Trifolium Species Research Articles

Diversity for resistance to a moderately virulent bluegreen aphid (Acyrthosiphon kondoi Shinji) population in Trifolium species

The resistance of a diverse range of Trifolium species (clovers) to a highly virulent bluegreen aphid (BGA, Acyrthosiphon kondoi Shinji) population (Urrbrae 2011) collected in South Australia was assessed in greenhouse and field experiments, with the aims of determining the potential impact of this insect pest on biomass and identifying resistant genotypes for future plant-breeding activities. Resistance to BGA was found in populations of clovers that show some level of outcrossing—white clover (T. repens L.), rose clover (T. hirtum All.), crimson clover (T. incarnatum L.) and red clover (T. pratense L.)—and in one entry of the inbreeding subspecies of subterranean clover, T. subterraneum L. subsp. subterraneum (Katzn. and Morley). Resistance was not found in T. s. brachycalycinum (Katzn. and Morley) or T. s. yanninicum (Katzn. and Morley). In a greenhouse experiment, damage from BGA resulted in forage yield penalties of 72–100% when aphids were inoculated at 14 days after sowing and 13−74% when inoculated at 42 days after sowing, showing that in optimum conditions BGA can be a serious pest of clovers. Observations of severe damage caused by BGA in two regenerating field trials in southern New South Wales confirmed that field damage could occur in seasons favourable to aphid growth and reproduction. The severe damage that BGA can cause to clovers, and the sources of resistance we found, suggest that breeding for BGA resistance in clovers is warranted and feasible.

Karyotype Analysis and Protein Profile for Three Trifolium Species

The aim of the present investigation was cytological and protein comparison among three species within the genus Trifolium. The results revealed, Idiogram of the haploid complements of T. alexandrinum, T. refeigratum and T. repens. Besides T. refeigratum and T. repens have 16 pairs of chromosomes with a pair of satellites located at the end of their short arm in chromosome 16. T. alexandrinum has 8 pairs of chromosomes whose karyotyping formula is 2 nsm (+), 10 nsm (-) + 4 nm. The T. alexandrinum was clustering alone as well as having polymorphic bands that were different from T. refeigratum and T. repens.

Evaluation of genetic variability in Algerian clover (Trifolium L.) based on morphological and isozyme markers

Medoukali I., Bellil I., Khelifi D. (2015): Evaluation of genetic variability in Algerian clover (Trifolium L.) based on morphological and isozyme markers. Czech J. Genet. Plant Breed., 51: 50–61. Genetic variation within and among fifteen Trifolium species represented by 157 accessions was assessed using morphological and isozyme markers. Most of morphometric characters contributed to the discrimination of the species. No significant relationship between the environment of the collection site and morphological features was detected. The two isozyme systems analysed, esterase (EST) and glutamate oxaloacetate transaminase (GOT), proved polymorphic. Phenotype diversity of isozyme markers ranged from 0.07 to 0.61 with an average of 0.31 based on the polymorphic information content. The pairwise Jaccard’s similarity coefficient ranged between 0.10 and 0.60, indicating that the collection represents genetically diverse species. A considerable number of species-specific zymograms were detected which can be used for the species identification. The clustering pattern of isozyme markers was incongruent with the groupings based on quantitative traits. The rich isozyme variability present among the Algerian clover species indicates that they can provide good gene resources for breeding.

Triterpenoid saponins from the aerial parts of Trifolium argutum Sol. and their phytotoxic evaluation

Four triterpenoid saponins (1–4) were isolated from the aerial parts of Trifolium argutum Sol. (sharp-tooth clover) and their structures were elucidated by comprehensive spectroscopic analysis, including 1D and 2D NMR techniques, mass spectrometry and chemical methods. Two of them are new compounds, characterized as 3-O-[α-l-rhamnopyranosyl-(1→2)-β-d-galactopyranosyl-(1→2)-β-d-glucuronopyranosyl]-3β,24-dihydroxyolean-12-ene-22-oxo-29-oic acid (1) and 3-O-[β-d-galactopyranosyl-(1→2)-β-d-glucuronopyranosyl]-3β,24-dihydroxyolean-12-ene-22-oxo-29-oic acid (2). The occurrence of 3β,24-dihydroxyolean-12-ene-22-oxo-29-oic acid (melilotigenin) in its natural form is reported for the first time as a triterpenoid aglycone within Trifolium species. The phytotoxicity of compounds was evaluated on four STS at concentration 1μM to 333μM. Compound 1 was the most active, showing more than 60% inhibition on the root growth of L. sativa at the higher dose, with IC50 (254.1μM) lower than that of Logran® (492.6μM), a commercial herbicide used as positive control. The structure–activity relationships indicated that both aglycones and glycosidic parts may influence the phytotoxicity of saponins.

