Estimating N transfers between N<Subscript>2</Subscript>-fixing actinorhizal species and the non-N<Subscript>2</Subscript>-fixing<Emphasis Type="Italic"> Prunus avium</Emphasis> under partially controlled conditions

Two methods of N transfer between plants—by litter decomposition and root-to-root exchange—were examined in mixed plantations of N-fixing and non-fixing trees. Nitrogen transfers from decaying litters were measured by placing ¹⁵N-labelled litters from four actinorhizal tree species around shoots of containerized Prunus avium. Nitrogen transfers by root-to-root exchanges were measured after foliar NO₃-¹⁵N fertilization of Alnus subcordata and Elaeagnus angustifolia growing in containers in association with P. avium. During the first 2 years of litter decomposition, from 5–20% of the N, depending on the litter identity, was released and taken up by P. avium. N availability in the different litters was strongly correlated with the amount of water-soluble N, which was highest in leaves of E. angustifolia. In the association between fixing and non-fixing plants, 7.5% of the A. subcordata N and 25% of E. angustifolia N was transferred to P. avium by root exchange. These results showed that the magnitude of N transfers by root exchange depended on the associated N₂-fixing species. Among the species investigated, E. angustifolia displayed the highest capacity for exudating N from roots as well as for releasing N from litters. These qualities make this tree a promising species for enhancing wood yields in mixed stands.     

Nodulation and N2 fixation of faba bean (Vicia faba L.) after soil amendment with crop residues

The effect of prior soil amendment with different N sources at 50 mg N (kg soil)^—1 on nodulation and N₂ fixation of faba bean (Vicia faba L. cv. Troy) using wheat (Triticum aestivum L. cv. Star) as reference crop was assessed in a pot experiment. Four treatments viz legume manure (LEGM) as clover shoots, cereal manure (CEREM) as barley straw, N fertilizer (FERT‐N) as Ca(NO₃)₂, and no‐manure control (NOMAN) were investigated consecutively at 45, 70, and 90 days after sowing (DAS). Faba bean nodulated profusely, with an increase on average from 629 nodules per pot at 45 DAS to nearly 2.3‐ and 3.3‐fold at 70 and 90 DAS, respectively. Low nodule numbers and nodule dry matter occurred under FERT‐N and CEREM, whereas high values were found for NOMAN and LEGM. Soil amendment affected percent N₂ fixation in relation to N source and plant age. Highest percent N₂ fixation (≥ 90 %) was found under the lowest N‐supplying amendments, no‐manure, and cereal manure, respectively. FERT‐N depressed N₂ fixation particularly at 45 DAS when N₂ fixation was reduced to as low as 23 %. The rise in N₂ fixation thereafter suggests that faba bean adjusted after depletion of mineral N in the soil. N₂ fixation was also decreased after cereal straw application, even though N concentration in faba bean plants was high. The results indicate that plant residues, both with high and low N concentration, applied to soil to raise its fertility may interfere with N₂ fixation of faba bean.     

Improved estimation of biological nitrogen fixation of soybean cultivars (<Emphasis Type="Italic">Glycine max</Emphasis> L. Merril) using <Superscript>15</Superscript>N natural abundance technique

Biological nitrogen fixation (BNF) of 17 soybean cultivars was comparatively estimated by the δ¹⁵N natural abundance technique using two non-nodulation soybeans (Clay and Chippewa) as reference plants. A field study was established on the experimental farm of the University of Abomey-Calavi, Benin on a typical “terre de barre” soil classified by Food and Agriculture Organization-United Nations Educational, Scientific and Cultural Organisation as Rhodic Ferralsol. A nitrogen-free pot trial was also carried out using soil substrate sampled from the Atlantic Ocean beach. In the N-free medium, N content of the whole soybean cultivars ranged from 2.6 to 8.1 mg N per plant compared with an average of 1.8 mg N per plant observed with the non-fixing soybeans. Plant δ¹⁵N of the nodulating soybeans ranged from −2.7756‰ (Jupiter) to 0.1951‰ (Conquista), while the non-nodulating cultivars Chippewa and Clay had 2.67‰ and 9.30‰, respectively. Percentage and amount of N derived from air (Ndfa) were significantly different (P < 0.01) among soybean cultivars, and values depended highly on the selected reference plants. When Clay was used as the reference plant, the average percentage Ndfa was 1.4 times higher than when Chippewa was the reference plant. Both reference plants consistently ranked promiscuous soybean cvs. TG× 1894 3F and TG× 1908 8F as the best cultivars and cv. TG× 1888 29F as the least in percentage Ndfa, suggesting that any of the reference plants could be used in δ¹⁵N method for assessing N₂-fixation. The two identified promiscuous soybean cultivars with greatest capacity to fix N could be included in a soybean extension program for West African farming systems.     

