Glasshouse and field experiments were conducted with chickpea (Cicer arietinum) rhizobia to determine the inoculation requirements of this highly Rhizobium-specific legume. There did not appear to be any host-strain specificity within the species. There was a strong nodulation response to inoculation with four strains (unaffected by time of sowing) even at a level of inoculant application below normal. In some field experiments, nodulation responses were not reflected in improved foliage dry matter production or seed yield. However, the correlations between degree of nodulation and plant growth and seed yield were significant. There was a distinct advantage in using solid inoculant applied in the row with the seed instead of conventional seed inoculation when fungicide-treated chickpea was being sown. Two strains, CB1189 and CC1192, were considered suitable for inoculants.
Sixteen field experiments were conducted to assess whether inoculant applied as a liquid or in solid form separately from the seed but into the seed bed could be used as a substitute for conventional methods of legume seed inoculation. The experiments were done over a period of 8 years, on several soil types, with both crop and pasture legumes. Criteria used to measure response included success of the applied strain of rhizobia in forming nodules, quality of nodulation, seedling establishment, foliage dry matter production, and seed yield. Under favourable conditions for sowing, solid and liquid inoculants were generally as good as seed inoculation at similar total rates of application in promoting root nodulation and plant growth. However, when conditions were unfavourable for survival of rhizobia, e.g. when seed was dusted with toxic fungicide or when germination was delayed owing to environmental circumstances, solid or liquid inoculant gave better nodulation and very often better plant growth and seed yield than seed inoculation. The implications of these findings on alternatives to conventional seed inoculation and how they might be applied to agricultural practice are discussed.
Nitrogen fixation by irrigated soybeans (Glycine max (L.) Merr. cv. Forrest) was studied in a field experiment on a grey clay soil at Trangie, N.S.W. during the summer of 1985-86. Cropping with oats during the previous winter diminished the concentration of plant-available nitrogen in the soil from 37.6 to 18.5 mg N kg-1 and induced differences in the natural abundance of 15N (S15N) in this nitrogen. Four rates of liquid inoculation with Bradyrhizobium japonicum strain CB 1809, interacted with soil nitrogen to produce a wide range of nodulation of the soybeans. The following main effects on growth and N2 fixation resulted: (a) Initially, growth and accumulation of plant nitrogen was lower in pre-cropped than in prefallowed soil but N2 fixation was higher. (b) Nitrogen fixation during seed development was high in pre-cropped soil and greatest at the highest rate of inoculation. It resulted in high yields of seed (3.5 t ha-1 with 100 times the normal inoculation) with significantly higher concentration of seed nitrogen than from plants grown in prefallowed soil. (c) With increasing rates of inoculation on the pre-fallowed soil, more uniform nodulation was associated with smaller variances in most of the parameters studied. Other findings included further validation of the S15N method of calculating the proportion (p) of plant nitrogen derived from N2 fixation, with good agreement between treatment effects based on such estimates and those based on the relative concentrations of ureides in vacuum-extracted xylem sap. The values of p from S15N measurements on shoot nitrogen were affected little by inclusion of root nitrogen, and similar values were obtained when uninoculated, unnodulated Forrest soybeans, a nonnodulating genotype (non-nod Clark 63) or extractable mineral nitrogen of soil were used to estimate the S15N of plant N assimilated from soil. More dry matter (flowers, young pods and older leaves) containing more nitrogen (23-26 kg N ha-1) fell from the canopy of plants during seed maturation on pre-fallowed soil (high nitrogen) than on pre-cropped soil (13-15 kg N ha-1). Several correlations between the various quantities measured were noted and are discussed. It is concluded that growing winter cereals on land newly broken from pasture, coupled with high rates of inoculation of the following soybeans, may be a profitable way of diminishing plant-available soil nitrogen, thus maximizing the contribution of nitrogen from N2 fixation with benefits in seed yield and protein content.
