National Rhizobium Program - Managing rhizobia to maximise Nitrogen fixation

Inoculation of legumes with rhizobia is fundamental to sustainable productivity of Australian agriculture. The National Rhizobium Program has specific aims of sustaining and increasing Nitrogen fixation by legumes in Australian agriculture.

Optimising nitrogen fixation of grain and forage legumes in the northern region

This project encompasses laboratory, glasshouse and field research to improve N fixation in grain and forage legumes in the northern region and assess compatability of rhizobial strains with current and new legume varieties.

Effects of secondary crops on bacterial growth and nitrogen removal in shrimp farm effluent treatment systems

Secondary crops provide a means of assimilating some effluent nitrogen from eutrophic shrimp farm settlement ponds. However, a more important role may be their stimulation of beneficial bacterial nitrogen removal processes. In this study, bacterial biomass, growth and nitrogen removal capacity were quantified in shrimp farm effluent treatment systems containing vertical artificial substrates and either the banana shrimp Penaeus merguiensis (de Man) or the grey mullet, Mugil cephalus L. Banana shrimp were found to actively graze biofilm on the artificial substrates and significantly reduced bacterial biomass relative to a control (24.5 ± 5.6mgCm−2 and 39.2 ± 8.7mgCm−2, respectively). Bacterial volumetric growth rates, however, were significantly increased in the presence of the shrimp relative to the control 45.2±11.3mgCm−2 per day and 22.0±4.3mgCm−2 per day, respectively). Specific growth rate, or growth rate per cell, of bacteria was therefore appreciably stimulated by the banana shrimp. Nitrate assimilation was found to be significantly higher on grazed substrate biofilm relative to the control (223±54 mgNm−2 per day and 126±36 mg Nm−2 per day, respectively), suggesting that increased bacterial growth rate does relate to enhanced nitrogen uptake. Regulated banana shrimp feeding activity therefore can increase the rate of newbacterial biomass production and also the capacity for bacterial effluent nitrogen assimilation. Mullet had a negligible influence on the biofilm associated with the artificial substrate but reduced sediment bacterial biomass (224 ± 92 mgCm−2) relative to undisturbed sediment (650 ± 254 mgCm−2). Net, or volumetric bacterial growth in the sediment was similar in treatments with and without mullet, suggesting that the growth rate per cell of bacteria in grazed sediments was enhanced. Similar rates of dissolved nitrogen mineralisation werefound in sediments with and without mullet but nitrificationwas reduced. Presence of mullet increased water column suspended solids concentrations, water column bacterial growth and dissolved nutrient uptake. This study has shown that secondary crops, particularly banana shrimp, can play a stimulatory role in the bacterial processing of effluent nitrogen in eutrophic shrimp effluent treatment systems.

Leaf chlorophyll concentration relates to transpiration efficiency in peanut

Two pot experiments were conducted in two different seasons at the University of Agricultural Science, Bangalore, India, to study (a) the relationship between chlorophyll concentration (by measuring the leaf light-transmittance characteristics using a SPAD metre) and transpiration efficiency (TE) and (b) the effect of leaf N on chlorophyll and TE relationship in peanut. In Experiment (Expt) I, six peanut genotypes with wide genetic variation for the specific leaf area (SLA) were used. In Expt II, three non-nodulating isogenic lines were used to study the effect of N levels on leaf chlorophyll concentration–TE relationship without potential confounding effects in biological nitrogen fixation. Leaf N was manipulated by applying N fertiliser in Expt II. Chlorophyll concentration, TE (g dry matter kg−1 of H2O transpired, measured using gravimetric method), specific leaf nitrogen (g N m−2, SLN), SLA (cm2 g−1), carbon isotope composition (Δ13C) were determined in the leaves sampled during the treatment period (35–55 days after sowing) in the two experiments. Results showed that the leaf chlorophyll concentration expressed as soil plant analytical development (SPAD) chlorophyll metre reading (SCMR) varied significantly among genotypes in Expt I and as a result of N application in Expt II. Changes in leaf N levels were strongly associated with changes in SCMR, TE and Δ13C. In both the experiments, a significant positive relationship between SCMR and TE with similar slopes but differing intercepts was noticed. However, correction of TE for seasonal differences in vapour pressure deficit (VPD) between the two experiments resulted in a single and stronger relationship between SCMR and TE. There was a significant inverse relationship between SCMR and Δ13C, suggesting a close linkage between chlorophyll concentration and Δ13C in peanut. This study provides the first evidence for a significant positive relationship between TE and leaf chlorophyll concentration in peanut. The study also describes the effect of growing environment on the relationships among SLA, SLN and SCMR.

