Distribution and remobilization of symbiotically fixed nitrogen in soybean (Glycine max)

Partitioning of nitrogen by soybeans ( Glycine max L. Merr. cv. Hodgson) grown in natural conditions was studied by successive exposures of root systems to ¹⁵N₂ and periodical measurements of ¹⁵N distribution. Nitrogen derived from the atmosphere was mainly found in the aerial parts of the plants, and the stage of development exerted a strong influence on the initial ¹⁵N distribution (measured one week after incorporation). Until day 69 after sowing, leaf blades contained 47 to 57% of the fixed N. After that, reproductive structures attracted increasing proportions, 10 to 60% between days 69 and 92. Around day 82, stems and petioles stored up to 30% of the newly fixed N. During pod development and pod filling and until maturity, fixed N was remobilized from vegetative tissues and pod walls to seeds. These transfers first concerned the newly incorporated N, but at maturity 80 to 90% of the total was recovered in the seeds. The high mobility of N originating from the atmosphere as compared to that coming from the soil (vegetative tissues exported only 50% of their total N) seems to indicate that fixed N was at least partially integrated in a special pool. This was certainly the case at the later stage of N₂ fixation, when a large portion of fixed N accumulated in the stems and petioles, probably in the form of storage compounds such as ureides for later transfer to the developing seeds. Further research is needed in order to investigate the nature and role of this pool in the nitrogen nutrition of soybeans.     

Growth and onto-morphogenesis of soybean (Glycine max Merril) in an open, naturally CO2-enriched environment

Springs emitting carbon dioxide are frequent in Central Italy and provide a way of testing the response of plants to CO₂ enrichment under natural conditions. Results of a CO₂ enrichment experiment on soybean at a CO₂ spring (Solfatara) are presented. The experimental site is characterized by significant anomalies in atmospheric CO₂ concentration produced by a large number of vents emitting almost pure CO₂ (93%) plus small amounts of hydrogen sulphide, methane, nitrogen and oxygen. Within the gas vent area, plants were grown at three sub-areas whose mean CO₂ concentrations during daytime were 350,652 and 2370 μmol mol^-1, respectively. Weekly harvests were made to measure biomass growth, leaf area and ontogenetic development. Biomass growth rate and seed yield were enhanced by elevated CO₂. In particular, onto-morphogenetic development was affected by elevated CO₂ with high levels of CO₂ increasing the total number of main stem leaf nodes and the area of the main stem trifoliolate leaves. Biochemical analysis of plant tissue suggested that there was no effect of the small amounts of H₂S on the response to CO₂ enrichment. Non-protein sulphydryl compounds did not accumulate in leaf tissues and the overall capacity of leaf extracts to oxidize exogenously added NADH was not decreased. The limitations and advantages of experimenting with crop plants at elevated CO₂ in the open and in the proximity of carbon dioxide springs are discussed.     

Inoculation with an enhanced N2‐fixing Bradyrhizobium japonicum strain (USDA110) does not alter soybean (Glycine max Merr.) response to elevated [CO2]

This study tested the hypothesis that inoculation of soybean (Glycine max Merr.) with a Bradyrhizobium japonicum strain (USDA110) with greater N₂ fixation rates would enhance soybean response to elevated [CO₂]. In field experiments at the Soybean Free Air CO₂ Enrichment facility, inoculation of soybean with USDA110 increased nodule occupancy from 5% in native soil to 54% in elevated [CO₂] and 34% at ambient [CO₂]. Despite this success, inoculation with USDA110 did not result in greater photosynthesis, growth or seed yield at ambient or elevated [CO₂] in the field, presumably due to competition from native rhizobia. In a growth chamber experiment designed to study the effects of inoculation in the absence of competition, inoculation with USDA110 in sterilized soil resulted in nodule occupation of >90%, significantly greater ¹⁵N₂ fixation, photosynthetic capacity, leaf N and total plant biomass compared with plants grown with native soil bacteria. However, there was no interaction of rhizobium fertilization with elevated [CO₂]; inoculation with USDA110 was equally beneficial at ambient and elevated [CO₂]. These results suggest that selected rhizobia could potentially stimulate soybean yield in soils with little or no history of prior soybean production, but that better quality rhizobia do not enhance soybean responses to elevated [CO₂].     

