Nano-silver treatments reduced bacterial colonization and biofilm formation at the stem-ends of cut gladiolus ‘Eerde’ spikes

Postharvest treatments with nano-silver (NS) alleviate bacteria-related stem blockage of some cut flowers to extend their longevity. Gladiolus (Gladiolus hybridus) is a commercially important cut flower species. For the first time, the effects of NS pulses on cut gladiolus ‘Eerde’ spikes were investigated towards reducing bacterial colonization of and biofilm formation on their stems. As compared with a deionized water (DIW) control, pulse treatments with NS at 10, 25 and 50 mg L−1 for 24 h significantly (P ≤ 0.05) prolonged the vase life of cut gladiolus spikes moved into vases containing DIW. The NS treatments enhanced floret ‘opening rate’ and ‘daily ornamental value’. Although there were no significant differences among NS treatments, a 25 mg L−1 NS pulse treatment tended to give the longest vase life and the best ‘display quality’. All NS pulse treatments significantly improved water uptake by and reduced water loss from flowering spikes, thereby delaying the loss of water balance and maintaining relative fresh weight. Fifty (50) mg L−1 NS pulse-treated cut gladiolus spikes tended to exhibit the most water uptake and highest water balance over the vase period. However, there was no significant difference between 25 and 50 mg L−1 NS pulse treatments. Observations of stem-end bacterial proliferation during the vase period on cut gladiolus spikes either with or without NS pulse treatments were performed by confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). As compared to the control treatment, they revealed that the 25 mg L−1 NS pulse treatment effectively inhibited bacterial colonization and biofilm formation on the stem-end cut surface and in the xylem vessels, respectively. In vitro culture of the bacterial microflora and analysis of biofilm architecture using CLSM revealed that NS treatment restricted bacterial biofilm formation. After static culture for 24 h at 35 °C with 25 mg L−1 NS in the medium, no biofilm form or structure was evident. Rather, only limited bacterial cell number and scanty extracellular polysaccharide (EPS) material were observed. In contrast, mature bacterial biofilm architecture comprised of abundant bacteria interwoven with EPS formed in the absence of NS.

Cellular stages of root formation, root system quality and survival of Pinus elliottii var. elliottii x P. caribaea var. hondurensis cuttings in different temperature environments.

Time to first root in cuttings varies under different environmental conditions and understanding these differences is critical for optimizing propagation of commercial forestry species. Temperature environment (15, 25, 30 or 35 +/- A 2A degrees C) had no effect on the cellular stages in root formation of the Slash x Caribbean Pine hybrid over 16 weeks as determined by histology. Initially callus cells formed in the cortex, then tracheids developed and formed primordia leading to external roots. However, speed of development followed a growth curve with the fastest development occurring at 25A degrees C and slowest at 15A degrees C with rooting percentages at week 12 of 80 and 0% respectively. Cutting survival was good in the three cooler temperature regimes (> 80%) but reduced to 59% at 35A degrees C. Root formation appeared to be dependant on the initiation of tracheids because all un-rooted cuttings had callus tissue but no tracheids, irrespective of temperature treatment and clone.

Cellular stages of root formation, root system quality and survival of Pinuselliottii var. elliottii x P. caribaea var. hondurensis cuttings in different temperature environments.

Time to first root in cuttings varies under different environmental conditions and understanding these differences is critical for optimizing propagation of commercial forestry species. Temperature environment (15, 25, 30 or 352C) had no effect on the cellular stages in root formation of the Slash * Caribbean Pine hybrid over 16 weeks as determined by histology. Initially callus cells formed in the cortex, then tracheids developed and formed primordia leading to external roots. However, speed of development followed a growth curve with the fastest development occurring at 25C and slowest at 15C with rooting percentages at week 12 of 80 and 0% respectively. Cutting survival was good in the three cooler temperature regimes (>80%) but reduced to 59% at 35C. Root formation appeared to be dependant on the initiation of tracheids because all un-rooted cuttings had callus tissue but no tracheids, irrespective of temperature treatment and clone.

Toxic effects of Pb2+ on growth of cowpea (Vigna unguiculata)

A concentration as low as 1 μM lead (Pb) is highly toxic to plants, but previous studies have typically related plant growth to the total amount of Pb added to a solution. In the present experiment, the relative fresh mass of cowpea (Vigna unguiculata) was reduced by 10% at a Pb2+ activity of 0.2 μM for the shoots and at a Pb2+ activity of 0.06 μM for the roots. The primary site of Pb2+ toxicity was the root, causing severe reductions in root growth, loss of apical dominance (shown by an increase in branching per unit root length), the formation of localized swellings behind the root tips (due to the initiation of lateral roots), and the bending of some root tips. In the root, Pb was found to accumulate primarily within the cell walls and intercellular spaces.

