BackgroundField pea (Pisum sativum L.) is a self-pollinating, diploid, cool-season food legume. Crop production is constrained by multiple biotic and abiotic stress factors, including salinity, that cause reduced growth and yield. Recent advances in genomics have permitted the development of low-cost high-throughput genotyping systems, allowing the construction of saturated genetic linkage maps for identification of quantitative trait loci (QTLs) associated with traits of interest. Genetic markers in close linkage with the relevant genomic regions may then be implemented in varietal improvement programs.ResultsIn this study, single nucleotide polymorphism (SNP) markers associated with expressed sequence tags (ESTs) were developed and used to generate comprehensive linkage maps for field pea. From a set of 36,188 variant nucleotide positions detected through in silico analysis, 768 were selected for genotyping of a recombinant inbred line (RIL) population. A total of 705 SNPs (91.7%) successfully detected segregating polymorphisms. In addition to SNPs, genomic and EST-derived simple sequence repeats (SSRs) were assigned to the genetic map in order to obtain an evenly distributed genome-wide coverage. Sequences associated with the mapped molecular markers were used for comparative genomic analysis with other legume species. Higher levels of conserved synteny were observed with the genomes of Medicago truncatula Gaertn. and chickpea (Cicer arietinum L.) than with soybean (Glycine max [L.] Merr.), Lotus japonicus L. and pigeon pea (Cajanus cajan [L.] Millsp.). Parents and RIL progeny were screened at the seedling growth stage for responses to salinity stress, imposed by addition of NaCl in the watering solution at a concentration of 18 dS m-1. Salinity-induced symptoms showed normal distribution, and the severity of the symptoms increased over time. QTLs for salinity tolerance were identified on linkage groups Ps III and VII, with flanking SNP markers suitable for selection of resistant cultivars. Comparison of sequences underpinning these SNP markers to the M. truncatula genome defined genomic regions containing candidate genes associated with saline stress tolerance.ConclusionThe SNP assays and associated genetic linkage maps developed in this study permitted identification of salinity tolerance QTLs and candidate genes. This constitutes an important set of tools for marker-assisted selection (MAS) programs aimed at performance enhancement of field pea cultivars.
Plant seeds have long been promoted as a production platform for novel fatty acids such as the ω3 long-chain (≥ C 20) polyunsaturated fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) commonly found in fish oil. In this article we describe the creation of a canola (Brassica napus) variety producing fish oil-like levels of DHA in the seed. This was achieved by the introduction of a microalgal/yeast transgenic pathway of seven consecutive enzymatic steps which converted the native substrate oleic acid to α-linolenic acid and, subsequently, to EPA, docosapentaenoic acid (DPA) and DHA. This paper describes construct design and evaluation, plant transformation, event selection, field testing in a wide range of environments, and oil profile stability of the transgenic seed. The stable, high-performing event NS-B50027-4 produced fish oil-like levels of DHA (9-11%) in open field trials of T 3 to T 7 generation plants in several locations in Australia and Canada. This study also describes the highest seed DHA levels reported thus far and is one of the first examples of a deregulated genetically modified crop with clear health benefits to the consumer.
There is a large gap between the refined approaches to characterise genotypes and the common use of location and season as a coarse surrogate for environmental characterisation of breeding trials. As a framework for breeding, the aim of this paper is quantifying the spatial and temporal patterns of thermal and water stress for field pea in Australia. We compiled a dataset for yield of the cv. Kaspa measured in 185 environments, and investigated the associations between yield and seasonal patterns of actual temperature and modelled water stress.Correlations between yield and temperature indicated two distinct stages. In the first stage, during crop establishment and canopy expansion before flowering, yield was positively associated with minimum temperature. Mean minimum temperature below~78C suggests that crops were under suboptimal temperature for both canopy expansion and radiationuse efficiency during a significant part of this early growth period. In the second stage, during critical reproductive phases, grain yield was negatively associated with maximum temperature over 258C.Correlations between yield and modelled water supply/demand ratio showed a consistent pattern with three phases: no correlation at early stages of the growth cycle, a progressive increase in the association that peaked as the crop approached the flowering window, and a progressive decline at later reproductive stages. Using long-term weather records and modelled water stress for 104 locations, we identified three major patterns of water deficit nation wide. Environment type 1 (ET1) represents the most favourable condition, with no stress during most of the preflowering phase and gradual development of mild stress after flowering. Type 2 is characterised by increasing water deficit between 400 degree-days before flowering and 200 degree-days after flowering and rainfall that relieves stress late in the season. Type 3 represents the more stressful condition with increasing water deficit between 400 degree-days before flowering and maturity. Across Australia, the frequency of occurrence was 24% for ET1, 32% for ET2 and 43% for ET3, highlighting the dominance of the most stressful condition. Actual yield averaged 2.2 t/ha for ET1, 1.9 t/ha for ET2 and 1.4 t/ha for ET3, and the frequency of each pattern varied substantially among locations. Shifting from a nominal (i.e. location and season) to a quantitative (i.e. stress type) characterisation of environments could help improving breeding efficiency of field pea in Australia.
Field pea (Pisum sativum L.) is an important grain legume consumed both as human food and animal feed. However, productivity in low rainfall regions can be significantly reduced by inferior soils containing high levels of boron and/or salinity. Furthermore, powdery mildew (PM) (Erysiphe pisi) disease also causes significant yield loss in warmer regions. Breeding for tolerance to these abiotic and biotic stresses are major aims for pea breeding programs and the application of molecular markers for these traits could greatly assist in developing improved germplasm at a faster rate. The current study reports the evaluation of a near diagnostic marker, PsMlo, associated with PM resistance and boron (B) tolerance as well as linked markers associated with salinity tolerance across a diverse set of pea germplasm. The PsMlo1 marker predicted the PM and B phenotypic responses with high levels of accuracy (>80%) across a wide range of field pea genotypes, hence offers the potential to be widely adapted in pea breeding programs. In contrast, linked markers for salinity tolerance were population specific; therefore, application of these markers would be suitable to relevant crosses within the program. Our results also suggest that there are possible new sources of salt tolerance present in field pea germplasm that could be further exploited.
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