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1

Riday, Heathcliffe, and Kenneth A. Albrecht. "Registration of Kura1 Trifolium ambiguum (M. Bieb) Germplasm." Journal of Plant Registrations 4, no. 1 (January 2010): 80–85. http://dx.doi.org/10.3198/jpr2009.03.0142crg.

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2

Williams, W. M. "Trifolium interspecific hybridisation: widening the white clover gene pool." Crop and Pasture Science 65, no. 11 (2014): 1091. http://dx.doi.org/10.1071/cp13294.

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White clover (Trifolium repens) is adapted to moist, fertile soils in temperate zones. Despite its heterozygous allotetraploid nature, it lacks useful genetic variation for survival and growth in semi-arid, infertile soils. Although white clover is apparently genetically isolated in nature, 11 other taxa have so far been found that can be artificially hybridised into the wider gene pool. These species range from annuals to long-lived, hardy perennials with adaptations to stress environments, and they potentially provide new traits for the breeding of more resilient varieties of white clover. The delineation of the secondary, tertiary and quaternary gene pools is described, along with a review of interspecific hybrids achieved to date. The results of large breeding programs to integrate traits from T. nigrescens and 4x T. ambiguum are reviewed, and schemes introduced for the use of T. uniflorum, T. occidentale, T. pallescens, 2x T. ambiguum and 6x T. ambiguum. Interspecific hybrid breeding of white clover has the potential to enable the development of resilient perennial clovers for seasonally dry, infertile grassland environments in many parts of the world.
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3

Sheaffer, C. C., R. D. Mathison, and P. Seguin. "Vegetative establishment of Kura clover." Canadian Journal of Plant Science 88, no. 5 (September 1, 2008): 921–24. http://dx.doi.org/10.4141/cjps08026.

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Kura clover (Trifolium ambiguum M. Bieb.) is a rhizomatous perennial legume that is often challenging to establish from seed. Our objectives were to evaluate the effect of planting date (early April, late April, and mid-May), plant material (rhizomes and rhizomes with buds), and rhizome planting density (11, 44, and 178 plants m-2) on establishment of Kura clover. Establishment of the clover was enhanced by early April planting using rhizomes with buds. Kura clover populations increased with increased rhizome planting densities. Establishment of Kura clover at a rhizome density of 178 rhizome m-2 has the potential to provide yield and stands similar to broadcast seeding. Key words: Kura Clover (Trifolium ambiguum M. Bieb.), vegetative propagation
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4

Dear, BS, and M. Zorin. "Persistence and productivity of Trifolium ambiguum M. Bieb. (Caucasian clover) in a high altitude region of south-eastern Australia." Australian Journal of Experimental Agriculture 25, no. 1 (1985): 124. http://dx.doi.org/10.1071/ea9850124.

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The persistence and productivity of 12 lines of Trifolium ambiguum M. Bieb., encompassing three ploidy groups, and one line of T. montanum subsp. hurnboldtianum were compared in an elevated environment (1150 m) over a 4-year period with one cultivar each of T. repens L. and T. pratense L. Trifolium ambiguum was found to be well adapted to the cold winters and dry summers, all lines persisting throughout the experimental period, whereas the T. repens and T. pratense cultivars disappeared by the fourth year. The T. arnbiguum lines 'Monaro' and 'Kirovakan' (both hexaploid) and 'Alpine' and Forest (diploids) had the highest dry matter production. The control cultivars of T. repens and T. pratense outyielded T. ambiguum in the first year but were substantially less productive in later years as their density declined. T. montanum persisted but produced less dry matter than T. ambiguum and did not produce rhizomes or daughter plants. Ploidy of T. ambiguum was found to affect the date of flowering, persistence, leaflet shape and leaflet area. Productivity was not closely related to ploidy; however, the two most productive lines were hexaploids. All the T. ambiguum lines remained well nodulated over the 4 years, but the diploids had higher leaf nitrogen contents than the other two groups (3.4% vs. 2.5 and 2.4% N). The number of daughter plants produced was positively correlated (r=0.84) with rhizome length, which varied between lines but was not influenced by ploidy. Two bred lines, 'Alpine' (diploid) and 'Monaro' (hexaploid) exhibited superior vigour and spreading ability and warrant further evaluation as possible pasture legumes.
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5

Seguin, P., P. H. Graham, C. C. Sheaffer, N. J. Ehlke, and M. P. Russelle. "Genetic diversity of rhizobia nodulating Trifolium ambiguum in North America." Canadian Journal of Microbiology 47, no. 1 (January 1, 2001): 81–85. http://dx.doi.org/10.1139/w00-121.

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Kura clover (Trifolium ambiguum M.B.) is a persistent rhizomatous forage legume, whose use in the U.S.A. is limited by establishment difficulties in part attributable to nodulation problems. In this study, soil was collected from established stands of Kura clover growing in 9 diverse North American environments. Rhizobia were plant-trapped using Kura clover cv. Endura as host, then rhizobia from nodules fingerprinted using BOX-PCR. The diversity of isolates from North America was then contrasted to that of rhizobia from a single Caucasian environment (Russia), the center of origin for this species. Populations were characterized using clustering methods, and genetic diversity estimated using the Shannon-Weaver diversity index. The genetic diversity of the North American populations was extremely limited, all isolates being closely related to two of the strains found in a locally available commercial inoculant. In contrast, Russian isolates formed a distinct cluster with significant internal genetic diversity. Genetic diversity indices for the North American and Russian populations were 3.5 and 10.76, respectively. The implication of this and other studies is that Kura clover is highly specific in Rhizobium requirement. If the performance of this legume in the U.S.A. is to be improved, either by modifying current establishment practices or plant breeding, it is essential that these studies be paralleled by more collections and evaluation of rhizobia from its center of origin, given the extremely limited diversity of rhizobia found in North America.Key words: genetic diversity, rhizobia, Kura clover, BOX-PCR.
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6

Speer, G. S., and D. W. Allinson. "Kura clover (Trifolium ambiguum): Legume for forage and soil conservation." Economic Botany 39, no. 2 (April 1985): 165–76. http://dx.doi.org/10.1007/bf02907841.

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7

Beauregard, M. S., P. Seguin, C. C. Sheaffer, and P. H. Graham. "Characterization and evaluation of North American Trifolium ambiguum -nodulating rhizobia." Biology and Fertility of Soils 38, no. 5 (September 1, 2003): 311–18. http://dx.doi.org/10.1007/s00374-003-0661-y.

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8

Zheng, W., P. Seguin, and M. S. Beauregard. "Diversity of Trifolium ambiguum ?nodulating rhizobia from the lower Caucasus." Biology and Fertility of Soils 40, no. 2 (July 1, 2004): 128–35. http://dx.doi.org/10.1007/s00374-004-0752-4.