Anatomical structure of stems of some species of Trifolium L. in correspondence with the systematics and ecology

The article discusses the results of a comparative study of the anatomy of the stems of 4 Trifolium L. species. Complexes of stem anatomical characteristics which can be applied for species diagnostic are described. Anatomy of T. arvense, T. medium and T. montanum shows xerophytic features aimed on desiccation resistance, manifested in three directions: increased functions of epidermal tissue, enhanced development of mechanical tissues, increased development of water conducting system.

Evidence That Chlorinated Auxin Is Restricted to the Fabaceae But Not to the Fabeae.

Auxin is a pivotal plant hormone, usually occurring in the form of indole-3-acetic acid (IAA). However, in maturing pea (Pisum sativum) seeds, the level of the chlorinated auxin, 4-chloroindole-3-acetic acid (4-Cl-IAA), greatly exceeds that of IAA. A key issue is how plants produce halogenated compounds such as 4-Cl-IAA. To better understand this topic, we investigated the distribution of the chlorinated auxin. We show for the first time, to our knowledge, that 4-Cl-IAA is found in the seeds of Medicago truncatula, Melilotus indicus, and three species of Trifolium. Furthermore, we found no evidence that Pinus spp. synthesize 4-Cl-IAA in seeds, contrary to a previous report. The evidence indicates a single evolutionary origin of 4-Cl-IAA synthesis in the Fabaceae, which may provide an ideal model system to further investigate the action and activity of halogenating enzymes in plants.

Free radical scavenging actions of three Trifolium species in the protection of blood plasma antioxidant capacity in vitro

Context: Three clover [Trifolium L. (Leguminosae)] species were selected on the basis of data from traditional medicine, phytochemical profiles, and agricultural significance.Objective: The in vitro evaluations of free radical scavenging properties, ferric reducing abilities, and antioxidant effects of extracts from T. pratense L. (crude extract and phenolic fraction), T. pallidum L., and T. scabrum L. (phenolic fractions) were performed.Materials and methods: Activities of the Trifolium extracts were determined at their final concentrations of 1.5–50 µg/ml. Free radical scavenging properties of methanol extract solutions were estimated by the reduction of DPPH• and ABTS• radicals. Measurements of the total antioxidant capacity (TAC) were carried out to assess the antioxidant activities of the extracts in human blood plasma under conditions of oxidative stress, induced by 200 μM peroxynitrite.Results: The phenolic fraction of T. pratense displayed the strongest ABTS• and DPPH• radical scavenging effects (EC50 value of 21.69 and 12.27 µg/ml, respectively). The EC50 value for T. pallidum extract attained 29.77 and 30.06 µg/ml. The two remaining extracts were less potent scavengers (EC50 value higher than 50 µg/ml). Similar differences were obtained during evaluation of the ferric reducing abilities. Analysis of antioxidant properties of the extracts in blood plasma did not provide such evident differences in their actions, however, it indicated that the T. pratense phenolic fraction displayed the strongest effect.Conclusions: The examined Trifolium extracts partly protected blood plasma and enhanced its non-enzymatic antioxidant defense against harmful action of peroxynitrite in vitro.