An asparaginyl endopeptidase mediates in vivo protein backbone cyclization

Proteases can catalyze both peptide bond cleavage and formation, yet the hydrolysis reaction dominates in nature. This presents an interesting challenge for the biosynthesis of backbone cyclized (circular) proteins, which are encoded as part of precursor proteins and require post-translational peptide bond formation to reach their mature form. The largest family of circular proteins are the plant-produced cyclotides; extremely stable proteins with applications as bioengineering scaffolds. Little is known about the mechanism by which they are cyclized in vivo but a highly conserved Asn (occasionally Asp) residue at the C terminus of the cyclotide domain suggests that an enzyme with specificity for Asn (asparaginyl endopeptidase; AEP) is involved in the process. Nicotiana benthamiana does not endogenously produce circular proteins but when cDNA encoding the precursor of the cyclotide kalata B1 was transiently expressed in the plants they produced the cyclotide, together with linear forms not commonly observed in cyclotide-containing plants. Observation of these species over time showed that in vivo asparaginyl bond hydrolysis is necessary for cyclization. When AEP activity was suppressed, either by decreasing AEP gene expression or using a specific inhibitor, the amount of cyclic cyclotide in the plants was reduced compared with controls and was accompanied by the accumulation of extended linear species. These results suggest that an AEP is responsible for catalyzing both peptide bond cleavage and ligation of cyclotides in a single processing event.

Phosphorus nutrition and tolerance of cotton to water stress I. Seed cotton yield and leaf morphology

Seed cotton yield and morphological changes in leaf growth were examined under drying soil with different phosphorus (P) concentrations in a tropical climate. Frequent soil drying is likely to induce a decrease in nutrients particularly P due to reduced diffusion and poor uptake, in addition to restrictions in available water, with strong interactive effects on plant growth and functioning. Increased soil P in field and in-ground soil core studies increased the seed cotton yield and related morphological growth parameters in a drying soil, with hot (daily maximum temperature >33°C) and dry conditions (relative humidity, 25% to 35%), particularly during peak boll formation and filling stage. The soil water content in the effective rooting zone (top 0.4 m) decreased to -1.5 MPa by day 5 of the soil drying cycle. However, the increased seed cotton yield for the high-P plants was closely related to increasing leaf area with increased P supply. Plant height, leaf fresh mass and leaf area per plant were positively related to the leaf P%, which increased with increasing P supply. Low P plants were lower in plant height, leaf area, and leaf tissue water in the drying soil. Individual leaf area and the water content of the fresh leaf (ratio of dry mass to fresh mass) were significantly dependent on leaf P%.

The influence of temperature on the foaming of milk

The effect of temperature (5-85 °C) on the foaming properties of cows' milk was investigated. The foaming properties of milk as a function of temperature varied considerably depending on fat content and the processing conditions used in manufacture. Skim milk foams were most stable when formed at 45 °C. Milk fat had a detrimental effect on foam formation and stability of whole milk especially in the temperature range 15-45 °C. The detrimental effects of milk fat on foaming properties were reduced by homogenization and ultra-high-temperature (UHT) treatment. No correlation was observed between foam formation and surface tension of whole milk in the temperature range 15-45 °C. There was a pronounced difference in the bubble size distributions of whole milk and skim milk especially at half-life of the foams. Bubbles in whole milk foams were smaller and showed a higher degree of rupture as a result of coalescence than those in skim milk foams.

Developing sustainable banana production systems: a case study from tropical Australia

The north Queensland banana industry is under pressure from government and community expectations to exhibit good environmental stewardship. The industry is situated on the high-rainfall north Queensland coast adjacent to 2 natural icons, the Great Barrier Reef to the east and World Heritage-listed rain forest areas to the west. The main environmental concern is agricultural industry pollutants harming the Great Barrier Reef. In addition to environmental issues the banana industry also suffers financial pressure from declining margins and production loss from tropical cyclones. As part of a broader government strategy to reduce land-based pollutants affecting the Great Barrier Reef, the formation of a pilot banana producers group to address these environmental and economic pressures was facilitated. Using an integrated farming systems approach, we worked collaboratively with these producers to conduct an environmental risk assessment of their businesses and then to develop best management practices (BMP) to address environmental concerns. We also sought input from technical experts to provide increased rigour for the environmental risk assessment and BMP development. The producers' commercial experience ensured new ideas for improved sustainable practices were constantly assessed through their profit-driven 'filter' thus ensuring economic sustainability was also considered. Relying heavily on the producers' knowledge and experience meant the agreed sustainable practices were practical, relevant and financially feasible for the average-sized banana business in the region. Expert input and review also ensured that practices were technically sound. The pilot group producers then implemented and adapted selected key practices on their farms. High priority practices addressed by the producers group included optimizing nitrogen fertilizer management to reduce runoff water nitrification, developing practical ground cover management to reduce soil erosion and improving integrated pest management systems to reduce pesticide use. To facilitate wider banana industry understanding and adoption of the BMP's developed by the pilot group, we conducted field days at the farms of the pilot group members. Information generated by the pilot group has had wider application to Australian horticulture and the process has been subsequently used with the north Queensland sugar industry. Our experiences have shown that integrated farming systems methodologies are useful in addressing complex issues like environmental and economic sustainability. We have also found that individual horticulture businesses need on-going technical support for change to more sustainable practices. One-off interventions have little impact, as farm improvement is usually an on-going incremental process. A key lesson from this project has been the need to develop practical, farm scale economic tools to clarify and demonstrate the financial impact of alternative management practices. Demonstrating continued profitability is critical to encourage widespread industry adoption of environmentally sustainable practices

