Strawberry clover (Trifolium fragiferum L.) is periodically raised as an alternative perennial pasture legume for temperate regions of Australia. Its tolerance of waterlogging is widely known, yet its ability to persist through periods of soil moisture deficit is often understated. Other desirable characteristics include its stoloniferous growth habit and tolerance of mildly saline conditions. Only four strawberry clover cultivars have been registered in Australia, and the most popular, cv. Palestine, is a direct introduction, released in 1938 and first certified in 1951. Furthermore, strawberry clover’s distribution has largely been confined to niche environments, particularly waterlogged and saline areas. This paper reviews the taxonomy and breeding system, morphology, distribution and ecology, and subsequent transfer of strawberry clover to Australia. It reviews and maps the suitability of strawberry clover for perennial pasture systems in the medium–-high rainfall and irrigated temperate zones of Australia, with reference to future climates. The paper also highlights the breeding focus, commercialisation and marketing required to supersede cv. Palestine and lists the germplasm available in the Australian Pastures Genebank, with origins. We conclude that, although strawberry clover is unlikely to become a dominant perennial pasture legume species in Australia, it could be used in a wider range of environments than just those affected by salinity and/or waterlogging stress.

The pasture legume Trifolium subterraneum ssp. yanninicum L. is waterlogging tolerant, but water-deficit (WD) susceptible. The interactive effect of waterlogged (WL) and WD (soil moisture fluctuation [SMF]) results in a severe stress impact. We studied three diverse ecotypes to identify traits associated with adaptation to SMF. Ecotypes were established in a glasshouse with two treatments imposed at 21 days: well-watered (WW-WW; 80% field capacity [FC]) and WL to WD (WL-WD). For WL-WD, pots were WL for 28 days (Harvest I), and then transitioned to WD (drained to 40% FC) and maintained for 10 days (Harvest II). For shoot relative growth rates (RGR) at Harvest I, WL had relatively little impact, although there was a greater reduction for Ecotype A (80% of WW) than Ecotypes B (92%) and C (87%). However, between Harvests I and II, the impact of WL-WD varied among ecotypes with Ecotype A being less affected (75% of WW-WW) than Ecotypes B (57%) and C (63%). For root RGR at Harvest I, WL resulted in a greater reduction for Ecotype A (52% of WW) than Ecotypes B (77%) and C (74%), while for WL-WD between Harvests I and II, Ecotype A showed a large increase (117% of WW-WW) compared to Ecotypes B (95%) and C (87%). In conclusion, the response to WD following WL varied among ecotypes, which demonstrates contrasting adaptation responses to SMF unrelated to WL tolerance. High yield under WL together with capacity for rapid growth post-WL in a drying soil profile could enhance adaptation to SMF.

The isoflavone formononetin (F) impacts livestock fertility and cultivars of the pasture legume Trifolium subterraneum L. (subclover) have been selected for F levels ≤0.2% of leaf dry weight. However, the impact of waterlogging (WL) on isoflavones is little studied. We investigated the response of isoflavones, biochanin A (BA), genistein (G) and F, to WL for: (1) Yarloop (high F) and eight low F cultivars each from subspecies subterraneum , brachycalycinum and yanninicum (Experiment 1); and (2) four cultivars and 12 ecotypes of ssp. yanninicum (Experiment 2). WL impacted F: estimated means increased from 0.19% (control) to 0.31% (WL) in Experiment 1 and from 0.61% to 0.97% in Experiment 2. Isoflavones under WL were highly heritable, particularly F (H 2 =95%). The proportions of BA, G and F were little changed by WL, with strong positive correlations between free-drained and waterlogged treatments. Isoflavone contents were not related to WL tolerance, as assessed by shoot relative growth rate. In conclusion, isoflavones varied among genotypes and increased with WL, but the proportion of individual isoflavones in each genotype was stable. High F under WL was unrelated with genotype tolerance to WL. Instead, it was a consequence of inherently high F for that particular genotype.