Declining soil phosphorus reserves on sustainable agricultural production systems

PhD scholarship investigating the relative sensitivity of nitrogen fixation in adapted grain and ley legume species to low soil phosphorus.

A convenient sample preparation protocol for scanning electron microscope examination of xylem-occluding bacterial biofilm on cut flowers and foliage

Microbes and their exopolysaccharides (EPS) can block xylem vessels, thereby increasing the hydraulic resistance and decreasing the vase life of cut flowers and foliage. Scanning electron microscopy (SEM) provides a powerful tool for investigation of bacteria-induced xylem occlusion. However, conventional preparation protocols for SEM involving chemicals can cause loss of hydrated EPS material, and thereby damage the bacterial biofilms during dehydration. A modified chemical fixation protocol involving pre-fixation with 75 mM lysine plus 2.5% glutaraldehyde followed by the normal fixation in 3% glutaraldehyde was, therefore, tested for improved preservation of bacterial biofilm at the stem-ends of cut Acacia holosericea foliage stems. Stem-end segments with different stages of bacterial growth were obtained from stems stood into water. The lysine-based protocol was compared with four other processing protocols of critical point drying (CPD) without fixation (control), freeze-drying (FD), conventional chemical fixation followed by drying with hexamethyldisilazane (HMDS), and conventional chemical fixation with CPD. The non-fixed control. FD and the glutaraldehyde fixation with HMDS drying gave poor preservation of hydrated material, including bacterial EPS. Conventional glutaraldehyde fixation followed by CPD was superior to these three methods in terms of better preserving the EPS. However, this fourth method gave condensation of biofilms during dehydration. In contrast, the modified lysine-based protocol resulted in superior preservation of EPS and biofilm structure. Thus, this fifth method was the most appropriate for examination of bacterial stem-end blockage in cut ornamentals. (C) 2012 Elsevier B.V. All rights reserved.

Volatile components associated with bacterial spoilage of tropical prawns

Analysis of headspace volatiles by gas chromatography/mass spectrometry from king (Penaeus plebejus), banana (P. merguiensis), tiger (P. esculentus/semisulcatus) and greasy (Metapenaeus bennettae) prawns stored in ice or ice slurry, which is effectively an environment of low oxygen tension, indicated the presence of amines at the early stages of storage (less than 8 days) irrespective of the nature of the storage media. Esters were more prevalent in prawns stored on ice (normal oxygen conditions) at the latter stages of storage (more than 8 days) and were only produced by Pseudomonas fragi, whereas sulphides and amines occurred whether the predominant spoilage organism was Ps.fragi or Shewanella putrefaciens. The free amino acid profiles of banana and king prawns were high in arginine (12–14%) and low in cysteine (0.1–0.17%) and methionine (0.1–0.2%). Filter sterilised raw banana prawn broth inoculated with a total of 15 cultures of Ps. fragi and S. putrefaciens and incubated for two weeks at 5°C, showed the presence of 17 major compounds in the headspace volatiles analysed using gas chromatography/mass spectrometry (GC/MS). These were mainly amines, sulphides, ketones and esters. Principal Component Analysis of the results for the comparative levels of the volatiles produced by pure cultures, inoculated into sterile prawn broth, indicated three subgroupings of the organisms; I, Ps. fragi from a particular geographic location; II, S. putrefaciens from another geographic location; and III, a mixture of Ps. fragi and S. putrefaciens from different geographic locations. The sensory impression created by the cultures was strongly related to the chemical profile as determined by GC/MS. Organisms, even within the same subgrouping classified as identical by the usual tests, produced a different range of volatiles in the same uniform substrate.