Breaking through the feed barrier: options for improving forage genetics

Pasture based on perennial ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.) is the foundation for production and profit in the Australasian pastoral sectors. The improvement of these species offers direct opportunities to enhance sector performance, provided there is good alignment with industry priorities as quantified by means such as the forage value index. However, the rate of forage genetic improvement must increase to sustain industry competitiveness. New forage technologies and breeding strategies that can complement and enhance traditional approaches are required to achieve this. We highlight current and future research in plant breeding, including genomic and gene technology approaches to improve rate of genetic gain. Genomic diversity is the basis of breeding and improvement. Recent advances in the range and focus of introgression from wild Trifolium species have created additional specific options to improve production and resource-use-efficiency traits. Symbiont genetic resources, especially advances in grass fungal endophytes, make a critical contribution to forage, supporting pastoral productivity, with benefits to both pastures and animals in some dairy regions. Genomic selection, now widely used in animal breeding, offers an opportunity to lift the rate of genetic gain in forages as well. Accuracy and relevance of trait data are paramount, it is essential that genomic breeding approaches be linked with robust field evaluation strategies including advanced phenotyping technologies. This requires excellent data management and integration with decision-support systems to deliver improved effectiveness from forage breeding. Novel traits being developed through genetic modification include increased energy content and potential increased biomass in ryegrass, and expression of condensed tannins in forage legumes. These examples from the wider set of research emphasise forage adaptation, yield and energy content, while covering the spectrum from exotic germplasm and symbionts through to advanced breeding strategies and gene technologies. To ensure that these opportunities are realised on farm, continuity of industry-relevant delivery of forage-improvement research is essential, as is sustained research input from the supporting pasture and plant sciences.

Ground cover species selection to manage common ragweed (Ambrosia artemisiifolia L.) in roadside edge of highway

In southern Quebec, Canada, Trifolium species are commonly used as supplement ground cover along roadside of major roads to assist turf recruitment. They often fail to establish particularly near roadside edges. The noxious weed, Ambrosia artemisiifolia, exploits the empty niches. Based on an assumption that heavy metal stress mainly drives plant species shift, we conducted a field experiment to test the emergence of four species near roadside edges along a metal gradient. The test species included a clover ground cover (Trifolium arvense), two candidate ground covers (Lotus corniculatus and Coronilla varia), and the weed (A. artemisiifolia). Two hundred wet-cold stratified seeds of each species were sown in plots (20 × 30 cm), assigned separately for one of four species treatments, replicated three times. We measured final emergence percentage and soil metal contents and analyzed their correlations. T. arvense emergence was negatively correlated with Zn, Pb, and Cu. In contrast, A. artemisiifolia emergence was positively correlated with Zn, Pb, and Cu. The experiment supports the hypothesis that A. artemisiifolia colonization along roadside edges may be attributing to its greater tolerance for Zn, Pb, and Cu than T. arvense. In evaluating the two legume candidates, L. corniculatus was positively correlated with Zn, Pb, and Cu like A. artemisiifolia, while C. varia emergence did not have significant relationship with Pb and Cu. The current finding implies that L. corniculatus can be a good candidate because of its emergence performance and its tolerance to heavy metal, similarly to that of A. artemisiifolia.

Evolutionary origin of highly repetitive plastid genomes within the clover genus (Trifolium).

BackgroundSome clover species, particularly Trifolium subterraneum, have previously been reported to have highly unusual plastomes, relative to closely related legumes, enlarged with many duplications, gene losses and the presence of DNA unique to Trifolium, which may represent horizontal transfer. In order to pinpoint the evolutionary origin of this phenomenon within the genus Trifolium, we sequenced and assembled the plastomes of eight additional Trifolium species widely sampled from across the genus.ResultsThe Trifolium plastomes fell into two groups: those of Trifolium boissieri, T. strictum and T. glanduliferum (representing subgenus Chronosemium and subg. Trifolium section Paramesus) were tractable, assembled readily and were not unusual in the general context of Fabeae plastomes. The other Trifolium species (“core Trifolium”) proved refractory to assembly mainly because of numerous short duplications. These species form a single clade, which we call the “refractory clade” (comprising subg, Trifolium sections Lupinaster, Trifolium, Trichocephalum, Vesicastrum and Trifoliastrum). The characteristics of the refractory clade are the presence of numerous short duplications and 7-15% longer genomes than the tractable species. Molecular dating estimates that the origin of the most recent common ancestor (MRCA) of the refractory clade is approximately 13.1 million years ago (MYA). This is considerably younger than the estimated MRCA ages of Trifolium (c. 18.6 MYA) and Trifolium subg. Trifolium (16.1 MYA).ConclusionsWe conclude that the unusual repetitive plastome type previously characterized in Trifolium subterraneum had a single origin within Trifolium and is characteristic of most (but not all) species of subgenus Trifolium. It appears that an ancestral plastome within Trifolium underwent an evolutionary change resulting in plastomes that either actively promoted, were permissive to, or were unable to control, duplications within the genome. The precise mechanism of this important change in the mode and tempo of plastome evolution deserves further investigation.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-014-0228-6) contains supplementary material, which is available to authorized users.

Exotic grassland species have stronger priority effects than natives regardless of whether they are cultivated or wild genotypes.

