Isoflavones of the red and Hungarian clover and possible impact on animal diet

The content of daidzein, genistein, formononetin, and biochanin A isoflavones was studied in natural populations of red and Hungarian clover, to estimate their impact on fodder quality and to determine directions in possible breeding programs. The study included 6 red clover (Trifolium pratense) and 6 Hungarian clover (Trifolium pannonicum) populations, collected in the central Balkans. The differences between the species and among the populations were analysed. The average content of total isoflavones was 1.393 mg g–1 and 0.487 mg g–1 of air dry matter in Hungarian clover, respectively. While the most prevailed isoflavone in red clover was biochanin A (46%), the Hungarian clover populations were rich in genistein (43%). The red clover leaves accumulated the highest content of isoflavones. The Hungarian clover flowers and leaves had an equal amount of isoflavones. The obtained values of the total isoflavones could not affect the overall nutrient quality and therefore, researched natural populations of two clover species could be considered for further breeding programs.

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Variety, time of harvest and conditions during growing season have impact on red clover isoflavone content

Agricultural and Food Science ◽

10.23986/afsci.69781 ◽

2018 ◽

Vol 27 (2) ◽

Author(s):

Eeva Arja Mustonen ◽

Mikko Tuori ◽

Päivi Kurki ◽

Mika Isolahti ◽

Juhani Taponen ◽

...

Keyword(s):

High Performance ◽

Trifolium Pratense ◽

Red Clover ◽

Weather Conditions ◽

Biochanin A ◽

Growing Seasons ◽

Crop Varieties ◽

Isoflavone Content ◽

Trifolium Pratense L ◽

Time Of Harvest

Red clover (Trifolium pratense L.) is the predominant legume used in northern European agriculture. Official red clover variety trials are conducted by Natural Resources Institute Finland (Luke) to determine the value of field crop varieties. The trials used for the current analysis were conducted in Luke units in southern Finland (Mikkeli) and northern Finland (Ruukki) in two consecutive years. Plant samples for isoflavone analyses were collected from four varieties grown as four replicates and harvested twice during both growing seasons. The four main isoflavones biochanin A, genistein, daidzein and formononetin were analysed using high performance liquid chromatography. Total phytoestrogen content in the varieties varied in the range of 11.2−14.8 mg g-1 dry matter (DM). The variety and the time of harvest had most effect on the isoflavone, especially formononetin, contents of red clover. A more northern growing area and challenging weather conditions were associated with increased isoflavone concentrations.

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Soluble Phenolic Compounds in Different Cultivars of Red Clover and Alfalfa, and their Implication for Protection against Proteolysis and Ammonia Production in Ruminants

Natural Product Communications ◽

10.1177/1934578x1501000732 ◽

2015 ◽

Vol 10 (7) ◽

pp. 1934578X1501000 ◽

Cited By ~ 1

Author(s):

Isabelle A. Kagan ◽

Ben M. Goff ◽

Michael D. Flythe

Keyword(s):

Phenolic Compounds ◽

Trifolium Pratense ◽

Phenolic Content ◽

Colorimetric Assay ◽

Red Clover ◽

Biochanin A ◽

Total Phenolic ◽

Ammonia Production ◽

Response Factors ◽

Soluble Phenolics

Red clover ( Trifolium pratense) contains soluble phenolic compounds with roles in inhibiting proteolysis and ammonia production. Alfalfa ( Medicago sativa) has been found to have a low phenolic content, but few alfalfa and red clover cultivars have been compared for phenolic content. Total soluble phenolics were quantified by a Folin-Ciocalteu colorimetric assay in nine red clover and 27 alfalfa cultivars. Mean total phenolic contents of red clover and alfalfa were 36.5 ± 4.3 mg/gdw and 15.8 ± 1.4 mg/gdw, respectively, with the greater standard deviation of red clover possibly indicating more diversity in phenolic content. Because different phenolic standards had different response factors in the colorimetric assay, the red clover and 11 alfalfa cultivars were analyzed by HPLC to determine if the differences in total soluble phenolics between genera reflected differences in the amounts of phenolics or in the classes of phenolics responding to the colorimetric assay. Two red clover cultivars differed in total phenolics and phaselic acid. Alfalfa produced different phenolic compounds from red clover, at lower concentrations. Extracts of two red clover cultivars were separated by thin-layer chromatography (TLC), and the bands were assayed for activity against Clostridium sticklandii, a bovine ruminal hyper ammonia-producing bacterium (HAB). Only biochanin A had anti-HAB activity. Inhibitory amounts indicated that five red clover cultivars could be suitable sources of anti-HAB activity.

