The process for implementation of a Quality Management System within a multi-functional cereal laboratory

The intent of this study was to design, document and implement a Quality Management System (QMS) into a laboratory that incorporated both research and development (R&D) and routine analytical activities. In addition, it was necessary for the QMS to be easily and efficiently maintained to: (a) provide documented evidence that would validate the system's compliance with a certifiable standard, (b) fit the purpose of the laboratory, (c) accommodate prevailing government policies and standards, and (d) promote positive outcomes for the laboratory through documentation and verification of the procedures and methodologies implemented. Initially, a matrix was developed that documented the standards' requirements and the necessary steps to be made to meet those requirements. The matrix provided a check mechanism on the progression of the system's development. In addition, it was later utilised in the Quality Manual as a reference tool for the location of full procedures documented elsewhere in the system. The necessary documentation to build and monitor the system consisted of a series of manuals along with forms that provided auditable evidence of the workings of the QMS. Quality Management (QM), in one form or another, has been in existence since the early 1900's. However, the question still remains: is it a good thing or just a bugbear? Many of the older style systems failed because they were designed by non-users, fiercely regulatory, restrictive and generally deemed to be an imposition. It is now considered important to foster a sense of ownership of the system by the people who use the system. The system's design must be tailored to best fit the purpose of the operations of the facility if maximum benefits to the organisation are to be gained.

New microsatellite loci for Carcharhinid sharks (Carcharhinus tilstoni and C. sorrah) and their cross-amplification in other shark species

Twelve microsatellite DNA markers were isolated in the spot-tail shark (Carcharhinus sorrah) and nine were isolated in Australian black-tip shark (Carcharhinus tilstoni). These loci plus 18 others developed for sharks from the genera Negaprion, Ginglymostoma, Carcharodon and Isurus were tested for amplification success on four species of Carcharhinus (including C. sorrah and C. tilstoni) and four other species representing three diverse families. Cross-amplification was most common within families. Five loci were subsequently tested for polymorphism on 50 C. sorrah and 60 C. tilstoni. The number of alleles per locus was two to 24 and the average heterozygosity was 0.54 (range 0.16-0.87) for C. sorrah and 0.64 (range 0.44-0.78) for C. tilstoni. These loci may be useful tools for genetic analyses of the Carcharhinidae.

The introduction and release of Chiasmia inconspicua and C. assimilis (Lepidoptera: Geometridae) for the biological control of Acacia nilotica in Australia

Two geometrid moths Chiasmia inconspicua and Chiasmia assimilis, identified as potential biological control agents for prickly acacia Acacia nilotica subsp. indica, were collected in Kenya and imported into quarantine facilities in Australia where laboratory cultures were established. Aspects of the biologies of both insects were studied and CLIMEX® models indicating the climatically favourable areas of Australia were developed. Host range tests were conducted using an approved test list of 74 plant species and no-choice tests of neonate larvae placed on both cut foliage and potted plants. C. inconspicua developed through to adult on prickly acacia and, in small numbers, Acacia pulchella. C. assimilis developed through to adult on prickly acacia and also in very small numbers on A. pulchella, A. deanei, A. decurrens, and A. mearnsii. In all experiments, the response on prickly acacia could be clearly differentiated from the responses on the non-target species. Both insects were approved for release in Australia. Over a three-year period releases were made at multiple sites in north Queensland, almost all in inland areas. There was no evidence of either insect's establishment and both colonies were terminated. A new colony of C. assimilis was subsequently established from insects collected in South Africa and releases of C. assimilis from this new colony were made into coastal and inland infestations of prickly acacia. Establishment was rapid at one coastal site and the insect quickly spread to other infestations. Establishment at one inland area was also confirmed in early 2006. The establishment in coastal areas supported a CLIMEX model that indicated that the climate of coastal areas was more suitable than inland areas.

Construction of microsatellite linkage maps for Corymbia

The genus Corymbia is closely related to the genus Eucalyptus, and like Eucalyptus contains tree species that are important for sub-tropical forestry. Corymbia's close relationship with Eucalyptus suggests genetic studies in Corymbia should benefit from transfer of genetic information from its more intensively studied relatives. Here we report a genetic map for Corymbia spp. based on microsatellite markers identified de novo in Corymbia sp or transferred from Eucalyptus. A framework consensus map was generated from an outbred F 2 population (n = 90) created by crossing two unrelated Corymbia torelliana x C. citriodora subsp. variegata F1 trees. The map had a total length of 367 cM (Kosambi) and was composed of 46 microsatellite markers distributed across 13 linkage groups (LOD 3). A high proportion of Eucalyptus microsatellites (90%) transferred to Corymbia. Comparative analysis between the Corymbia map and a published Eucalyptus map identified eight homeologous linkage groups in Corymbia with 13 markers mapping on one or both maps. Further comparative analysis was limited by low power to detect linkage due to low genome coverage in Corymbia, however, there was no convincing evidence for chromosomal structural differences because instances of non-synteny were associated with large distances on the Eucalyptus map. Segregation distortion was primarily restricted to a single linkage group and due to a deficit of hybrid genotypes, suggesting that hybrid inviability was one factor shaping the genetic composition of the F2 population in this inter-subgeneric hybrid. The conservation of microsatellite loci and synteny between Corymbia and Eucalyptus suggests there will be substantial value in exchanging information between the two groups.

