Changing climates and associated increased variability pose risks to alfalfa (Medicago sativa L.) cultivation, with the requirement to establish, survive, and maintain production under water stress. Crop wild relatives (CWR) of alfalfa include populations that have evolved to survive in a number of different, extreme environments, but until recently have had limited use in breeding programs. Here we report on the phenotypic diversity of alfalfa crop wild relatives that were selected to represent extremes in drought tolerance (by sourcing germplasm from environments with extremes in low rainfall, high temperature, shallow soils, and winter freezing) with the aim of providing germplasm with drought tolerance and improved forage yield traits for breeding programs in both warm and cool dry temperate environments. Newly formed hybrids created between M. sativa, M. arborea L. (a woody shrub), and M. truncatula Gaertn. (an annual species from the Mediterranean region) were developed or acquired to introduce new genetic diversity from the tertiary genepool. Preliminary characterization and evaluation was used for taxonomic classification, and to identify wild accessions and pre‐bred (hybrid) lines that offer new diversity for growth habit, seed size, fall dormancy, and forage yield. The accessions and pre‐breeding lines described have been donated to the Australian Pastures Genebank for conservation and distribution.
In the rain-fed mixed-farming systems of southern Australia, the consistent supply of high-value forage is limited by a range of climatic, edaphic and systems constraints. Over 2 years, we compared biomass production and nutritional value of 30 accessions of perennial legumes, and predicted intake, grazing days and growth of ewes and lambs. There was significant variation in nutritional value and biomass production between and within species. Lucerne (Medicago sativa) and sulla (Hedysarum coronarium) produced the greatest amount of biomass and energy. There was variability among accessions in digestibility (DMD), acid detergent fibre (ADF) and crude protein, and the rate of change in these traits as plants matured. Trifolium species had the highest DMD across all growth stages. Hairy canary clover (Dorycnium hirsutum), erect canary clover (Dorycnium rectum), greater birdsfoot trefoil (Lotus uliginosus), Australian trefoil (Lotus australis) and running postman (Kennedia prostrata) had energy levels that would not maintain liveweight of mature sheep. In the second year, species differed in response to harvesting treatments. Lucerne and sainfoin (Onobrychis viciifolia) were more productive under a frequent cutting regime. Accessions of white clover (Trifolium repens), red clover (Trifolium pratense), alsike clover (Trifolium hybridum), cullen (Cullen australasicum), strawberry clover (Trifolium fragiferum), sainfoin and birdsfoot trefoil (Lotus corniculatus) showed some promise, while Tedera (Bituminaria bituminosa) and milkvetch (Astragalus cicer) performed poorly under the experimental conditions. We conclude by discussing additional agronomic and nutritional factors that need consideration when developing novel perennial legumes for mixed-farming systems in the context of a changing climate.
BackgroundMedicago truncatula Gaertn. (barrel medic) is cultivated as a pasture legume for its high protein content and ability to improve soils through nitrogen fixation. Toxic concentrations of the micronutrient Boron (B) in agricultural soils hamper the production of cereal and leguminous crops. In cereals, the genetic analysis of B tolerance has led to the development of molecular selection tools to introgress and maintain the B tolerance trait in breeding lines. There is a comparable need for selection tools in legumes that grow on these toxic soils, often in rotation with cereals.ResultsGenetic variation for B tolerance in Medicago truncatula was utilised to generate two F2 populations from crosses between tolerant and intolerant parents. Phenotyping under B stress revealed a close correlation between B tolerance and biomass production and a segregation ratio explained by a single dominant locus. M. truncatula homologues of the Arabidopsis major intrinsic protein (MIP) gene AtNIP5;1 and the efflux-type transporter gene AtBOR1, both known for B transport, were identified and nearby molecular markers screened across F2 lines to verify linkage with the B-tolerant phenotype. Most (95%) of the phenotypic variation could be explained by the SSR markers h2_6e22a and h2_21b19a, which flank a cluster of five predicted MIP genes on chromosome 4. Three CAPS markers (MtBtol-1,-2,-3) were developed to dissect the region further. Expression analysis of the five predicted MIPs indicated that only MtNIP3 was expressed when leaf tissue and roots were assessed. MtNIP3 showed low and equal expression in the roots of tolerant and intolerant lines but a 4-fold higher expression level in the leaves of B-tolerant cultivars. The expression profile correlates closely with the B concentration measured in the leaves and roots of tolerant and intolerant plants. Whereas no significant difference in B concentration exists between roots of tolerant and intolerant plants, the B concentration in the leaves of tolerant plants is less than half that of intolerant plants, which further supports MtNIP3 as the best candidate for the tolerance trait-defining gene in Medicago truncatula.ConclusionThe close linkage of the MtNIP3 locus to B toxicity tolerance provides a source of molecular selection tools to pasture breeding programs. The economical importance of the locus warrants further investigation of the individual members of the MIP gene cluster in other pasture and in grain legumes.
Tetraploid plants, including those induced from diploid barrel medic (Medicago truncatula Gaertn.) and the snail medic [Medicago scutellata (L.) Mill.], have the potential to increase vigor and leaf size to improve adaptation to environments with low and variable rainfall in Mediterranean climates. The growth of tetraploid lines induced from barrel medic cultivar Sultan‐SU was evaluated against the diploid parent line and a natural tetraploid snail medic cultivar Sava in field conditions in Adelaide, South Australia. In field conditions, tetraploid lines had up to 106% greater autumn biomass than Sultan‐SU, 58% larger seed weight, and a 127% increase in leaf area. In a controlled environment experiment, snail medic Sava was the only entry to show improved early season (assessed at 6 wk) shoot biomass and canopy cover under short days and low temperatures that were designed to simulate a late start to the growing season. An examination of fresh flower buds showed that the fertility and stability of induced tetraploid lines varied between lines and generations. Hardseed breakdown patterns differed among tetraploid mutant lines, with two lines having much harder seed than Sultan‐SU. Some tetraploid Medicago spp. produced greater early biomass and larger leaves than diploids. Increasing ploidy level provides plant breeders with a promising tool in the development of new cultivars better suited to future climate scenarios.
The identification of the trait-defining gene and the development of a diagnostic marker enable efficient introgression of this economically important trait in annual medic improvement programs.
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