Heritability estimates for growth in the tropical abalone Haliotis asinina using microsatellites to assign parentage

The tropical abalone Haliotis asinina is a wild-caught and cultured species throughout the Indo-Pacific as well as being an emerging model species for the study of haliotids. H. asinina has the fastest recorded natural growth rate of any abalone and reaches sexual maturity within one year. As such, it is a suitable abalone species for selective breeding for commercially important traits such as rapid growth. Estimating the amount of variation in size that is attributable to heritable genetic differences can assist the development of such a selective breeding program. Here we estimated heritability for growth-related traits at 12 months of age by creating a single cohort of 84 families in a full-factorial mating design consisting of 14 sires and 6 dams. Of 500 progeny sampled, 465 were successfully assigned to their parents based on shared alleles at 5 polymorphic microsatellite loci. Using an animal model, heritability estimates were 0.48 ± 0.15 for shell length, 0.38 ± 0.13 for shell width and 0.36 ± 0.13 for weight. Genetic correlations were > 0.98 between shell parameters and weight, indicating that breeding for weight gains could be successfully achieved by selecting for shell length.

Quantitative and molecular genetics of juvenile wood traits in radiata and slash/Caribbean pines.

The Juvenile Wood Initiative (JWI) project has been running successfully since July 2003 under a Research Agreement with FWPA and Letters of Association with the consortium partners STBA (Southern Tree Breeding Association), ArborGen and FPQ (Forestry Plantations Queensland). Over the last five and half years, JWI scientists in CSIRO, FPQ, and STBA have completed all 12 major milestones and 28 component milestones according to the project schedule. We have made benchmark progress in understanding the genetic control of wood formation and interrelationships among wood traits. The project has made 15 primary scientific findings and several results have been adopted by industry as summarized below. This progress was detailed in 10 technical reports to funding organizations and industry clients. Team scientists produced 16 scientific manuscripts (8 published, 1 in press, 2 submitted, and several others in the process of submission) and 15 conference papers or presentations. Primary Scientific Findings. The 15 major scientific findings related to wood science, inheritance and the genetic basis of juvenile wood traits are: 1. An optimal method to predict stiffness of standing trees in slash/Caribbean pine is to combine gravimetric basic density from 12 mm increment cores with a standing tree prediction of MoE using a time of flight acoustic tool. This was the most accurate and cheapest way to rank trees for breeding selection for slash/Caribbean hybrid pine. This method was also recommended for radiata pine. 2. Wood density breeding values were predicted for the first time in the STBA breeding population using a large sample of 7,078 trees (increment cores) and it was estimated that selection of the best 250 trees for deployment will produce wood density gains of 12.4%. 3. Large genetic variation for a suite of wood quality traits including density, MFA, spiral grain, shrinkage, acoustic and non-acoustic stiffness (MoE) for clear wood and standing trees were observed. Genetic gains of between 8 and 49% were predicted for these wood quality traits with selection intensity between 1 to 10% for radiata pine. 4. Site had a major effect on juvenile-mature wood transition age and the effect of selective breeding for a shorter juvenile wood formation phase was only moderate (about 10% genetic gain with 10% selection intensity, equivalent to about 2 years reduction of juvenile wood). 5. The study found no usable site by genotype interactions for the wood quality traits of density, MFA and MoE for both radiata and slash/Caribbean pines, suggesting that assessment of wood properties on one or two sites will provide reliable estimates of the genetic worth of individuals for use in future breeding. 6. There were significant and sizable genotype by environment interactions between the mainland and Tasmanian regions and within Tasmania for DBH and branch size. 7. Strong genetic correlations between rings for density, MFA and MoE for both radiata and slash/Caribbean pines were observed. This suggests that selection for improved wood properties in the innermost rings would also result in improvement of wood properties in the subsequent rings, as well as improved average performance of the entire core. 8. Strong genetic correlations between pure species and hybrid performance for each of the wood quality traits were observed in the hybrid pines. Parental performance can be used to identify the hybrid families which are most likely to have superior juvenile wood properties of the slash/Caribbean F1 hybrid in southeast Queensland. 9. Large unfavourable genetic correlations between growth and wood quality traits were a prominent feature in radiata pine, indicating that overcoming this unfavourable genetic correlation will be a major technical issue in progressing radiata pine breeding. 10. The project created the first radiata pine 18 k cDNA microarray and generated 5,952 radiata pine xylogenesis expressed sequence tags (ESTs) which assembled into 3,304 unigenes. 11. A total of 348 genes were identified as preferentially expressed genes in earlywood or latewood while a total of 168 genes were identified as preferentially expressed genes in either juvenile or mature wood. 12. Juvenile earlywood has a distinct transcriptome relative to other stages of wood development. 13. Discovered rapid decay of linkage disequilibrium (LD) in radiata pine with LD decaying to approximately 50% within 1,700 base pairs (within a typical gene). A total of 913 SNPS from sequencing 177,380 base pairs were identified for association genetic studies. 14. 149 SNPs from 44 genes and 255 SNPs from a further 51 genes (total 95 genes) were selected for association analysis with 62 wood traits, and 30 SNPs were shortlisted for their significant association with variation of wood quality traits (density, MFA and MoE) with individual significant SNPs accounting for between 1.9 and 9.7% of the total genetic variation in traits. 15. Index selection using breeding objectives was the most profitable selection method for radiata pine, but in the long term it may not be the most effective in dealing with negative genetic correlations between wood volume and quality traits. A combination of economic and biological approaches may be needed to deal with the strong adverse correlation.