Genetic variation within two sympatric spotted gum eucalypts exceeds between taxa variation

Population substructure and hybridization, among other factors, have the potential to cause erroneous associations in linkage disequilibrium (LD) mapping. Two closely related spotted gum eucalypts, Corymbia variegata and C. henryi (Myrtaceae) occur in sympatry in the east coast of Australia and potentially interbreed. They are morphologically similar but are distinguished as separate species based on capsule and foliage size. To determine whether they hybridize in nature and its implications for LD mapping, we investigated the level of molecular divergence between the two species at two sympatric locations separated by 300 kilometres. Very few individuals of intermediate morphology were identified, despite the two species occurring only metres apart. Analysis of genetic structure using 12 microsatellite loci showed that genetic differentiation between populations of the same species at different locations (FST = 0.07 for both species; p = 0.0001) was significantly higher than that observed between species at each location (mean FST = 0.02 and 0.04 for Cherry tree and Bunyaville respectively; p = 0.0001; all Mann-Whitney U-test p ≤ 0.01). No species-specific alleles or significant allele frequency differences were detected within a site, suggesting recurrent local gene flow between the two species. The lack of significant allele frequency differences implies no population stratification along taxonomic lines. This suggested that there is little concern for cryptic hybridization when sampling from sites of sympatry for LD mapping.

In vitro propagation of Corymbia torelliana x C. citriodora (Myrtaceae) via cytokinin-free node culture

Hybrids between Corymbia torelliana (F.Muell.) K.D.Hill & L.A.S.Johnson and C. citriodora subsp. variegata (F.Muell.) A.R.Bean & M.W.McDonald are used extensively to establish forestry plantations in subtropical Australia. Methods were developed for in vitro seed germination, shoot multiplication and plantlet formation that could be used to establish in vitro and ex vitro clone banks of juvenile Corymbia hybrids. Effects of sodium hypochlorite concentration and exposure time on seed contamination and germination, and effects of cytokinin and auxin concentrations on shoot multiplication and subsequent rooting, were assessed. A two-step surface sterilisation procedure, involving 70% ethanol followed by 1% sodium hypochlorite, provided almost no contamination and at least 88% germination. A novel method of cytokinin-free node culture proved most effective for in vitro propagation. Lateral bud break of primary shoots was difficult to induce by using cytokinin, but primary shoots rooted prolifically, elongated rapidly and produced multiple nodes in the absence of exogenous cytokinin. Further multiplication was obtained either by elongating lateral shoots of nodal explants in cytokinin-free medium or by inducing organogenic callus and axillary shoot proliferation with 2.2 µm benzyladenine. Plantlets were produced using an in vitro soil-less method that provided extensive rooting in sterile propagation mixture. These methods provide a means for simultaneous laboratory storage and field-testing of clones before selection and multiplication of desired genotypes.

Salinity tolerance of twelve hybrid Bermudagrass (Cynodon Dactylon (L.) Pers. x C. Transvaalensis Burtt Davy) genotypes

Salinity is an increasingly important issue in both rural and urban areas throughout much of Australia. The use of recycled/reclaimed water and other sources of poorer quality water to irrigate turf is also increasing. Hybrid Bermudagrass (Cynodon dactylon (L.) Pers. x C. transvaalensis Burtt Davey), together with the parent species C. dactylon, are amongst the most widely used warm-season turf grass groups. Twelve hybrid Bermudagrass genotypes and one accession each of Bermudagrass (C. dactylon), African Bermudagrass (C. transvaalensis) and seashore paspalum (Paspalum vaginatum Sw.) were grown in a glasshouse experiment with six different salinity treatments applied hydroponically through the irrigation water (ECW = <0.1, 6, 12, 18, 24 or 30 dSm-1) in a flood-and-drain system. Each pot was clipped progressively at 2-weekly intervals over the 12-week experimental period to determine dry matter production; leaf firing was rated visually on 3 occasions during the last 6 weeks of salinity treatment. At the end of the experiment, dry weights of roots and crowns below clipping height were also determined. Clipping yields declined sharply after about the first 6 weeks of salinity treatment, but then remained stable at substantially lower levels of dry matter production from weeks 8 to 12. Growth data over this final 4-week experimental period is therefore a more accurate guide to the relative salinity tolerance of the 15 entries than data from the preceding 8 weeks. Based on these data, the 12 hybrid Bermudagrass genotypes showed moderate salinity tolerance, with FloraDwarfM, 'Champion Dwarf', NovotekM and 'TifEagle' ranking as the most salt tolerant and 'Patriot', 'Santa Ana', 'Tifgreen' and TifSport M the least tolerant within the hybrid group. Nevertheless, Santa Ana, for example, maintained relatively strong root growth as salinity increased, and so may show better salt tolerance in practice than predicted from the growth data alone. The 12 hybrid Bermudagrasses and the single African Bermudagrass genotype were all ranked above FloraTeXM Bermudagrass in terms of salt tolerance. However, seashore paspalum, which is widely acknowledged as a halophytic species showing high salt tolerance, ranked well above all 14 Cynodon genotypes in terms of salinity tolerance.