With a growing population and changing climate, there is an urgent need to increase global agricultural yields, while minimizing environmental impacts and land use. A component of this “sustainable intensification” process is the shift from synthetic fertilizer to nitrogen-fixing legumes. Subterranean clover ( Trifolium subterraneum L.) is the world's most widely grown annual pasture legume and in Australia its widespread adoption over the past century has led to increases in soil fertility and productivity, enabling greater animal production and crop yields. Subterranean clover is an unusual plant, burying its seed to protect it from grazing animals, which complicates efforts to harvest the seed from below the soil surface. The challenge of harvesting this seed has driven innovative farmers and inventors in Australia to develop specialized machinery and techniques, turning Australia into the largest subterranean clover seed producer in the world. In this chapter we review literature, patents, primary sources, personal interviews, and family histories to collate this story, deliver an integrated history of subterranean clover seed harvesting and make recommendations for the future of the industry. The history of this unique, farmer-led industry provides a number of lessons for the development of successful agricultural systems and the factors that influence agricultural innovation. We analyze these lessons from the past in order to provide insights for sustainable intensification in the future.

The pasture legume Trifolium subterraneum ssp. yanninicum exhibits waterlogging tolerance. This study investigates diversity for waterlogging tolerance within ssp. yanninicum. We tested the hypotheses that (1) variation for waterlogging tolerance exists within ssp. yanninicum and (2) is related to phenotypic and growth trait differences, which (3) reflect eco-geographic variables at site of origin. Twenty-eight diverse ssp. yanninicum ecotypes collected from the Mediterranean region and four cultivars were grown in a controlled environment glasshouse. Seedling traits were measured at 14 and 21 days after sowing. Waterlogged and free-draining (control) treatments were then imposed for 28 days. Relative distance and multivariate plasticity indices were calculated. Under waterlogging, shoot (87–108% of controls) and root (80–116% of controls) relative growth rates (RGRs) differed significantly among ssp. yanninicum. Waterlogging tolerance, as assessed by shoot RGR, had strong positive correlations with root RGR (r = 0.86; P < 0.001), petiole length (r = 0.59; P < 0.001) and leaf size (r = 0.55; P < 0.01) under waterlogging. The proportion of biomass as leaf increased under waterlogging, due to leaf size being maintained (mean 102% of controls), but petiole length decreased (mean 84% of controls). Petiole length was the most plastic trait. Seed size, seedling traits, maturity duration and eco-geographic variables at site of origin were not related to waterlogging tolerance. Wide variation in waterlogging tolerance exists within ssp. yanninicum. Petiole length reduction, an easy-to-measure and non-destructive indicator, could be used as a preliminary selection tool when screening large numbers of ssp. yanninicum for waterlogging tolerance in a breeding program.

Seed retention has not been evaluated for subterranean clover (Trifolium subterraneumL.), because its geocarpic seed-bearing burrs are currently harvested by suction systems. Development of improved harvest methods requires knowledge of subterranean clover seed retention characteristics and their changes with plant development. This study evaluates burr attachment and peduncle tensile strength during burr maturity until plant senescence across the three subspecies:subterraneum(cv. Dalkeith),yanninicum(cv. Monti) andbrachycalycinum(cv. Mawson). Peduncle tensile strength was greater than burr-to-peduncle attachment strength for each subspecies, with peak mean peduncle strength 30–130% greater than peak mean burr-to-peduncle attachment strength. Both strength measurements decreased significantly (greater than 50% for each subspecies) as the plant senesced, which was associated with reductions in burr moisture content, and burr and peduncle diameters. Microscopy indicated a ductile to brittle transition as peduncles senesced, reducing energy absorption and increasing the likelihood of failure at defects. These results are important for the commercial production of subterranean clover seed and suggest it may be possible to harvest seed before plant senescence with dig-invert machinery, similar to that used for peanut harvesting. However, this approach would require harvesting prior to maximum seed development and the implications for seed viability and yield need to be further evaluated.

Farmers are often overlooked and undervalued as sources of innovation, but can be powerful drivers of ingenuity and development. We evaluate historical developments in the Australian subterranean clover seed-production industry as a case study of farmer-driven innovation. Subterranean clover seed machinery patents (75% of which were patented by farmers) are analysed using conventional innovation frameworks, such as the theory of inventive problem solving (TRIZ), to extract lessons for supporting farmer-driven innovation. The small scale of this industry, compared with mainstream cereal-cropping industries and the isolation of farmers, provides analogous lessons for agriculture in developing countries. Economic drivers are important in enabling farmer innovation and the value proposition for developing new inventions must be clear to justify the time and expense. Farmers are different from firms and their on-farm knowledge and experience can form an essential part of innovation. Drivers of innovation also differ, with farmers less likely to attempt to commercialize inventions. Farmers can also be hesitant to share their inventions, instead holding them as trade secrets in competitive industries. Support and collaboration are needed from government and researchers to assist in commercialization or dissemination of useful innovations and to prevent knowledge from being confined to a localized farmer or region. Advances in agriculture require farmer input in research and development, but the benefits will be greater if farmers are enabled to be drivers of innovation.