Nodulation, N2 fixation (estimated by 15N natural abundance methods) and dry matter production were studied in a lucerne (Medicago sativa) crop managed for hay production at Ginninderra Experiment Station, A.C .T. Measurements were taken in the year of establishment and during two subsequent growing seasons. There were three treatments: (1) no inoculation and no annual fertilizer applied, (2) initial inoculation and superphosphate applied annually, (3) no inoculation, superphosphate applied annually and ammonium sulfate periodically. Before planting and after each growth season, soil was analysed for extractable mineral nitrogen, total nitrogen and the 15N natural abundance of this nitrogen, to the depth explored by lucerne roots. Before planting, no appropriate root-nodule bacteria (Rhizobium meliloti) were detected in the soil and initially plants were nodulated only in the inoculated treatment. Thereafter nodulation increased on the other treatments. Eight months after sowing there were no differences between treatments in numbers of R. meliloti g-l soil or in nodulation. In the third growing season, almost 30 kg ha-1 (dry wt) of nodules were recovered to a depth of 25 cm. These nodules were primarily located on fine, ephemeral roots and many appeared to be renewed after cutting of the lucerne. In the year of establishment, dry matter yields (0% moisture) totalled 3 to 4 t ha-1 in three hay cuts. In succeeding years, total yields were in the range 10 to 13 t ha-1 in four or five cuts per season. Nitrogen removed in the harvested lucerne reached 340 to 410 kg N ha-lyr-l in the second and third years and between 65 and 96% of this N arose from N2 fixation, depending on the method of calculation used. Poorer dry matter production and N2 fixation in treatment 1 in the third growing season was attributed to an insufficient supply of available phosphorus. Fixed N removed in Lucerne hay from treatment 2 totalled at least 640 kg N ha-1 in the three years of the experiment. Also, there were substantial increases in soil nitrogen due to lucerne growth. Although soil compaction made the quantification difficult, at the end of the experiment it was estimated that there was at least an extra 800 kg N ha-1 in the total soil nitrogen under lucerne compared to strips of Phalaris aquatica grown between the lucerne plots. It was concluded that lucerne contributed at least the same amount of fixed nitrogen to the soil as was being removed in the harvested hay.
In a field experiment at Leeton, N.S.W., Chaffey soybeans were grown with irrigation at various plant spacings and with various inoc~ilation treatments and two pre-planting soil treatments. Uninoculated plants were almost completely non-nodulated. Measurements of the natural abundance of 15N (S15N) in the total nitrogen of the plants were made at all stages of growth and in the grain at harvest. The 6lSN in all nodulated treatments declined progressively with time in comparison with un-nodulated plants, due to the incorporation of atmospheric N2 of lower 15N concentration than the soil nitrogen. This enabled calculation of the proportions of plant-nitrogen obtained from the soil and by symbiotic N2-fixation. The main findings were as follows: There was a gradient of S15N in plant-available nitrogen across the experimental area. Therefore, treatments were compared by using the nearest non-nodulated plot for the estimate of S15N in plantavailable soil nitrogen. Despite large differences in plant size due to plant spacing, S15N in mature nonnodulated plants did not differ significantly, indicating that the natural abundance of 15N in plantavailable soil nitrogen was uniform in root zones of different sizes. In well-nodulated plants, the proportion (p) of shoot nitrogen derived from N2-fixation increased with time, reaching approximately 70% and 90% in previously fallowed and previously cropped soil respectively, during a period of rapid growth between 78 and 98 d& after planting. The fixed N, in the best-nodulated treatments at (114 days) was 143 and 244 kg N ha-1 respectively for previously fallowed and previously cropped soil. There were consistent trends for increased N2 fixation with increased inoculation rates. In non-nodulated plants, nitrogen recovered in the grain represented most of that present in the shoots at maturity. In well-nodulated treatments, grain nitrogen, although similar in S15N to that of shoots + fruits, represented only 47 and 59% of the 406 and 348 kg N ha-1 present at maturity in shoots + fruits from previously fallowed and previously cropped soils respectively. After harvesting more than 3 t ha-1 of grain, the nitrogen balance in the previously cropped soil, if all of the residual nitrogen in the soybeans could have been retained in the soil, was positive. In the previously fallowed soil there could have been a net depletion of soil nitrogen.
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