DArT markers: Diversity analyses and mapping in Sorghum bicolor

The sequential nature of gel-based marker systems entails low throughput and high costs per assay. Commonly used marker systems such as SSR and SNP are also dependent on sequence information. These limitations result in high cost per data point and significantly limit the capacity of breeding programs to obtain sufficient return on investment to justify the routine use of marker-assisted breeding for many traits and particularly quantitative traits. Diversity Arrays Technology (DArT™) is a cost effective hybridisation-based marker technology that offers a high multiplexing level while being independent of sequence information. This technology offers sorghum breeding programs an alternative approach to whole-genome profiling. We report on the development, application, mapping and utility of DArT™ markers for sorghum germplasm. Results: A genotyping array was developed representing approximately 12,000 genomic clones using PstI+BanII complexity with a subset of clones obtained through the suppression subtractive hybridisation (SSH) method. The genotyping array was used to analyse a diverse set of sorghum genotypes and screening a Recombinant Inbred Lines (RIL) mapping population. Over 500 markers detected variation among 90 accessions used in a diversity analysis. Cluster analysis discriminated well between all 90 genotypes. To confirm that the sorghum DArT markers behave in a Mendelian manner, we constructed a genetic linkage map for a cross between R931945-2-2 and IS 8525 integrating DArT and other marker types. In total, 596 markers could be placed on the integrated linkage map, which spanned 1431.6 cM. The genetic linkage map had an average marker density of 1/2.39 cM, with an average DArT marker density of 1/3.9 cM. Conclusion: We have successfully developed DArT markers for Sorghum bicolor and have demonstrated that DArT provides high quality markers that can be used for diversity analyses and to construct medium-density genetic linkage maps. The high number of DArT markers generated in a single assay not only provides a precise estimate of genetic relationships among genotypes, but also their even distribution over the genome offers real advantages for a range of molecular breeding and genomics applications.

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.

Genetic erosion and changes in distribution of sorghum (Sorghum bicolor L. (Moench)) landraces in north-eastern Ethiopia

Ethiopia is believed to be the centre of origin and domestication for sorghum, where sorghum remains one of the main staple crops. Loss of biodiversity is occurring at an alarming rate in Ethiopia and crops, including sorghum, have long been recognized as vulnerable to genetic erosion. A major collection of sorghum germplasm was made in 1973 by Gebrekidan and Ejeta from north-eastern Ethiopia. A new collection of landraces was made in 2003, and these were field evaluated at Sirinka in 2004 along with representative samples from the 1973 collection. Farmer surveys and soil and climate surveys were also performed. Preliminary analysis demonstrated that some important landraces have disappeared either locally or regionally in the past 30 years and many other landraces have become marginalized. Landraces which are less preferred in terms of agronomic value and end use, and introductions, have become increasingly important. Late maturing landraces were found to be particularly vulnerable, with a number disappearing altogether. Farmers have become more risk averse, and factors such as declining soil fertility, more frequent drought and unreliable rainfall, and increased pest infestation have contributed to a change in farmer landrace selection. Data are presented on the variability and unique characters of some of the Ethiopian landraces, and implications for conservation are discussed.

A detached leaf bioassay to screen Durian cultivars for susceptibility to Phytophthora palmivora

A detached-leaf bioassay was developed and used to screen five durian (Durio zibethinus) cultivars against Phytophthora palmivora isolates from a trunk canker, root and fruit. The fruit isolate was less aggressive than the canker and root isolates. The bioassay using the canker isolate was later used to determine the variation in resistance of D. macarantha and nineteen cultivars of D. zibethinus. The cultivars displayed a range of responses with Parung and Gob being most tolerant, with Gaan Yaow, Chanee and Penang 88 being susceptible. The remaining germplasm fell between Gaan Yaow and Penang 88 in susceptibility. The leaf bioassay was found to be a rapid and reliable method for assessing the susceptibility of durian cultivars.

Zoysia japonica, Zoysia Grass 'Palisades'

Open-pollination: originated as a chance seedling from Z44 (maternal clonal parent), obtained from Beltsville MD in 1981, with an unknown pollen source from a zoysia grass germplasm field nursery at the Texas Agricultural Experiment Station in Dallas. ‘Palisades’ was selected over the parent Z44 on the basis of its lower tendency to produce thatch, its excellent lateral growth habit and its superior mowing qualities. ‘Palisades’ has been vegetatively propagated, and is uniform in growth expression. No seedling establishment from ‘Palisades’ has been noticed in either greenhouse or field studies. Selection criteria: rapid regrowth and spread by, and/or from, stolons and rhizomes; turf colour and density; tolerance to low mowing; winter hardiness; shade tolerance; low water use requirements. Propagation: vegetative. Breeder: Milton C. Engelke, Dallas, USA. PBR Certificate Number 2594, Application Number 2001/199, granted 26 October 2004.

