Effect of vesicular-arbuscular mycorrhizae on the growth and yield of rice-stubble cultured soybeans

Summary Inoculation with vesicular-arbuscular (VA) mycorrhizal fungiGlomus fasciculatus, G. mosseae, G. etunicatus orAcaulospora scrobiculatus, increased plant dry weight and seed yields of pot-grown soybean plants in sterilized soil. Inoculation with a mixture ofG. fasciculatus, G. mosseae andG. etunicatus, orG. fasciculatus alone, increased seed yields and other agronomic traits of soybean plants grown in a no-tillage, rice-stubble field.     

Selection for improved nitrogen fixation inGlycine max (L.) Merr. andPhaseolus vulgaris L.

Summary While symbiotic nitrogen (N₂) fixation byG. max andP. vulgaris reduces their need for combined N, N₂ fixation under field conditions is rarely maximized. This paper reviews constraints to N₂ fixation in these species, then examines the genetic variability recorded for traits affecting N₂ fixation and the further work needed in this area. It considers emerging programs for the improvement of N₂ fixation inG. max andP. vulgaris and pays particular attention to methodological considerations.Scientific Journal Series, Minnesota Agri. Exp. Station. No 14190.     

Drought tolerance processes in soybeans

Fifteen soybean cultivars were evaluated in two water supply conditions, inducing or not a drought stress. Main canopy traits were measured several times during the reproductive period and, at maturity date, the yield components were estimated. Using principal components analysis, the main physiological functions involved in soybean drought tolerance are described: leaf cells enlargement and assimilates transport. These processes could be a good basis on which to define new selection criteria for soybean drought tolerance.     

Prevalence of 1-aminocyclopropane-1-carboxylate deaminase in Rhizobium spp

This is the first report documenting the presence of 1-aminocyclopropane-1-carboxylate (ACC) deaminase in Rhizobium. This enzyme, previously found in free-living bacteria, yeast and fungi, degrades ACC, the immediate precursor of ethylene in higher plants. Thirteen different rhizobial strains were examined by Southern hybridization, Western blots and ACC deaminase enzyme assay. Five of them tested positive for ACC deaminase. Induction of the expression of ACC deaminase was examined in one of the positively tested strains, Rhizobium leguminosarum bv. viciae 128C53K. This rhizobial ACC deaminase had a trace basal level of expression without ACC, but could be induced by a concentration of ACC as low as 1 μM. The more ACC added to this Rhizobium the higher the expression level of the ACC deaminase. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effect of soil pH and sewage sludge on VA mycorrhizal infection of soybeans

Summary Small plots were amended in 1976 or 1978 with four kinds of sewage sludge. The sludges represented samples considered to be relatively free of heavy metals as well as sludges highly contaminated with heavy metals. Sludges were added to a silt loam soil at rates of 224 or 448 Mgha⁻¹. The soils were maintained at a high or low pH regime. In 1984, soybeans (Glycine max L. Merril. var. ‘Clark’) were planted and grown to the R4 stage. After harvest, roots were removed from the soil, washed, and examined for VA mycorrhizal infection. It was found that the heavy metal content of the sludge alone was generally not related to determining the extent of mycorrhizal infection. A heat treated sludge, high in heavy metals, exhibited the highest degree of mycorrhizal infection when the soil was maintained at a pH of 6.2. With this treatment, 52% of the root segments examined were infected by mycorrhiza. When the same sludge was added to a soil with a slightly lower pH (5.7) none of the roots examined were infected by mycorrhiza. When soybean roots were examined from soils that received no sludge and were maintained at either a low (5.6) or high (6.2) pH, there was no significant difference in mycorrhizal infection between the pH regimes. These results therefore indicate that sewage sludge may inhibit mycorrhizal infection if the sludge contains a high concentration of heavy metals and the sludge is applied to the soil with a low pH. Scientific Article No. A-4093 and Contribution No 7078 of the Maryland Agric. Exp. Stn., Dept. of Agronomy, University of Maryland, College Park, MD 20742.     