Successful fertility experiments with cryopreserved spermatozoa of barramundi, Lates calcarifer (Bloch), using dimethylsulfoxide and glycerol as cryoprotectants

The fertility of cryopreserved Lates calcarifer sperm was studied to increase the availability of semen for routine fertilization of stripped eggs and to provide a tool for selective breeding. Semen diluted (1:4 v/v) and frozen (-196 degrees C) with 5% dimethylsulfoxide (DMSO) or 10% glycerol (final concentration) as cryoprotectants was used to inseminate freshly stripped ova. Frozen-thawed sperm were motile for about 4 min after being mixed with seawater. In the DMSO medium, post-thaw sperm activation was immediate after dilution with seawater, but in the glycerol medium maximum motility intensity was delayed for up to 1 min. When eggs and sperm were mixed before the addition of seawater, semen frozen with DMSO as cryoprotectant gave a mean hatch rate (84.1%) no different (P > 0.05) from that of unfrozen semen diluted with Ringer's solution (80.7%) or with DMSO (83.7%), but higher (P < 0.05) than that of semen frozen with glycerol (60.9%). Adding sperm to seawater 30 s before mixing with eggs did not improve the fertility of sperm cryopreserved with glycerol. Eggs inseminated with glycerol-cryoprotected sperm showed higher mortality during incubation than those inseminated with DMSO-cryoprotected sperm. Sperm held in liquid nitrogen for 90 days with DMSO as cryoprotectant yielded acceptable fertilization and hatching rates with semen-to-ova ratios of up to 1:100 (v/v) , and produced fish with no apparent abnormalities over a 29-day period after hatch. These results show that cryopreservation of L. calcarifer sperm is feasible and well suited to a variety of hatchery purposes.

Increasing seedstock production of domesticated giant tiger prawns

DEEDI has been approached to perform an important outdoor pond experiment using the heated nursery ponds at Bribie Island Aquaculture Research Centre to get tiger prawn broodstock out of expensive clear-water tank systems and into ponds. This approach will enable a faster and more cost-effective adoption of existing selective breeding technology. Additional benefits will involve increased profitability and sustainability of genetic selection programs, improved understanding of prawn maturation benefiting other tiger prawn research and improved collaboration between DEEDI and CSIRO.

Developmental and growth controls of tillering and water-soluble carbohydrate accumulation in contrasting wheat (Triticum aestivum L.) genotypes: can we dissect them?