Factors contributing to wear tolerance of Bermudagrass (Cynodon dactylon (L.) Pers., C. dactylon x C. transvaalensis Burtt-Davey) on a sand-based profile under simulated sports field conditions

Wear resistance and recovery of 8 Bermudagrass (Cynodon dactylon (L.) Pers.) and hybrid Bermudagrass (C. Dactylon x C. transvaalensis Burtt-Davey) cultivars grown on a sandbased soil profile near Brisbane, Australia, were assessed in 4 wear trials conducted over a two year period. Wear was applied on a 7-day or a 14-day schedule by a modified Brinkman Traffic Simulator for 6-14 weeks at a time, either during winter-early spring or during summer-early autumn. The more frequent wear under the 7-day treatment was more damaging to the turf than the 14-day wear treatment, particularly during winter when its capacity for recovery from wear was severely restricted. There were substantial differences in wear tolerance among the 8 cultivars investigated, and the wear tolerance rankings of some cultivars changed between years. Wear tolerance was associated with high shoot density, a dense stolon mat strongly rooted to the ground surface, high cell wall strength as indicated by high total cell wall content, and high levels of lignin and neutral detergent fiber. Wear tolerance was also affected by turf age, planting sod quality, and wet weather. Resistance to wear and recovery from wear are both important components of wear tolerance, but the relative importance of their contributions to overall wear tolerance varies seasonally with turf growth rate.

Morphological and developmental comparisons of seven greens quality hybrid bermudagrass (Cynodon dactylon (L.)Pers.x C transvaalensis Burtt-Davy

Fine-textured hybrid bermudagrass [Cynodon dactylon (L.) Pers. x C. transvaalensis Burtt-Davy] cultivars have been widely used for golf putting greens and lawn bowls greens in warm-climate areas for more than 40 years. During the past decade, the choice of cultivar for professional turfgrass managers has been expanded by a range of secondgeneration hybrid bermudagrasses, which differ from the first-generation cultivars ‘Tifgreen’ and ‘Tifdwarf ’ in their management requirements. In this paper, we present comparative morphological and developmental data for seven cultivars (Champion Dwarf, FloraDwarf, MS-Supreme, Novotek, Tifdwarf, TifEagle, Tifgreen) grown in spaced plant and sward experiments at Cleveland, Australia (27º32’S lat, 153º15’E long, 25 masl). The four ‘ultradwarf ’ cultivars (Champion Dwarf, MS-Supreme, FloraDwarf, TifEagle) showed slower vertical extension and produced fewer inflorescences than Tifdwarf, Tifgreen, and Novotek. However, in terms of the length of stolon internodes and their overall rate of lateral spread, Champion Dwarf, FloraDwarf, and TifEagle were comparable to Tifdwarf; MS-Supreme (with longer internodes) spread faster laterally, though slower than Tifgreen (which had the longest stolon internodes). In unmown swards, the four ultradwarfs produced shorter leaves than Tifgreen, Tifdwarf, and Novotek, but only Champion Dwarf produced significantly narrower leaves than Tifgreen, Tifdwarf, and Novotek, with TifEagle leaves also significantly narrower than those of Tifgreen and Novotek. Minimum threshold temperatures for growth were approximately 9° to 10°C (air temperature) and 15° to 16°C at 10 cm soil depth.

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.

