Effect of root zone temperature on oilseed rape (Brassica napus) response to boron

Oilseed rape (Brassica napus) is sensitive to low boron (B) supply, and its growth response to B may be influenced by soil temperature. To test the relationship between B and temperature, oilseed rape (cv. Hyola 42) seedlings were grown at 10 degrees C (low) root zone temperature (RZT) with B supply from deficient to adequate B levels until growth of low B plants just began to slow down. Half of the pots were then transferred to 20 degrees C (warm) RZT for 11 days before they were moved back to 10 degrees C RZT for the final 4 days. Both plant dry mass and B uptake increased after plants were exposed to warm RZT. However, plant B deficiency was exacerbated by warm RZT in low B plants because of increased relative growth rate and shoot-root ratio without a commensurate increase in B uptake rate. It is concluded that RZT above the critical threshold for chilling injury in oilseed rape can nevertheless affect the incidence of B deficiency by altering shoot-root ratio and hence the balance between shoot B demand and B uptake.

Improved beef production from supplementation of Hereford, Brahman and crossbred cattle grazing low and medium quality pastures in the subtropics of Australia

Subtropical grasslands are low in organic matter digestibility (OMD) (0.60) and nitrogen (N) (15 g/kg) for much of the year and this limits cattle production which is characterized by low calving rates and low weaning weights. Production has been based on Bos taurus British breeds of cattle but this is changing and now many breeding herds comprise B, indicus cows and their crosses. This change has increased some aspects of production, but low calving rates persist. A 4-year study was undertaken with a view to improve calving rates and weaner output by supplementing cows grazing either native or improved pastures with a high protein oilseed meal (cottonseed meal; CSM) on four sites. These sites were subdivided into a total of 36 paddocks to allow for two replications in a 3 breeds X 3 supplementation rates X 2 pastures factorial design. Selected cows (no. = 216) from Hereford (H), Brahman (B) and Brahman X Hereford (BH) breed types were set to graze either native pastures (0.45 to 0.62 OMD, 8 to 15 g N per kg; low quality) or improved pastures (0.47 to 0.67 OA ID, 10 to 22 g N per kg; medium quality). Cows were given either 0, 750 or 1500 g/day of CSM for 130 days from calving until 4 weeks into a 12- to 13-week mating period. The CSM was given as two meals per week. Live weight at mating of cows on the low quality pasture was increased (P < 0.01) over those not supplemented by feeding either 750 g CSM per day (H and B cows) or 1500 g CSM per day tall cows). There was no significant effect of supplementation on the mating weights of B cows grazing the medium quality sites. Calving rate of B cows was not increased by their supplementation on either low (4-year mean 58.3 %) or medium quality pastures (66.8%) but did tend to be higher in H cows when supplemented at 1500 g CSM per day on the low (66.7 v. 78.0 (s.e. 6.09) %; P < 0.1) and medium quality pastures (70.5 v. 93.5 (s.e. 4.72) %). An increased calving rate (65.8 (s.e. 6.6) % to 83.2 (s.e. 5.82) % in supplemented BH cows grazing low quality pastures approached significance (P < 0.1) when given CSM at 1500 g/day but there was no increased trend in calving rate when this breed type was supplemented on medium quality pastures. Weaning weights of calves from and B and BH cows were increased (P < 0.05) by supplementation of their darns at 750 g/day and for calves weaned from H cows supplemented at 1500 g/day of CSM. Supplementation at 1500 g/day on low quality pastures increased weaner output per cow mated by 120% for H, by 65% for BH cows and by 50% for B cows. Weaner output was increased by 34 and 40%, respectively, for B and H cows when supplemented at 750 g/day and grazing medium quality pastures but there teas no significant effect of supplementation on output from BH cows. Responses in many parameters differed between years. These results were interpreted as a response to the protein in the oilseed meal supplement by B, taurus and B. taurus X B. indicus cross cows grazing on the subtropical pastures. The study also highlighted that responses to the meal differed between breed types, between the quality of the grazed pasture and between the years of supplementation.