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Forage yield and persistence of red clover progenies in two environments

Scientia Agricola ◽

10.1590/s0103-90162003000300005 ◽

2003 ◽

Vol 60 (3) ◽

pp. 447-452 ◽

Cited By ~ 3

Author(s):

Daniel Portella Montardo ◽

Miguel Dall'Agnol ◽

Nilton Rodrigues Paim

Keyword(s):

Dry Matter ◽

Recurrent Selection ◽

Trifolium Pratense ◽

Red Clover ◽

Forage Yield ◽

Winter Period ◽

Forage Production ◽

Growing Seasons ◽

Trifolium Pratense L ◽

Physiographic Regions

Red clover (Trifolium pratense L.) is one of the most important temperate legume species, used to lessen the lack of forage during the critical fall-winter period in Rio Grande do Sul, RS, Brazil. However, its utilization has been restricted mainly because of its lack of persistence. This work evaluates the dry matter yield and persistence of red clover half-sib progenies in two physiographic regions of RS: "Depressão Central", in Eldorado do Sul and "Encosta Superior do Nordeste", in Veranópolis. Experiments were carried out for two growing seasons and results were compared to two red clover commercial checks, Qüiñequeli and Estanzuela 116. The region of Veranópolis was more adequate for red clover forage production, enabling better yield and persistence. The best check for both locations was cultivar Estanzuela 116. At Eldorado do Sul persistence was highly affected and some progenies were superior to the best check. At Veranópolis the best check was very productive, with good persistence. The best progenies at both locations were selected to be propagated and submitted to additional recurrent selection cycles.

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Single-Laboratory Validation of UHPLC-MS/MS Assays for Red Clover Isoflavones in Human Serum and Dietary Supplements

Journal of AOAC International ◽

10.1093/jaoacint/qsaa033 ◽

2020 ◽

Vol 103 (4) ◽

pp. 1160-1166 ◽

Cited By ~ 1

Author(s):

Ruth N Muchiri ◽

Richard B van Breemen

Keyword(s):

Quantitative Analysis ◽

Human Serum ◽

Dietary Supplements ◽

Trifolium Pratense ◽

Red Clover ◽

Biochanin A ◽

Internal Standards ◽

Selective Reaction ◽

Coefficients Of Variation ◽

Trifolium Pratense L

Abstract Background Extracts of red clover (Trifolium pratense L.) containing estrogenic and pro-estrogenic isoflavones are used in dietary supplements primarily for the management of menopausal symptoms in women. Objective A UHPLC-MS/MS assay was developed and validated for the quantitative analysis of the six major red clover isoflavones in dietary supplements and in human serum in support of clinical trials. Methods Enzymatic deconjugation of isoflavone glucuronides and sulfate conjugates in human serum specimens was carried out followed by protein precipitation. Isoflavones in red clover dietary supplements were acid hydrolyzed to release aglycons from glycosides. UHPLC separations (< 4 min) were combined with MS/MS using collision-induced dissociation, selective reaction monitoring and deuterated internal standards to measure biochanin A, formononetin, daidzein, genistein, irilone, and prunetin. Results The method was validated with respect to selectivity, specificity, accuracy, linearity, precision, LOD, and LOQ. The calibration curves for all analytes were linear (R2 > 0.998). The mean recovery for low-, medium- and high-quality control standards ranged between 80% and 108%. The precision of the method was assessed using coefficients of variation, which were <15%. Conclusions The UHPLC-MS/MS method is fast, precise, sensitive, selective, accurate, and applicable to the quantitative analysis of red clover isoflavones in different matrices. Highlights This validated UHPLC-MS/MS assay is applicable to the rapid quantitative analysis of red clover isoflavones in human serum and in dietary supplements.

