In vitro breeding strategies in the development of Australian native plants

Plant tissue culture is a technique that exploits the ability of many plant cells to revert to a meristematic state. Although originally developed for botanical research, plant tissue culture has now evolved into important commercial practices and has become a significant research tool in agriculture, horticulture and in many other areas of plant sciences. Plant tissue culture is the sterile culture of plant cells, tissues, or organs under aseptic conditions leading to cell multiplication or regeneration or organs and whole plants. The steps required to develop reliable systems for plant regeneration and their application in plant biotechnology are reviewed in countless books. Some of the major landmarks in the evolution of in vitro techniques are summarised in Table 5.1. In this chapter the current applications of this technology to agriculture, horticulture, forestry and plant breeding are briefly described with specific examples from Australian plants when applicable.

A note on inbreeding in dairy cattle breeding

Starting from the study at the beginning of the East German "Heterosisfeldversuch", where PANICKE et al. (1975) considered the possibilities of a targeted use of inbreeding and heterotic effects, we show and discuss results of inbreeding studies in the USA dairy cattle breeding. Several research groups worldwide presented effective tools for managing inbreeding in dairy cattle. Their efforts underline the need of inbreeding studies. Contemplating inbreeding is necessary for any breeding decision to avoid inbreeding depression and for improved genetic analyses, e.g. in QTL- estimation. A novel methodology (HERNANDEZ-SANCHEZ et al., 2004a and b) is suggested for estimating inbreeding at the three levels of population, individual and locus.

Effect of coffee combining green coffee bean constituents with typical roasting products on the Nrf2/ARE pathway in vitro and in vivo

This study investigated Nrf2-activating properties of a coffee blend combining raw coffee bean constituents with 5-O-caffeoylquinic acid (CGA) as a lead component with typical roasting products such as N-methylpyridinium (NMP). In cell culture (HT29) the respective coffee extract (CN-CE) increased nuclear Nrf2 translocation and enhanced the transcription of ARE-dependent genes as exemplified for NAD(P)H:quinone oxidoreductase and glutathione-S-transferase (GST)A1, reflected in the protein level by an increase in GST enzyme activity. In a pilot human intervention study (29 healthy volunteers), daily consumption of 750 mL of CN-coffee for 4 weeks increased Nrf2 transcription in peripheral blood lymphocytes on average. However, the transcriptional response pattern of Nrf2/ARE-dependent genes showed substantial interindividual variations. The presence of SNPs in the Nrf2-promoter, reported recently, as well as the detection of GSTT1*0 (null) genotypes in the study collective strengthens the hypothesis that coffee acts as a modulator of Nrf2-dependent gene response in humans, but genetic polymorphisms play an important role in the individual response pattern.

Intron-mediated improvement of a selectable marker gene for plant transformation using Agrobacterium tumefaciens

In binary vectors, the antibiotic resistance gene used for selection of transformed plant cells is also usually expressed in the transforming Agrobacterium cells. This expression gives the bacterium antibiotic resistance, an unnecessary advantage on selective medium containing the antibiotic. Insertion of a castor bean catalase-1 (CAT-1) gene intron or a Parasponia andersonii haemoglobin gene intron into the coding region of the selectable marker gene, hph, completely abolished the expression of the gene in Agrobacterium, rendering it susceptible to hygromycin B. Use of these modified binary vectors minimized the overgrowth of Agrobacterium during plant transformation. Both of the introns were correctly spliced in plant cells and significantly enhanced hph gene expression in transformed rice tissue. The presence of these introns in the hph coding sequence not only maintained the selection efficiency of the hph gene, but with the CAT-1 intron also substantially increased the frequency of rice transformation. Transgenic lines with an intron-hph gene generally contained fewer gene copies and produced substantially more mRNA of the predicted size. Our results also indicate that transgenic plants with many copies of the transgene were more likely to show gene silencing than plants with 1-3 copies.

Wheat breeding nurseries, target environments, and indirect selection for grain yield

Use of appropriate nursery environments will maximize gain from selection for yield of wheat (Triticum aestivum L.) in the target population of environments of a breeding program. The objective of this study was to investigate how well-irrigated (low-stress) nursery environments predict yield of lines in target environments that varied in degree of water limitation. Fifteen lines were sampled from the preliminary yield evaluation stage of the Queensland wheat breeding program and tested in 26 trials under on-farm conditions (Target Environments) across nine years (1985 to 1993) and also in 27 trials conducted at three research stations (Nursery Environments) in three years (1987 to 1989). The nursery environments were structured to impose different levels of water and nitrogen (N) limitation, whereas the target environments represented a random sample of on-farm conditions from the target population of environments. Indirect selection and pattern analysis methods were used to investigate selection for yield in the nursery environments and gain from selection in the target environments. Yield under low-stress nursery conditions was an effective predictor of yield under similar low-stress target environments (r = 0.89, P < 0.01). However, the value of the low-stress nursery as a predictor of yield in the water-limited target environments decreased with increasing water stress (moderate stress r = 0.53, P < 0.05, to r = 0.38, P > 0.05; severe stress r = -0.08, P > 0.05). Yield in the stress nurseries was a poor predictor of yield in the target environments. Until there is a clear understanding of the physiological-genetic basis of variation for adaptation of wheat to the water-limited environments in Queensland, yield improvement can best be achieved by selection for a combination of yield potential in an irrigated low-stress nursery and yield in on-farm trials that sample the range of water-limited environments of the target population of environments.