In wheat, tillering and water-soluble carbohydrates (WSCs) in the stem are potential traits for adaptation to different environments and are of interest as targets for selective breeding. This study investigated the observation that a high stem WSC concentration (WSCc) is often related to low tillering. The proposition tested was that stem WSC accumulation is plant density dependent and could be an emergent property of tillering, whether driven by genotype or by environment. A small subset of recombinant inbred lines (RILs) contrasting for tillering was grown at different plant densities or on different sowing dates in multiple field experiments. Both tillering and WSCc were highly influenced by the environment, with a smaller, distinct genotypic component; the genotypeenvironment range covered 350750 stems m(2) and 25210mg g(1) WSCc. Stem WSCc was inversely related to stem number m(2), but genotypic rankings for stem WSCc persisted when RILs were compared at similar stem density. Low tilleringhigh WSCc RILs had similar leaf area index, larger individual leaves, and stems with larger internode cross-section and wall area when compared with high tilleringlow WSCc RILs. The maximum number of stems per plant was positively associated with growth and relative growth rate per plant, tillering rate and duration, and also, in some treatments, with leaf appearance rate and final leaf number. A common threshold of the red:far red ratio (0.390.44; standard error of the difference0.055) coincided with the maximum stem number per plant across genotypes and plant densities, and could be effectively used in crop simulation modelling as a ocut-off' rule for tillering. The relationship between tillering, WSCc, and their component traits, as well as the possible implications for crop simulation and breeding, is discussed.

Breeding and fecundity of the endemic Australian gudgeon, sleepy cod Oxyeleotris lineolatus (Steindachner 1867) (Eleotridae)

Sleepy cod (Oxyeleotris lineolatus Steindachner) is a tropical species of eleotrid native to northern Australia. A related species, sand or marbled goby, is the highest priced freshwater fish in Asia, and a market for a similar fish exists in expatriate Chinese communities. Sleepy cod breed when minimum temperatures reach 24 °C for more than 3 days. During the breeding season the genital papilla is broad and flattened in females compared to the triangular papilla of males and juveniles. Spawning pairs were usually of approximately equal size. Females could spawn up to 10 times during one breeding season. Wet weather increased the frequency of spawning. Eggs were usually laid hanging from the underside of a surface. Most spawning occurred between 05:00 and 10:00 h. Females attended egg masses immediately after spawning, after which males cared for eggs until hatching, 3–5 days later. Agitation of the egg mass was essential for development. The mean number of eggs per spawning was 43 130. Larvae commenced feeding 2–5 days after hatching, on plankton from 100 to 250 m in size. A spawning trap used to collect egg masses is described. The breeding biology of sleepy cod is considered to be an adaptation to the monsoonal tropics.

Selective herbicide strategies for use in Australian desmanthus seed crops

Grass and broad-leaved weeds can reduce both yields and product marketability of desmanthus (Desmanthus virgatus) seed crops, even when cultural control strategies are used. Selective herbicides might economically control these weeds, but, prior to this study, the few herbicides tolerated by desmanthus did not control key weed contaminants of desmanthus seed crops. In this study, the tolerance of desmanthus cv. Marc to 55 herbicides used for selective weed control in other leguminous crops was assessed in 1 pot trial and 3 Queensland field trials. One field trial assessed the tolerance of desmanthus seedlings to combinations of the most promising pre-emergent and post emergent herbicides. The pre-emergent herbicides, imazaquin, imazethapyr, pendimethalin, oryzalin and trifluralin, gave useful weed control with very little crop damage. The post-emergent herbicides, haloxyfop, clethodim, propyzamide, carbetamide and dalapon, were safe for controlling grass weeds in desmanthus. Selective post-emergence control of broad-leaved weeds was achieved using bentazone, bromoxynil and imazethapyr. One trial investigated salvaging second-year desmanthus crops from mature perennial weeds, and atrazine, terbacil and hexazinone showed some potential in this role. Overall, our results show that desmanthus tolerates herbicides which collectively control a wide range of weeds encountered in Queensland. These, in combination with cultural weed control strategies, should control most weeds in desmanthus seed crops.