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Yield Performance and Content of Selected Organic Compounds in Trifolium pratense Treated with Various Biostimulants against the Background of Nitrogen Fertilisation

Legume Research - An International Journal ◽

10.18805/lr-522 ◽

2020 ◽

Cited By ~ 1

Author(s):

A. Godlewska ◽

G. A. Ciepiela

Keyword(s):

Chlorophyll Content ◽

Dry Matter ◽

Trifolium Pratense ◽

Red Clover ◽

Application Rate ◽

Soluble Carbohydrates ◽

Soluble Carbohydrate ◽

Yield Performance ◽

Biomass Yields ◽

Nitrogen Application Rate

The present work is an attempt to assess the effect of biostimulants of various origins and nitrogen regime on yield performance, chlorophyll content, protein content and soluble carbohydrate in red clover. A field experiment was arranged as a randomized subblock design (split-plot) in Poland in August 2013. The following factors were examined: type of biostimulant; nitrogen application rate: 0 (control) and 30 kg. ha-1. Biostimulants significantly increased red clover biomass yields. All the examined biostimulants contributed to an increase in the amount of organic components in red clover dry matter. Nitrogen applied at the rate of 30 kg. ha-1 significantly increased red clover yield, chlorophyll content in red clover leaves and protein compounds but it also significantly reduced soluble carbohydrates in the dry matter of test plants.

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Biological estimation of oestrogenic activity in red clover (Trifolium pratense): relative potencies of parts of plant and changes with storage

Journal of Dairy Research ◽

10.1017/s0022029900011432 ◽

1963 ◽

Vol 30 (2) ◽

pp. 243-249 ◽

Cited By ~ 3

Author(s):

D. S. Flux ◽

R. E. Munford ◽

G. F. Wilson

Keyword(s):

Room Temperature ◽

Trifolium Pratense ◽

Red Clover ◽

Biochanin A ◽

Reduced Pressure ◽

Ovariectomized Mice ◽

Fresh Material ◽

Freeze Dried ◽

Relative Potencies ◽

Different Parts

SummaryImmature ovariectomized mice were used to compare the oestrogenic activities of different parts of the same clover plants; to estimate the relative potencies of biochanin A, genistein and diethylstilboestrol; and to compare the effects of different methods of storage on the oestrogenic potency of red clover leaf and petiole. Test materials were incorporated in the diet fed to the mice and the uterine weight response was used to measure oestrogenic activity.With material harvested at four different times, the leaf or leaf and petiole had the highest activity; small and large stem fractions of the plant were both relatively less active, and blossom and seed head were inactive.Comparison of successive estimates of the relative potency of isoflavones and diethylstilboestrol indicated that the relative responsiveness to the two types of oestrogen did not remain constant. Thus comparisons of estimates of oestrogenic activity of plant material, obtained in terms of diethylstilboestrol in different experiments, could be invalid.The potencies of alcohol, acetone and freeze-dried preparations were compared after storage for 18, 115, 212 and 230 days. Of the procedures tested, the most satisfactory was that in which fresh material was placed in 95% alcohol and the liquor and residue dried and stored at room temperature over concentrated sulphuric acid at reduced pressure.

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Single-cut red clover combined with late timothy may be valuable in short-term rotations

Canadian Journal of Plant Science ◽

10.4141/p99-072 ◽

2000 ◽

Vol 80 (2) ◽

pp. 309-313 ◽

Cited By ~ 2

Author(s):

H. T. Kunelius ◽

K. B. McRae ◽

S. A. E. Fillmore ◽

G. Dürr

Keyword(s):

Dry Matter ◽

Trifolium Pratense ◽

Red Clover ◽

Phleum Pratense ◽

Water Soluble ◽

Nutrient Concentrations ◽

Soluble Carbohydrates ◽

In Vitro Digestibility ◽

Short Term

Late-maturing cultivars of red clover (Trifolium pratense L.) and timothy (Phleum pratense L.) may be grown in short-term rotations with other crops and harvested for hay and silage. Harvesting forage in mid-summer is often advantageous for field curing, but the quality of mature forage may be low. We studied how harvests at different developmental stages affected yield, composition of herbage, and species persistence. The late-maturing single-cut red clover cultivar Altaswede and late timothy cultivar Farol were grown alone and in combination with and without applied N. Total forage, and red clover and timothy dry matter yields, increased between the first (27 June) and third (11 July) dates of harvest. Timothy grown alone or combined with red clover and fertilized with N produced greater forage yields than red clover alone or red clover + timothy without applied N. The in vitro digestibility of dry matter declined from 670 to 625 g kg−1 and crude protein from 134 to 109 g kg−1 between 27 June and 11 July. Water soluble carbohydrates in forage were low (36–66 g kg−1) in cuts 1 and 2. Macronutrient and micronutrient concentrations were generally highest at the first date of harvest on 27 June. Nutrient concentrations in cut 2 were similar for the three harvest schedules. We conclude that growing late-maturing single-cut red clover combined with late timothy produced high yields with good nutritional quality even at advanced stages of maturity. Single-cut red clover persisted for the first production year making this combination suited for short-term rotations that involve late harvesting of the primary growth. Key words: Red clover, Trifolium pratense, timothy, Phleum pratense, composition