During community assembly, early arriving exotic species might suppress other species to a greater extent than do native species. Because most exotics were intentionally introduced, we hypothesize there was human selection on regeneration traits during introduction. This could have occurred at the across- or within-species level (e.g. during cultivar development). We tested these predictions by seeding a single species that was either native, exotic 'wild-type' (from their native range), or exotic 'cultivated' using 28 grassland species in a glasshouse experiment. Priority effects were assessed by measuring species' effect on establishment of species from a seed mix added 21 d later. Exotic species had higher germination and earlier emergence dates than native species, and differences were found in both 'wild' and 'cultivated' exotics. Exotic species reduced biomass and species diversity of later arriving species much more than native species, regardless of seed source. Results indicate that in situations in which priority effects are likely to be strong, effects will be greater when an exotic species arrives first than when a native species arrives first; and this difference is not merely a result of exotic species cultivation, but might be a general native-exotic difference that deserves further study.

Clovers (Trifolium spp.).

Clovers (Trifolium spp.) constitute one of the major forage legumes widely grown for its rich protein content and its major role in maintaining environmental sustainability by improving the soil fertility. Gene technology can assist plant improvement efforts in clovers (Trifolium spp.), aiming to improve forage quality, yield, and adaptation to biotic and abiotic stresses. An efficient and reproducible protocol for Agrobacterium-mediated transformation of a range of Trifolium species, using cotyledonary explants and different selectable marker genes, is described. The protocol is robust and allows for genotype and Agrobacterium strain-independent transformation of clovers. Stable meiotic transmission of transgenes has been demonstrated for selected transgenic clovers carrying single T-DNA inserts recovered from Agrobacterium-mediated transformation. This methodology can also be successfully used for "isogenic transformation" in clovers: the generation of otherwise identical plants with and without the transgene from the two cotyledons of a single seed. Stable transgenes may be used in further functional genomics, develop new traits and profile gene expression using reporters, and facilitate purification of tissue or single cells.

Endopolyploidy Patterns in Organs of Trifolium Species (Fabaceae)

Abstract The pattern of endopolyploidy in the genus Trifolium was studied in mature organs of T. montanum and T. repens at reproductive stage, with comparative data for T. pratense, all from natural populations. Endopolyploidy in root, stem, petiole, leaf, inflorescence stalk, sepal, petal, stamen and carpel was detected by flow cytometry. 2C, 4C and 8C nuclei were found in organs of T. montanum and T. repens, and additionally 16C nuclei in organs of T. repens. The organs of T. montanum and T. repens differed in degree of endopolyploidy based on cycle values calculated from flow cytometry data; it was lowest in leaf and sepal in T. montanum and T. repens, and highest in T. montanum in petal and carpel and in T. repens in petiole and inflorescence stalk. These results are also seen in the two or more peaks of interphase nuclei in the flow cytometry histograms. There were significant correlations between the organs of T. pratense and T. repens as well as substantial differences between Trifolium species in the degree of endopolyploidy. T. pratense showed higher absolute endopolyploidy than T. montanum and T. repens. Principal component analysis showed that individuals of T. repens and T. montanum are more similar to each other than to individuals of T. pratense in degree of endopolyploidy. The observed variation between species might be explained by phylogenetic relationships and genome size differences.

Biological activity of clovers – Free radical scavenging ability and antioxidant action of six Trifolium species

Context: Clovers were chosen on the basis of traditional medicine recommendations, agricultural value, or available information on their promising chemical profiles.Objective: This study evaluates and compares free radical scavenging and antioxidant properties of six clover species: Trifolium alexandrinum L. (Leguminosae), Trifolium fragiferum L., Trifolium hybridum L., Trifolium incarnatum L., Trifolium resupinatum var. majus Boiss., and Trifolium resupinatum var. resupinatum L.Materials and methods: Free radical scavenging activity of the extracts (1.5–50 µg/ml) was estimated by reduction of 1,1-diphenyl-2-picrylhydrazyl (DPPH•) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic) acid (ABTS•) radicals. The Trifolium extract effects on total antioxidant capacity of blood plasma were determined by the reduction of ABTS•+ and DPPH• radicals, as well as with the use of the ferric reducing ability of plasma (FRAP) assay.Results: The UPLC analysis of chemical profiles of the examined extracts showed the presence of three or four groups of phenolic substances, including phenolic acids, clovamides, isoflavones, and other flavonoids. The measurements of free radical scavenging and ferric reducing ability of the examined clover extracts revealed the strongest effect for T. alexandrinum. Furthermore, antioxidant activity assays in human plasma have shown protective effects of all extracts against peroxynitrite-induced reduction of total antioxidant capacity.Conclusions: Trifolium plants may be a rich source of bioactive substances with antioxidant properties. The examined extracts displayed free radical scavenging action and partly protected blood plasma against peroxynitrite-induced oxidative stress; however, the beneficial effects of T. alexandrinum and T. incarnatum seem to be slightly higher.