Cynodon dactylon, Green Couch Grass, Bermuda Grass 'Grand Prix'

‘Grand Prix’ is a selection from a cross between ‘Wintergreen’ and ‘Couch 5’ (also designated C5). ‘Couch 5’ was a selection from an earlier series of crosses by the breeder between ‘Wintergreen’ and a number of Cynodon dactylon accessions, which were collected by the breeder from the Mornington Peninsula area of Victoria between 1986 and 1990. C5 was an experimental breeding line, and was not subsequently reserved as vegetative germplasm. Living material of C5 is no longer in existence. Following the crossing of ‘Couch 5’ and ‘Wintergreen’ in 1998, the resultant seed was germinated on moist blotting paper. Individual seedlings, a total of 150 in number, were planted into 150mm pots and these plants observed during 1998 and 1999. During the summer of 1999-2000, the majority of the seedling plants were culled on the basis of their shoot density, leaf texture, internode length, and colour. In the spring of 2000, the remaining 20 potted seedlings were planted individually into 4m2 plots at the Evergreen Turf farm at Pakenham (Victoria), and allowed to expand fully across these plots. The final selection of Seedling 12 (later designated DN12) in late 2002 was based on shoot density, leaf colour, turf quality, and reduced thatch accumulation as expressed in these plots. Propagation: the original plant has been multiplied through four (4) vegetative expansions prior to PBR application without showing any discernible off types. Breeder: David Nickson, Frankston, VIC. PBR Certificate Number 3133, Application Number 2005/291, granted 12 September 2006.

Cynodon dactylon, Green Couch Grass, Bermuda Grass 'Winter Gem'

‘Winter Gem’ is a selection from a cross between ‘Wintergreen’ and Couch 5 (also designated C5). Couch 5 was a selection from an earlier series of crosses by the breeder between ‘Wintergreen’ and a number of Cynodon dactylon accessions, which were collected by the breeder from the Mornington Peninsula area of Victoria between 1986 and 1990. C5 was an experimental breeding line, and was not subsequently reserved as vegetative germplasm. Living material of C5 is no longer in existence. Following the crossing of Couch 5 and ‘Wintergreen’ in 1998, the resultant seed was germinated on moist blotting paper. Individual seedlings, a total of 150 in number, were planted into 150mm pots and these plants observed during 1998 and 1999. During the summer of 1999-2000, the majority of the seedling plants were culled on the basis of their shoot density, leaf texture, internode length, and colour. In the spring of 2000, the remaining 20 potted seedlings were planted individually into 4m2 plots at the Evergreen Turf farm at Pakenham (Victoria), and allowed to expand fully across these plots. The final selection of Seedling 9 (later designated DN9) in late 2002 was based on shoot density, leaf texture, and retention of winter colour as expressed in these plots. Propagation: The original plant had been multiplied through four (4) vegetative expansions prior to PBR application without showing any discernible off types. Breeder: David Nickson, Frankston, VIC. PBR Certificate Number 3132, Application Number 2005/290, granted 11 September 2006.

QTL mapping of multiple foliar disease and root-lesion nematode resistances in wheat.

A genetic linkage map, based on a cross between the synthetic hexaploid CPI133872 and the bread wheat cultivar Janz, was established using 111 F1-derived doubled haploid lines. The population was phenotyped in multiple years and/or locations for seven disease resistance traits, namely, Septoria tritici blotch (Mycosphaeralla graminicola), yellow leaf spot also known as tan spot (Pyrenophora tritici-repentis), stripe rust (Puccinia striiformis f. sp. tritici), leaf rust (Puccinia triticina), stem rust (Puccinia graminis f. sp. tritici) and two species of root-lesion nematode (Pratylenchyus thornei and P. neglectus). The DH population was also scored for coleoptile colour and the presence of the seedling leaf rust resistance gene Lr24. Implementation of a multiple-QTL model identified a tightly linked cluster of foliar disease resistance QTL in chromosome 3DL. Major QTL each for resistance to Septoria tritici blotch and yellow leaf spot were contributed by the synthetic hexaploid parent CPI133872 and linked in repulsion with the coincident Lr24Sr24/ locus carried by parent Janz. This is the first report of linked QTL for Septoria tritici blotch and yellow leaf spot contributed by the same parent. Additional QTL for yellow leaf spot were detected in 5AS and 5BL. Consistent QTL for stripe rust resistance were identified in chromosomes 1BL, 4BL and 7DS, with the QTL in 7DS corresponding to the Yr18Lr34/ region. Three major QTL for P. thornei resistance (2BS, 6DS, 6DL) and two for P. neglectus resistance (2BS, 6DS) were detected. The recombinants combining resistance to Septoria tritici blotch, yellow leaf spot, rust diseases and root-lesion nematodes from parents CPI133872 and Janz constitute valuable germplasm for the transfer of multiple disease resistance into new wheat cultivars.