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.     

Biological nitrogen fixation in faba bean (Vicia faba L.) in the Ethiopian highlands as affected by P fertilization and inoculation

 N₂ fixation by leguminous crops is a relatively low-cost alternative to N fertilizer for small-holder farmers in developing countries. N₂ fixation in faba bean (Vicia faba L.) as affected by P fertilization (0 and 20 kg P ha^–1) and inoculation (uninoculated and inoculated) with Rhizobium leguminosarium biovar viciae (strain S-18) was studied using the ¹⁵N isotope dilution method in the southeastern Ethiopian highlands at three sites differing in soil conditions and length of growing period. Nodulation at the late flowering stage was significantly influenced by P and inoculation only at the location exhibiting the lowest soil P and pH levels. The percentage of N derived from the atmosphere ranged from 66 to 74%, 58 to 74% and 62 to 73% with a corresponding total amount of N₂ fixed ranging from 169 to 210 kg N ha^–1, 139 to 184 kg N ha^–1 and 147 to 174 kg N ha^–1 at Bekoji, Kulumsa and Asasa, respectively. The total N₂ fixed was not significantly affected by P fertilizer or inoculation across all locations, and there was no interaction between the factors. However, at all three locations, N₂ fixation was highly positively correlated with the dry matter production and total N yield of faba bean. Soil N balances after faba bean were positive (12–58 kg N ha^–1) relative to the highly negative N balances (–9–44 kg N ha^–1) following wheat (Triticum aestivum L.), highlighting the importance of rotation with faba bean in the cereal-based cropping systems of Ethiopia. Received: 13 January 2000     

Biological nitrogen fixation by common beans (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) increases with bio-char additions

This study examines the potential, magnitude, and causes of enhanced biological N₂ fixation (BNF) by common beans (Phaseolus vulgaris L.) through bio-char additions (charcoal, biomass-derived black carbon). Bio-char was added at 0, 30, 60, and 90 g kg⁻¹ soil, and BNF was determined using the isotope dilution method after adding ¹⁵N-enriched ammonium sulfate to a Typic Haplustox cropped to a potentially nodulating bean variety (CIAT BAT 477) in comparison to its non-nodulating isoline (BAT 477NN), both inoculated with effective Rhizobium strains. The proportion of fixed N increased from 50% without bio-char additions to 72% with 90 g kg⁻¹ bio-char added. While total N derived from the atmosphere (NdfA) significantly increased by 49 and 78% with 30 and 60 g kg⁻¹ bio-char added to soil, respectively, NdfA decreased to 30% above the control with 90 g kg⁻¹ due to low total biomass production and N uptake. The primary reason for the higher BNF with bio-char additions was the greater B and Mo availability, whereas greater K, Ca, and P availability, as well as higher pH and lower N availability and Al saturation, may have contributed to a lesser extent. Enhanced mycorrhizal infections of roots were not found to contribute to better nutrient uptake and BNF. Bean yield increased by 46% and biomass production by 39% over the control at 90 and 60 g kg⁻¹ bio-char, respectively. However, biomass production and total N uptake decreased when bio-char applications were increased to 90 g kg⁻¹. Soil N uptake by N-fixing beans decreased by 14, 17, and 50% when 30, 60, and 90 g kg⁻¹ bio-char were added to soil, whereas the C/N ratios increased from 16 to 23.7, 28, and 35, respectively. Results demonstrate the potential of bio-char applications to improve N input into agroecosystems while pointing out the needs for long-term field studies to better understand the effects of bio-char on BNF.     