Host genetic control of symbiosis in soybean (Glycine max L.)

Genes controlling nitrogen-fixing symbioses of legumes with specialized bacteria known as rhizobia are presumably the products of many millions of years of evolution. Different adaptative solutions evolved in response to the challenge of survival in highly divergent complexes of symbionts. Whereas efficiency of nitrogen fixation appears to be controlled by quantitative inheritance, genes controlling nodulation are qualitatively inherited. Genes controlling nodulation include those for non-nodulation, those that restrict certain microsymbionts, and those conditioning hypernodulation, or supernodulation. Some genes are naturally occurring polymorphisms, while others were induced or were the result of spontaneous mutations. The geographic patterns of particular alleles indicate the role of coevolution in determining symbiont specificites and compatibilities. For example, the Rj4 allele occurs with higher frequency (over 50%) among the soybean (G. max) from Southeast Asia. DNA homology studies of strains of Bradyrhizobium that nodulate soybean indicated two groups so distinct as to warrant classification as two species. Strains producing rhizobitoxine-induced chlorosis occur only in Group II, now classified as B. elkanii. Unlike B. japonicum, B. elkanii strains are characterized by (1) the ability to nodulate the rj1 genotype, (2) the formation of nodule-like structures on peanut, (3) a relatively high degree of ex planta nitrogenase activity, (4) distinct extracellular polysaccharide composition, (5) distinct fatty acid composition, (6) distinct antibiotic resistance profiles, and (7) low DNA homology with B. japonicum. Analysis with soybean lines near isogenic for the Rj4 versus rj4 alleles indicated that the Rj4 allele excludes a high proportion of B. elkanii strains and certain strains of B. japonicum such as strain USDA62 and three serogroup 123 strains. These groups, relatively inefficient in nitrogen fixation with soybean, tend to predominate in soybean nodules from many US soils. The Rj4 allele, the most common allelic form in the wild species, has a positive value for the host plants in protecting them from nodulation by rhizobia poorly adapted for symbiosis.     

Distribution of two isoforms of NADP-dependent isocitrate dehydrogenase in soybean (Glycine max)

Two different cDNAs that encode NADP-specific isocitrate dehydrogenase (NADP-IDH) isozymes of soybean (Glycine max) were characterized. The nucleotide sequences of the coding regions of these cDNAs have 74% identity to each other and give predicted amino acid sequences that have 83% identity to each other. Using PCR techniques, their coding regions were subcloned into a protein overexpression vector, pQE32, to yield pIDH4 and pIDH1, respectively. Both IDH4 and IDH1 enzymes were expressed in Escherichia coli as catalytically active His6 tagged proteins, purified to homogeneity by affinity chromatography on nickel chelate resin and rabbit polyclonal antibodies to each were generated. Surprisingly, antiserum to IDH4 did not react with IDH1 protein and IDH1 antiserum reacted only very weakly with IDH4 protein. IDH4 antibody reacts with a protein of expected molecular weight in cotyledon, young leaf, young root, mature root and nodules but the reaction with mature leaf tissue was low compared to other tissues. Western blot results show that IDH1 was not expressed in young roots but a protein that reacts with the IDH1 antibody was highly expressed in leaves, showing that there was tissue-specific accumulation of NADP-IDH isozymes in soybean.     