Location of major effect genes in sorghum (Sorghum bicolor (L.) Moench).

Major effect genes are often used for germplasm identification, for diversity analyses and as selection targets in breeding. To date, only a few morphological characters have been mapped as major effect genes across a range of genetic linkage maps based on different types of molecular markers in sorghum (Sorghum bicolor (L.) Moench). This study aims to integrate all available previously mapped major effect genes onto a complete genome map, linked to the whole genome sequence, allowing sorghum breeders and researchers to link this information to QTL studies and to be aware of the consequences of selection for major genes. This provides new opportunities for breeders to take advantage of readily scorable morphological traits and to develop more effective breeding strategies. We also provide examples of the impact of selection for major effect genes on quantitative traits in sorghum. The concepts described in this paper have particular application to breeding programmes in developing countries where molecular markers are expensive or impossible to access.

Environment characterization as an aid to wheat improvement: interpreting genotype-environment interactions by modelling water-deficit patterns in North-Eastern Australia.

Genotype-environment interactions (GEI) limit genetic gain for complex traits such as tolerance to drought. Characterization of the crop environment is an important step in understanding GEI. A modelling approach is proposed here to characterize broadly (large geographic area, long-term period) and locally (field experiment) drought-related environmental stresses, which enables breeders to analyse their experimental trials with regard to the broad population of environments that they target. Water-deficit patterns experienced by wheat crops were determined for drought-prone north-eastern Australia, using the APSIM crop model to account for the interactions of crops with their environment (e.g. feedback of plant growth on water depletion). Simulations based on more than 100 years of historical climate data were conducted for representative locations, soils, and management systems, for a check cultivar, Hartog. The three main environment types identified differed in their patterns of simulated water stress around flowering and during grain-filling. Over the entire region, the terminal drought-stress pattern was most common (50% of production environments) followed by a flowering stress (24%), although the frequencies of occurrence of the three types varied greatly across regions, years, and management. This environment classification was applied to 16 trials relevant to late stages testing of a breeding programme. The incorporation of the independently-determined environment types in a statistical analysis assisted interpretation of the GEI for yield among the 18 representative genotypes by reducing the relative effect of GEI compared with genotypic variance, and helped to identify opportunities to improve breeding and germplasm-testing strategies for this region.

Potential gains through selecting for resistance in spotted gum to Quambalaria pitereka

Abstract Quambalaria shoot blight, caused by the fungus Quambalaria pitereka, is a serious disease affecting the expanding eucalypt plantation estate in subtropical and tropical eastern Australia. Trees that are severely infected are often multi-stemmed and stunted and infection of young trees may give rise to poor form in mature trees. A spotted gum clonal trial provided the opportunity to investigate the impact of the disease on tree growth and factors influencing tree architecture (tree form), which affects wood quality. We measured the effect that Q. pitereka infection during plantation establishment (up to 6 months old) has on growth and tree architecture and productivity to age 3 years. Our results show that the pathogen has a significant impact on trees at plantation establishment, which results in a negative impact on wood quality, potentially reducing merchantable value at final harvest. Tree growth and form was significantly improved where germplasm with low susceptibility to Q. pitereka infection was used.

Identification and analysis in sweet corn

Identification and analysis of allelic variation in carotenoid biosynthesis genes present in sweet corn germplasm for eye health.

PI09004 New Generation Pineapple Breeding Project

To work with a major industry production/marketing unit to develop new pineapple varieties with good plant vigour, high yields and post-harvest attributes and eating quality equal to or better than the standard industry varieties 73-50 and MD2 under commercial production systems. The project uses previously developed germplasm as parental material.

National Citrus Triploid Breeding

This project advances commercially desirable citrus selections that have resilient seedlessness. It builds on existing expertise and develops germplasm resources in the utilisation of interploid crossing for the production of new triploid hybrids with outstanding fruit quality. Advanced germplasm will progress toward commercialisation with fruit displays, and the production of a final generation of trees for semi-commercial plantings. At the opposite end of the breeding spectrum, new triploid hybrids will be produced. Growers will see triploid citrus from their national breeding project for the first time, providing a window on future new varieties that will emerge from the pipe-line of germplasm that has been developed through past project investment.

Barley Grains Defects (Blackpoint, Pre-harvest sprouting, Kernal staining)

Screening new and existing breeding germplasm and cultivars for grain defect tolerance for breeding programs, evaluate new methods and technologies to screen more effectively for the barley grains defects - pre-harvest sprouting, blackpoint, kernel discolouration, and investigate genetic mechanisms involved in controlling barley grain defect tolerance.