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9

Baturo-Ciesniewska, A., J. Andrzejewska, K. A. Albrecht, Cz Sadowski, and L. Lenc. "First Report of Sclerotinia Stem Blight Caused by Sclerotinia trifoliorum on Trifolium ambiguum in Poland." Plant Disease 97, no. 1 (January 2013): 142. http://dx.doi.org/10.1094/pdis-07-12-0624-pdn.

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Kura clover (Trifolium ambiguum M. Bieb.), a rhizomatous, persistent legume native to the Caucasus region, has received recent attention in North America and New Zealand as a pasture and silage crop. It is reported to be resistant to most pathogens affecting other clovers, including Sclerotinia trifoliorum Eriks. (3,4), one of the most destructive pathogens of clovers in northern Europe. Kura clover (cv. KTA202) was established in May 2009 near Mochełek, Poland (53° 13′ N, 17° 51′ E) on a Luvisol soil. By May 2011, 70% of plants grown in an experimental field (350 m2) had died, and 20% of the remaining plants were yellow and wilted. At crowns and the lower stem regions, wet, brown lesions with delicate white mycelium were observed. Lesion development was followed by death of the entire plant in a few days. By early June, only a few asymptomatic plants per square meter remained in the field. Tissue fragments of 20 symptomatic plants were surface-sterilized with 1% NaOCl for 1 min and plated on potato dextrose agar (PDA). A fungus with morphological characteristics of S. trifoliorum was consistently isolated. DNA isolation from sclerotia was performed with the DNeasy Plant Mini Kit (Qiagen, USA). Amplification and sequencing of the ITS region of rDNA was performed with primers ITS1/ITS4. NCBI Blast analysis of the 542-bp segment showed a 99% homology with most of S. trifoliorum and S. sclerotiorum strains in GenBank (e.g., AY547267.1 and EU082466.1). Sequence of isolate St0211TA was deposited in GenBank (Accession No. JQ743329). To determine growth rate of hyphae, morphology, and dimensions of sclerotia, colonies were grown in three replications on PDA at 20 ± 1°C in the dark. S. trifoliorum (CBS 122377) and S. sclerotiorum from our local collection were used as controls. Mean growth rate of S. trifoliorum isolates (20.5 mm/day) was slower compared to S. sclerotiorum (32.3 mm/day). Sclerotia began to form on delicate and smooth mycelium of S. trifoliorum on the entire surface of the plate in 7 to 8 days. Sclerotial size on day 28 was 2.0 to 9.0 × 2.0 to 7.0 mm (average 4.2 × 3.6 mm). Ultimately, the identification of S. trifoliorum was confirmed on the basis of ascospore morphology. Apothecia grew from sclerotia in wet sand at 12°C after 12 weeks. Asci contained dimorphic ascospores: four larger 13.0 to 16.0 × 6.0 to 9.0 mm (average 14.1 × 7.4 μm) and four smaller 10.0 to 12.0 × 5.4 to 6.0 mm (average 10.6 × 6.0 μm), typical for this species (1). Isolate St0511TA, which most intensively produced apothecia, was deposited in CBS (No. 133234). Koch's postulates were fulfilled by pathogenicity tests carried out on 2-week-old T. ambiguum seedlings grown in pots (6 × 30 plants), sprayed with a mycelial fragment suspension, and incubated at 15°C (2). Brown, wet spots with delicate white mycelium were observed on cotyledons after 3 days. After 5 days, approximately 10% of cotyledons were killed and mycelium appeared on stems and leaves, and after 10 days, 73% of seedlings were dead. S. trifoliorum was reisolated from all symptomatic tissues. To our knowledge, this is the first report of S. trifoliorum stem blight on T. ambiguum in the field. References: (1) E. N. Njambere et al. Plant Dis. 92:917, 2008. (2) L. H. Rhodes, Sclerotinia Crown and Stem Rot Resistance, http://www.naaic.org/stdtests/scleroti.htm , 1991. (3) A. K. Slesaravichyus et al., Selektsiya i Semenovodstvo Moskva 6:21, 1988. (4) N. L. Taylor and R. R. Smith, Adv. Agron. 63:153, 1998.
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10

Taylor, N. L., J. A. Anderson, and E. G. Williams. "Registration of Trifolium ambiguum ✕ T. repens Hexaploid Germplasm HBC/F2‐C." Crop Science 38, no. 1 (January 1998): 286–87. http://dx.doi.org/10.2135/cropsci1998.0011183x003800010065x.

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11

Abberton, M. T., T. P. T. Michaelson-Yeates, A. H. Marshall, K. Holdbrook-Smith, and I. Rhodes. "Morphological characteristics of hybrids between white clover, Trifolium repens L., and Caucasian clover, Trifolium ambiguum M. Bieb." Plant Breeding 117, no. 5 (November 1998): 494–96. http://dx.doi.org/10.1111/j.1439-0523.1998.tb01981.x.

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12

Abberton, M. T., J. H. MacDuff, S. Vagg, A. H. Marshall, and T. P. T. Michaelson-Yeates. "Nitrogen Fixation in Hybrids of White Clover (Trifolium repens L.) and Caucasian Clover (Trifolium ambiguum M. Bieb)." Journal of Agronomy and Crop Science 185, no. 4 (December 2000): 241–47. http://dx.doi.org/10.1046/j.1439-037x.2000.00438.x.

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13

Pryor, H. N., R. M. Elliot, W. L. Lowther, and C. W. Ronson. "Effect of rhizobia from caucasian clover(Trifolium ambiguum)on nodulation and nitrogen fixation of white clover(Trifolium repens)." New Zealand Journal of Agricultural Research 47, no. 1 (January 2004): 75–83. http://dx.doi.org/10.1080/00288233.2004.9513573.

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14

Hill, MJ, C. Mulcahy, and GG Rapp. "Perennial legumes for the high rainfall zone of eastern Australia. 1. Evaluation in single rows and selection of Caucasian clover (Trifolium ambiguum M. Bieb.)." Australian Journal of Experimental Agriculture 36, no. 2 (1996): 151. http://dx.doi.org/10.1071/ea9960151.