The soybean (Glycine max) nodulation‐suppressive CLE peptide,GmRIC1, functions interspecifically in common white bean (Phaseolus vulgaris), but not in a supernodulating line mutated in the receptor PvNARK

Legume plants regulate the number of nitrogen‐fixing root nodules they form via a process called the Autoregulation of Nodulation (AON). Despite being one of the most economically important and abundantly consumed legumes, little is known about the AON pathway of common bean (Phaseolus vulgaris). We used comparative‐ and functional‐genomic approaches to identify central components in the AON pathway of common bean. This includes identifying PvNARK, which encodes a LRR receptor kinase that acts to regulate root nodule numbers. A novel, truncated version of the gene was identified directly upstream of PvNARK, similar to Medicago truncatula, but not seen in Lotus japonicus or soybean. Two mutant alleles of PvNARK were identified that cause a classic shoot‐controlled and nitrate‐tolerant supernodulation phenotype. Homeologous over‐expression of the nodulation‐suppressive CLE peptide‐encoding soybean gene, GmRIC1, abolished nodulation in wild‐type bean, but had no discernible effect on PvNARK‐mutant plants. This demonstrates that soybean GmRIC1 can function interspecifically in bean, acting in a PvNARK‐dependent manner. Identification of bean PvRIC1, PvRIC2 and PvNIC1, orthologues of the soybean nodulation‐suppressive CLE peptides, revealed a high degree of conservation, particularly in the CLE domain. Overall, our work identified four new components of bean nodulation control and a truncated copy of PvNARK, discovered the mutation responsible for two supernodulating bean mutants and demonstrated that soybean GmRIC1 can function in the AON pathway of bean.     

Effect of ammonium- and nitrate-nitrogen nutrition on aluminium tolerance of soybean (Glycine max L.)

Ammonium-nitrogen supply increased Al tolerance (parameter root elongation rate) of soybean (Glycine max L.) plants compared to nitrate-nitrogen supply when grown at constant pH in solution culture. This protective effect of ammonium against Al could only partially be attributed to lowered activity of monomeric aluminium species in the ammonium solution. For ammonium and nitrate-grown plants the relationship between Al concentration in the root tips and total length could be described by the same regression equation. The higher Al tolerance of soybean plants grown in the presence of ammonium was due to restricted ad/absorption of Al which resulted from competition with positively charged Al species for binding sites in the apoplast. Induction of higher symplastic Al tolerance is unlikely because preculture with ammonium decreased rather than increased aluminium tolerance of the plants.     

Soybean (Glycine max) modulation and N2-fixation as affected by exposure to a low root-zone temperature

Low root-zone temperatures (RZTs) are known to reduce soybean N₂-fixation. However, the relative sensitivity of the various stages of symbiosis establishment and function (N₂-fixation) to suboptimal RZTs is unresolved. We conducted experiments to examine the effect of exposure to a RZT of 15°C on nodulation. The control RZT was 25°C. Root temperatures were controlled by circulating cooled water around pots on a growth bench. Soybean seedlings [ Glycine max (L.) Merr. cv. Maple Arrow] were inoculated with 1 ml of a log-phase culture (approximately 10⁻⁸ cells) of Bradyhizobium japonicum strain 532C. They were then (1) maintained continuously at RZTs of 15 or 25°C, transferred to 15 or 25°C from the alternate temperature 7 days after inoculation (DAI), or transferred to 15 or 25°C at 14 DAI, and (2) maintained at 15 or 25°C, or transferred at either 1, 4 or 7 DAI. When seedlings were maintained at a RZT of 25°C nodule primordia (<1 mm) were visible at 7 DAI and N₂-fixation commenced at 14 DAI. Nodule function (N₂-fixation) appeared to be relatively insensitive to low RZTs since exposure of plants to 15°C following the onset of N₂-fixation (14 DAI) resulted in 68% of the N fixed and 78% of the dry weight of the 25°C RZT, although N partitioning to shoot tissues was reduced. In contrast, exposure to the low RZT shortly after inoculation declayed the onset of N₂-fixation for 4 to 6 weeks, primarily by inhibiting the early stages of nodulation. This resulted in fixed N and dry weight levels of 9% and 22% of controls, respectively.     