Differing mechanisms of simple nitrile formation on glucosinolate degradation in Lepidium sativum and Nasturtium officinale seeds.

Glucosinolates are sulphur-containing glycosides found in brassicaceous plants that can be hydrolysed enzymatically by plant myrosinase or non-enzymatically to form primarily isothiocyanates and/or simple nitriles. From a human health perspective, isothiocyanates are quite important because they are major inducers of carcinogen-detoxifying enzymes. Two of the most potent inducers are benzyl isothiocyanate (BITC) present in garden cress (Lepidium sativum), and phenylethyl isothiocyanate (PEITC) present in watercress (Nasturtium officinale). Previous studies on these salad crops have indicated that significant amounts of simple nitriles are produced at the expense of the isothiocyanates. These studies also suggested that nitrile formation may occur by different pathways: (1) under the control of specifier protein in garden cress and (2) by an unspecified, non-enzymatic path in watercress. In an effort to understand more about the mechanisms involved in simple nitrile formation in these species, we analysed their seeds for specifier protein and myrosinase activities, endogenous iron content and glucosinolate degradation products after addition of different iron species, specific chelators and various heat treatments. We confirmed that simple nitrile formation was predominantly under specifier protein control (thiocyanate-forming protein) in garden cress seeds. Limited thermal degradation of the major glucosinolate, glucotropaeolin (benzyl glucosinolate), occurred when seed material was heated to >120 degrees C. In the watercress seeds, however, we show for the first time that gluconasturtiin (phenylethyl glucosinolate) undergoes a non-enzymatic, iron-dependent degradation to a simple nitrile. On heating the seeds to 120 degrees C or greater, thermal degradation of this heat-labile glucosinolate increased simple nitrile levels many fold.

Reduced efficacy of macrocyclic lactone treatments in controlling gastrointestinal nematode infections of weaner dairy calves in subtropical eastern Australia.

Faecal Egg Count Reduction Tests (FECRTs) for macrocyclic lactone (ML) and levamisole (LEV) drenches were conducted on two dairy farms in the subtropical, summer rainfall region of eastern Australia to determine if anthelmintic failure contributed to severe gastrointestinal nematode infections observed in weaner calves. Subtropical Cooperia spp. were the dominant nematodes on both farms although significant numbers of Haemonchus placei were also present on Farm 2. On Farm 1, moxidectin pour-on (MXD) drenched at 0.5 mg kg-1 liveweight (LW) reduced the overall Cooperia burden by 82% (95% confidence limits, 37-95%) at day 7 post-drench. As worm burdens increased rapidly in younger animals in the control group (n = 4), levamisole was used as a salvage drench and these calves withdrawn from the trial on animal welfare grounds after sample collection at day 7. Levamisole (LEV) dosed at 6.8 mg kg-1 LW reduced the worm burden in these calves by 100%, 7 days after drenching. On Farm 2, MXD given at 0.5 mg kg-1 LW reduced the faecal worm egg count of cooperioids at day 8 by 96% (71-99%), ivermectin oral (IVM) at 0.2 mg kg-1 LW by 1.6% (-224 to 70%) and LEV oral at 7.1 mg kg-1 LW by 100%. For H. placei the reductions were 98% (85-99.7%) for MXD, 0.7% (-226 to 70%) for IVM and 100% for LEV. This is the first report in Australia of the failure of macrocyclic lactone treatments to control subtropical Cooperia spp. and suspected failure to control H. placei in cattle.

Ageing delays the cellular stages of adventitious root formation in pine.