Spoilage pattern of five species of Australian prawns: Deterioration is influenced by environment of capture and mode of storage

Five species of commercial prawns Penaeus plebejus, P. merguiensis, P. semisulcatus/P. esculentus and M. bennettae, were obtained from South-East and North Queensland, chilled soon after capture and then stored either whole or deheaded on ice and ice slurry, until spoilage. Total bacterial counts, total volatile nitrogen, K-values and total demerit scores were assessed at regular intervals. Their shelf lives ranged from 10-17 days on ice and >20 days on ice slurry. Initial bacterial flora on prawns from shallower waters (4-15m) were dominated by Gram-positives and had lag periods around 7 days, whereas prawns from deeper waters (100m) were dominant in Pseudomonas spp. with no lag periods in bacterial growth. The dominant spoiler in ice was mainly Pseudomonas fragi whereas the main spoiler in ice slurry was Shewanella putrefaciens. Bacterial interactions seem to play a major role in the patterns of spoilage in relation to capture environment and pattern of storage

Effect of nitrogen on the skin colour and other quality attributes of ripe ‘Kensington Pride’ mango (Mangifera indica L.) fruit

Near-ripe ‘Kensington Pride’ mango (Mangifera indica L.) fruit with green skin colour generally return lower wholesale and retail prices. Pre-harvest management, especially nitrogen (N) nutrition, appears to be a major causal factor. To obtain an understanding of the extent of the problem in the Burdekin district (dry tropics; the major production area in Australia), green mature ‘Kensington Pride’ mango fruit were harvested from ten orchards and ripened at 20 ± 0.5 O C. Of these orchards, 70% produced fruit with more than 25% of the skin surface area green when ripe. The following year, the effect of N application on skin colour and other quality attributes was investigated on three orchards, one with a high green (HG) skin problem and two with a low green (LG) skin problem. N was applied at pre-flowering and at panicle emergence at the rate of 0,75,150,300 g per tree (soil applied) or 50 g per tree as foliar N for the HG orchard, and 0,150,300,450 g per tree (soil applied) or 50 g per tree (foliar) for the LG orchards. In all orchards the proportion of green colour on the ripe fruit was significantly (P<0.05) higher with soil applications of 150 g N or more per tree. Foliar sprays resulted in a higher proportion of green colour than the highest soil treatment in the HG orchard, but not in the LG orchards. Anthracnose disease severity was significantly (P<0.05) higher with 300 g of N per tree or foliar treatment in the HG orchard, compared with no additional N. Thus, N can reduce mango fruit quality by increasing green colour and anthracnose disease in ripe fruit.

Sustaining productivity of a Vertosol at Warra, Queensland, with fertilisers, no-tillage or legumes. 7. Yield, nitrogen and disease-break benefits from lucerne in a two-year lucerne–wheat rotation

Continuous cultivation and cereal cropping of southern Queensland soils previously supporting native vegetation have resulted in reduced soil nitrogen supply, and consequently decreased cereal grain yields and low grain protein. To enhance yields and protein concentrations of wheat, management practices involving N fertiliser application, with no-tillage and stubble retention, grain legumes, and legume leys were evaluated from 1987 to 1998 on a fertility-depleted Vertosol at Warra, southern Queensland. The objective of this study was to examine the effect of lucerne in a 2-year lucerne–wheat rotation for its nitrogen and disease-break benefits to subsequent grain yield and protein content of wheat as compared with continuous wheat cropping. Dry matter production and nitrogen yields of lucerne were closely correlated with the total rainfall for October–September as well as March–September rainfall. Each 100 mm of total rainfall resulted in 0.97 t/ha of dry matter and 26 kg/ha of nitrogen yield. For the March–September rainfall, the corresponding values were 1.26 t/ha of dry matter and 36 kg/ha of nitrogen yield. The latter values were 10% lower than those produced by annual medics during a similar period. Compared with wheat–wheat cropping, significant increases in total soil nitrogen were observed only in 1990, 1992 and 1994 but increases in soil mineralisable nitrogen were observed in most years following lucerne. Similarly, pre-plant nitrate nitrogen in the soil profile following lucerne was higher by 74 kg/ha (9–167 kg N/ha) than that of wheat–wheat without N fertiliser in all years except 1996. Consequently, higher wheat grain protein (7 out of 9 seasons) and grain yield (4 out of 9 seasons) were produced compared with continuous wheat. There was significant depression in grain yield in 2 (1993 and 1995) out of 9 seasons attributed to soil moisture depletion and/or low growing season rainfall. Consequently, the overall responses in yield were lower than those of 50 kg/ha of fertiliser nitrogen applied to wheat–wheat crops, 2-year medic–wheat or chickpea–wheat rotation, although grain protein concentrations were higher following lucerne. The incidence and severity of the soilborne disease, common root rot of wheat caused by Bipolaris sorokiniana, was generally higher in lucerne–wheat than in continuous wheat with no nitrogen fertiliser applications, since its severity was significantly correlated with plant available water at sowing. No significant incidence of crown rot or root lesion nematode was observed. Thus, productivity, which was mainly due to nitrogen accretion in this experiment, can be maintained where short duration lucerne leys are grown in rotations with wheat.