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A range of perennial legume species, including Trifolium ambiguum, T medium, Astragalus cicer, Coronilla varia, Lotus corniculatus, L. pedunculatus, and Lespedeza cuneata, were evaluated in glasshouse and field trials in 1989-92 at Armidale, New South Wales. Hexaploid germplasm of T. ambiguum was subjected to selection for increased seedling vigour, early biomass production and profuse flowering in glasshouse screening and spaced plant field nurseries. On an acid, solodic soil, T. ambiguum and T. medium grew modestly, Lotus spp. grew vigorously initially, but died out within 3 years, A. cicer and C. varia failed to persist beyond 1 year, but isolated plants of Lespedeza cuneata thrived. On an acid, freedraining, red-basalt soil, all accessions grew vigorously until early 1991. During the succeeding drought, Lotus sp. died out, A. cicer thinned out on the acid soil, but T. ambiguum, T medium, and C. varia remained persistent and productive. A large nursery of A. cicer accessions on the same soil limed to 5 t/ha before planting remained vigorous to the present. Superior hexaploid material of T ambiguum selected from spaced plants, originated chiefly from CPI 43909 and the ARS-2678 germplasm collection from Utah, USA.
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15

Woodman, R. F. "Trifolium ambiguum(Caucasian clover) in montane tussock grasslands, south island, New Zealand." New Zealand Journal of Agricultural Research 42, no. 3 (January 1999): 207–22. http://dx.doi.org/10.1080/00288233.1999.9513372.

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16

Seguin, P., P. H. Graham, C. C. Sheaffer, N. J. Ehlke, and M. P. Russelle. "Genetic diversity of rhizobia nodulating Trifolium ambiguum in North America." Canadian Journal of Microbiology 47, no. 1 (2001): 81–85. http://dx.doi.org/10.1139/cjm-47-1-81.

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17

Anderson, J. A., C. Mousset-Déclas, E. G. Williams, and N. L. Taylor. "An in vitro chromosome doubling method for clovers (Trifolium spp.)." Genome 34, no. 1 (February 1, 1991): 1–5. http://dx.doi.org/10.1139/g91-001.

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This research reports a new technique for chromosome doubling of clover (Trifolium sp.) axillary meristems via in vitro colchicine application. Plant material utilized included T. pratense (red clover) cv. Kenstar clones, and three interspecific hybrids: T. ambiguum (kura clover) × T. repens (white clover); T. alpestre × T. pratense; and T. sarosiense × T. pratense. Vegetative axillary meristems were excised from plants, surface sterilized, and trimmed to a length of 0.5–1 mm. Meristems were placed on the surface of a shoot proliferation medium (ML8) containing colchicine (0.1%) for 48 or 72 h and then transferred back to ML8. Alternative treatments were to preculture meristems on ML8 for 7 days prior to colchicine treatment. Plantlets with two or three trifoliolate leaves were induced to root on CR2 or RL rooting media. Preculturing of meristems on ML8 prior to colchicine exposure resulted in the highest chromosome doubling frequencies among the different genotypes, although there was apparent genotype × treatment interaction. Chromosome doubling frequencies were as high as 81 and 44% for initial root tips and mature shoots, respectively. To make rapid assessments of ploidy level of flowering plants, pollen shape was examined. Chromosome doubling increased the pollen stainability of the T. ambiguum × T. repens hybrid from 2.5 to 33.6%, but did not result in fertility in the other two interspecific hybrids.Key words: Trifolium, colchicine, chromosome doubling, interspecific hybrids.
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Hill, MJ, and C. Mulcahy. "Seedling vigour and rhizome development in Trifolium ambiguum M. Bieb. (Caucasian clover) as affected by density of companion grasses, fertility, drought and defoliation in the first year." Australian Journal of Agricultural Research 46, no. 4 (1995): 807. http://dx.doi.org/10.1071/ar9950807.

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Trifolium ambiguum was grown from seed in a mixture with Festuca arundinacea and Phalaris aquatica at four grass densities under high fertility, and at two intermediate grass densities under low fertility, with frequent or infrequent defoliation imposed from week 15 to week 31, in large pots for 1 year. Selective defoliation of the clover at week 35, and 3-4 weeks drought were imposed on high and low fertility treatments in the post-cutting period. The impact of these treatments was assessed by destructive harvest of shoots, roots and rhizomes. Plant densities of T. ambiguum and grasses were unaffected by the treatments; however, rhizome numbers were reduced by increasing density of grasses, by frequent defoliation, and at high fertility compared with low fertility where grass vigour was very poor. Selective defoliation of T. ambiguum and short-term drought temporarily reduced the contribution of T. ambiguum to harvested herbage, but had no permanent effect on growth rate. Final root and rhizome biomass of T. ambiguum was reduced by increasing density of grass, by frequent defoliation and at high fertility where grass competition was vigorous. We conclude that early growth of T. ambiguum, in particular root and rhizome development, may be better where the density of companion grasses is low, or grass vigour is low due to low fertility, provided soil P and S are not limiting for clover growth. Defoliation in the presence of vigorous grass competition reduces root and rhizome development and may not be effective in minimizing competition. Options for inclusion of T. ambiguum in mixed swards are discussed.
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19

Pryor, Heather N., W. L. Lowther, H. J. McIntyre, and C. W. Ronson. "An inoculantRhizobiumstrain for improved establishment and growth of hexaploid Caucasian clover (Trifolium ambiguum)." New Zealand Journal of Agricultural Research 41, no. 2 (January 1998): 179–89. http://dx.doi.org/10.1080/00288233.1998.9513301.

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20

Allinson, D. W., G. S. Speer, R. W. Taylor, and K. Guillard. "Nutritional characteristics of kura clover (Trifolium ambiguum Bieb.) compared with other forage legumes." Journal of Agricultural Science 104, no. 1 (February 1985): 227–29. http://dx.doi.org/10.1017/s0021859600043161.

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Kura or Caucasian clover (Trifolium ambiguum Bieb.) is a perennial, rhizomatous legume indigenous to Caucasian Russia and adjacent regions (Kannenberg & Elliott,1962; Bryant, 1974). Those agronomic characteristics that lend it potential significance, namely an extensive root-rhizome system, winter-hardiness, resistance to numerous and serious clover pests, and the production of nectar-rich flowers, were enumerated by Parker & Allen (1952). These authors also pointed out that usage of kura clover was hampered by ineffective nodulation. However, in recent years commercial and effective inoculant preparations have become available. Consequently, it has been suggested that kura clover could have potential as a pasture legume and for soil conservation purposes (Bryant, 1974; Spencer et al.1975). Pertinent literature on kura clover is scanty. The present study evaluated the nutritional characteristics of kura clover relative to otherforage legumes.
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Meredith, M. R., T. P. T. Michaelson-Yeates, H. J. Ougham, and H. Thomas. "Trifolium ambiguum as a source of variation in the breeding of white clover." Euphytica 82, no. 2 (1995): 185–91. http://dx.doi.org/10.1007/bf00027065.

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22

Lane, Ian, Eric Watkins, and Marla Spivak. "Turfgrass Species Affect the Establishment and Bloom of Kura Clover (Trifolium ambiguum) in Lawns." HortScience 54, no. 5 (May 2019): 824–28. http://dx.doi.org/10.21273/hortsci13779-18.