Efficient transformation and regeneration of diverse cultivars of peanut (Arachis hypogaea L.) by particle bombardment into embryogenic callus produced from mature seeds

Peanut, one of the world's most important oilseed crops, has a narrow germplasm base and lacks sources of resistance to several major diseases. The species is considered recalcitrant to transformation, with few confirmed transgenic plants upon particle bombardment or Agrobacterium treatment. Reported transformation methods are limited by low efficiency, cultivar specificity, chimeric or infertile transformants, or availability of explants. Here we present a method to efficiently transform cultivars in both botanical types of peanut, by (1) particle bombardment into embryogenic callus derived from mature seeds, (2) escape-free (not stepwise) selection for hygromycin B resistance, (3) brief osmotic desiccation followed by sequential incubation on charcoal and cytokinin-containing media; resulting in efficient conversion of transformed somatic embryos into fertile, non-chimeric, transgenic plants. The method produces three to six independent transformants per bombardment of 10 cm(2) embryogenic callus. Potted, transgenic plant lines can be regenerated within 9 months of callus initiation, or 6 months after bombardment. Transgene copy number ranged from one to 20 with multiple integration sites. There was ca. 50% coexpression of hph and luc or uidA genes coprecipitated on separate plasmids. Reporter gene (luc) expression was confirmed in T-1 progeny from each of six tested independent transformants. Insufficient seeds were produced under containment conditions to determine segregation ratios. The practicality of the technique for efficient cotransformation with selected and unselected genes is demonstrated using major commercial peanut varieties in Australia (cv. NC-7, a virginia market type) and Indonesia (cv. Gajah, a spanish market type).

Purification and characterization of a plant antimicrobial peptide expressed in Escherichia coli

MiAMP1 is a low-molecular-weight, cysteine-rich, antimicrobial peptide isolated from the nut kernel of Macadamia integrifolia. A DNA sequence encoding MiAMP1 with an additional ATG: start codon was cloned into a modified pET vector under the control of the T7 RNA polymerase promoter. The pET vector was cotransformed together with the vector pSB161, which expresses a rare arginine tRNA. The peptide was readily isolated in high yield from the insoluble fraction of the Escherichia coil extract. The purified peptide was shown to have an identical molecular weight to the native peptide by mass spectroscopy indicating that the N-terminal methionine had been cleaved. Analysis by NMR spectroscopy indicated that the refolded recombinant peptide had a similar overall three-dimensional structure to that of the native peptide. The peptide inhibited the growth of phytopathogenic fungi in vitro in a similar manner to the native peptide. To our knowledge, MiAMP1 is the first antimicrobial peptide from plants to be functionally expressed in E. coil. This will permit a detailed structure-function analysis of the peptide and studies of its mode of action on phytopathogens. (C) 1999 Academic Press.

Dry matter production and boron concentrations of vegetative and reproductive tissues of canola and sunflower plants grown in nutrient solution

Canola (Brassica napus L.) and sunflower (Helianthus annuus L.), two important oilseed crops, are sensitive to low boron (B) supply. Symptoms of B deficiency are often more severe during the reproductive stage, but it is not known if this is due to a decreased external B supply with time or an increased sensitivity to low B during this stage. Canola and sunflower were grown for 75 days after transplanting (DAT) in two solution culture experiments using Amberlite (IRA-743) B-specific resin to maintain constant B concentration in solution over the range 0.6 - 53 muM. Initially, the vegetative growth of both crops was good in all treatments. With the onset of the reproductive stage, however, severe B deficiency symptoms developed and growth of canola and sunflower was reduced with less than or equal to 0.9 and less than or equal to 0.7 muM B, respectively. At these concentrations, reproductive parts failed to develop. The critical B concentration (i.e. 90% of maximum shoot dry matter yield) in the youngest opened leaf was 18 mg kg(-1) in canola and 25 mg kg(-1) in sunflower at 75 DAT. The results of this study indicate that the reproductive stage of these two oilseed crops is more sensitive than the vegetative stage to low B supply.