Evidence of N<Subscript>2</Subscript>O emission and gaseous nitrogen losses through nitrification-denitrification induced by rice plants (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

A greenhouse experiment was conducted with a specially designed apparatus consisting of an upper and lower chamber where the treatment with rice was carried out (treatment 1). The apparatus also had a single chamber where treatment 2, without rice plants, was carried out. The scope of this study was to elucidate the influence of rice plant growth on gaseous N losses as N₂ and N₂O produced by nitrification-denitrification in a flooded soil fertilized with (NH₄)₂SO₄ (with 56.50 atom% ¹⁵N). Gas samples were withdrawn weekly and analyzed for (N₂ + N₂O)-¹⁵N losses by mass spectrometer and for N₂O by gas chromatograph. The gaseous (N₂ + N₂O)-¹⁵N losses of the treatment with rice plants were significantly (P =0.01) higher than those of the treatment without rice plants, as were the amounts of N₂O emitted. Rice plants facilitate the efflux of N₂ and N₂O from soil to atmosphere, as about half of the total gaseous ¹⁵N loss as N₂ and N₂O was found in the upper chamber. The proportion of N₂O-¹⁵N to (N₂ + N₂O)-¹⁵N in the upper chamber was 10.56%, much higher than that of the lower chamber in treatment 1 and the headspace of treatment 2.     

<Emphasis Type="Italic">Calamintha nepeta</Emphasis> (L.) Savi and <Emphasis Type="Italic">Micromeria thymifolia</Emphasis> (Scop.) Fritsch cultivated in Italy

Calamintha nepeta and Micromeria thymifolia have been traditionally used in the Mediterranean area as condiments and medicinal plants for a long time. Whereas in parts of Italy C. nepeta (special recipes have been developed in Lazio and Tuscany) is also an established garden plant showing different evolutionary products and their interaction among each other and the wild progenitor, M. thymifolia is being developed into a new crop plant. Both plants and their uses are described with regard to Italy. There is a marked tendency to broaden the use of condiments and spices which results in new crop plants which have to be documented and elaborated in further studies. Many species of Labiatae are predisposed to use by man and new items can be found even in areas which have to be considered as well studied.     

<Emphasis Type="Italic">Ziziphus </Emphasis><Emphasis Type="Italic">spina-christi</Emphasis> (L.) Willd.: a multipurpose fruit tree

Neglected and underutilized species often play a vital role in securing food and livestock feed, income generation and energy needs of rural populations. In spite of their great potential little attention has been given to these species. This increases the possibility of genetic erosion which would further restrict the survival strategies of people in rural areas. Ziziphus spina-christi is a plant species that has edible fruits and a number of other beneficial applications that include the use of leaves as fodder, branches for fencing, wood as fuel, for construction and furniture making, and the utilization of different parts e.g. Fruits, leaves, roots and bark in folk medicine. Moreover, the plant is adapted to dry and hot climates which make it suitable for cultivation in an environment characterized by increasing degradation of land and water resources. Lack of research in Z. spina-christi hinders its successful improvement and promotion. Therefore, studies are needed to fully exploit this species. This article aims at summarizing information on different aspects of Z. spina-christi to stimulate interest in this crop which is of importance in Sudan and other countries of the semi-arid tropics.

Salt tolerance in <Emphasis Type="Italic">Zea mays</Emphasis> (L). following inoculation with <Emphasis Type="Italic">Rhizobium</Emphasis> and <Emphasis Type="Italic">Pseudomonas</Emphasis>