Towards better management of Australia’s shark fishery: genetic analyses reveal unexpected ratios of cryptic blacktip species Carcharhinus tilstoni and C. limbatus

The common blacktip shark (Carcharhinus limbatus) and the Australian blacktip shark (C. tilstoni) are morphologically similar species that co-occur in subtropical and tropical Australia. In striking contrast to what has been previously reported, we demonstrate that the common blacktip shark is not rare in northern Australia but occurs in approximately equal frequencies with the Australian blacktip shark. Management of shark resources in northern Australia needs to take account of this new information. Species identification was performed using nucleotide sequences of the control, NADH dehydrogenase subunit 4 (ND4) and cytochrome oxidase I (COI) regions in the mitochondrial genome. The proportion of overall genetic variation (FST) between the two species was small (0.042, P < 0.01) based on allele frequencies at five microsatellite loci. We confirm that a third blacktip species (C. amblyrhynchoides, graceful shark) is closely related to C. tilstoni and C. limbatus and can be distinguished from them on the basis of mtDNA sequences from two gene regions. The Australian blacktip shark (C. tilstoni) was not encountered among 20 samples from central Indonesia that were later confirmed to be common blacktip and graceful sharks. Fisheries regulators urgently need new information on life history, population structure and morphological characters for species identification of blacktip shark species in Australia.

Soil C stocks in grains and sugar farming systems

Quantify soil C stocks in grains and sugarcane cropping systems of Queensland, including impacts of management practices.

Evaluation of wood characteristics of tropical post-mid rotation plantation Eucalyptus cloeziana and E. pellita: Part (c) Wood quality and structural properties.

This study provides information about wood quality, structural properties, processing characterists and product suitability of wood harvested from fast-grown hardwood plantations. Wood quality attributes tested included density, extractive content, unit shrinkage, heartwood proportion and sapwood width. Structural properties tested included small clear and full section strength and stiffness, hardness, joint group, visual grade assessment and natural vibration-based grade assessment. The variation between the inner, intermediate and outer heartwood zones and the variation between provenances was also tested. Overall, the wood qualtiy attributes measured for 19 year-old E. cloeziana and 15 year-old E. pellita plantation material fall between those expected from the wood of mature, native forest trees and those found in younger plantation material of the same species.

Reciprocal and advanced generation hybrids between Corymbia citriodora and C-torelliana: forestry breeding and the risk of gene flow

Corymbia F1 hybrids have high potential for plantation forestry; however, little is known of their reproductive biology and potential for genetic pollution of native Corymbia populations. This study aims to quantify the influence of reproductive isolating barriers on the success of novel reciprocal and advanced generation Corymbia hybrids. Two maternal taxa, Corymbia citriodora subsp. citriodora and Corymbia torelliana, were pollinated using five paternal taxa, C. citriodora subsp. citriodora, C. torelliana, one C. torelliana x C. citriodora subsp. citriodora hybrid and two C. torelliana x C. citriodora subsp. variegata hybrids. Pollen tube, embryo and seed development were assessed. Reciprocal hybridisation between C. citriodora subsp. citriodora and C. torelliana was successful. Advanced generation hybrids were also created when C. citriodora subsp. citriodora or C. torelliana females were backcrossed with F1 hybrid taxa. Prezygotic reproductive isolation was identified via reduced pollen tube numbers in the style and reduced numbers of ovules penetrated by pollen tubes. Reproductive isolation was weakest within the C. citriodora subsp. citriodora maternal taxon, with two hybrid backcrosses producing equivalent capsule and seed yields to the intraspecific cross. High hybridising potential was identified between all Corymbia species and F1 taxa studied. This provides opportunities for advanced generation hybrid breeding, allowing desirable traits to be amplified. It also indicates risks of gene flow between plantation and native Corymbia populations.

Alginate encapsulation of shoot tips and nodal segments for short-term storage and distribution of the eucalypt Corymbia torelliana × C. citriodora

A protocol was developed for short-term preservation and distribution of the plantation eucalypt, Corymbia torelliana × C. citriodora, using alginate-encapsulated shoot tips and nodes as synthetic seeds. Effects of sowing medium, auxin concentration, storage temperature and planting substrate on shoot regrowth or conversion into plantlets were assessed for four different clones. High frequencies of shoot regrowth (76–100%) from encapsulated explants were consistently obtained in hormone-free half- and full-strength Murashige and Skoog (MS) sowing media. Conversion into plantlets from synthetic seeds was achieved on half-strength MS medium by treating shoot tips or nodes with 4.9–78.4 μM IBA prior to encapsulation. Pre-treatment with 19.6 μM IBA provided 62–100% conversion, and 95–100% of plantlets survived after acclimatisation under nursery conditions. Synthetic seeds containing explants pre-treated with IBA were stored for 8 weeks much more effectively at 25°C than at 4°C, with regrowth frequencies of 50–84% at 25°C compared with 0–4% at 4°C. To eliminate the in vitro culture step after encapsulation, synthetic seeds were allowed to pre-convert before sowing directly onto a range of ex vitro non-sterile planting substrates. Highest frequencies (46–90%) of plantlet formation from pre-converted synthetic seeds were obtained by transferring shoot tip-derived synthetic seeds onto an organic compost substrate. These plantlets exhibited almost 100% survival in the nursery without mist irrigation. Pre-conversion of non-embryonic synthetic seeds is a novel technique that provides a convenient alternative to somatic embryo-derived artificial seeds.