Conditional lethality involving nuclear and cytoplasmic chlorophyll mutants in soybeans

Summary A conditionally lethal phenotype occurred when a nuclear chlorophyll mutant (y ₂₀-k ₂) was present with a cytoplasmic chlorophyll mutant (cyt-Y ₂) in soybean (Glycine max [L.] Merr.). Nuclear mutant y ₂₀-k ₂, Genetic Type Collection Number T253, has yellow foliage, tan-saddle-pattern seed and is viable. The y ₂₀-k ₂ mutant cannot be separated by classical genetic tests into two separate components, y ₂₀ (yellow foliage) and k ₂ (tan-saddle-pattern seed). Mutant cyt-Y ₂, T275, is inherited cytoplasmically, has yellow foliage, and is viable. The genotype cyt-Y ₂ y ₂₀-k ₂/ y ₂₀-k ₂ is a conditional lethal; the genotype is lethal under field conditions, but plants survive under greenhouse conditions. This interaction is unique to y ₂₀-k ₂. This conditionally lethal genotype may be useful in molecular studies on the interaction between nuclear and plastid genomes.This is a joint contribution of North Central Region, USDA ARS, and Journal Paper No. J-11429 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa, and the Agriculture Experiment Station, Univ. of Puerto Rico, Mayaguez Campus, Mayaguez, Puerto Rico 00708. Projects 2471 and 2475. The research was supported in part by the Iowa Soybean Promotion Board.     

Effects of N-serve on soybeans and soil nitrogen transformations

Summary Plant dry weight, total N, and total Ca was increased at 0.1 and 1 ppm N-serve. At greater 10 ppm the plants showed visual symptoms of a stunted growth, stem elongation, flowers, and pods failed to form or were aborted, young leaves were curled, and roots were club shaped with many branches. These symptoms were increasingly evident with increasing N-serve application rates. The reason was attributed to an auxin effect. Dry wt and total N in the plant was less than the control at the higher N-serve applications. There was little effect on nitrogenase activity at less than 10 ppm N-serve. Nodulation tended to increase at 0.1 and 1 ppm N-serve.Nitrification was inhibited up to 104 days at 20 ppm N-serve. The soil pH of the high N-serve rates was decreased at 104 days probably due to nitrification. Generally there were little detectable differences among treatments in soil organic N. The average soil organic N from 0 to 104 days decreased by 0.01%. Average increase in total N within each pot at harvest was equivalent to about 138 kg N/ha.     

Siderophore production byBradyrhizobium spp. strains nodulating groundnut

EighteenBradyrhizobium spp. strains, fourRhizobium spp. strains and oneAzorhizobium caulinodans strain were grown under Fe limitation and assayed for siderophore production. It was further assessed if Fe accumulation in two groundnut cultivars was influenced by inoculant strain or nitrate fertilisation. Growth ofBradyrhizobium spp. strains nodulating groundnut was slow with mean generation times from 11–24 h. All strains, except MAR 967, showed a reduced growth rate when deprived of Fe; none of the strains showed starvation at 1 M Fe. In the CAS (chrome azurol S)-agar assay, all strains, which formed colonies, produced siderophores as visualised by orange halos around the colonies on blue plates.Bradyrhizobium strains produced much smaller halos than the referenceRhizobium meliloti strain. In the CAS-supernatant assay, all strains, except MAR 967, gave positive responses (measured as absorbance at 630 nm) when supernatants of Fe-depleted cultures were assayed with CAS-indicator complex in comparison with Fe-supplemented cultures. Responses of all fourRhizobium spp. strains were large, while responses of allBradyrhizobium strains, exceptB. japonicum MAR 1491 (USDA 110), were small and mostly insignificant. A small response, i.e. a low Fe-scavenging ability, implies either the production of small quantities of siderophores or the production of low affinity siderophores. Among theBradyrhizobium strains, MAR 1574 and MAR 1587 gave the largest responses taken over the two assays. Fe accumulation in groundnut cultivar Falcon was seven times larger than in cultivar Natal Common. No correlation was found between the quantity of nodule tissue and Fe accumulation, making it unlikely that bacteroids are involved in Fe acquisition by groundnuts. Nitrate-fertilised plants accumulated significantly more Fe, suggesting involvement of nitrate reductase in Fe assimilation in groundnut. The two most successful Fe-scavengingBradyrhizobium spp. strains were also the most effective in nodulating groundnut, the reverse also being true. Strain MAR 967, with the lowest Fe requirement, produced the largest nodule dry weight. These data indicate that improved Fe scavenging properties and/or reduced Fe requirement improve rhizospheric growth and with that nodulation effectiveness.     