In Vitro Conditions for a Successful Biolistics transformation System of Pineapples (ananas comosus merr.), in 'Contributing to a Sustainable Future'

In order to develop an efficient and reliable biolistics transformation system for pineapples parameters need to be optimised for growth, survival and development of explants pre- and post transformation. We have optimised in vitro conditions for culture media for the various stages of plant and callus initiation and development, and for effective selection of putative transgenic material. Shoot multiplication and proliferation is best on medium containing MS basic nutrients and vitamins with the addition of 0.1 mg/L myo-inositol, 20 g/L sucrose, 2.5 mg/L BAP and 3 g/L Phytagel, followed by transfer to basic MS medium for further development. Callus production on leaf base explants is best on MS nutrients and vitamins, to which 10 mg/L of BAP and NAA each was added. Optimum explant age for bombardment is 17-35 week old callus, while a pre-bombardment osmoticum treatment in the medium is not required. By comparing several antibiotics as selective agent, it has been established that a two-step selection of 2 fortnightly sub-cultures on 50 μg/mL of geneticin in the culture medium, followed by monthly sub-cultures on 100 μg/mL geneticin is optimal for survival of transgenic callus. Shoot regeneration from callus cultures is optimal on medium containing MS nutrients and vitamins, 5% coconut water and 400 mg/L casein hydrolysate. Plants can be readily regenerated and multiplied from transgenic callus through organogenesis. Rooting of shoots does not require any additional plant hormones to the medium. A transformation efficiency of 1 – 3.5% can be achieved, depending on the gene construct applied.

Standardising bull breeding soundness evaluations and reporting in Australia

There is substantial variation in bull breeding soundness evaluation procedures and reports in Australia; the situation is compounded by difficulties in interpretation and the validity of many reports. In an effort to overcome this, the scientific literature was reviewed [Fordyce G. In: Fordyce G, editor. Bull fertility: selection and management in Australia. Eight Mile Plains, Australia: Australian Cattle Vets; 2002] and the needs of stakeholders were considered in preparing a manual, Evaluating and Reporting Bull Fertility [Entwistle KW, Fordyce G. Evaluating and reporting bull fertility. Eight Mile Plains, Australia: Australian Cattle Vets; 2003.] that outlined standards for assessing and reporting bull breeding soundness. A new recording and reporting system, called Bull Reporter, is based on standards from this manual and groups bull fertility traits into five summary categories: Scrotum, Physical, Crush-side Semen, Sperm Morphology, and Serving. The client will generally select which categories they wish to have included in the evaluation to suit their specific purposes. While there is adequate room for comments, the veterinarian is not required to make an overall judgment of whether the bull has normal capacity to sire calves under natural mating management, but ensures the standards for each selected category are met. Professional, standardised, easy-to-read reports are produced either electronically [Entwistle KW, Fordyce G. Evaluating and reporting bull fertility. Eight Mile Plains, Australia: Australian Cattle Vets; 2003.] or manually. A bull owner or their agent signs the certificate to affirm that bulls have not undergone procedures to rectify faults which may have otherwise caused them to fail the standards. An accreditation system for assessing sperm morphology was established because of its demonstrated relationship with pregnancy rates and because of the difficulties in achieving consistent and accurate assessments among laboratories. It is considered that Bull Reporter is applicable to beef and dairy bulls across all levels of management, genotypes and environments throughout Australia, with substantial potential for application elsewhere in the world.

Applications of pedigree-based genome mapping in wheat and barley breeding programs.

The aim of the pedigree-based genome mapping project is to investigate and develop systems for implementing marker assisted selection to improve the efficiency of selection and increase the rate of genetic gain in breeding programs. Pedigree-based whole genome marker application provides a vehicle for incorporating marker technologies into applied breeding programs by bridging the gap between marker-trait association and marker implementation. We report on the development of protocols for implementation of pedigree-based whole genome marker analysis in breeding programs within the Australian northern winter cereals region. Examples of applications from the Queensland DPI&F wheat and barley breeding programs are provided, commenting on the use of microsatellites and other types of molecular markers for routine genomic analysis, the integration of genotypic, phenotypic and pedigree information for targeted wheat and barley lines, the genomic impacts of strong selection pressure in case study pedigrees, and directions for future pedigree-based marker development and analysis.