Enhanced quantitative resistance to Leptosphaeria maculans conferred by expression of a novel antimicrobial peptide in canola (Brassica napus L.)

The novel antimicrobial peptide MiAMP1, originally isolated from the seeds of Macadamia integrifolia, was constitutively expressed in transgenic tobacco and canola plants to test its effect on disease resistance. Analysis of plants transformed with 35S-MiAMP1 construct by northern and western blot analyses demonstrated the presence of MiAMP1 mRNA and the mature peptide in the transgenic plants. The MiAMP1 purified from the leaves of transgenic plants was biologically active with the same in vitro antifungal activity as native MiAMP1 purified from the seeds of macadamia. The effect of MiAMP1 expression on the economically important canola pathogen Leptosphaeria maculans (causal agent of blackleg disease) was evaluated in comparison with an untransformed control line and an azygous segregant derived from one of the transgenic lines. Lesion development on the cotyledons of the inoculated canola seedlings was significantly reduced in the T-2 progeny of seven independently transformed transgenic lines. These results suggested that, transgenic canola expressing MiAMP1 may be useful for the management of blackleg disease.

The Three-dimensional Solution Structure of NaD1, a New Floral Defensin from Nicotiana alata and its Application to a Homology Model of the Crop Defense Protein alfAFP

NMR spectroscopy and simulated annealing calculations have been used to determine the three-dimensional structure of NaD1, a novel antifungal and insecticidal protein isolated from the flowers of Nicotiana alata. NaD1 is a basic, cysteine-rich protein of 47 residues and is the first example of a plant defensin from flowers to be characterized structurally. Its three-dimensional structure consists of an a-helix and a triple-stranded anti-parallel beta-sheet that are stabilized by four intramolecular disulfide bonds. NaD1 features all the characteristics of the cysteine-stabilized up motif that has been described for a variety of proteins of differing functions ranging from antibacterial insect defensins and ion channel-perturbing scorpion toxins to an elicitor of the sweet taste response. The protein is biologically active against insect pests, which makes it a potential candidate for use in crop protection. NaD1 shares 31% sequence identity with alfAFP, an antifungal protein from alfalfa that confers resistance to a fungal pathogen in transgenic potatoes. The structure of NaD1 was used to obtain a homology model of alfAFP, since NaD1 has the highest level of sequence identity with alfAFP of any structurally characterized antifungal defensin. The structures of NaD1 and alfAFP were used in conjunction with structure - activity data for the radish defensin Rs-AFP2 to provide an insight into structure-function relationships. In particular, a putative effector site was identified in the structure of NaD1 and in the corresponding homology model of alfAFP. (C) 2002 Elsevier Science Ltd. All rights reserved.

Contrary effects of jasmonate treatment of two closely related plant species on attraction of and oviposition by a specialist herbivore

Elevated jasmonic acid (JA) concentrations in response to herbivory can induce wounded plants to produce defences against herbivores. In laboratory and field experiments we compared the effects of exogenous JA treatment to two closely related cabbage species on the host-searching and oviposition preference of the diamondback moth (DBM), Plutella xylostella. JA-treated Chinese cabbage (Brassica campestris) was less attractive than untreated Chinese cabbage to ovipositing DBM, while JA-treatment of common cabbage (B. oleracea) made plants more attractive than untreated controls for oviposition by this insect. Similar effects were observed when plants of the two species were damaged by DBM larvae. In the absence of insect-feeding, or JA application, Chinese cabbage is much more attractive to DBM than common cabbage. Inducible resistance therefore appears to occur in a more susceptible plant and induced susceptibility appears to occur in a more resistant plant, suggesting a possible balance mechanism between constitutive and inducible defences to a specialist herbivore.