Platte Valley Yellows,a chlorotic condition of soybeans: Symptoms and preliminary chemical analyses

Summary A chlorotic condition of soybeans in the river valleys of eastern Nebraska was studied. The pattern of chlorosis, presence of petiolar necrosis, reductions in rhizobial and mycorrhizal infection, and plant and soil chemical analyses suggest that the condition is more complex than normal iron deficiency chlorosis or other mineral deficiencies or toxicities. Nodulation is almost totally inhibited in affected plants. Mycorrhizal fungi, while able to infect early in the season, do not persist on affected plants. Breakdown of endodermis was observed in the roots of affected plants. The factors contributing to this condition have not yet been determined. Published with the approval of the director as Paper No. 5672, Journal Series, Nebraska Agricultural Experiment Station.     

A non-destructive field assay for soybean nitrogen fixation by acetylene reduction

Summary A system for employing open-ended root chambers to measurein situ acetylene reduction rates under field conditions is described. Gas mixtures containing about 2 mbar acetylene were continuously flowed through the chambers providing a continuous record of acetylene reduction. These chambers have been used to measure acetylene reduction rates of soybeans during three growing seasons. The system has proved to be reliable with a high degree of precision. The large amount of plant-to-plant variability observed in N₂ fixation research has been confirmed by the data collected with this system. However, such variability in physiological studies can be reduced by using a non-destructive system to compare the response of an individual plant with its rates before treatment.     

Development of resistant transgenic soybeans with inverted repeat-coat protein genes of soybean dwarf virus

In an attempt to generate soybean plants resistant to soybean dwarf virus (SbDV), we transformed a construct containing inverted repeat-SbDV coat protein (CP) genes spaced by β-glucuronidase (GUS) sequences into soybean somatic embryos via microprojectile bombardment. Three T₀ plants with an introduced CP gene were obtained, and one generated T₁ seeds. The presence of the transgene in T₁ plants was confirmed by PCR and Southern blot hybridization analysis, but expression of CP was not detected by northern blot hybridization analysis. Two months after inoculation of SbDV by aphid, T₂ plants contained little SbDV-specific RNA and remained symptomless. These plants contained SbDV-CP-specific siRNA. These results suggest that the T₂ plants achieved resistance to SbDV by an RNA-silencing-mediated process.     

Occurrence ofDinophysis spp. and toxic shellfish in the Northern Adriatic

The dynamics of the toxicity of the musselMytilus galloprovincialis was compared between two different shellfish farms, 5 km apart, but using the same cultivation technique. The main differences concerned the freshwater influx and the open aspect to the Gulf of Trieste. It is suggested that a deep closed bay and abundant fresh water inflow are the two main conditions for the low toxicity levels in mussels and for shorter periods of danger. A detailed study of the phytoplankton samples revealed the presence of eight species ofDinophysis in the area of both shellfish farms. During the period of the DSP outbreak in Slovenia (autumn and winter 1989).D. fortii andD. acuminata were the most frequentDinophysis species. There was a high positive correlation between the onset of mussel toxicity and the appearance ofDinophysis spp.     

Intersubgeneric hybridization of soybeans with a wild perennial species,Glycine clandestina Wendl

Summary The exploitation of wild perennial species of subgenus Glycine has been formidable in soybean breeding programs because of extremely poor crossability and an early pod abortion. The combination of gibberellic acid application to hybridized gynoecia and improved seed culture media formulations resulted in a new intersubgeneric hybrid between Glycine max (L.) Merr. (2n=40) and G. clandestina Wendt. (2n=40). Of the 31 immature seeds cultured, 1 regenerated 21 plants through organogenesis while the remaining 30 failed to germinate. All the regenerated plants were similar morphologically, carried expected 2n=40, possessed hybrid isozyme patterns and were completely sterile. Complete absence of chromosome pairing was observed in 40.9% sporocytes. The occurrence of 1 to 6 loosely paired rod bivalents suggests some possibilities of allosyndetic pairing. Hybrid plants set aborted pods after backcrossing to G. max.     

Establishment of early instar Heliothis zea on soybeans

Early instar Heliothis zea (Boddie) (Lepidoptera: Noctuidae) established on all stages of soybean (Glycine max [L.] Merrill) tested when larvae were confined to plants, although establishment decreased after podset stage and as seeds matured. In tests where larvae were allowed to disperse freely on plants, establishment also decreased as maturity increased, but was much lower in each stage than when confined to plants. Neonate larvae settled more quickly on terminals and expanding trifoliates than on mature trifoliates based on initiation of feeding and movement away from initial sites of placement. The ability of neonate larvae placed a short distance from soybean plants to reach a host was affected by high soil and ambient temperature. The results of these experiments suggest that larval host establishment and spin-down behavior play a major role in the late instar population distribution among soybean fields and that maturity of the host strongly influences that behavior.