QTL mapping of domestication-related traits in soybean (Glycine max)

BACKGROUND AND AIMS: Understanding the genetic basis underlying domestication-related traits (DRTs) is important in order to use wild germplasm efficiently for improving yield, stress tolerance and quality of crops. This study was conducted to characterize the genetic basis of DRTs in soybean (Glycine max) using quantitative trait locus (QTL) mapping. METHODS: A population of 96 recombinant inbred lines derived from a cultivated (ssp. max) x wild (ssp. soja) cross was used for mapping and QTL analysis. Nine DRTs were examined in 2004 and 2005. A linkage map was constructed with 282 markers by the Kosambi function, and the QTL was detected by composite interval mapping. KEY RESULTS: The early flowering and determinate habit derived from the max parent were each controlled by one major QTL, corresponding to the major genes for maturity (e1) and determinate habit (dt1), respectively. There were only one or two significant QTLs for twinning habit, pod dehiscence, seed weight and hard seededness, which each accounted for approx. 20-50 % of the total variance. A comparison with the QTLs detected previously indicated that in pod dehiscence and hard seededness, at least one major QTL was common across different crosses, whereas no such consistent QTL existed for seed weight. CONCLUSIONS: Most of the DRTs in soybeans were conditioned by one or two major QTLs and a number of genotype-dependent minor QTLs. The common major QTLs identified in pod dehiscence and hard seededness may have been key loci in the domestication of soybean. The evolutionary changes toward larger seed may have occurred through the accumulation of minor changes at many QTLs. Since the major QTLs for DRTs were scattered across only six of the 20 linkage groups, and since the QTLs were not clustered, introgression of useful genes from wild to cultivated soybeans can be carried out without large obstacles.     

Effects of exogenous application and stem infusion of ascorbate on soybean (Glycine max) root nodules

Numerous biochemical and physiological studies have demonstrated the importance of ascorbate (ASC) as a reducing agent and antioxidant in higher plant metabolism. Of special note is the capacity of ASC to eliminate damaging activated oxygen species (AOS) including O₂^−· and H₂O₂. N₂-fixing legume nodules are especially vulnerable to oxidative damage because they contain large amounts of leghaemoglobin which produces AOS through spontaneous autoxidation; thus, ASC and other components of the ascorbate–reduced glutathione (ASC–GSH) pathway are critical antioxidants in nodules. In order to establish a meaningful correlation between concentrations of ASC and capacity for N₂ fixation in legume root nodules, soybean ( Glycine max ) plants were treated with excess ASC via exogenous irrigation or continuous intravascular infusion through needles inserted directly into plant stems. Treatment with ASC led to striking increases in nitrogenase activity (acetylene reduction), nodule leghaemoglobin content, and activity of ASC peroxidase, a key antioxidant enzyme. The concentration of lipid peroxides, which are indicators of oxidative damage and onset of senescence, was decreased in ASC-treated nodules. These results support the conclusion that ASC is critical for N₂ fixation and that elevated ASC allows nodules to maintain a greater capacity to fix N₂ over longer periods.     

Application of gibberellic acid to the surface of soybean seed (t Glycine max (L.) Merr.) and symbiotic nodulation,plant development,final grain and protein yield under short season conditions

In short-season soybean production areas, low soil temperature is the major factor limiting soybean establishment, nodulation and nitrogen fixation. Gibberellic acid (GA) pretreatment of crop seeds can overcome low soil temperature inhibition of seed germination and seedling development. However, previous studies have found that the application of GAs decreased legume nodulation and nitrogen fixation under optimal growth conditions. A field experiment was conducted under short season conditions in eastern Canada to determine whether the application of GA₃ to soybean seed could accelerate germination, and increase plant nodulation and nitrogen fixation. The results indicated that GA₃ application accelerated seedling emergence but decreased plant nodulation and nitrogen accumulation at early plant growth stages. However, these initial negative effects were overcome as the plants developed. Gibberellic acid applied to soybean seed at the time of planting did not influence final grain and protein yield.     