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Lawns represent one of the largest cultivated areas in urban landscapes, and in the Upper Midwest of the United States, lawns are typically composed of a small number of cool-season turfgrass species. There is increased interest in enhancing areas dedicated to lawns using flowering species for conservation purposes—for example, to support pollinators. In this study we used a model flowering forb, Kura clover (Trifolium ambiguum M. Bieb.), because—like many flowering species of conservation interest—it is slow to establish and is sensitive to grass competition. We varied the Kura clover seeding rate into four different turfgrass species treatments: kentucky bluegrass (Poa pratensis L.), hard fescue (Festuca brevipila Tracy), tall fescue [Schedonorus arundinaceum (Schreb.) Darbysh.], and perennial ryegrass (Lolium perenne L.) in two separate trials. Establishment and bloom of Kura clover was significantly greater in trial 1 for kentucky bluegrass and hard fescue than tall fescue and perennial ryegrass. In trial 2, Kura clover established significantly greater in kentucky bluegrass compared with tall fescue and perennial ryegrass, whereas Kura establishment in hard fescue was not significantly different from the other treatments. The seeding rate of Kura clover did not affect establishment in either trial. The results from this study suggest kentucky bluegrass and hard fescue are promising turf companion grasses for future forb/turf interseeding research.
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Williams, E. G., N. L. Taylor, J. Van Den Bosch, and W. M. Williams. "Registration of Tetraploid Hybrid Clover Germplasm from the Cross of Trifolium Ambiguum ✕ T. Repens." Crop Science 30, no. 2 (1990): 427. http://dx.doi.org/10.2135/cropsci1990.0011183x003000020050x.

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Anderson, J. A., N. L. Taylor, and E. G. Williams. "Cytology and Fertility of the Interspecific Hybrid Trifolium ambiguum ✕ T. repens and Backcross Populations." Crop Science 31, no. 3 (May 1991): 683–87. http://dx.doi.org/10.2135/cropsci1991.0011183x003100030027x.

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Taylor, N. L., J. A. Anderson, E. G. Williams, and W. M. Williams. "Registration of Octoploid Hybrid Clover Germplasm from the Cross of Trifolium Ambiguum ✕ T. Repens." Crop Science 31, no. 5 (September 1991): 1395. http://dx.doi.org/10.2135/cropsci1991.0011183x003100050088x.

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YAMADA, Toshihiko, and Hisao FUKUOKA. "Production of interspecific hybrids between Trifolium ambiguum M.Bieb. and T.repens L. by ovule culture." Ikushugaku zasshi 36, no. 3 (1986): 233–39. http://dx.doi.org/10.1270/jsbbs1951.36.233.

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Wijkstra, G. S., and P. L. Guy. "DISEASE NOTES OR NEW RECORDS: Trifolium ambiguum is not resistant to alfalfa mosaic virus." Australasian Plant Pathology 25, no. 3 (1996): 213. http://dx.doi.org/10.1071/ap96037.

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28

Patrick, H. N., and W. L. Lowther. "Influence of the number of rhizobia on the nodulation and establishment of Trifolium ambiguum." Soil Biology and Biochemistry 27, no. 4-5 (April 1995): 717–20. http://dx.doi.org/10.1016/0038-0717(95)98654-7.

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29

Lang, Jaroslav, and K. Vejražka. "Yields and quality of forage legumes under imbalanced year precipitation conditions on south Moravia." Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis 60, no. 6 (2012): 217–24. http://dx.doi.org/10.11118/actaun201260060217.

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In this paper, yield and quality of forage following species of forage legumes were evaluated with regard to precipitations: lucerne (Medicago sativa L.), red clover (Trifolium pratense L.), white clover (Trifolium repens L.), kura clover (Trifolium ambiguum M.), alsike clover (Trifolium hybridum L.) and birdsfoot trefoil (Lotus corniculatus L.). The trial was sown in the spring of 2008, evaluated in the period 2009–2011. Analysis of samples was performed with the apparatus NIRS 6500. Following parameters were evaluated: production of dry matter, energy concentration (NEL), contents of fibre and crude protein. The highest three year yield average was measured for lucerne (15.01 t.ha−1), followed by red clover group (9.3–11.8 t.ha−1). Kura clover gained the lowest yield (1.97 t.ha−1). The average crude protein contents (g.kg−1) were: lucerne 211.47, red clover group (184.3–194.8), white clover group (229.1–238.7) and birdsfoot trefoil (204.2). The obtained results indicated that lucerne responded at best to periods of drought. Although the production of dry matter decreased in periods of drought, the canopy of stands remained to be complete in contradistinction to white clover, which partly disappeared from the stand. Red clover and alsike clover disappeared from the stand during the trial.
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30

Sheaffer, C. C., and G. C. Marten. "Kura clover forage yield, forage quality, and stand dynamics." Canadian Journal of Plant Science 71, no. 4 (October 1, 1991): 1169–72. http://dx.doi.org/10.4141/cjps91-163.

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Persistent legumes are needed for livestock production in the northern U.S.A. and Canada. We compared forage yield, forage quality and stands of kura clover (Trifolium ambiguum M. Bieb.), a rhizomatous perennial legume, with commonly grown forage legumes. Kura clover had lower initial stands and yields than most other legumes, but was ultimately among the highest yielding legumes and had greater stand persistence and higher forage digestibility than other legumes when subjected to a diversity of cutting schedules. Key words: Kura clover, forage yield, cutting schedules, persistence
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31

Widdup, K. H., S. W. Hussain, W. M. Williams, W. L. Lowther, H. N. Pryor, and B. L. Sutherland. "The development and plant characteristics of interspecific hybrids between white and caucasian clover." NZGA: Research and Practice Series 11 (January 1, 2003): 143–48. http://dx.doi.org/10.33584/rps.11.2003.3004.

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A series of Trifolium ambiguum x T. repens hybrid populations has been developed at the hexaploid (four T. repens and two T. ambiguum genomes) and pentaploid (four T. repens and one T. ambiguum genome) level. When tested for effectiveness of symbiotic nitrogen fixation, the 6x and 5x hybrids nodulated with either T. repens or T. ambiguum strains of rhizobia but only formed an effective N- fixing symbiosis with a mix of T. repens rhizobia strains. When grown in the field, the 6x and 5x hybrid plants had a similar morphology to white clover in that the hybrids grew surface stolons (fewer numbers than white clover) and no underground rhizomes. The advantage of the 6x hybrid was deeper roots and a greater proportion of root, a characteristic considered important for greater drought tolerance and persistence than for white clover. The 6x hybrid indicated 55% of the seed production potential of white clover, but there was large variation between plants in all reproductive traits. The 5x hybrid showed poor levels of seed set. The growth pattern of the 6x hybrids in the field indicated lower herbage yield in the first year but improved performance compared with white clover into the second year. At this early stage, the hybrid breeding populations consist of unselected and novel hybrid combinations. Variations in growth and reproductive characteristics exist between hybrid plants, thus providing scope for improvement through selection and breeding. Key words: caucasian clover, interspecific hybrid, morphology, nitrogen fixation, seed production, white clover
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32

Seguin and A. F. Mustafa, P. "Chemical composition and ruminal nutrient degradabilities of fresh and ensiled Kura clover (Trifolium ambiguum M.B.)." Canadian Journal of Animal Science 83, no. 3 (September 1, 2003): 577–82. http://dx.doi.org/10.4141/a03-032.