Sustaining productivity of a Vertosol at Warra, Queensland, with fertilisers, no-tillage or legumes. 8. Effect of duration of lucerne ley on soil nitrogen and water, wheat yield and protein.

Soil nitrogen (N) supply in the Vertosols of southern Queensland, Australia has steadily declined as a result of long-term cereal cropping without N fertiliser application or rotations with legumes. Nitrogen-fixing legumes such as lucerne may enhance soil N supply and therefore could be used in lucerne-wheat rotations. However, lucerne leys in this subtropical environment can create a soil moisture deficit, which may persist for a number of seasons. Therefore, we evaluated the effect of varying the duration of a lucerne ley (for up to 4 years) on soil N increase, N supply to wheat, soil water changes, wheat yields and wheat protein on a fertility-depleted Vertosol in a field experiment between 1989 and 1996 at Warra (26degrees 47'S, 150degrees53'E), southern Queensland. The experiment consisted of a wheat-wheat rotation, and 8 treatments of lucerne leys starting in 1989 (phase 1) or 1990 (phase 2) for 1,2,3 or 4 years duration, followed by wheat cropping. Lucerne DM yield and N yield increased with increasing duration of lucerne leys. Soil N increased over time following 2 years of lucerne but there was no further significant increase after 3 or 4 years of lucerne ley. Soil nitrate concentrations increased significantly with all lucerne leys and moved progressively downward in the soil profile from 1992 to 1995. Soil water, especially at 0.9-1.2 m depth, remained significantly lower for the next 3 years after the termination of the 4 year lucerne ley than under continuous wheat. No significant increase in wheat yields was observed from 1992 to 1995, irrespective of the lucerne ley. However, wheat grain protein concentrations were significantly higher under lucerne-wheat than under wheat wheat rotations for 3-5 years. The lucerne yield and soil water and nitrate-N concentrations were satisfactorily simulated with the APSIM model. Although significant N accretion occurred in the soil following lucerne leys, in drier seasons, recharge of the drier soil profile following long duration lucerne occurred after 3 years. Consequently, 3- and 4-year lucerne-wheat rotations resulted in more variable wheat yields than wheat-wheat rotations in this region. The remaining challenge in using lucerne-wheat rotations is balancing the N accretion benefits with plant-available water deficits, which are most likely to occur in the highly variable rainfall conditions of this region.

Influence of nitrogen based supplements on live weight, fertility and mortality of heifers grazing dry season native pasture

Supplements containing urea or biuret were fed in the dry season to yearling and two year old pregnant heifers grazing native spear grass pastures in north Queensland. Liveweight change and survival during the dry season and fertility in the following year were measured. In the first experiment during a relatively favourable dry season, supplementation significantly (P<0.01) reduced liveweight loss in yearling heifers (5 vs. 32 kg). In the following year during a drought, supplement significantly (P<.01) reduced liveweight loss in yearling heifers (32 vs. 41 kg) and significantly (P <0.01) reduced mortalities (23.5% vs. 5.2%) in pregnant and lactating heifers. The supplement had no significant effect on subsequent fertility in either experiment. 14th Biennial Conference.

Piggery pond sludge as a nitrogen source for crops. 1. Mineral N supply estimated from laboratory incubations and field application of stockpiled and wet sludge