Boron nutrition and chilling tolerance of warm climate crop species

Background Field observations and glasshouse studies have suggested links between boron (B)-deficiency and leaf damage induced by low temperature in crop plants, but causal relationships between these two stresses at physiological, biochemical and molecular levels have yet to be explored. Limited evidence at the whole-plant level suggests that chilling temperature in the root zone restricts B uptake capacity and/or B distribution/utilization efficiency in the shoot, but the nature of this interaction depends on chilling tolerance of species concerned, the mode of low temperature treatment (abrupt versus gradual temperature decline) and growth conditions (e.g. photon flux density and relative humidity) that may exacerbate chilling stress. Scope This review explores roles of B nutrition in chilling tolerance of continual root or transient shoot chills in crop species adapted to warm season conditions. It reviews current research on combined effects of chilling temperature (ranging from > 0 to 20 degrees C) and B deficiency on growth and B nutrition responses in crop species differing in chilling tolerance. Conclusion For subtropical/tropical species (e.g. cucumber, cassava, sunflower), root chilling at 10-17 degrees C decreases B uptake efficiency and B utilization in the shoot and increases the shoot : root ratio, but chilling-tolerant temperate species (e.g. oilseed rape, wheat) require much lower root chill temperatures (2-5 degrees C) to achieve the same responses. Boron deficiency exacerbates chilling injuries in leaf tissues, particularly under high photon flux density. Suggested mechanisms for B x chilling interactions in plants are: (a) chilling-induced reduction in plasmalemma hydraulic conductivity, membrane fluidity, water channel activity and root pressure, which contribute to the decrease in root hydraulic conductance, water uptake and associated B uptake; (b) chilling-induced stomatal dysfunction affecting B transport from root to shoot and B partitioning in the shoot; and (c) B deficiency induced sensitivity to photo-oxidative damage in leaf cells. However, specific evidence for each of the mechanisms is still lacking. Impacts of B status on chilling tolerance in crop species have important implications for the management of B supply during sensitive stages of growth, such as early growth after planting and early reproductive development, both of which can coincide with the occurrence of chilling temperatures in the field.

Apparent ileal digestibility of amino acids in dietary ingredients for broiler chickens

The apparent ileal digestibility coefficients of amino acids in 107 samples representing 22 food ingredients were determined using 6-week-old broiler chickens. The ingredients assayed included five cereals ( barley, maize, sorghum, triticale and wheat), two cereal by-products ( rice polishings and wheat middlings), four oilseed meals ( canola, cottonseed, soyabean and sunflower meals), full-fat canola, maize gluten meal, four grain legumes ( chickpeas, faba beans,field peas and lupins) and five animal protein sources ( blood, feather,fish, meat and meat and bone meals). The mean ileal digestibility coefficients of amino acids in wheat and maize were higher than those in sorghum, triticale and barley. However, variations observed in individual amino acid digestibilities among samples within cereal type were greater than those determined between cereals. Threonine and lysine were the least digestible indispensable amino acids in the five cereals evaluated. The most digestible indispensable amino acid was phenylalanine in wheat and, leucine in maize and sorghum. In the case of the wheat middlings and rice polishings, threonine was the least digestible indispensable amino acid and arginine was the best digested. In the oilseed meals assayed, amino acid digestibility was highest for soya-bean and sunflower meals, intermediate for canola meal and lowest for cottonseed meal. Ileal digestibility coefficients of amino acids in lupins were found to be slightly lower than those in soya-bean meal. The amino acid digestibilities of field peas, faba beans and chickpeas were considerably lower than those of lupins. Digestibility of arginine was the highest and that of threonine was the lowest of the indispensable amino acids in oilseed meals and grain legumes, except in cottonseed meal. Lysine was the least digestible amino acid in cottonseed meal. In the animal protein sources assayed, digestibility coefficients of amino acids in blood meal were high, intermediate in fish meal, and low in meat meal, meat and bone meal and feather meal. Variation in amino acid digestibility coefficients determined for blood meal samples was small. However, wide variations in amino acid digestibilities were observed for other animal protein sources, highlighting significant batch-to-batch differences. In particular, marked variations were determined for meat meal and meat and bone meal samples. Cystine was the least digested amino acid in animal protein meals, with the exception of blood meal in which isoleucine had the lowest digestibility. The limitations of using apparent digestibility values in diet formulations and the concept of the standardized digestibility system to overcome these limitations are discussed.