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Résumé Des expériences ont été réalisées dans deux types de conditions (avec ou sans possibilité de dispersion) pour déterminer si le taux d'installation des chenilles de premier stade de H. zea Bod. dépendait d'une mortalité liée au stade développement de Glycine max L. L'installation était considérée comme réussie quand les chenilles avaient atteint le second stade. Bien qu'il fût possible de distinguer les variétés de soja tant par le taux d'installation des chenilles que par les dégâts provoqués, les plantes les plus jeunes étaient plus colonisées par des chenilles que les témoins plus âgés. Toutes les variétés de soja à n'importe quel stade étaient acceptées lorsque la dispersion des chenilles était impossible. Avec possibilité de dispersion, les chenilles ont aussi accepté tous les stades phénologiques, mais en moins grands nombres que lorsque la dispersion était impossible. Le comportement de dispersion était beaucoup plus fréquent sur les plantes à port indéterminé que sur celles à port déterminé. Les possibilités de réinstallation après dispersion des chenilles de premier stade ont été limitées.

     

Identification of quantitative trait loci for plant height, lodging, and maturity in a soybean population segregating for growth habit

The use of molecular markers to identify quantitative trait loci (QTLs) has the potential to enhance the efficiency of trait selection in plant breeding. The purpose of the present study was to identify additional QTLs for plant height, lodging, and maturity in a soybean, Glycine max (L.) Merr., population segregating for growth habit. In this study, 153 restriction fragment length polymorphisms (RFLP) and one morphological marker (Dt1) were used to identify QTLs associated with plant height, lodging, and maturity in 111 F₂-derived lines from a cross of PI 97100 and Coker 237. The F₂-derived lines and two parents were grown at Athens, Ga., and Blackville, S.C., in 1994 and evaluated for phenotypic traits. The genetic linkage map of these 143 loci covered about 1600 cM and converged into 23 linkage groups. Eleven markers remained unlinked. Using interval-mapping analysis for linked markers and single-factor analysis of variance (ANOVA), loci were tested for association with phenotypic data taken at each location as well as mean values over the two locations. In the combined analysis over locations, the major locus associated with plant height was identified as Dt1 on linkage group (LG) L. The Dt1 locus was also associated with lodging. This locus explained 67.7% of the total variation for plant height, and 56.4% for lodging. In addition, two QTLs for plant height (K007 on LG H and A516b on LG N) and one QTL for lodging (cr517 on LG J) were identified. For maturity, two independent QTLs were identified in intervals between R051 and N100, and between B032 and CpTI, on LG K. These QTLs explained 31.2% and 26.2% of the total variation for maturity, respectively. The same QTLs were identified for all traits at each location. This consistency of QTLs may be related to a few QTLs with large effects conditioning plant height, lodging, and maturity in this population.     

The effect of silicon on the manganese nutrition of soybeans (Glycine max (L.) Merrill)

Summary Silicon during the early vegetative stage did not affect the oven dry weight of any of the various tissues of the soybean plant. Silicon did, however, decrease the Mn concentration in the youngest fully mature leaf at intermediate levels of Mn. This effect did not occur at the lowest or highest Mn levels. Deficiency and toxicity symptoms were moderated to a slight degree by Si except at the highest level of Mn.     

The role of active Bradyrhizobium japonicum in iron stress response of soybeans

The objective of this study was to identify the sites of H-ion exudation and Fe(III) reduction along both inoculated and non-inoculated roots of A7 and T203 soybeans. A split-root system was used in which half the roots of each plant were inoculated and actively fixing nitrogen and the other half were not. Expectedly, the Fe-stress response was strong on both sides of the split-root system in the +N-Fe treatment of variety A7 (inactive nodules) but not of variety T203. The Fe-stress response of A7 was enhanced by the presence of active nodules. Variety T203 is Fe inefficient and normally fails to produce any Fe-stress response, but in the absence of nitrogen and iron (–N–Fe), inoculated roots responded to Fe stress with exudation of both H-ions and reductants. Intact split-root systems were embedded in agar to determine the location of H-ion exudation and Fe(III) reduction. On the inoculated side of the –N–Fe and –N+Fe treatments (active nodules) of both soybean varieties, H-ion production was associated mainly with the active nodules. However, quantities of H-ion release were much greater under Fe stress (–N–Fe) than with adequate Fe (–N+Fe). Reduction of Fe(III) to Fe(II) was found only on the nodulated side with T203, but on both sides with A7. In variety T203 the Fe reduction was associated with younger roots located just below the nodule clusters on the inoculated side of the –N treatments. Active nodules appear to play a key role in the Fe-deficiency stress response of T203 soybean.     