Is lek breeding rare in bats?

Bats (Mammalia: Chiroptera) are among the most successful mammals and likely display the widest range of mating systems within the Class. One mating system that is underrepresented in the Chiroptera is lek breeding, which is characterized by aggregations of sexually displaying males that are visited by receptive females who appraise male displays and actively choose mates, yet receive no direct benefits such as assistance in parenting. Leks are thought to form when males can defend neither resources nor females, making it more economical to establish small breeding territories and self-advertise through sexual displays. Lekking is rare in mammals, and it has been suggested that a lack in the mobility required by females to economically seek out aggregations of sexually displaying males may explain this rarity. Bats, like birds, do not suffer reduced mobility and yet out of over a thousand described species, only one has been confirmed to breed in leks. We examine the rarity of lekking in bats by providing an overview on the current state of knowledge of their mating systems and discuss the ecological and social determinants for the observed trends, contrasted with the prerequisites of lek-breeding behaviour. We use the breeding behaviour of New Zealand's lesser short-tailed bat Mystacina tuberculata, which is believed to be a lek breeder, as a case study for the examination of potential lekking behaviour in bats, and highlight the importance of such research for the development of effective conservation strategies.

Females as mobile resources: communal roosts promote the adoption of lek breeding in a temperate bat

Males of lek-breeding species defend clustered territories from which they display to visiting females. However, the mechanisms leading to the adoption of clustered male display sites are often unknown. In this study, we examined the possibility of a resource-based lek in New Zealand’s lesser short-tailed bat (Mystacina tuberculata) (Mammalia: Chiroptera), by assessing the placement of “singing roosts” used by males in relation to communal roosting sites used by females. The “resource-based lek” model posits that males settle near resources required by females to increase female encounter rates. For most bat species, where females are highly mobile and widely dispersed across landscapes while foraging, communal daytime roosts dominated by females may represent such a resource. Through use of video footage, spatial analyses of singing-roost locations, and passive-integrated transponder tags we confirmed that M. tuberculata employs a lek mating system. We found that male singing roosts were significantly clustered in space, were defended by resident individuals, and were visited by females (who did not receive resources from males) for mating purposes. Transponder records also indicated that some singing roosts were shared between multiple males. Spatial logistic regression indicated that singing-roost locations were associated with communal roosting sites. Communal roosts are selected based on criteria independent of the locations of singing roosts, suggesting that males responded to the location of communal roosts and not the reverse. Mystacina tuberculata thus provides evidence of a resource-based lek, and is only the second bat species worldwide confirmed to use a lek-mating system.

Identification of amino groups in the carbohydrate binding activity of winged bean acidic agglutinin

Chemical modification studies reveal that the modification of amino groups in WBA II leads to a complete loss in the hemagglutinating and saccharide binding activities. Since WBA II is a dimeric molecule and contains two binding sites, one amino group in each of the binding sites is inferred to be essential for its activity. The presence of amino group which has a potential to form hydrogen bonded interactions with the ligand, substantiates our observation regarding the forces involved in WBA II-receptor and WBA II-simple sugar interactions.

Fluorescence-polarization studies on binding of 4-methylumbelliferyl beta-D-galactopyranoside to Ricinus communis (castor-bean) agglutinin

The binding of Ricinus communis (castor-bean) agglutinin 1 to saccharides was studied by equilibrium dialysis and fluorescence polarization by using the fluorescently labelled sugar 4-methylumbelliferyl beta-D-galactopyranoside. No appreciable change in ligand fluorescence of 4-methylumbelliferyl beta-D-galactopyranoside was considerably polarized on its binding to the lectin. The association constants obtained by Scatchard analysis of equilibrium-dialysis and fluorescence-polarization data do not differ much from each other, and at 25 degrees C, Ka = 2.4 (+/- 0.2) X 10(4)M-1. These values agree reasonably well with that reported in the literature for Ricinus agglutinin 1. The number of binding sites obtained by the different experimental procedures is 1.94 +/- 0.1 per molecule of 120 000 daltons and is equal to the reported value of 2. The consistency in the values of Ka and number of binding sites indicate the absence of additional subsites on Ricinus agglutinin 1 for its specific sugars. In addition, the excellent agreement between the binding parameters obtained by equilibrium dialysis and fluorescence polarization indicate the potential of ligand-fluorescence-polarization measurements in the investigation of lectin-sugar interactions.

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.

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.