Comparison of apparent ileal amino acid digestibility of feed ingredients measured with broilers, layers, and roosters

The apparent ileal digestibility of amino acids in 7 feed ingredients was determined using broilers, layers, and roosters. The ingredients included 3 cereals (wheat, sorghum, and corn), 3 oilseed meals (canola, cottonseed, and soybean meals), and 1 animal protein meal (meat and bone meal). Dietary protein in the assay diets was supplied solely by the test ingredient. All diets contained 20 g/kg of acid-insoluble ash as an indigestible marker, and each diet was offered ad libitum in mash form to 5 replicate pens of broilers and layers, and 4 replicate pens of roosters. The digestibility coefficients of individual amino acids for wheat, corn, and sorghum were higher (P < 0.05) in broilers than in layers and roosters. The digestibility of most amino acids for corn and sorghum was higher (P < 0.05) in roosters compared with those in layers, whereas the digestibility for wheat in layers and roosters was similar. In general, the digestibility of amino acids in canola meal, cottonseed meal, and meat and bone meal was similar among the 3 classes of chickens. The digestibility of amino acids in soybean meal was higher (P < 0.05) for layers compared with those for broilers and roosters but similar between broilers and roosters. These results suggest that the class of chickens significantly influenced the apparent ileal digestibility of amino acids in some feed ingredients.

Do predators aggregate in response to pest density in agroecosystems? Assessing within-field spatial patterns

1. The spatial heterogeneity of predator populations is an important component of ecological theories pertaining to predator-prey dynamics. Most studies within agricultural fields show spatial correlation (positive or negative) between mean predator numbers and prey abundance across a whole field over time but generally ignore the within-field spatial dimension. We used explicit spatial mapping to determine if generalist predators aggregated within a soybean field, the size of these aggregations and if predator aggregation was associated with pest aggregation, plant damage and predation rate. 2. The study was conducted at Gatton in the Lockyer Valley, 90 km west of Brisbane, Australia. Intensive sampling grids were used to investigate within-field spatial patterns. The first row of each grid was located in a lucerne field (10 m from interface) and the remaining rows were in an adjacent soybean field. At each point on the grid the abundance of foliage-dwelling and ground-dwelling pests and predators was measured, predation rates [using sentinel Helicoverpa armigera (Hubner) egg cards] and plant damage were estimated. Eight grids were sampled across two summer cropping seasons (2000/01, 2001/02). 3. Predators exhibited strong spatial patterning with regions of high and low abundance and activity within what are considered to be uniform soybean fields. Ground-dwelling and foliage-dwelling predators were often aggregated in patches approximately 40 m across. 4. Lycosidae (wolf spiders) displayed aggregation and were consistently more abundant within the lucerne, with a decreasing trap catch with distance from the lucrene/soybean interface. This trend was consistent between subsequent grids in a single field and between fields. 5. The large amount of spatial variability in within-field arthropod abundance (pests and predators) and activity (egg predation and plant damage) indicates that whole field averages were misleading. This result has serious implications for sampling of arthropod abundance and pest management decision-making based on scouting data. 6. There was a great deal of temporal change in the significant spatial patterns observed within a field at each sampling time point during a single season. Predator and pest aggregations observed in these fields were generally not stable for the entire season. 7. Predator aggregation did not correlate consistently with pest aggregation, plant damage or predation rate. Spatial patterns in predator abundance were not associated consistently with any single parameter measured. The most consistent positive association was between foliage-dwelling predators and pests (significant in four of seven grids). Inferring associations between predators and prey based on an intensive one-off sampling grid is difficult, due to the temporal variability in the abundance of each group. 8. Synthesis and applications. This study demonstrated that generalist predator populations are rarely distributed randomly and field edges and adjacent crops can have an influence on within-field predator abundance. This must be considered when estimating arthropod (pest and predator) abundance from a set of samples taken at random locations within a field.