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Kura clover is a perennial rhizomatous forage legume mainly used for grazing in permanent pastures. A study was conducted to determine the ensiling potential of Kura clover by comparing the chemical composition and ruminal nutrient degradabilities of fresh and ensiled forage of two cultivars (Cossack and Endura). Ensiling characteristics were determined by ensiling forages in mini-silos for 50 d. Ruminal nutrient degradabilities of fresh and ensiled forages were determined by the nylon bag technique using two lactating cows fitted with ruminal cannulas. Results showed that silages of both cultivars were well preserved as indicated by low pH and high lactic acid concentration. Cultivar had little effect on the chemical composition of Kura clover silage. However, ensiling increased (P < 0.05) soluble protein and non-protein nitrogen (NPN) and reduced (P < 0.05) neutral and acid detergent insoluble protein fractions. Ensiling increased NPN and reduced true protein by 87 and 29%, respectively. Results of the in situ study indicated that cultivar had no effect on effective ruminal degradability of dry matter (DM), crude protein (CP), an d neutral detergent fiber (NDF). Relative to fresh forage, ensiled Kura clover had similar effective ruminal degradability of DM (712 g kg-1 of DM) and NDF (417 g kg-1 of NDF) but a higher (P < 0.05) effective ruminal degradability of CP (806 vs. 725 g kg-1 of CP). It was concluded that Kura clover may be preserved as silage with minimal alterations in forage quality. These findings will increase the acceptability of Kura clover by widening possible utilizations. Key words: Forage quality, Kura clover (Trifolium ambiguum M.B.), protein fractions, ruminal degradability, silage
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33

Hussain, S. W., and W. M. Williams. "Development of a fertile genetic bridge between Trifolium ambiguum M. Bieb. and T. repens L." Theoretical and Applied Genetics 95, no. 4 (September 1997): 678–90. http://dx.doi.org/10.1007/s001220050612.

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34

Monk, S., D. J. Moot, B. Belgrave, M. P. Rolston, and J. R. Caradus. "Availability of seed for hill country adapted forage legumes." NZGA: Research and Practice Series 16 (January 1, 2016): 257–67. http://dx.doi.org/10.33584/rps.16.2016.3237.

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New Zealand hill and high country are marginal environments for perennial ryegrass and white clover. Other pasture species, particularly legumes, provide more productive and persistent options for the range of soil climate and topography found in these environments. On cultivatable hill country, lucerne (alfalfa) has been successfully introduced to dryland areas with imported seed increasing five-fold to 210 t per annum over the last decade. This has led to the first release of a New Zealand selected cultivar in 20 years. For subterranean clover reliance on hardseeded Australian cultivars that frequently fail to meet New Zealand biosecurity standards means seed supply is inconsistent. The potential exists to create a niche seed market through selection of locally adapted material grown for seed in New Zealand. Lotus pedunculatus is available commercially but seed often fails to meet certification standards and is predominantly used in forestry. Lotus corniculatus requires agronomic research to overcome some management constraints and the re-establishment of seed supply before it would be a viable option for drier hill country. Seed production for perennial lupins in New Zealand is predominantly for an export ornamental market, with some direct relationships with growers allowing onfarm use. Caucasian clover seed production has ceased in New Zealand and the demand for seed particularly from high country farmers is no longer met. The smallseeded annual balansa clover is being integrated into farm systems and its prolific seeding has enabled some on-farm production of seed for personal use. For it and arrowleaf and Persian clovers, imported cultivars are available and a local market is unlikely to thrive until agronomic and hard seed issues are addressed. Seed supply of forage legumes to satisfy demand for hill country is problematic and will require development of different models including grower co-ops, regional seed retailers, and on-farm production for niche markets. Keywords: alfalfa, Lolium perenne, Lupinus polyphyllus, Medicago sativa, perennial ryegrass, Sitona lepidus, Trifolium ambiguum, Trifolium michelianum, Trifolium repens, Trifolium resupinatum, Trifolium subterraneum, Trifolium tumens, Trifolium vesiculosum, white clover
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35

Hay, F. R., R. D. Smith, R. H. Ellis, and L. H. Butler. "Developmental changes in the germinability, desiccation tolerance, hardseededness, and longevity of individual seeds of Trifolium ambiguum." Annals of Botany 105, no. 6 (March 12, 2010): 1035–52. http://dx.doi.org/10.1093/aob/mcq037.

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36

Hall, E. J., R. Reid, B. Clark, and R. Dent. "The search for alternative temperate perennial pasture species for low to medium rainfall environments in Tasmania." NZGA: Research and Practice Series 16 (January 1, 2016): 275–79. http://dx.doi.org/10.33584/rps.16.2016.3239.

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In response to the need to find better adapted and more persistent perennial pasture plants for the dryland pastures in the cool-temperate low to medium rainfall (500-700 mm) regions, over 1000 accessions representing 24 species of perennial legumes and 64 species of perennial grasses, were introduced, characterised and evaluated for production and persistence under sheep grazing at sites throughout Tasmania. The work has identified four alternative legume species in Talish Clover (Trifolium tumens). Caucasian Clover (T. ambiguum), Stoloniferous Red Clover (T. pratense var. stoloniferum), Lucerne x Yellow Lucerne Hybrid (Medicago sativa x M.sativa subsp. falcata); and two grass species in Coloured Brome (Bromus coloratus) and Hispanic Cocksfoot (Dactylis glomerata var hispanica). Keywords: persistence, perennial grass, perennial legume
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Hill, MJ, C. Mulcahy, and GG Rapp. "Perennial legumes for the high rainfall zone of eastern Australia. 2. Persistence and potential adaptation zones." Australian Journal of Experimental Agriculture 36, no. 2 (1996): 165. http://dx.doi.org/10.1071/ea9960165.