Antimony accumulation and toxicity tolerance mechanisms in Trifolium species

Mining and industrial activity is increasing soil Sb concentrations worldwide. In contrast to animals and humans, plants seem to be highly tolerant of Sb. Asymptomatic accumulation of Sb in plants may cause problems with high exposure of humans and cattle through inadverted Sb intake with food or feed. The mechanisms of Sb tolerance in plants are poorly explored. At polluted sites Sb frequently is associated with high arsenic in the soil and both metalloids share some chemical properties. As phytochelatins play a major role in arsenic tolerance in plants, the present study aimed to investigate the possible role of phytochelatins (PCs) in Sb tolerance of clover, a fodder species that can contribute to Sb intake by cattle. For this purpose two clover species (Trifolium pratense L. and Trifolium repens L.) were exposed to different antimoniate (Sb(V)) concentrations (0, 50, 100 and 200μM) in the form of KSb(OH)6. Tolerance to Sb was assessed using relative root elongation, vital staining, and malondialdehyde (MDA) production as stress markers. Antimony tolerance was also analysed in plants treated with L-buthionine-[S,R]-sulphoximine (BSO), an inhibitor of PC synthesis. In spite of huge Sb accumulation in this fodder species (up to 380 and 770μgg−1 dry weight of Sb in shoots of T. pratense and T. repens, respectively), no negative effects on growth, lipid peroxidation or cell viability were observed. BSO treatment did not enhance the susceptibility of clover to Sb toxicity. In conclusion, Sb pollution of soils used for fodder production is dangerous because, due to the absence of visual toxicity symptoms in plants, Sb can be inadvertently incorporated into the trophic chain. Results here do not support a role for PCs in the high Sb tolerance of clover.

Ultraperformance Liquid Chromatography Tandem Mass Spectrometry Determination of Cyanogenic Glucosides in Trifolium Species

Cyanogenic glucosides were analyzed by ultra-high-performance liquid chromatography combined with mass spectrometry in 88 Trifolium species grown at the same site. On the basis of the occurrence of cyanogenic glucosides and the linamarin/lotaustralin ratio species could be grouped into five clusters. Cluster C1 included 37 species, which did not contain cyanogens. Cluster C2 (22 species) included plants containing only lotaustralin. In clusters C3 (14 species), C4 (13 species), and C5 (2 species) both linamarin and lotaustralin were present but at different ratios. In C3 and C4 the linamarin/lotaustralin ratio was below 1, whereas in cluster C5 the ratio was much higher. Generally, the total content of cyanogens was below 500 μg/g dry matter. Only in Trifolium repens var. biasoletti and Trifolium montanum extremely high cyanogen concentrations were observed. There was no general rule of occurrence of cyanogens. Samples of the same species from different countries accumulated cyanogens or could be free of these compounds.

Flooding and fragment size interact to determine survival and regrowth after fragmentation in two stoloniferous Trifolium species.

Clonal plants, which reproduce by means of stolons and rhizomes, are common in frequently flooded habitats. Resilience to disturbance is an important trait enabling plants to survive in such highly disturbed habitats. Resource storage is thought to enable clonal plants to resume growth after clonal fragmentation caused by disturbance. Here we investigated if submergence prior to disturbance reduces survival and regrowth of clonal fragments and whether or not genotypes originating from highly disturbed riverine habitats are more resistant to mechanical disturbance than genotypes from less disturbed coastal dune slack habitats. We further tested if variation in survival and regrowth was affected by internode size. Clones from contrasting habitats of two closely related Trifolium species were first genotypically characterized by amplification fragment length polymorphism and then subjected to soil flooding and subsequent clonal fragmentation. These species differ with respect to their abundance in riverine and dune slack habitats, with Trifolium repens mainly occurring in riverine grasslands and Trifolium fragiferum in coastal dune slacks. Soil flooding decreased survival and regrowth by up to 80 %. Plants originating from riverine grasslands were less negatively affected by fragmentation than plants from dune slack habitats. Surprisingly, ramets did not always benefit from being attached to a larger internode, as internode size was often negatively correlated with survival after fragmentation. Regrowth, on the other hand, was generally positively correlated with internode size. These unexpected results indicate that there may be contrasting selection pressures on internode size in stoloniferous species growing in severely disturbed habitats.