Vegetative propagation programs internationally are affected by the significant decline of rooting success as trees mature. This study compared the cellular stages of root formation in stem cuttings from 15-week-old (juvenile) and 9-y-old (mature) stock plants of the slash x Caribbean pine hybrid (Pinus elliottii var. elliottii x P. caribaea van hondurensis). The cellular stages of root formation were the same in both juvenile and mature cuttings, beginning with cell divisions of the vascular cambium forming callus tissue. Within the callus, tracheids differentiated and elongated to form root primordia. Roots in juvenile cuttings developed faster than those in mature cuttings and the juvenile cuttings had a much higher rooting percent at the end of the study (92% and 26% respectively). Cuttings of the two juvenile genotypes had more primary roots (5.5 and 3.3) than the three mature genotypes (0.96, 0.18 and 0.07). The roots of juvenile cuttings were more evenly distributed around the basal circumference when compared with those on cuttings from the mature genotypes. Further work is needed to improve understanding of physiological changes with maturation so that the rooting success and the speed of development in cuttings from mature stock plants can be optimised, hence improving genetic gain.

New Modified Dipping Method Using 4-hexylresorcinol for Preventing Blackspot Formation in Prawns.

Industry acceptance of alternative sulphite treatments for preventing blackspot in prawns is limited. This study demonstrated that dipping prawns (Penaeus monodon and Fenneropenaeus merguiensis) in a less concentrated solution for longer times provides better control of blackspot formation than the recommended 2 minute dip in 50 mg/L 4-hexylresorcinol. Protection improved as modified dipping times and solution concentrations increased. Blackspot protection provided by most of the modified dip methods was more effective than a 1 minute dip in 1% sodium metabisulphite. The recommended 2 minute dip method should be limited to 125 kg when used on consecutive 25-kg batches of prawns. Yields increased by less than 10% even after 72-hour exposure in the modified dip. The new dipping method meets most international residue standards when applying the modified treatments evaluated.

Relationships between Indole-3-butyric Acid, Photoinhibition and Adventitious rooting of Corymbia torelliana, C. citriodora and F1 Hybrid Cuttings

Rooted cutting propagation is widely used for maximising tree yield, quality and uniformity in conjunction with clonal selection. Some eucalypt species are deployed as rooted cuttings but many are considered to difficult to root. This study examined IBA effects on photoinhibition, root formation, mortality and root and shoot development in cuttings of Corymbia torelliana, C. citriodora and their hybrids. IBA had little or no effect on photoinhibition but it had strong, dose-dependent effects on root formation and mortality. IBA frequently increases primary root number of rooted cutting but it did not increase total root weight, length, surface area or volume, possibly because the highest doe (8g IBA/kg IBA/kg powder) caused leaf abscission and sometimes reduced leaf area (by 55-79%)or shoot dry weight (by 40-58%). An intermediate dose (3g IBA/kg powder) most consistnely improved root formation with little or no effect on mortality or shoot development. Across the F1 hybrid families this treatment increased the number of rooted cuttings by 72-121% and more than ddoubled the number of primary roots per rooted cutting (from 1.1-1.7 roots to 3.5-4.1 roots). This simple treatment will facilitate commercial multiplication of superior individuals or selected families of C. torelliana x C. citriodora through a vegetative propagation system.

Infection and disease development of Quambalaria spp. on Corymbia and Eucalyptus spp.

Quambalaria spp. are eucalypt leaf and shoot pathogens of growing global importance, yet virtually nothing is known regarding the manner in which they infect and colonize their hosts. A study of the infection process of Q. pitereka and Q.eucalypti on Corymbia and Eucalyptus species was thus undertaken using light, scanning and transmission electron microscopy after artificial inoculation. Conidial germination was triggered when relative humidity levels exceeded 90% and commenced within 2 h in the presence of free water. Light reduced germination but did not prevent germination from occurring. Conidial germination and hyphal growth occurred on the upper and lower leaf surfaces with penetration occurring via the stomata or wounds on the leaf surface or juvenile stems. There was no evidence of direct penetration of the host. Following penetration through the stomata, Q. pitereka and Q. eucalypti hyphae grew only intercellularly without the formation of haustoria or interaction apparatus, which is characteristic of the order Microstromatales. Instead, the presence of an interaction zone is demonstrated in this paper. Conidiophores arose through stomatal openings producing conidia 7 days after infection.

Reduced risk of acute poisoning in Australian cattle from used motor oils after introduction of lead-free petrol.

Lead (Pb) poisoning of cattle has been relatively common in Australia and sump oil has been identified as an important cause of Pb toxicity for cattle because they seem to have a tendency to drink it. Lead-free petrol has been available in Australia since 1975, so the aim of this study was to assess the current risk to cattle from drinking used automotive oils. Sump or gear box oil was collected from 56 vehicles being serviced. The low levels of Pb found suggest that the removal of leaded petrol from the Australian market as a public health measure has benefited cattle by eliminating the risk of acute poisoning from used engine oil.