.M. Abstract Harvesting subterranean clover seed is technically challenging and is reliant on suction harvesters developed in the early 1960s. Current challenges with the cost, difficulty and environmental impacts of seed harvesting threaten the future of the seed production industry. To overcome these challenges, we are investigating the use of peanut harvesting technology to harvest subterranean clover seed. A two-row peanut digger was tested on two subterranean clover seed crops to evaluate its applicability and efficacy. Preliminary tests show this approach has the potential to harvest subterranean clover seed and address some of the negative impacts of the current system. Future work should evaluate machinery modifications, plant agronomy, physiology and viability.

Messina (Melilotus siculus) cv. Neptune, an annual pasture legume native to the Mediterranean Basin, has recently been released for saltland pastures in southern Australia following demonstration of biomass production and persistence superior to other commercial pasture legumes in saline environments prone to winter waterlogging. Self-regenerating annual pasture legumes also require seed adaptations for both tolerating and avoiding salinity at germination in these environments. This study examined diversity within Neptune and 20 other messina accessions for salt tolerance at germination, recovery of germinability from temporary salt stress, and timing and extent of hardseed softening, compared with balansa clover (Trifolium michelianum) cv. Frontier, burr medic (Medicago polymorpha) cv. Scimitar and white melilot (Melilotus albus) cv. Jota. Germination rates after 14 days at 300 mM NaCl relative to 0 mM NaCl were ≥99% for Neptune and 18 other messina accessions, 66% for Scimitar, 21% for Jota and 11% for Frontier. No genotype germinated at 600 mM NaCl; however, when transferred to 0 mM NaCl after 14 days at 600 mM, all genotypes except Scimitar and Jota recovered partial germination, ranging from 13% to 93% of controls (0 mM NaCl for 28 days). The softening rate of hard (impermeable) seeds in the field varied among genotypes, with deferral of hardseed softening until late autumn–early winter, when rainfall is more likely, indicating greater persistence. The months in which the hardseed level first became significantly lower than the initial level (in freshly harvested seeds) after placement on the soil surface in December were: March for Frontier; April for Scimitar, Jota and Neptune; and March–July for all messina accessions. This study confirmed that messina has high salt tolerance and several avoidance mechanisms at germination that contribute to its adaptation to saline soils in southern Australia. Several messina accessions were superior to Neptune for individual traits which could be exploited for plant breeding. These results also have implications for saltland pastures in other regions of the world with Mediterranean-type climates.

Background and AimsIn the annual pasture legume Trifolium subterraneum , ssp. yanninicum exhibits higher waterlogging tolerance than ssp. brachycalycinum and ssp. subterraneum . This study investigates waterlogging tolerance within ssp. yanninicum ecotypes and explores correlations with seedling phenotypic traits and site of origin eco-geographic variables.MethodsTwenty eight diverse ssp. yanninicum ecotypes collected from the Mediterranean region and four cultivars were grown in a controlled environment glasshouse. After 14 days of growth seedling traits were measured. After 21 days of growth, free-drained (control) and waterlogged treatments were imposed for 28 days. Eco-geographic variables were generated from ‘WorldClim’ using collection site locations.ResultsUnder waterlogging, shoot relative growth rate (RGR) ranged from 87–108% and root RGR ranged from 80–116% of controls. Waterlogging reduced shoot dry weight (DW) in four of 32 genotypes, while root DW was reduced in 13 genotypes. Leaf size was maintained, or even increased, under waterlogging in 31 genotypes. However, petiole length was more affected by waterlogging and has value as a waterlogging tolerance indicator. Waterlogging tolerance was not significantly correlated with seedling DW, flowering time or precipitation at the site of origin, while shoot growth under waterlogging had a positive correlation with summer temperatures at origin.ConclusionsGenotypes of ssp. yanninicum tolerated transient waterlogging and greater tolerance was observed among ecotypes, rather than cultivars. An easy-to-measure indicator of tolerance was found in petiole length reduction. This study highlights untapped genotypic variability for breeders to improve the productivity and persistence of ssp. yanninicum under waterlogging.