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Cultivars and accessions of a range of perennial legumes were evaluated in small plot field trials at 4 on-farm locations on the Northern Tablelands of New South Wales. Variable establishment from seed resulted in good stands at 2 sites, and moderate to poor stands at the other 2 sites. In spite of this, trends in behaviour of individual species were apparent even where stands were very poor. This was exemplified by the gradual improvement in frequency of Trifolium medium at the most montane site. Trifolium ambiguum and Coronilla varia were most persistent across all sites. Good stands of Lotus corniculatus, L. pedunculatus, T. pratense and Medicago sativa thinned out within several years with M. sativa most persistent. Astragalus cicer thinned out rapidly on the relatively acidic soils but maintained a good stand at 1 site on a deep, free-draining granite. From these results and known climatic responses, simple logical rules were constructed to predict potential legume adaptation zones for eastern Australia. The zones of adaptation for the range of species are compared with a nominal zone for T. repens to identify areas where legume persistence might be improved with additional species.
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38

Walker, J. A., and J. R. King. "Above and below-ground competition between Kura clover (Trifolium ambiguum) and meadow bromegrass (Bromus biebersteinii): A greenhouse study." Canadian Journal of Plant Science 89, no. 1 (January 1, 2009): 21–27. http://dx.doi.org/10.4141/cjps08010.

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Kura clover (Trifolium ambiguum) is a perennial legume that shows potential for use in pasture mixtures in western Canada. Previous studies have shown that early growth of kura clover is reduced when grown in mixtures with grass species. A greenhouse study examined the relative importance of above- and below-ground competition on growth and development of kura clover when grown with a grass species. Kura clover plants were grown in pots with meadow bromegrass (Bromus biebersteinii) and barriers were put in place to remove all competition, shoot competition, root competition, or to allow full competition. After 12 wk, plants were harvested. Height, leaf area, leaf number, leaf dry weight, and root/crown dry weight were measured. Kura clover leaf number and leaf area were greatest when roots did not interact with meadow bromegrass roots. Vegetative biomass of kura clover doubled when there was no root competition. Shoot competition did not alter leaf number, leaf area, or leaf biomass. Successful establishment of kura clover is dependent on the reduction of root competition during the seedling phase. Measures taken to minimize the below-ground interaction should positively affect the yield potential of both species. Key words: Kura clover, meadow bromegrass, shoot competition, root competition
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39

Williams, Warren M., Isabelle M. Verry, Helal A. Ansari, S. Wajid Hussain, Ihsan Ullah, Michelle L. Williamson, and Nicholas W. Ellison. "Eco-geographically divergent diploids, Caucasian clover (Trifolium ambiguum) and western clover (T. occidentale), retain most requirements for hybridization." Annals of Botany 108, no. 7 (August 31, 2011): 1269–77. http://dx.doi.org/10.1093/aob/mcr226.

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40

Granatstein, David, Joan R. Davenport, and Elizabeth Kirby. "Growing Legumes in Orchard Alleys as an Internal Nitrogen Source." HortScience 52, no. 9 (September 2017): 1283–87. http://dx.doi.org/10.21273/hortsci12121-17.

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The drive alley in modern apple (Malus ×domestica Bork.) orchards often receives enough light to grow plants other than the typical perennial grass cover. By planting leguminous species in this area, it is possible to produce a portion of the nitrogen needs of the orchard by mowing the vegetation and blowing it onto the tree row where it mineralizes and releases available N over the tree roots. Four perennial legume species [alfalfa (Medicago sativa L.), ladino white clover (Trifolium repens L.), birdsfoot trefoil (Lotus corniculatus L.), kura clover (Trifolium ambiguum L.)] were compared with the resident grass cover crop in a mature apple orchard. All legumes were direct-seeded into the alley to avoid any soil disturbance and were successfully established. Legume biomass and tissue N were monitored, along with biweekly monitoring of tree row soil nitrogen with both soil sampling and ion exchange resins using Plant Root Simulator® probes. Four mowings of alfalfa contained ≈43 kg total N/ha that was added to the tree row during the second season (2009), with a dry matter C:N of 10.8. Economically, legume nitrogen appears to be less expensive than other sources of organic N and may be cost competitive with synthetic fertilizer N when prices are high.
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Nguyen, Tuan Dung, Mitchell Andrews, Tommy W. S. Ley, Daniel Dash, Megan Petterson, Bevan S. Weir, James D. Morton, Alistair D. Black, and Richard J. Lucas. "Caucasian clover (Trifolium ambiguum) specific rhizobia persist in low and high fertility soils in the South Island of New Zealand." New Zealand Journal of Agricultural Research 63, no. 3 (March 24, 2019): 332–40. http://dx.doi.org/10.1080/00288233.2019.1589538.

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42

Kim, SungUn, Kenneth Albrecht, Craig Sheaffer, Dokyoung Lee, Senthil Subramanian, and Vance Owens. "Biomass Production of Prairie Cordgrass (Spartina pectinata Link.) Using Urea and Kura Clover (Trifolium ambiguum Bieb.) as a Source of Nitrogen." BioEnergy Research 13, no. 4 (June 1, 2020): 1095–107. http://dx.doi.org/10.1007/s12155-020-10139-2.

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Abstract Optimizing nitrogen (N) management is an important factor for sustainable perennial biomass systems. However, N application is costly, both financially and environmentally. Our objectives were to determine: (1) N rate and plant spacing effects on yield and yield components of prairie cordgrass swards and (2) fertilizer N replacement value (FNRV) of kura clover in prairie cordgrass-kura clover binary mixtures. Plots were established in Illinois, Minnesota, South Dakota, and Wisconsin, USA, in 2010. Kura clover was transplanted on 30-cm centers in all treatments in which it was a component; prairie cordgrass seedlings were transplanted within the kura clover on 60- and 90-cm centers. Monoculture prairie cordgrass stands were established at the same population densities of mixed stands and fertilized with 0, 75, 150, or 225 kg N ha-1. Biomass was harvested in the autumn from 2011 to 2013. N (urea), year, plant spacing, and year × plant spacing affected prairie cordgrass production at all locations. Prairie cordgrass yield increased with N application, but the response varied by location. N application tended to increase prairie cordgrass tiller density and consistently increased tiller mass. Prairie cordgrass yield with 0 N was equal to or less than the yield of prairie cordgrass/kura clover mixtures at all locations in 2011 and 2012; however, kura clover provided a FNRV of 25–82 kg N ha-1 to prairie cordgrass in 2013. Kura clover has potential to provide N to prairie cordgrass in binary mixtures of these two species and on land that may not be easily farmed due to wetness.
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43

KRAWUTSCHKE, M., J. KLEEN, N. WEIHER, R. LOGES, F. TAUBE, and M. GIERUS. "Changes in crude protein fractions of forage legumes during the spring growth and summer regrowth period." Journal of Agricultural Science 151, no. 1 (March 30, 2012): 72–90. http://dx.doi.org/10.1017/s002185961200024x.