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CULTIVAR-ENVIRONMENT INTERACTIONS IN RED CLOVER

Canadian Journal of Plant Science ◽

10.4141/cjps84-017 ◽

1984 ◽

Vol 64 (1) ◽

pp. 139-144 ◽

Cited By ~ 1

Author(s):

T. M. CHOO ◽

N. N. COULSON ◽

J. E. LANGILLE ◽

A. F. RAYMENT ◽

J. S. BUBAR ◽

...

Keyword(s):

Dry Matter ◽

Trifolium Pratense ◽

Red Clover ◽

The Other ◽

Environment Interaction ◽

Dry Matter Yield ◽

Population Average ◽

Trifolium Pratense L ◽

Yield Difference ◽

Interaction Variance

The performance of nine double-cut and three single-cut red clover (Trifolium pratense L.) cultivars was studied at five locations in Atlantic Canada. The single-cut clovers survived relatively well at all locations while the double-cut clovers did not survive well at one location during the first winter. An analysis of data for total dry-matter yield from five environments, i.e., location-year combinations, showed that the single-cut clovers outyielded the double-cut clovers in Newfoundland but yielded lower at the other locations. Among the double-cut cultivars, Florex, Lakeland, Tapiopoly, and Violetta were found to be desirable cultivars because they had a high mean yield and did not yield lower than the population average at any of the five environments. A large cultivar-environment interaction variance was detected for the double-cut cultivars; therefore, at least 15 test environments are required in future trials in order to detect a yield difference of 6% by multiple comparison procedures.Key words: Trifolium pratense L., stability

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Dry-matter and Nitrogen Yields of Legumes Interseeded into Sweet Corn

HortScience ◽

10.21273/hortsci.31.2.206 ◽

1996 ◽

Vol 31 (2) ◽

pp. 206-208 ◽

Cited By ~ 12

Author(s):

Steven J. Guldan ◽

Charles A. Martin ◽

Jose Cueto-Wong ◽

Robert L. Steiner

Keyword(s):

Dry Matter ◽

Lens Culinaris ◽

Trifolium Pratense ◽

Sweet Corn ◽

Red Clover ◽

Corn Yield ◽

Hairy Vetch ◽

Vicia Villosa ◽

Barrel Medic ◽

Nitrogen Yields

Five legumes [hairy vetch (Vicia villosa Roth.), barrel medic (Medicago truncatula Gaerth.), alfalfa (Medicago sativa L.), black lentil (Lens culinaris Medik.), and red clover (Trifolium pratense L.)] were interseeded into sweet corn (Zea mays L.) at last cultivation when sweet corn was at about the V9 (early) or blister (late) stage. The effect of legume interseeding on sweet corn yield, and late-season dry-matter and N yields of aboveground portions of the legumes was determined. Sweet corn yield was not affected by legume interseeding. In 1993, legume dry-matter yields were 1420 kg·ha–1 interseeded early and 852 kg·ha–1 interseeded late. Nitrogen yields were 49 kg·ha–1 interseeded early and 33 kg·ha–1 interseeded late. In 1994, dry-matter yields were 2760 kg·ha–1 interseeded early and 1600 kg·ha–1 interseeded late. Nitrogen yields were 83 kg·ha–1 interseeded early and 50 kg·ha–1 interseeded late. In 1993, barrel medic was the highest-yielding legume with dry matter at 2420 kg·ha–1 and N at 72 kg·ha–1 interseeded early, while red clover yielded the lowest with dry matter at 340 kg·ha–1 and N at 12 kg·ha–1 interseeded late. In 1994, dry-matter and N yields ranged from 4500 and 131 kg·ha–1, respectively, for early interseeded barrel medic to 594 kg·ha–1 and 16 kg·ha–1, respectively, for late interseeded red clover.

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