Rice molecular breeding laboratories in the genomics era: Current status and future considerations

Using DNA markers in plant breeding with marker-assisted selection (MAS) could greatly improve the precision and efficiency of selection, leading to the accelerated development of new crop varieties. The numerous examples of MAS in rice have prompted many breeding institutes to establish molecular breeding labs. The last decade has produced an enormous amount of genomics research in rice, including the identification of thousands of QTLs for agronomically important traits, the generation of large amounts of gene expression data, and cloning and characterization of new genes, including the detection of single nucleotide polymorphisms. The pinnacle of genomics research has been the completion and annotation of genome sequences for indica and japonica rice. This information-coupled with the development of new genotyping methodologies and platforms, and the development of bioinformatics databases and software tools-provides even more exciting opportunities for rice molecular breeding in the 21st century. However, the great challenge for molecular breeders is to apply genomics data in actual breeding programs. Here, we review the current status of MAS in rice, current genomics projects and promising new genotyping methodologies, and evaluate the probable impact of genomics research. We also identify critical research areas to "bridge the application gap" between QTL identification and applied breeding that need to be addressed to realize the full potential of MAS, and propose ideas and guidelines for establishing rice molecular breeding labs in the postgenome sequence era to integrate molecular breeding within the context of overall rice breeding and research programs.

The accuracy of varietal selection using factor analytic models for multi-environment plant breeding trials

Modeling of cultivar x trial effects for multienvironment trials (METs) within a mixed model framework is now common practice in many plant breeding programs. The factor analytic (FA) model is a parsimonious form used to approximate the fully unstructured form of the genetic variance-covariance matrix in the model for MET data. In this study, we demonstrate that the FA model is generally the model of best fit across a range of data sets taken from early generation trials in a breeding program. In addition, we demonstrate the superiority of the FA model in achieving the most common aim of METs, namely the selection of superior genotypes. Selection is achieved using best linear unbiased predictions (BLUPs) of cultivar effects at each environment, considered either individually or as a weighted average across environments. In practice, empirical BLUPs (E-BLUPs) of cultivar effects must be used instead of BLUPs since variance parameters in the model must be estimated rather than assumed known. While the optimal properties of minimum mean squared error of prediction (MSEP) and maximum correlation between true and predicted effects possessed by BLUPs do not hold for E-BLUPs, a simulation study shows that E-BLUPs perform well in terms of MSEP.

Strawberry breeding in a subtropical environment

Strawberry breeding aims to provide cultivars that maximise consumer satisfaction and producer profitability in a changing environment. In this paper some concepts of profitability, consumer satisfaction and sustainability are explored for a subtropical climate using Queensland Australia, and Florida USA, as examples. The typical production environment is annual autumn planting of bare rooted runners into polythene covered raised beds at about 40000 plants/ha. Harvesting is late autumn to early spring, with fruit arriving at the major markets up to 2000km away from the production area within 1-4 days of harvest. The basic premise in the breed-big work is that consumers must enjoy the experience of eating strawberries, and that perceived flavour, sweetness, and juiciness are the major contributors to this experience. Using market chain information, we developed a basic value model comprised of costs, returns, and sustainability of market. To this basic outline are applied operational descriptors, such as 'speed of harvest', and associated plant characteristics, such as 'fruit display'. The expression of each plant characteristic is ascribed a value or level and together numerically describe the phenotype. This description is mathematically manipulated to provide a 'value index' for the cultivar. Nine cultivars including 'Strawberry Festival', 'Kabarla', 'DPI Rubygem' and 'Sweet Charlie' are described, and environmental issues that may impact on the subtropical strawberry breeding objectives are discussed. Product differentiation and the use of exotic germplasm as a new source of genes for flavour and resistance to disease and environmental stress will likely be the cornerstones of future progress in subtropical strawberry breeding. This approach should satisfy both consumers and producers.