This study aimed to investigate the effect of inoculation with plant growth-promoting Rhizobium and Pseudomonas species on NaCl-affected maize. Two cultivars of maize (cv. Agaiti 2002 and cv. Av 4001) selected on the basis of their yield potential were grown in pots outdoors under natural conditions during July. Microorganisms were applied at seedling stage and salt stress was induced 21 days after sowing and maintained up to 50% flowering after 120 days of stress. The salt treatment caused a detrimental effect on growth and development of plants. Co-inoculation resulted in some positive adaptative responses of maize plants under salinity. The salt tolerance from inoculation was generally mediated by decreases in electrolyte leakage and in osmotic potential, an increase in osmoregulant (proline) production, maintenance of relative water content of leaves, and selective uptake of K ions. Generally, the microbial strain acted synergistically. However, under unstressed conditions, Rhizobium was more effective than Pseudomonas but under salt stress the favorable effect was observed even if some exceptions were also observed. The maize cv. Agaiti 2002 appeared to be more responsive to inoculation and was relatively less tolerant to salt compared to that of cv. Av 4001.     

<Emphasis Type="Italic">Rhizobium</Emphasis>,<Emphasis Type="Italic"> Bradyrhizobium</Emphasis> and<Emphasis Type="Italic"> Agrobacterium</Emphasis> strains isolated from cultivated legumes

The present study was conducted to isolate and characterize rhizobial strains from root nodules of cultivated legumes, i.e. chickpea, mungbean, pea and siratro. Preliminary characterization of these isolates was done on the basis of plant infectivity test, acetylene reduction assay, C-source utilization, phosphate solubilization, phytohormones and polysaccharide production. The plant infectivity test and acetylene reduction assay showed effective root nodule formation by all the isolates on their respective hosts, except for chickpea isolate Ca-18 that failed to infect its original host. All strains showed homology to a typical Rhizobium strain on the basis of growth pattern, C-source utilization and polysaccharide production. The strain Ca-18 was characterized by its phosphate solubilization and indole acetic acid (IAA) production. The genetic relationship of the six rhizobial strains was carried out by random amplified polymorphic DNA (RAPD) including a reference strain of Bradyrhizobium japonicum TAL-102. Analysis conducted with 60 primers discriminated between the strains of Rhizobium and Bradyrhizobium in two different clusters. One of the primers, OPB-5, yielded a unique RAPD pattern for the six strains and well discriminated the non-nodulating chickpea isolate Ca-18 from all the other nodulating rhizobial strains. Isolate Ca-18 showed the least homology of 15% and 18% with Rhizobium and Bradyrhizobium, respectively, and was probably not a (Brady)rhizobium strain. Partial 16S rRNA gene sequence analysis for MN-S, TAL-102 and Ca-18 strains showed 97% homology between MN-S and TAL-102 strains, supporting the view that they were strains of B. japonicum species. The non-infective isolate Ca-18 was 67% different from the other two strains and probably was an Agrobacterium strain.     

Benefits of inoculation of the common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>) crop with efficient and competitive<Emphasis Type="Italic"> Rhizobium tropici</Emphasis> strains

Cropping in low fertility soils, especially those poor in N, contributes greatly to the low common bean (Phaseolus vulgaris L.) yield, and therefore the benefits of biological nitrogen fixation must be intensively explored to increase yields at a low cost. Six field experiments were performed in oxisols of Paraná State, southern Brazil, with a high population of indigenous common bean rhizobia, estimated at a minimum of 10³ cells g^–1 soil. Despite the high population, inoculation allowed an increase in rhizobial population and in nodule occupancy, and further increases were obtained with reinoculation in the following seasons. Thus, considering the treatments inoculated with the most effective strains (H 12, H 20, PRF 81 and CIAT 899), nodule occupancy increased from an average of 28% in the first experiment to 56% after four inoculation procedures. The establishment of the selected strains increased nodulation, N₂ fixation rates (evaluated by total N and N-ureide) and on average for the six experiments the strains H 12 and H 20 showed increases of 437 and 465 kg ha^–1, respectively,in relation to the indigenous rhizobial population. A synergistic effect between low levels of N fertilizer and inoculation with superior strains was also observed, resulting in yield increases in two other experiments. The soil rhizobial population decreased 1 year after the last cropping, but remained high in the plots that had been inoculated. DGGE analysis of soil extracts showed that the massive inoculation apparently did not affect the composition of the bacterial community.     