Early seedling and seasonal N2-fixing symbiotic activity of two soybean [Glycine max (L.) Merr.] cultivars inoculated with Bradyrhizobium strains of diverse origin

In areas with a short growing season the poor adaptability of soybean [Glycine max Meer. (L.)] to cool soil conditions is considered the primary yield limiting factor. Soybean requires temperatures in the 25 to 30°C range for optimum N₂-fixation and yield. Field studies were conducted in 1990 and 1991 at Montreal, Quebec to determine whether adaptability to cool soil conditions, with respect to earlier symbiosis establishment and function, existed among either Bradyrhizobium strains or soybean genotypes. An early maturing isoline of the soybean cultivar Evans and the cultivar Maple Arrow were inoculated with one of four strains isolated from the cold soils of Hakkaido, northern Japan, or the commercially used strains 532C or USDA110, at two planting dates. Plot biomass and nodulation were assessed at seedling (V2), and flowering(R2) growth stages and harvest maturity. Soybean genotypes did not differ for pre-flowering nodulation or N₂-fixation in the cool spring conditions of the first year. Seasonal N₂-fixation rates were also determined at the final harvest by the N-balance and ¹⁵N-isotope dilution methods. Significantly higher symbiotic activity was found for two of the four Hakkaido strains and was reflected in higher final soybean seed yield and total N₂-fixation for the growing season, as compared to the two commercial strains. Planting 14 days earlier resulted in greater early vegetative and total seasonal N₂ fixation and yield in the second year when soil temperatures were warmer, emphasizing the need for the development of soybean-Bradyrhizobium combinations superior in nodule development and function under cool soil conditions.     

Growth comparisons of a supernodulating soybean (Glycine max) mutant and its wild-type parent

The growth of a supernodulating, nitrate-tolerant soybean [ Glycine max (L.) Merr.] mutant nts 382 (nitrate-tolerant symbiosis) was compared to that of its wild-type parent, cv. Bragg, over the first 50 days after sowing. Plants were grown either inoculated in the absence of an external nitrogen source or uninoculated in the presence of 5 m M KNO₃. For both treatments, nts 382 growth up to 13 days after planting was faster than that of cv. Bragg. Thereafter, supernodulation of inoculated nts 382 occurred and growth of cv. Bragg was faster; shoot and root dry weight increments and leaf area were greater in cv. Bragg, but the N content of nts 382 was higher. Relative growth and net assimilation rates were lower in nts 382, which had faster shoot and root respiration rates. Shoot growth of uninoculated plants was similar for both mutant and wild-type but roots of nts 382 were slightly smaller than those of cv. Bragg. Total plant N content was similar in uninoculated cv. Bragg and nts 382 but the latter had a higher leaf N content. Early lateral root formation (prior to nodule emergence) was greater in nts 382 regardless of whether rhizobia or KJNO₃ were present. We conclude that nts 382 has some inherent differences from its parent but that supernodulation significantly retards plant growth.     

Development ofBradyrhizobium infections in supernodulating and non-nodulating mutants of soybean (Glycine max [L.] Merrill)

Summary The early events in the development of nodules induced byBradyrhizobium japonicum were studied in serial sections of a wild type (cv. Bragg), a supernodulating mutant (nts 382) and four non-nodulating mutants (nod49, nod139, nod772, andrj ₁) of soybean (Glycine max [L.] Merrill). Cultivar Bragg responded to inoculation in a similar manner to that described previously for cv. Williams; centres of sub-epidermal cell divisions were observed both with and without associated infection threads and most infection events were blocked before the formation of a nodule meristem. The non-nodulating mutants (nod49, nod772, andrj ₁) had, at most, a few centres of sub-epidermal cell divisions. In general, these were devoid of infection threads and did not develop beyond the very early stages of nodule ontogeny. Sub-epidermal cell divisions or infection threads were never observed on mutant nodl39. This mutant is not allelic to the other non-nodulating mutants and represents a defect in a separate complementation group or gene that is required for nodulation. The supernodulating mutant nts382, which is defective in autoregulation of nodulation, had a similar number of sub-epidermal cell divisions as the wild-type Bragg, but a much greater proportion of these developed to an advanced stage of nodule ontogeny. Mutant nts382, like Bragg, possessed other infection events that were arrested at an early stage of development. The results are discussed in the context of the progression of events in nodule formation and autoregulation of nodulation in soybean.Abbreviations nts nitrate tolerant symbiosis - RT root tip (i.e., position of the tap root tip at the time of inoculation) - SERH shortest emerging root hair (i.e., position of the shortest emerging root hair on the tap root at the time of inoculation) - SCD subepidermal cell divisions     