Use of part records in Merino breeding programs - The inheritance of wool growth and fibre traits during different times of the year to determine their value in Merino breeding programs

Fibre diameter can vary dramatically along a wool staple, especially in the Mediterranean environment of southern Australia with its dry summers and abundance of green feed in spring. Other research results have shown a very low phenotypic correlation between fibre diameter grown between seasons. Many breeders use short staples to measure fibre diameter for breeding purposes and also to promote animals for sale. The effectiveness of this practice is determined by the relative response to selection by measuring fibre traits on a full 12 months wool staple as compared to measuring them only on part of a staple. If a high genetic correlation exists between the part record and the full record, then using part records may be acceptable to identify genetically superior animals. No information is available on the effectiveness of part records. This paper investigated whether wool growth and fibre diameter traits of Merino wool grown at different times of the year in a Mediterranean environment, are genetically the same trait, respectively. The work was carried out on about 7 dyebanded wool sections/animal.year, on ewes from weaning to hogget age, in the Katanning Merino resource flocks over 6 years. Relative clean wool growth of the different sections had very low heritability estimates of less than 0.10, and they were phenotypically and genetically poorly correlated with 6 or 12 months wool growth. This indicates that part record measurement of clean wool growth of these sections will be ineffective as indirect selection criteria to improve wool growth genetically. Staple length growth as measured by the length between dyebands, would be more effective with heritability estimates of between 0.20 and 0.30. However, these measurements were shown to have a low genetic correlation with wool grown for 12 months which implies that these staple length measurements would only be half as efficient as the wool weight for 6 or 12 months to improve total clean wool weight. Heritability estimates of fibre diameter, coefficient of variation of fibre diameter and fibre curvature were relatively high and were genetically and phenotypically highly correlated across sections. High positive phenotypic and genetic correlations were also found between fibre diameter, coefficient of variation of fibre diameter and fibre curvature of the different sections and similar measurements for wool grown over 6 or 12 months. Coefficient of variation of fibre diameter of the sections also had a moderate negative phenotypic and genetic correlation with staple strength of wool staples grown over 6 months indicating that coefficient of variation of fibre diameter of any section would be as good an indirect selection criterion to improve stable strength as coefficient of variation of fibre diameter for wool grown over 6 or 12 months. The results indicate that fibre diameter, coefficient of variation of fibre diameter and fibre curvature of wool grown over short periods of time have virtually the same heritability as that of wool grown over 12 months, and that the genetic correlation between fibre diameter, coefficient of variation of fibre diameter and fibre curvature on part and on full records is very high (rg > 0.85). This indicates that fibre diameter, coefficient of variation of fibre diameter and fibre curvature on part records can be used as selection criteria to improve these traits. However, part records of greasy and clean wool growth would be much less efficient than fleece weight for wool grown over 6 or 12 months because of the low heritability of part records and the low genetic correlation between these traits on part records and on wool grown for 12 months.

Breeding Ananas for the cut-flower and garden markets

Only small quantities of Ananas have been marketed as cut flowers or as potted plants for garden use in Australia. Worldwide there have, until very recent times, been no breeding programs to develop ornamental characteristics and hence the choice of cultivars has been limited mainly to semi-domesticated selections or those developed by amateur enthusiasts. Interest in developing Ananas selections specifically for the ornamental market is now increasing. A small program has operated in Australia since 1995. In this program, a total of 4,700 seedlings were generated over three generations using various parental combinations of Ananas comosus var. comosus, A. comosus var. bracteatus, A. comosus var. ananassoides 'FRF223', A. comosus var. erectifolious 'Selvagem 6' and Ananas macrodontes 'I.26-803'. Several selections have been developed for the garden and or cut-flower market. Characteristics represented include a pink or red syncarp, dark red-brown foliage and a dwarf, clumping growth habit. While a surprising display of ornamental diversity exists within Ananas, the genus is limited in comparison to the other bromeliad genera. Opportunity might exist however to introgress characteristics such as additional foliage colours, plant morphology and syncarp colours from other genera into Ananas.