Source of variation of isoflavone concentrations in perennial clover species.

Introduction:Clover has attracted considerable interest not only as a valuable livestock forage plant, but also as an important source of isoflavones. The current study was aimed to assess the variation of concentration of three isoflavones in clover species grown under a cool temperate climate environment in Lithuania.Materials and Methods:Isoflavone contents were quantified in the plant parts of 21 accessions belonging to five perennial species of genus Trifolium (T. pratense, T. repens, T. medium, T. rubens, and T. pannonicum). Daidzein, formononetin, and genistein concentrations were quantified in separate plant parts by reversed-phase high-performance liquid chromatography. The analyte extraction was performed from dried and ground leaves, stems, flowers, and roots. The procedure included acid hydrolysis of isoflavone glycosides to aglycones.Results:According to the averaged sum of the three isoflavones quantified in leaves-stems-flowers, the five clover species ranked as follows: T. medium (7.54-3.62-2.31 mg/g) >T. pratense> T. rubens> T. pannonicum> T. repens (0.191-0.204-0.171 mg/g). The contribution of individual compound to the total isoflavone content depended on the species, accession, and plant part. The major part of the isoflavones is concentrated in leaves or stems; however, there is a great variation also.Conclusion:There exists a large variation in the total as well as in individual concentration of isoflavones among the clover species and plant parts and within species. With regard to isoflavone concentration and variability within species, some accessions of T. medium and T. pratense can be considered a highly promising source of phytoestrogens.

Genome sequence of Rhizobium leguminosarum bv trifolii strain WSM1689, the microsymbiont of the one flowered clover Trifolium uniflorum.

Rhizobium leguminosarum bv. trifolii is a soil-inhabiting bacterium that has the capacity to be an effective N2-fixing microsymbiont of Trifolium (clover) species. R. leguminosarum bv. trifolii strain WSM1689 is an aerobic, motile, Gram-negative, non-spore-forming rod that was isolated from a root nodule of Trifolium uniflorum collected on the edge of a valley 6 km from Eggares on the Greek Island of Naxos. Although WSM1689 is capable of highly effective N2-fixation with T. uniflorum, it is either unable to nodulate or unable to fix N2 with a wide range of both perennial and annual clovers originating from Europe, North America and Africa. WSM1689 therefore possesses a very narrow host range for effective N2 fixation and can thus play a valuable role in determining the geographic and phenological barriers to symbiotic performance in the genus Trifolium. Here we describe the features of R. leguminosarum bv. trifolii strain WSM1689, together with the complete genome sequence and its annotation. The 6,903,379 bp genome contains 6,709 protein-coding genes and 89 RNA-only encoding genes. This multipartite genome contains six distinct replicons; a chromosome of size 4,854,518 bp and five plasmids of size 667,306, 518,052, 341,391, 262,704 and 259,408 bp. This rhizobial genome is one of 20 sequenced as part of a DOE Joint Genome Institute 2010 Community Sequencing Program.

Genome sequence of the clover-nodulating Rhizobium leguminosarum bv. trifolii strain SRDI565.

Rhizobium leguminosarum bv. trifolii SRDI565 (syn. N8-J) is an aerobic, motile, Gram-negative, non-spore-forming rod. SRDI565 was isolated from a nodule recovered from the roots of the annual clover Trifolium subterraneum subsp. subterraneum grown in the greenhouse and inoculated with soil collected from New South Wales, Australia. SRDI565 has a broad host range for nodulation within the clover genus, however N2-fixation is sub-optimal with some Trifolium species and ineffective with others. Here we describe the features of R. leguminosarum bv. trifolii strain SRDI565, together with genome sequence information and annotation. The 6,905,599 bp high-quality-draft genome is arranged into 7 scaffolds of 7 contigs, contains 6,750 protein-coding genes and 86 RNA-only encoding genes, and is one of 100 rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project.