Subterranean clover (Trifolium subterraneum L.) is Australia’s most widely sown annual pasture legume. Its widespread use as a pasture plant requires a well-functioning seed production industry, and Australia is the only significant producer of subterranean clover seed globally. However, the sustainability of this industry is under threat due to its reliance on ageing harvest equipment and the resultant environmental impacts. In order to evaluate seed harvesting practices, technology, and issues, we report on case studies, workshops, and a survey of seed producers across southern Australia. The Horwood Bagshaw Clover Harvester, designed in the 1950s, remains the most popular subterranean clover seed harvester. We discuss its use and modifications, and document several contemporary issues facing the seed production industry. Issues are primarily soil erosion and degradation; the expensive, slow and labour-intensive harvest process; and poor reliability and maintainability of harvesters that are now at least 30 years old. We conclude the root cause of these issues is the suction harvest technology utilised by the Horwood Bagshaw Clover Harvester. Analysis of the current harvest system is provided to support the development of new approaches to harvest subterranean clover seeds.

Tolerance to waterlogging and recovery ability was compared among the three subspecies of subterranean clover (Trifolium subterraneum L.) to identify tolerance mechanisms, in order to guide future subclover breeding activities. Three cultivars each of ssp. yanninicum, subterraneum and brachycalycinum were grown in a controlled environment glasshouse with T. michelianum as a waterlogging-tolerant control. After 28 days of growth two treatments were imposed for 35 days: free-draining (control) and waterlogged. A 21-day recovery period followed. After 35 days, waterlogging reduced shoot dry weight (DW) to 58–27% and root DW to 35–21% of respective controls in each subspecies. Shoot relative growth rate (RGR) of yanninicum was least affected (78–104% of control), compared to subterraneum (51–100%) and brachycalycinum (45–69%). The subterraneum cv. Denmark had a similar response to subspecies yanninicum. Shoot RGR of T. michelianum was 138% of its control. After recovery, all traits (except tap root length) recovered to control values. Up to 21 days, the main traits associated with waterlogging tolerance (shoot and root growth) were high petiole length, less petiole anthocyanin pigmentation and high leaf chlorophyll content, but by 35 days these were a high proportion of leaf biomass, high stomatal conductance and enhanced root porosity. Average root diameter and nodulation rates were unrelated to waterlogging tolerance. Subspecies yanninicum had superior waterlogging tolerance. Priority traits for easily-assessed indicators of waterlogging tolerance are less reduction of leaf size and high stomatal conductance.

Glasshouse and field investigations of the phenotypic expressions of resistance of a 97-member World Core Collection of subterranean clover (Trifolium subterraneum) collected from its native Mediterranean habitat and representing approximately 80% of the total genetic diversity within the known 10,000 accessions of the species against the most important damping-off and root rot (Phytophthora clandestina, and Pythium irregulare) and foliar (Kabatiella caulivora, Uromyces trifolii-repentis, and Erysiphe trifoliorum) pathogens were performed. An additional 28 diverse cultivars were also included. Associations of these genotypes among 18 disease parameters and 17 morphological traits, and among these disease parameters and 24 climatic and eco-geographic variables from their collection sites, were examined. Many genotypes showed strong phenotypic expression of novel host disease resistance against one or more pathogens, enabling their potential deployment as disease-resistant parents in subterranean clover breeding programs. These new sources of resistance enable future "pyramiding" of different resistance genes to improve resistance against these pathogens. Of particular value were genotypes with multiple disease-resistance across soilborne and/or foliar diseases, because many of these pathogens co-occur. All diseases had some parameters significantly correlated with one or more morphological traits and with one or more sites of origin variables. In particular, there were significant negative correlations between damping-off (i.e., germination) and 8 of the 17 morphological characters. The outcomes of these studies provide crucial information to subterranean clover breeding programs, enabling them to simultaneously select genotypes with multiple resistance to co-occurring soilborne and foliar diseases and desirable traits to offer renewed hope for re-establishing a more productive subterranean clover livestock feedbase despite multiple diseases prevailing widely.