Micronutrient uptake and distribution in mycorrhizal or phosphorus-fertilized soybeans

Summary Soybean plants were grown in a soil very low in available P. Seedlings were inoculated with two vesicular-arbuscular mycorrhizal (VAM) fungi or were left non-inoculated and fertilized with P. Assimilation and allocation of micronutrients (Fe, Mn, Zn, and Cu) were determined during host development, and the uptake of trace elements in VAM plants was compared to P-fertilized, non-VAM plants of similar weight, growth stage, and P status. Copper and zinc concentrations were always higher in VAM plants, while iron and manganese concentrations were lower than in the equivalent P-fertilized soybeans. Differences in the micronutrient content of fully-mature soybean pods reflected differences in the leaves and roots. Thus, for trace elements, seed quality can be altered by VAM colonization in a fashion not duplicated by P fertilizer. Contribution from the Western Regional Research Center, USDA-ARS (CRIS No. 5325-20580-003).     

The use of the two T-DNA binary system to derive marker-free transgenic soybeans

Summary A binary vector, pPTN133, was assembled that harbored two separate T-DNAs. T-DNA one contained a bar cassette, while T-DNA two carried a GUS cassette. The plasmid was mobilized into the Agrobacterium tumefaciens strain EHA101. Mature soybean cotyledonary node explants were inoculated and regenerated on medium amended with glufosinate. Transgenic soybeans were grown to maturity in the greenhouse. Fifteen primary transformants (T₀) representing 10 independent events were characterized. Seven of the 10 independent T₀ events co-expressed GUS. Progeny analysis was conducted by sowing the T₁ seeds and monitoring the expression of the GUS gene after 21 d. Individual T₁ plants were subsequently scored for herbicide tolerance by leaf painting a unifoliate leaf with a 100 mgl⁻¹ solution of glufosinate and scoring the leaf 5 d post application. Herbicide-sensitive and GUS-positive individuals were observed in four of the 10 independent events. Southern blot analysis confirmed the absence of the bar gene in the GUS positive/herbicide-sensitive individuals. These results demonstrate that simultaneous integration of two T-DNAs followed by their independent segregation in progeny is a viable means to obtain soybeans that lack a selectable marker.     

Phosphorus fertilization and tillage effect on dinitrogen fixation in soybeans

Summary Tillage has been shown to affect the uptake of phosphorus (P) and yield of soybeans, [Glycine max (L.) Merr.], but there is little information concerning the effects of P fertilization on nitrogen (N₂) fixation in soybeans under no-tillage. Two field experiments were conducted in 1980 and 1981 to determine the effects of soil P on N₂ fixation under no-tillage and to study the interaction of P fertilization and tillage of N₂ fixation, nutrient uptake, and yield of soybeans. In Exp. I, P was applied in 1977 at five rates up to 384 kg P ha⁻¹ and the effects of residual soil P were evaluated in 1980 and 1981 under no-tillage management. Nitrogen fixation rates, as measured by acetylene reduction assay, were significantly affected by soil P in Exp. I, but the assay proved to be a poor technique for estimating total plant N in these tests. Acetylene reduction rates and plant P increased rapidly as soil P increased from 2 to 20 mg kg⁻¹, with little additional increase above 20 mg P kg⁻¹. In Exp. II, rates (0, 32, 64, and 128 kg P ha⁻¹) and time (fall, spring and fall plus spring) of P application were compared under conventional tillage and no tillage. However, plant P increased with increasing levels of applied P. Applied P had no affect on acetylene reduction rates but rates were greater for no-tillage than conventional tillage at the V9 and R5 stages of growth in 1981. Plant uptake of P was more efficient under no-tillage than under conventional tillage in 1980 and 1981. Application of 64 kg P ha⁻¹ under no-tillage resulted in equivalent plant P levels as the 128 kg P ha⁻¹ applied under conventional tillage.