Accounting for variability in the detection and use of markers for simple and complex traits

There are many sources of variability in gene–phenotype associations. During the measurement of genotype and phenotype and during selection, researchers must deal with experimental error in trials; gene-gene interaction (epistasis) for sub-traits and observed traits; trait-trait interaction (pleiotropy) and gene- or genotype-by-environment interaction. These effects can be structured in a framework that allows simulation of the entire gene-environment ‘landscape’. Studies of these landscapes have been published by others. Here we aim to explain with simple examples some of the types of insights that can be made. A current challenge for breeders working with simple marker–phenotype associations is to design selection strategies that can rapidly create new combinations of multiple marker-based traits. For a real-world example in wheat, we have used simulation to show how gene enrichment during early generations (selection of homozygotes and heterozygotes with desirable alleles) can greatly reduce resource requirements when combining 9 genes into one genotype through marker-assisted selection. Another wheat example compares phenotypic and QTL-based selection for coleoptile length where the QTL also had a pleiotropic association with plant height. These simulations show the relative negative effects of either low heritability, or less than complete detection of QTL associated with traits. Finally, we revisit a marker-assisted selection (MAS) example whereby a QTL study is undertaken on a population for a complex trait, and then those QTL are used in selection. This process is subject to all sources of error described above. If the trait is complex, then interactions among sub-traits; between sub-traits and the environment; or between the chromosomal locations of controlling genes, create an extremely ‘rugged’ selection landscape that slows breeding progress. In this situation, a detailed understanding of some of these interactions is required if MAS is to be able to exceed the progress of conventional breedin

Phenotypic evaluation and genetic analysis of seedling emergence in a global collection of wheat genotypes (Triticum aestivum L.) under limited water availability

The challenge in establishing an early-sown wheat crop in southern Australia is the need for consistently high seedling emergence when sowing deep in subsoil moisture (>10 cm) or into dry top-soil (4 cm). However, the latter is strongly reliant on a minimum soil water availability to ensure successful seedling emergence. This study aimed to: (1) evaluate 233 Australian and selected international wheat genotypes for consistently high seedling emergence under limited soil water availability when sown in 4 cm of top-soil in field and glasshouse (GH) studies; (2) ascertain genetic loci associated with phenotypic variation using a genome-wide association study (GWAS); and (3) compare across loci for traits controlling coleoptile characteristics, germination, dormancy, and pre-harvest sprouting. Despite significant (P 85%) across nine environments. Moreover, 21 environment-specific quantitative trait loci (QTL) were detected in GWAS analysis on chromosomes 1B, 1D, 2B, 3A, 3B, 4A, 4B, 5B, 5D, and 7D, indicating complex genetic inheritance controlling seedling emergence. We aligned QTL for known traits and individual genes onto the reference genome of wheat and identified 16 QTL for seedling emergence in linkage disequilibrium with coleoptile length, width, and cross-sectional area, pre-harvest sprouting and dormancy, germination, seed longevity, and anthocyanin development. Therefore, it appears that seedling emergence is controlled by multifaceted networks of interrelated genes and traits regulated by different environmental cues

Phenotypic Evaluation and Genetic Analysis of Seedling Emergence in a Global Collection of Wheat Genotypes (Triticum aestivum L.) Under Limited Water Availability

The challenge in establishing an early-sown wheat crop in southern Australia is the need for consistently high seedling emergence when sowing deep in subsoil moisture (>10 cm) or into dry top-soil (4 cm). However, the latter is strongly reliant on a minimum soil water availability to ensure successful seedling emergence. This study aimed to: (1) evaluate 233 Australian and selected international wheat genotypes for consistently high seedling emergence under limited soil water availability when sown in 4 cm of top-soil in field and glasshouse (GH) studies; (2) ascertain genetic loci associated with phenotypic variation using a genome-wide association study (GWAS); and (3) compare across loci for traits controlling coleoptile characteristics, germination, dormancy, and pre-harvest sprouting. Despite significant (P 85%) across nine environments. Moreover, 21 environment-specific quantitative trait loci (QTL) were detected in GWAS analysis on chromosomes 1B, 1D, 2B, 3A, 3B, 4A, 4B, 5B, 5D, and 7D, indicating complex genetic inheritance controlling seedling emergence. We aligned QTL for known traits and individual genes onto the reference genome of wheat and identified 16 QTL for seedling emergence in linkage disequilibrium with coleoptile length, width, and cross-sectional area, pre-harvest sprouting and dormancy, germination, seed longevity, and anthocyanin development. Therefore, it appears that seedling emergence is controlled by multifaceted networks of interrelated genes and traits regulated by different environmental cues

A tillering inhibition gene influences root-shoot carbon partitioning and pattern of water use to improve wheat productivity in rainfed environments