Trifolium subterraneum L. subsp. yanninicum is a pasture legume that is widely grown in medium and high rainfall areas of southern Australia and shows waterlogging tolerance. This study investigated diversity within subsp. yanninicum corresponding to eco-geographic variables, which may help to identify adapted parents with new traits for genetic improvement. Diversity for 10 morphological traits, flowering time and leaf isoflavone content was investigated using 108 ecotypes derived from wild Mediterranean populations and 10 cultivars, grown as spaced plants. Among the ecotypes, the range of flowering time was 94–149 days after sowing, and contents of formononetin, genistein and biochanin A were 0.05–1.38%, 0.73–2.33% and 0.15–2.10% of dry matter, respectively. Leaf markings also varied considerably. Leaf size and petiole length were correlated at each growth stage. Later flowering genotypes had larger leaves, longer petioles, longer internodes and thicker stems at flowering, but smaller leaves and shorter petioles at both 63 and 88 days after sowing. Contents of genistein and biochanin A were unrelated, but both were negatively associated with formononetin. Flowering time had a weak positive influence on genistein and biochanin A, but a weak negative influence on formononetin. All traits among the ecotypes (except stem diameter and leaf mark crescent size) were significantly correlated with at least one of 22 eco-geographic variables from their collection sites. Precipitation and altitude were more influential than temperature. The study found sufficient diversity to broaden the narrow genetic base of current subsp. yanninicum cultivars; however, other agronomically important traits also need to be considered. Further diversity may result from targeted collection, particularly in areas not represented in annual legume genebanks.

Part of the Plant Sciences Commons, and the Soil Science Commons This document is available at https://uknowledge.uky.edu/igc/22/2-11/7 The 22nd International Grassland Congress (Revitalising Grasslands to Sustain Our Communities) took place in Sydney, Australia from September 15 through September 19, 2013. Proceedings Editors: David L. Michalk, Geoffrey D. Millar, Warwick B. Badgery, and Kim M. Broadfoot Publisher: New South Wales Department of Primary Industry, Kite St., Orange New South Wales, Australia

A remotely sensed pasture management technology, Pasture Growth Rate (PGR) has been developed by a research consortium of the Department of Agriculture Western Australia (DAWA), Commonwealth Scientific and Research Organisation (CSIRO), and Department of Land Information (DLI). A pilot study was funded by Australian Wool Innovation in 2003 to deliver the PGR information to a number of producers; four cooperative groups in the Great Southern and two groups in Dandaragan. Six producers from these groups were studied during the pasture growing season to determine how they used the technology and assess improvement in profitability, if any of their farming business. Key Findings • In all cases the use of PGR improved the profitability of the sheep enterprise.The increase in gross margin ranged from $23 to $334/per winter grazed hectare (WGHA). • The increased margin resulted from better utilisation of pasture, more effective feed budgeting and the introduction of new management techniques into the farming system. • The information on PGR provided timely information that was not discernible from observation of either the animals or pasture in the paddock until two to three weeks later. • The information on PGR improved confidence in decision making and helped reduce stress levels.

Subterranean clover (Trifolium subterraneum L.) is a diploid self-pollinated annual pasture legume native to the Mediterranean region and widely sown in southern Australia and other countries with Mediterranean-type climates. This study utilised a core collection of 97 lines, representing around 80% of the genetic diversity of the species, to examine morphological diversity within subterranean clover. A total of 23 quantitative agro-morphological and 13 semi-quantitative morphological marker traits were assayed on the core collection and 28 diverse Australian cultivars as spaced plants in a replicated common garden experiment. Relationships between these traits and 24 climatic and edaphic parameters at their sites of origin were also examined within the core collection. Significant diversity was present for all traits. The Australian cultivars had similar levels of diversity to the core collection for several traits. Among the agro-morphological traits, time to flowering, leaf size and petiole diameter in mid-winter, plant area in late winter, maximum stem length, content of the oestogenic isoflavone biochanin A and total isoflavone content, were correlated with seven or more environmental variables. These can be considered highly adaptive, being the result of strong environmental selection pressure over time. For the first time in a clover species, morphological markers, including leaf mark, anthocyanin pigmentation and pubescence traits, have been associated with rainfall and soil parameters. This suggests they either have an adaptive role or the genes controlling them may be linked to other genes controlling adaptive traits. This study demonstrated the value of core collections to examine diversity within much larger global collections. It also identified adaptive traits from wild plants that can be utilised to develop more productive and persistent subterranean clover cultivars. The high heritability of these traits indicates that selection gains can be readily made.