could be calculated. Detached nodules produced more hydrogen relative to the acetylene reduced than did isolated bacteroids and intact plants.Results obtained indicate that the regulation of nitrogenase activity and the efficiency of substrate consumption depend on environmental conditions.     

Spatial variability of symbiotic N2 fixation in grass-white clover pastures estimated by the 15N isotope dilution method and the natural 15N abundance method

Aiming at estimating the spatial variability in N₂ fixation, and to evaluate the appropriateness of the ¹⁵N isotope dilution (ID) method and the natural¹⁵N abundance (NA) method in reflecting spatial variability under the influence of cattle grazing, the symbiotic N₂ fixation in grass–white clover mixture was studied. At the Foulum site, where the ID method was used, differences in the climatic conditions between the two years of investigations caused a considerable difference in plant growth rates and proportion of clover. Consequently, the total N₂ fixation in ungrazed reference plots was significantly less in 1998 than in 1997, being 5.9 and 12.5 g N m^–2, respectively. In both years there was a wide range in concentration of inorganic N in the soil with coefficients of variance of approximately 60–190% for ammonium and 70–340% for nitrate. Significant negative correlations between pNdfa, determined by the ID method, and the log-transformed values of inorganic N and total N in grass were found. The NA method was applied on three nearby commercial dairy farms. They also showed high coefficients of variation. The coefficient of variance for NO₃ ^–-N ranged from 37 to 282% and for NH₄ ⁺-N from 29 to 237%. Average estimates of pNdfa values, which in the NA method were calculated using apparent B values ranging from –2.10 to –2.59, were generally lower (0.7–0.87) for these farms than for the Foulum site (0.89–0.95) using the ID method. For the NA method the ¹⁵N values, i.e. deviation in ¹⁵N concentration from atmospheric N₂, ranged from –7.0 to 5.7 for the grass N, which in several cases was lower than for clover N. Due to this high variability of the ¹⁵N values, probably caused by deposition and plant assimilation of ¹⁵N depleted urinary N in the pastures, the NA method was marginal for accurate determination of pNdfa. Consequently no significant correlation between the pNdfa determined by this method, and the log-transformed values of inorganic N in soil or total N in grass were found.     

Effects of arsenate and phosphate on their accumulation by an arsenic-hyperaccumulator Pteris vittata L.

Arsenate and phosphate interactions are important for better understanding their uptake and accumulation by plant due to their similarities in chemical behaviors. The present study examined the effects of arsenate and phosphate on plant biomass and uptake of arsenate and phosphate by Chinese brake (Pteris vittata L.), a newly-discovered arsenic hyperaccumulator. The plants were grown for 20 weeks in a soil, which received the combinations of 670, 2670, or 5340 mol kg^–1 arsenate and 800, 1600, or 3200 mol kg^–1 phosphate, respectively. Interactions between arsenate and phosphate influenced their availability in the soil, and thus plant growth and uptake of arsenate and phosphate. At low and medium arsenate levels (670 and 2670 mol kg^–1), phosphate had slight effects on arsenate uptake by and growth of Chinese brake. However, phosphate substantially increased plant biomass and arsenate accumulation by alleviating arsenate phytotoxicity at high arsenate levels (5340 mol kg^–1). Moderate doses of arsenate increased plant phosphate uptake, but decreased phosphate concentrations at high doses because of its phytotoxicity. Based on our results, the minimum P/As molar ratios should be at least 1.2 in soil solution or 1.0 in fern fronds for the growth of Chinese brake. Our findings suggest that phosphate application may be an important strategy for efficient use of Chinese brake to phytoremediate arsenic contaminated soils. Further study is needed on the mechanisms of interactive effects of arsenate and phosphate on Chinese brake in hydroponic systems.     

Effect of plant genotype and nitrogen fertilizer on symbiotic nitrogen fixation by soybean cultivars

Summary Isotopic as well as non-isotopic methods were used to assess symbiotic nitrogen fixation within eight soybean [Glycine max (L.) Merr.] cultivars grown at 20 and 100 kg N/ha levels of nitrogen fertilizer under field conditions.The¹⁵N methodology revealed large differences between soybean cultivars in their abilities to support nitrogen fixation. In almost all cases, the application of 100 kg N/ha resulted in lower N₂ fixed in soybean than at 20 kg N/ha in the first year of the study. However, N₂ fixed in one cultivar, Dunadja, was not significantly affected by the higher rate of N fertilizer application. These results were confirmed by measurements of acetylene reduction activity, nodule dry weight and N₂ fixed as measured by the difference method. Further proof of differences in N₂ fixed within soybean cultivars and the ability of Dunadja to fix similar amounts of N₂ at 20 and 100 kg N/ha was obtained during a second year experiment. Dunadja yield was affected by N fertilizer and produced larger yield at 100 kg N/ha than at 20 kg N/ha. This type of cultivar could be particularly useful in situations where soil N levels are high or where there is need to apply high amounts of N fertilizer.The present study reveals the great variability between legume germplasms in the ability to fix N₂ at different inorganic N levels, and also the potential that exists in breeding for nitrogen fixation associative traits. The¹⁵N methodology offers a unique tool to evaluate germplasms directly in the field for their N₂ fixation abilities at different N fertilizer levels.