Screening and selection of faba beans (<Emphasis Type="Italic">Vicia faba</Emphasis> L.) for cold tolerance and comparison to wild relatives

Since autumn-sown faba beans possess several advantages including higher seed yield over the spring cultivars, the study was aimed to screen and select cold tolerant accessions of faba beans (Vicia faba L.) and compare these to wild species in the highland of the west Mediterranean region, Turkey. A total of 114 accessions of Vicia species including 109 accessions of faba bean, three accessions of narbon bean (V. narbonensis L.) and two accessions of V. montbretii Fisch. et C.A. Mey. were screened for cold tolerance at seedling stage in two successive years, 2005–2006 and 2006–2007 growth seasons. Accessions were evaluated for cold tolerance using a 1 (Highly cold tolerant)-5 (Highly cold susceptible) visual scale. Considerable variation was found for cold tolerance and some agronomical characteristics in faba beans. Wild relatives of faba bean were found to be more tolerant to cold than those of cultivated faba beans. Although some pigmented accessions were free from freezing damage at −9.6°C without snow cover, accessions with white flowers were damaged. The proposed screening technique could easily be used to evaluate many faba bean accessions for cold tolerance. To increase yield, it was concluded that the cold tolerant accessions with high yield could be grown as autumn-sown crop in the target environment.     

Comparative effects of applying leguminous and non-leguminous green manures and inorganic N on biomass yield and nitrogen uptake in flooded rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The beneficial role of green manures in rice production is generally ascribed to their potential of supplying plant nutrients, particularly nitrogen (N). However, the mechanisms through which green manures enhance the crop productivity are poorly understood. Pot experiments were conducted using a ¹⁵N-tracer technique: (1) to compare the biomass production potential of sesbania (Sesbania aculeata Pers.) and maize (Zea mays L.) as green manuring crops for lowland rice and (2) to compare the effect of the two types of green manure and inorganic N on the dry matter accumulation and N uptake by two rice (Oryza sativa L.) cultivars, viz. IR-6 and Bas-370. Although maize produced three times higher shoot biomass compared with sesbania, the latter showed higher N concentration; and thus the total N yield was similar in the two types of plants. Applying the shoot material of the two plants to flooded rice significantly enhanced the dry matter yield and N uptake by the two rice cultivars, the positive effects generally being more pronounced with sesbania than with maize amendment. The difference in the growth-promoting potential of the two plant residues was related more to an increased uptake of the native soil N rather than to their direct role as a source of plant-available N. A positive added nitrogen interaction (ANI) was observed due to both plant residues, the effect was much more pronounced with the application of sesbania than with maize residues. In both rice cultivars, inorganic N also caused a substantial ANI, particularly at higher application rate. Losses from the applied N were 2–3 times lower from sesbania, compared with maize treatment. Green manuring with sesbania also caused much lower N losses than the inorganic N applied at equivalent or higher rates. The overall benefit of green manuring to rice plants was higher than inorganic N applied at comparable rates. The two rice cultivars differed in their response to green manuring, IR-6 generally being more responsive than Bas-370.     

Extent and Pattern of Genetic Diversity for Morpho-agronomic Traits in Ethiopian Highland Pulse Landraces II. Faba Bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.)

A field experiment was conducted in 2001 at Holetta and Kulumsa, Ethiopia, to study genetic diversity in Ethiopian faba bean (Vicia faba L.) landraces. One hundred sixty random germplasm accessions were grown in an alpha lattice design with two replications. Data on 12 traits were collected and analyzed. Significant differences were observed among the accessions for most of the traits (except number of pods/podding nodes) at each location even though differences pooled over location were mostly non-significant. Cluster analysis distinguished seven diversity classes of different sizes. Accessions from the northern half of the country (North and South Wello, North Gonder and North Shewa) were closely related while those from the southern part of the country (Arsi) were highly diverse. Cumulative effects of a number of characters dictated differentiation of the accessions into clusters. Some overlapping were encountered between accessions from the northern and those from the southern parts of the country. The study revealed that accessions from different regions might have similar genetic background and those from the same origin might also have different genetic background. Therefore, geographic diversity should not necessarily be used as an index of genetic diversity and parental selection should be based on a systematic study of genetic diversity in a specific population. Genetic distances between most of the clusters were significant that crosses between parents selected out of them are expected to generate desirable progenies. Future germplasm collection, conservation and utilization strategies should put more focus not only on inter-regional diversity in the country as a whole but also on intra-regional diversity in Arsi.     