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SUMMARYOnly a few previous studies have analysed the crude protein (CP) fractions of the Cornell Net Carbohydrate and Protein System during the growth period of forage legumes. The objective of the present study was to investigate the changes in CP fractions during the spring growth and summer–autumn regrowth period of five forage legume species (alfalfa (also known as lucerne, Medicago sativa L.), birdsfoot trefoil (Lotus corniculatus L.), kura clover (Trifolium ambiguum M.B.), red clover (Trifolium pratense L.) and white clover (Trifolium repens L.)) grown in binary mixtures with perennial ryegrass (Lolium perenne L.) and also in pure stands (two red clover cultivars). Additionally, the specific polyphenol oxidase (PPO) activity was measured photometrically in the leaves of pure red clover swards. In both pure and mixed cropping, CP fraction A increased with advancing maturity, except for the legumes from mixed cropping in the summer–autumn growth period 2004 and 2005. The variation of CP fraction A was mostly positively related to the N yield and the amount of dinitrogen fixation. Although CP fraction A of pure red clover was negatively correlated with the specific PPO activity in the spring growth period, the specific PPO activity was less relevant for the variation of CP fraction A with respect to the whole growing season. CP fraction B generally made up the largest proportion of the CP. Pure red clover stands showed reducing amounts of CP fraction C during the growth period, whereas in legumes grown with ryegrass an increase was usually observed. Despite these differences, there was generally an increase of CP fraction C when the content of non-structural carbohydrates decreased. Red clover and birdsfoot trefoil herbage contained the highest proportions of CP fraction C in the CP, regardless of growth period and year. In conclusion, red clover and birdsfoot trefoil had a more favourable CP composition for ruminant nutrition compared to the other legume species, and in red clover this could not be clearly attributed to the specific PPO activity.
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Sekaran, Udayakumar, Jesus Raul Loya, Gandura Omar Abagandura, Senthil Subramanian, Vance Owens, and Sandeep Kumar. "Intercropping of kura clover (Trifolium ambiguum M. Bieb) with prairie cordgrass (Spartina pectinata link.) enhanced soil biochemical activities and microbial community structure." Applied Soil Ecology 147 (March 2020): 103427. http://dx.doi.org/10.1016/j.apsoil.2019.103427.

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45

Bykova, T. O., A. V. Ivashov, S. P. Ivanov, W. N. Sattarow, and L. P. ,. Vahrusheva. "MELITTOPHILIC COMPLEX OF PLANTS PROVIDING FORAGE BASE FOR HONEY BEES (APIS MELLIFERA) IN THE MOUNTAIN-FOREST ZONE OF CRIMEA." Ekosistemy -, no. 21 (2020): 123–41. http://dx.doi.org/10.37279/2414-4738-2020-21-123-141.

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For the first time, in the mountain-forest ecosystems of Crimea of parcel organization level adjacent to the beehives of honey bees (Apis mellifera L.), a species composition of plants representing the melittophilic complex has been identified which provides honey bees with pollen and nectar during the entire warm season. The complex includes 57 species of plants where the honey bees that collect pollen or nectar have been registered. The core of the melittophilic complex consisted of 35 species of plants, the proportion of simultaneously flowering (in any period of the season) flowers of each one exceeded 2.5 % of the total number of flowers of all flowering species at this time. In early spring, the list of such species included 15 plant species belonging to 14 genera and 12 families. In this period, the decisive forage value (species whose flower share exceeded 10 % of all flowering flowers) was represented by 4 plant species: Cornus mas L., Corylus avellana L., Dentaria quinquefolia M. B. and Prunus divaricata Ledeb., the important value (the flowers share made <10 %, but >2.5 %) had 4 species: Ficaria verna Huds., Galanthus plicatus M. Bieb., Primula vulgaris Huds. and Scilla bifolia L. In spring, the forage base made up 20 plant species belonging to 18 genera and 7 families. Four species had decisive forage value: Cerasus avium L., Malus sylvestris L., Prunus spinosa L. and Pyrus elaeagrifolia Jacq., 7 species had important value: Crataegus monogyna Jacq., Fragaria vesca L., Lamium purpureum L., Crepis pulchra L., Prunus domestica L., Thlaspi arvense L. and Trifolium ambiguum M. Most of the species belonged to the Rosaceae family (10 species). In summer period, the forage basis comprised 20 species of melittophilic plants belonging to 19 genera and 9 families. 5 species had decisive value: Cichorium intybus L., Cirsium arvense (L.) Scop., Echium vulgare L., Medicago sativa L. and Trifolium ambiguum M., the important value had 7 species: Achillea millefolium L., Betonica officinalis L., Ballota nigra L., Centaurea diffusa Lam., Clematis vitalba L., Linaria vulgaris Mill. and Onobrychis sativa Lam. The representatives of the Asteraceae family (7 species) dominated; the representatives of Fabaceae (5 species) and Lamiaceae (4 species) also played a significant role. In autumn, there were the following food sources for honey bees: Diplotaxis tenuifolia (L.) DC., Centaurea diffusa Lam., Erýngium campéstre L., Carduus crispus L., and on the steppe slopes of the mountains: Scilla autumnalis L. and Crocus speciosus M. B. It has been established that the greatest number of melittophilic plant species in the studied mountain-forest biogeocenoses grow on parcels of forest edges and glades, steppe slopes of the mountains in their natural state. Oak-hornbeam and floodplain tree-shrub parcels have a smaller variety of species, but are essential for bees in early spring and spring..
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46

Black, A. D., A. J. Harvey, J. L. Moir, and D. J. Moot. "Caucasian clover responses to fertiliser, lime and rhizobia inoculation at Lake Heron Station, Canterbury." Proceedings of the New Zealand Grassland Association 76 (January 1, 2014): 105–10. http://dx.doi.org/10.33584/jnzg.2014.76.2967.

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Abstract The agronomic performance of Caucasian clover in high country grasslands was the subject of two experiments at Lake Heron Station, Canterbury. In the first experiment, Caucasian clover was direct drilled into an undeveloped pasture (soil pH = 5.5; Olsen P = 7 mg/ litre) with fertilisers containing similar P and S but ± 20 kg/ha of N, in December 2011. After 11 months, shoots of Caucasian clover were small (3 mg), indicating an inoculation failure, and effects of fertilisers were not biologically meaningful. Therefore, the influence of rhizobia inoculant, superphosphate and lime on early growth of Caucasian clover was assessed using the same soil in a glasshouse. Un-inoculated plants were 13-24% the size of inoculated plants and this lack of vigour was not overcome by fertilisers. In the second field experiment, an established Caucasian clover/ browntop pasture (soil pH = 5.5; Olsen P = 5 mg/litre) received 0, 100, 200 and 400 kg/ha of superphosphate with 0 or 5 t/ha of lime in February 2012. Despite the high inputs, spring pasture yields 9 and 21 months after fertiliser application were low (1260-2400 kg DM/ ha), but the contribution of Caucasian clover was high (66-76%). Keywords: high country, nitrogen, phosphorus, Trifolium ambiguum
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47

Baker, John M. "Vegetative propagation of kura clover: A field-scale test." Canadian Journal of Plant Science 92, no. 7 (November 2012): 1245–51. http://dx.doi.org/10.4141/cjps2012-014.