Genetic modification of shoot and root morphology has potential to improve water and nutrient 19 uptake of wheat crops in rainfed environments. Near-isogenic lines (NILs) varying for a tillering 20 inhibition (tin) gene and representing multiple genetic backgrounds were investigated in contrasting 21 controlled environments for shoot and root growth. Leaf area, shoot and root biomass were similar 22 until tillering whereupon reduced tillering in tin-containing NILs produced reductions of up to 60% in 23 total leaf area and biomass, and increases in total root length of up to 120% and root biomass to 24 145%. Together, root-to-shoot ratio increased two-fold with the tin gene. The influence of tin on shoot 25 and root growth was greatest in the cv. Banks genetic background, particularly in the biculm-selected 26 NIL, and was typically strongest in cooler environments. A separate de-tillering study confirmed 27 greater root-to-shoot ratios with regular tiller removal in non-tin containing genotypes. In validating 28 these observations in a rainfed field study, the tin allele had a negligible effect on seedling growth but 29 was associated with significantly (P<0.05) reduced tiller number (-37%), leaf area index (-26%) and 30 spike number (-35%) to reduce plant biomass (-19%) at anthesis. Root biomass, root-to-shoot ratio at 31 early stem elongation and root depth at maturity were increased in tin-containing NILs. Soil water use 32 was slowed in tin-containing NILs resulting in greater water availability, greater stomatal 33 conductance, cooler canopy temperatures and maintenance of green leaf area during grain-filling. 34 Together these effects contributed to increases in harvest index and grain yield. In both the controlled 35 and field environments, the tin gene was commonly associated with increased root length and biomass 36 but the significant influence of genetic background and environment suggests careful assessment of 37 tin-containing progeny in selection for genotypic increases in root growth

Adaptive responses of wild mungbean (Vigna radiata ssp. sublobata) to photo-thermal environment. II. Growth, biomass, and seed yield

The leaf growth, dry matter production, and seed yield of 11 wild mungbean (Vigna radiata ssp. sublobata) accessions of diverse geographic origin were observed under natural and artificial photoperiod–temperature conditions, to determine the extent to which genotypic differences could be attributed to adaptive responses to photo-thermal environment. Environments included serial sowings in the field in SE Queensland, complemented by artificial photoperiod extension and controlled-environment growth rooms. Photo-thermal environment influenced leaf growth, total dry matter production (TDM), and seed yield directly, through effects of (mainly cool) temperature on growth, and indirectly, through effects on phenology. In terms of direct effects, leaf production, leaf expansion, and leaf area were all sensitive to temperature, with implied base temperatures higher than usually observed in cultivated mungbean (V. radiata ssp. radiata). Genotypic sensitivity to temperature varied systematically with accession provenance and appeared to be of adaptive significance. In terms of the indirect effects of photo-thermal environment, genotypic and environmental effects on TDM were positively related to changes in total growth duration, and harvest index was negatively related to the period from sowing to flowering, similar to cultivated mungbean. However, seed yield was positively related to the duration of reproductive growth, reflecting the indeterminate growth habit of the wild accessions. As a consequence, the wild accessions are more responsive to favourable environments than typically observed in cultivated mungbean, which is determinate in habit. It is suggested that the introduction of the indeterminate trait into mungbean from the wild subspecies would increase the responsiveness of mungbean to favourable environments, analogous to that of black gram (V. mungo). Although the wild subspecies appeared more sensitive to cool temperature than cultivated mungbean, it may provide a source of tolerance to the warmer temperatures experienced during the wet season in the tropics

The influence of shoot and root size on nitrogen uptake in wheat is affected by nitrate affinity in the roots during early growth

Shoot and root system size influences N uptake in wheat (Triticum aestivum L.). Previously, we showed that four wheat genotypes with different biomass had similar N uptake at tillering. In the present study, we determined whether the similarity in N uptake in these genotypes was associated with genotypic differences in the affinity of the root system for NO3 - uptake. Kinetic parameters of NO3 - uptake were measured in hydroponic seedlings of vigorous and nonvigorous early growth wheat genotypes by exposing them to solutions with differing concentrations of K15NO3 for 15min. In the low concentration range, the high-affinity transport system of the nonvigorous cultivar Janz showed a higher maximum influx rate than the three vigorous lines and a higher affinity than two of the three vigorous lines. At high NO3 - concentrations, where the low-affinity transport system was functional, the responsiveness of NO3 - uptake to external concentrations was greater in Janz than in the vigorous lines. Both the high- and low-affinity transport systems were inducible. The genotypic variation in the kinetic parameters of NO3 - uptake was large enough to offset differences in morphological traits and should be considered in efforts to improve N uptake. In a field trial, the growth and N uptake performance of the four wheat genotypes was investigated over the winter-spring growing season (June-November of 2010). The field trial showed that although early N uptake was disproportionately large relative to biomass accumulation, the differences in uptake at tillering can be changed by subsequent patterns of uptake

Wheat genotypes with high early vigour accumulate more nitrogen and have higher photosynthetic nitrogen use efficiency during early growth

Genotypic differences in early growth and nitrogen (N) uptake among 24 wheat (Triticum aestivum L.) genotypes were assessed in a field trial. At late tillering, large genetic variation was observed for shoot biomass (23-56 gm(-2) ground area) and N uptake (1.1-1.8 gm(-2) ground area). A strong correlation between aboveground biomass and N uptake was observed. Variation around this relationship was also found, with some genotypes having similar N uptake but large differences in aboveground biomass. A controlled environment experiment was conducted to investigate the underlying mechanisms for this variation in aboveground biomass using three vigorous genotypes (38-19, 92-11 and CV97) and a non-vigorous commercial cultivar (Janz). Vigorous genotypes had lower specific leaf N in the youngest fully expanded leaf than Janz. However, there was no difference in chlorophyll content, maximum Rubisco activity or the rate of electron transport per unit area. This suggests that Janz invested more N in non-photosynthetic components than the vigorous lines, which could explain the higher photosynthetic N use efficiency of the vigorous genotypes. The results suggest that the utilisation of wheat genotypes with high early vigour could improve the efficiency of N use for biomass production in addition to improving N uptake during early growth