Clustering of Chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) Accessions

The variation in 20 morphological and agronomical traits has been evaluated in a set of chickpea genetic resources from four countries. Data indicated differences between accessions in leaf, flower, pod, and seed traits and characteristics, as well as in vegetation period. Multivariate analyses of these data segregated chickpeas into groups. Polymorphism in seed glutelines was absent, while variation in seed prolamines was very limited. DNA amplification patterns have been analyzed by semi-arbitrary primers and by specific microsatellite primers. Only some of semi-arbitrary primers generated usable DNA banding patterns, moreover interpretation of these patterns can be more or less difficult. On the contrary specific primers amplifying microsatellites at the specific loci generated unambiguous and reproducible differences between chickpeas.     

Mitigation of salt stress in wheat seedlings by a <Emphasis Type="Italic">gfp</Emphasis>-tagged <Emphasis Type="Italic">Azospirillum lipoferum</Emphasis>

Root colonization and mitigation of NaCl stress on wheat seedlings were studied by inoculating seeds with Azospirillum lipoferum JA4ngfp15 tagged with the green fluorescent protein gene (gfp). Colonization of wheat roots under 80 and 160 mM NaCl stress was similar to root colonization with this bacterial species under non-saline conditions, that is, single cells and small aggregates were mainly located in the root hair zone. These salt concentrations had significant inhibitory effects on development of seedlings, but not on growth in culture of gfp-A. lipoferum JA4ngfp15. Reduced plant growth (height and dry weight of leaves and roots) under continuous irrigation with 160 mM NaCl was ameliorated by bacterial inoculation with gfp-A. lipoferum JA4ngfp15. Inoculation of plants subjected to continuous irrigation with 80 mM NaCl or to a single application of either NaCl concentration (80 or 160 mM NaCl) did not mitigate salt stress. This study indicates that, under high NaCl concentration, inoculation with modified A. lipoferum reduced the deleterious effects of NaCl; colonization patterns on roots were unaffected and the genetic marker did not induce undesirable effects on the interaction between the bacterium and the plants.     

Transfer of leaf rust and stripe rust resistance from <Emphasis Type="Italic">Aegilops umbellulata</Emphasis> Zhuk. to bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Aegilops umbellulata acc. 3732, an excellent source of resistance to major wheat diseases, was used for transferring leaf rust and stripe rust resistance to cultivated wheat. An amphiploid between Ae. umbellulata acc. 3732 and Triticum durum cv. WH890 was crossed with cv. Chinese Spring Ph ^I to induce homoeologous pairing between Ae. umbellulata and wheat chromosomes. The F₁ was crossed to the susceptible Triticum aestivum cv. ‘WL711’ and leaf rust and stripe rust resistant plants were selected among the backcross progenies. Homozygous lines were selected and screened against six Puccinia triticina and four Puccinia striiformis f. sp. tritici pathotypes at the seedling stage and a mixture of prevalent pathotypes of both rust pathogens at the adult plant stage. Genomic in situ hybridization in some of the selected introgression lines detected two lines with complete Ae. umbellulata chromosomes. Depending on the rust reactions and allelism tests, the introgression lines could be classified into two groups, comprising of lines with seedling leaf rust resistance gene Lr9 and with new seedling leaf rust and stripe rust resistance genes. Inheritance studies detected an additional adult plant leaf rust resistance gene in one of the introgression lines. A minimum of three putatively new genes—two for leaf rust resistance (LrU1 and LrU2) and one for stripe rust resistance (YrU1) have been introgressed into wheat from Ae. umbellulata. Two lines with no apparent linkage drag have been identified. These lines could serve as sources of resistance to leaf rust and stripe rust in breeding programs.     