Yields and nitrogen nutrition of intercropped maize and ricebean (Vigna umbellata [Thumb.] Ohwi and Ohashi)

Maize (Zea mays L.) and ricebean (Vigna umbellata [Thumb.] Ohwi and Ohashi) were grown in intercrop and monoculture on Tropaqualf soils under rainfed conditions in Northern Thailand yearly from 1983 to 1986. De Wit's replacement design was used to compare intercrops and monocultures with a constant plant density equivalent to 80 000 maize or 160 000 ricebean plants ha⁻¹. Combined nitrogen was applied at varying levels to 200 kg N ha⁻¹. In the final two seasons the intercrop ratio of maize: ricebean was also varied. At the time of maize maturity intercrops yielded upt 49 kg ha⁻¹ more N in the above ground plant parts than the best monoculture. Dry matter, grain and nitrogen yield of maize and ricebean in intercrop relative to their monoculture yields (RY, relative yield) were significantly greater than their respective share of the plant population. Relative yield totals (RYT) for grain, dry matter and nitrogen were always greater than 1. Nitrogen uptake per maize plant increased with progressive replacement of maize by ricebean plants. This increase was similar to that obtained by applying combined N. Available soil nitrogen tended to decrease with increasing maize:ricebean ratio. Increasing the maize:ricebean ratio increased the % of nitrogen derived from fixation in ricebean, the increase being equivalent to that obtained by decreasing combined nitrogen application. Approximately the same amount of fertilizer and soil nitrogen was taken up by maize plus ricebean in intercrop as the maize monoculture. The results suggest that the improved nitrogen economy of the intercrop resulted from the strong competitiveness of maize in the use of mineral nitrogen and the enhancement of nitrogen fixation in intercropped ricebean which made it less dependent on the depleted pool of soil nitrogen.     

Plant responses to nutrient addition and predictive ability of vegetation N:P ratio in an austral fen

1. Previous studies of the N:P ratio in wetland plants have been carried out in northern hemisphere wetlands where atmospheric nitrogen deposition is higher. There is little research on foliar N:P ratio as a potential indicator of nutrient limitation in vegetation communities in southern hemisphere wetlands. This study aimed to redress this knowledge gap and answer the following questions: how well does the plant tissue nitrogen to phosphorus (N:P) ratio predict wetland plant community nutrient limitation, as indicated by vegetation standing stocks and below-ground biomass, in southern hemisphere fens? Secondly, what are the impacts of realistic upper levels of farm nutrient run-off on natural montane fen vegetation?
2. Low (35 kg ha⁻¹ year⁻¹) and high (70 kg ha⁻¹ year⁻¹) levels of nitrate-N or ammonium-N with and without P (20 kg ha⁻¹ year⁻¹) were added to 81 vegetation plots over a period of 2.75 years. Species composition, plant nutrient status, and above-ground live vegetation standing stocks were assessed after 3 years, and below-ground biomass after 2 years.
3. Plant tissue analysis suggested the community was N limited or N and P co-limited; we found greater standing stocks of vegetation in plots treated with 70 kg ha⁻¹ year⁻¹ ammonium-N, indicating N limitation. No difference between other treatments was found in above-ground standing stocks or below-ground biomass. Plant species cover increased in both high N treatments, consistent with N limitation. These changes in plant species cover were accompanied by significant decreases in species richness in both high N treatments. Native species dominated the vegetation and this was unaffected by nutrient addition (90% cover).
4. This is one of the first studies to test and find support for the N:P ratio in southern hemisphere wetlands. Observed declines in species richness after N fertilisation in an N-limited fen suggests increased N may pose risks to austral wetlands. Responses by plant communities (changes in composition, biomass) to lower levels of nutrient addition may require longer periods of fertilisation to be apparent in slow growing ecosystems.