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Baker, J. M. 2012. Vegetative propagation of kura clover: a field-scale test. Can. J. Plant Sci. 92: 1245–1251. Kura clover (Trifolium ambiguum M. Bieb.) is a potentially valuable forage legume, but it has been underutilized. A major reason is the difficulty of establishing it from seed. Since kura is rhizomatous, there have been attempts to propagate it vegetatively, but no reports of success at the field scale. Two harvesting methods were tested to transplant material from a mature 17-ha field to a newly tilled 17-ha field: a bermudagrass sprigger that harvests bare rhizome sprigs, and a potato digger that harvests crowns and rhizomes, along with soil. The harvested propagules were distributed over the new field in July 2010 with a manure spreader, then disked and packed. Survival and growth were observed for the remainder of 2010 and through 2011, and recovery of the source field was also monitored. The material harvested with the sprigger did not compete well with weeds and had virtually disappeared by midsummer 2011, but the material harvested with the potato digger thrived, steadily increasing to nearly 80% of the biomass in the new field by the 3rd cutting in summer 2011. Properly done, vegetative propagation is a viable option for kura clover establishment.
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Marshall, A. H., T. A. Williams, M. T. Abberton, T. P. T. Michaelson-Yeates, P. Olyott, and H. G. Powell. "Forage quality of white clover (Trifolium repens L.) x Caucasian clover (T. ambiguum M. Bieb.) hybrids and their grass companion when grown over three harvest years." Grass and Forage Science 59, no. 1 (March 2004): 91–99. http://dx.doi.org/10.1111/j.1365-2494.2004.00409.x.

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49

Ginakes, Peyton, Julie M. Grossman, John M. Baker, and Thanwalee Sooksa-nguan. "Living Mulch Management Spatially Localizes Nutrient Cycling in Organic Corn Production." Agriculture 10, no. 6 (June 23, 2020): 243. http://dx.doi.org/10.3390/agriculture10060243.

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Kura clover (Trifolium ambiguum) is a perennial living mulch species that can be used in conjunction with zone tillage to reduce nitrogen pollution, maintain ground cover, and provide nitrogen to crops. In such systems, kura clover is maintained between crop rows by limiting tillage only to within-row areas. However, the effect of zone-tilled living mulches on soil quality and nutrient cycling in these distinct regions is relatively unexplored. We examined three pools of labile soil organic matter (SOM): microbial biomass, particulate organic matter (POM), and permanganate oxidizable carbon (POXC). Soil samples were collected from both within-row and between-row locations of a zone-tilled kura clover living mulch at three time points per year: before spring zone tillage, approximately ten days after spring zone tillage and corn (Zea mays) planting, and at corn harvest in 2015 and 2016. In 2016, POM and POXC decreased within rows relative to between-row regions after tillage, suggesting that zone till management stimulated decomposition of readily available SOM to effectively localize nutrient cycling in this region and slow mineralization between rows where living kura clover remained. This work shows that zone-tilled living mulches may be a promising avenue for enhancing the synchrony of nutrient mineralization specifically within crop rows, while maintaining year-round ground cover between rows.
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Piñeyro, M. J., K. A. Albrecht, A. M. Mondjana, and C. R. Grau. "First Report of Alfalfa mosaic virus in Kura Clover (Trifolium amgibuum) in Wisconsin." Plant Disease 86, no. 6 (June 2002): 695. http://dx.doi.org/10.1094/pdis.2002.86.6.695a.

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Abstract:
Kura clover (Trifolium ambiguum M. Bieb.) has been reported to be resistant to several viruses, including Alfalfa mosaic virus (AMV), Bean yellow mosaic virus (BYMV), Clover yellow vein virus (CYVV), Peanut stunt virus, Red clover vein mosaic virus (RCVMV), and White clover mosaic virus (WCMV) (2). Furthermore, 54 of 61 kura clover plants were resistant to Clover yellow mosaic virus (CYMV). Field-grown kura clover plants had no visual symptoms of virus infection, but a small proportion of plant introductions tested positive for CYVV, WCMV, and RCVMV (1). These and similar studies have given kura clover the reputation of being highly resistant to most viruses that affect other forage legumes. Viral-like symptoms, characterized by mosaic, crinkling, and reduced size of leaflets were observed on 53 kura clover plants in an 88-plant collection (plant introductions and cultivars) growing in the field. A 20-plant subset was screened for AMV, Bean pod mottle virus, BYMV, Soybean mosaic virus, Tobacco ring spot virus, and Tobacco streak virus using enzyme-linked immunosorbent assay (ELISA). Only AMV was found, and it was detected in nine plants. To our knowledge, this is the first report of AMV in kura clover. The remaining 68 plants were tested for AMV. In total, 70 plants were positive, and 18 plants were negative. AMV was detected in leaves and rhizomes of kura clover. Simultaneously, plants were scored on two occasions for interveinal mosaic, yellowing, curling of leaves, and general chlorosis. There was no correlation between visual symptoms and ELISA results. Results from ELISA were confirmed with a local lesion assay on ‘Bountiful’ beans (Phaseolus vulgaris L.). Leaves from three AMV-positive plants were bulked and ground in phosphate buffer (pH 7.0). Leaves of challenge plants were dusted with Carborundum, and infected sap was rubbed on the youngest leaf of each plant. This procedure was repeated with leaves from three AMV-negative plants. All 12 bean plants inoculated with sap from AMV-positive kura clover developed local lesions or systemic reactions. None of the 12 negative controls developed local lesions. The transmission of AMV from one kura clover plant to another was attempted with the inoculation procedure described above, except that a phosphate-sulfite buffer was used and with soybean aphids (Aphis glycines Matsamura). Seven virus-free ‘Endura’ kura clover plants were inoculated with sap from AMV-positive kura clover plants, and five negative control plants were included. Ten other plants were inoculated with AMV using soybean aphids. Aphids were allowed to feed for 3 min on AMV-positive kura clover plants, then allowed to feed, 10 per plant, on AMV-negative plants. There were six negative controls for this treatment. Three weeks after inoculation, top-growth was clipped, and 9-week-old regrowth was tested for AMV. Two of the mechanically inoculated plants tested positive for AMV using ELISA, and infection was further confirmed by the local lesion assay described above. Therefore, it is demonstrated that AMV can be mechanically transmitted to kura clover. AMV was not transmitted by the colony of soybean aphids, which previously transmitted AMV to soybean (3). This suggests virus-strain aphid specificity and possibly host specificity for phid transmission of AMV to kura clover. References: (1) R. Alconero. Plant Dis. 67:1270, 1983. (2) O. W. Barnett and P. B. Gibson. Crop Sci. 15:32, 1975. (3) J. H. Hill et al. Plant Dis. 85:561, 2001.
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