Piggery pond sludge (PPS) was applied, as-collected (Wet PPS) and following stockpiling for 12 months (Stockpiled PPS), to a sandy Sodosol and clay Vertosol at sites on the Darling Downs of Queensland. Laboratory measures of N availability were carried out on unamended and PPS-amended soils to investigate their value in estimating supplementary N needs of crops in Australia's northern grains region. Cumulative net N mineralised from the long-term (30 weeks) leached aerobic incubation was described by a first-order single exponential model. The mineralisation rate constant (0.057/week) was not significantly different between Control and PPS treatments or across soil types, when the amounts of initial mineral N applied in PPS treatments were excluded. Potentially mineralisable N (No) was significantly increased by the application of Wet PPS, and increased with increasing rate of application. Application of Wet PPS significantly increased the total amount of inorganic N leached compared with the Control treatments. Mineral N applied in Wet PPS contributed as much to the total mineral N status of the soil as did that which mineralised over time from organic N. Rates of C02 evolution during 30 weeks of aerobic leached incubation indicated that the Stockpiled PPS was more stabilised (19-28% of applied organic C mineralised) than the WetPPS (35-58% of applied organic C mineralised), due to higher lignin content in the former. Net nitrate-N produced following 12 weeks of aerobic non-leached incubation was highly correlated with net nitrate-N leached during 12 weeks of aerobic incubation (R^2 = 0.96), although it was <60% of the latter in both sandy and clayey soils. Anaerobically mineralisable N determined by waterlogged incubation of laboratory PPS-amended soil samples increased with increasing application rate of Wet PPS. Anaerobically minemlisable N from field-moist soil was well correlated with net N mineralised during 30 weeks of aerobic leached incubation (R^2 =0.90 sandy soil; R^2=0.93 clay soil). In the clay soil, the amount of mineral N produced from all the laboratory incubations was significantly correlated with field-measured nitrate-N in the soil profile (0-1.5 m depth) after 9 months of weed-free fallow following PPS application. In contrast, only anaerobic mineralisable N was significantly correlated with field nitrate-N in the sandy soil. Anaerobic incubation would, therefore, be suitable as a rapid practical test to estimate potentially mineralisable N following applications of different PPS materials in the field.

Spectral and thermal sensing for nitrogen and water status in rainfed and irrigated wheat environments

Variable-rate technologies and site-specific crop nutrient management require real-time spatial information about the potential for response to in-season crop management interventions. Thermal and spectral properties of canopies can provide relevant information for non-destructive measurement of crop water and nitrogen stresses. In previous studies, foliage temperature was successfully estimated from canopy-scale (mixed foliage and soil) temperatures and the multispectral Canopy Chlorophyll Content Index (CCCI) was effective in measuring canopy-scale N status in rainfed wheat (Triticum aestivum L.) systems in Horsham, Victoria, Australia. In the present study, results showed that under irrigated wheat systems in Maricopa, Arizona, USA, the theoretical derivation of foliage temperature unmixing produced relationships similar to those in Horsham. Derivation of the CCCI led to an r2 relationship with chlorophyll a of 0.53 after Zadoks stage 43. This was later than the relationship (r2 = 0.68) developed for Horsham after Zadoks stage 33 but early enough to be used for potential mid-season N fertilizer recommendations. Additionally, ground-based hyperspectral data estimated plant N (g kg)1) in Horsham with an r2 = 0.86 but was confounded by water supply and N interactions. By combining canopy thermal and spectral properties, varying water and N status can potentially be identified eventually permitting targeted N applications to those parts of a field where N can be used most efficiently by the crop.

Modeling chickpea growth and development: Nitrogen accumulation and use

Quantitative information regarding nitrogen (N) accumulation and its distribution to leaves, stems and grains under varying environmental and growth conditions are limited for chickpea (Cicer arietinum L.). The information is required for the development of crop growth models and also for assessment of the contribution of chickpea to N balances in cropping systems. Accordingly, these processes were quantified in chickpea under different environmental and growth conditions (still without water or N deficit) using four field experiments and 1325 N measurements. N concentration ([N]) in green leaves was 50 mg g-1 up to beginning of seed growth, and then it declined linearly to 30 mg g-1 at the end of seed growth phase. [N] in senesced leaves was 12 mg g-1. Stem [N] decreased from 30 mg g-1 early in the season to 8 mg g-1 in senesced stems at maturity. Pod [N] was constant (35 mg g-1), but grain [N] decreased from 60 mg g-1 early in seed growth to 43 mg g-1 at maturity. Total N accumulation ranged between 9 and 30 g m-2. N accumulation was closely linked to biomass accumulation until maturity. N accumulation efficiency (N accumulation relative to biomass accumulation) was 0.033 g g-1 where total biomass was -2 and during early growth period, but it decreased to 0.0176 g g-1 during the later growth period when total biomass was >218 g m-2. During vegetative growth (up to first-pod), 58% of N was partitioned to leaves and 42% to stems. Depending on growth conditions, 37-72% of leaf N and 12-56% of stem N was remobilized to the grains. The parameter estimates and functions obtained in this study can be used in chickpea simulation models to simulate N accumulation and distribution.