     

DNA changes involving repeated sequences in senescing soybean (Glycine max) cotyledon nuclei

Age-related DNA changes in soybean ( Glycine max L. cv. Ransom) cotyledon nuclei were investigated by Feulgen cytophotometry and cloned DNA probes. The amount of nuclear DNA in 17-day-old senescing cotyledons was 23% lower than that of 5-day-old young cotyledons. In order to understand the nature of senescence-related DNA loss, fragments of repetitive DNA from young cotyledons were cloned into Escherichia coli HB101 cells by DNA recombination. The cloned DNA probed changes in specific repeated nucleotide sequences in senescing cotyledons. The colony hybridization between cloned DNA and [³²P]-labeled total DNA from 5- and 17-day cotyledons indicated loss of specific repeated sequences. The selected sequences of repeated DNA further showed a loss in their copy numbers. The study suggested that some repetitive DNA sequences were degraded selectively in the genome of senescing cotyledons.     

Effects of N fertilization on N2 fixation and N balances of soybean grown after lowland rice

We report a study in northern Thailand to examine the effects of fertilizer N, applied both to paddy rice and to a subsequent soybean crop on symbiotic and yield characteristics of soybean and on the differences between inputs of fixed N₂ and the removal of N as harvested product. Treatments were a factorial arrangement of 0, 100 and 300 kg N ha^-1 applied to the rice (designated R0, R100 and R300, respectively), and 0,25 and 50 kg N ha^-1, applied as starter fertilizer to the soybean (S0, S25 and S50, respectively).Nitrogen applied to the rice increased rice yields by up to 74% but proportions recovered by the rice were low (45% [R100] and 14% [R300]). The rice N treatments had only marginal effects on soybean nodulation (up to 17% reduction in early growth) and above-ground dry matter (up to 9% increase). Effects on soybean seed yield and total N₂ fixed were insignificant. Starter N, applied to the soybean at sowing, also marginally reduced nodulation and enhanced above-ground dry matter. Total N₂ fixed was unaffected but seed yield was increased by up to 6%. For all treatments, total above-ground N ranged from 145 to 179 kg ha^-1 with 72 to 85% (122 and 140 kg ha^-1) derived from N₂ fixation. When harvested product consisted of seed only, differences between inputs of fixed N₂ and removals of seed N were close to zero (-10 to+9 kg N ha^-1) with little effect of fertilizer N. The N balances were reduced by an average of 18 kg N ha^-1 when straw was included as harvested product. We concluded that N applied to the rice and to the following soybean was inefficiently used by those crops and had only marginal effects of symbiotic activity of the soybean. Furthermore, the benefit of the N₂ fixing soybean in this system was to slow the decline of, rather than enhance, the N fertility of the soil     

Mapping QTLs for seed yield and drought susceptibility index in soy-bean（Glycine max L.） across different environments

Drought stress has long been a major constraint in maintaining yield stability of soybean （Glycine max （L.） Merr.） in rainfed ecosystems. The identification of consistent quantitative trait loci （QTL） involving seed yield per plant （YP） and drought susceptibility index （DSI） in a population across different environments would therefore be important in molecular marker-assisted breeding of soybean cultivars suitable for rainfed regions. The YP of a recombinant line population of 184 F2：7：11 lines from a cross of Kefengl and Nannong1138-2 was studied under water-stressed （WS） and well-watered （WW） conditions in field （F） and greenhouse （G） trials, and DSI for yield was calculated in two trials. Nineteen QTLs associated with YP-WS and YP-WW, and 10 QTLs associated with DSI, were identi- fied. Comparison of these QTL locations with previous findings showed that the majority of these regions control one or more traits re- lated to yield and other agronomic traits. One QTL on molecular linkage group （MLG） K for YP-F, and two QTLs on MLG C2 for YP-G, remained constant across different water regimes. The regions on MLG C2 for YP-WW-F and MLG H for YP-WS-F had a pleiotropic effect on DSI-F, and MLG A1 for YP-WS-G had a pleiotropic effect on DSI-G. The identification of consistent QTLs for YP and DSI across different environments will significantly improve the efficiency of selecting for drought tolerance in soybean.