Estimating N2 fixation by field-grown lupins (Lupinus angustifolius L.) using soil and plant 15N enrichment

Summary Biological N₂ fixation was estimated in a field experiment following the addition of NH₄Cl or KNO₃ to unconfined microplots (1.5 m²) at 2.5 g N m^-2 (10 atom% ¹⁵N). A model of total N and ¹⁵N accumulation in lupins and decreasing ¹⁵N enrichment in the KCl-extractable soil-N pool (0–0.15 m depth) was used to estimate the proportion of N in lupins derived from biological N₂ fixation. Estimates of N₂ fixation derived from the model were compared with ¹⁵N isotope-dilution estimates obtained using canola, annual ryegrass, and wheat as nonfixing reference plants. Biomass, total N accumulation, or ¹⁵N enrichment in the lupin and reference crops did not differ whether NH _inf4 ^sup+ or NO _inf3 ^sup- was added as the labelled inorganic-N source. The decrease in soil ¹⁵N enrichment was described by first-order kinetics, whereas total N and ¹⁵N accumulation in the lupins were described by logistical equations. Using these equations, the uptake of soil N by lupins was estimated and was then used to calculate fixed N₂. Estimates of N₂ fixation derived from the model increased from 0 at 50 days after sowing to a maximum of 0.79 at 190 days after sowing. Those based on the ¹⁵N enrichment of the NO _inf3 ^sup- pool were 10% higher than those based on the mineral-N pool. ¹⁵N isotope-dilution estimates of N₂ fixation ranged from 0.37 to 0.55 at 68 days after sowing and from 0.71 to 0.77 at 190 days after sowing. Reference plant-derived values of N₂ fixation were all higher than modelled estimates during the early states of growth, but were similar to modelled estimates at physiological maturity. The use of the model to estimate N₂ derived from the atmosphere has the intrinsic advantage that the need for a non-fixing reference plant is avoided.     

Evaluation and utilization of <Emphasis Type="Italic">Aegilops</Emphasis> and wild <Emphasis Type="Italic">Triticum</Emphasis> species for enhancing iron and zinc content in wheat

Grains of 80 accessions of nine species of wild Triticum and Aegilops along with 15 semi-dwarf cultivars of bread and durum wheat grown over 2 years at Indian Institute of Technology, Roorkee, were analyzed for grain iron and zinc content. The bread and durum cultivars had very low content and little variability for both of these micronutrients. The related non-progenitor wild species with S, U and M genomes showed up to 3–4 folds higher iron and zinc content in their grains as compared to bread and durum wheat. For confirmation, two Ae. kotschyi Boiss. accessions were analyzed after ashing and were found to have more than 30% higher grain ash content than the wheat cultivars containing more than 75% higher iron and 60% higher zinc than that of wheat. There were highly significant differences for iron and zinc contents among various cultivars and wild relatives over both the years with very high broad sense heritability. There was a significantly high positive correlation between flag leaf iron and grain iron (r = 0.82) and flag leaf zinc and grain zinc (r = 0.92) content of the selected donors suggesting that the leaf analysis could be used for early selection for high iron and zinc content. ‘Chinese Spring’ (Ph ^I ) was used for inducing homoeologous chromosome pairing between Aegilops and wheat genomes and transferring these useful traits from the wild species to the elite wheat cultivars. A majority of the interspecific hybrids had higher leaf iron and zinc content than their wheat parents and equivalent or higher content than their Aegilops parents suggesting that the parental Aegilops donors possess a more efficient system for uptake and translocation of the micronutrients which could ultimately be utilized for wheat grain biofortification. Partially fertile to sterile BC₁ derivatives with variable chromosomes of Aegilops species had also higher leaf iron and zinc content confirming the possibility of transfer of required variability. Some of the fertile BC₁F₃ and BC₂F₂ derivatives had as high grain ash and grain ash iron and zinc content as that of the donor Aegilops parent. Further work on backcrossing, selfing, selection of fertile derivatives, leaf and grain analyses for iron and zinc for developing biofortified bread and durum wheat cultivars is in progress. Nidhi Rawat, Vijay K. Tiwari, and Neelam Singh have contributed equally to the work.