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The non-turning or only superficial turning of soil is considered to be a gentle tillage method. Nevertheless, conventional ploughs are widely used in organic farming for crop production reasons. For the further development of reduced tillage, and up to no tillage, the effects of three cover crop species and their incorporation with different tillage intensities on nitrogen (N) dynamics, weed emergence and the yield of the subsequent main crop, oats, were examined in a repeated organic one-year trial. Sinapis alba, Trifolium resupinatum, Vicia sativa and bare fallow were tested and incorporated using (1) a plough (PL), (2) reduced tillage (RT), (3) mulching + drilling (MD) and (4) direct drilling (DD). V. sativa was the most promising cover crop in combination with RT, MD and DD. In Trial 1, the soil mineral N content and oat yields after the introduction of V. sativa were on a similar level as those in the PL treatments, and weeds were not yield-limiting there. In Trial 2, the biomass production of V. sativa was only about half of that of Trial 1 and did not offer sufficient weed control, but V. sativa was still successful in the RT treatments. In both trials, the yield differences were more pronounced between the cover crop treatments after RT than after PL. RT, therefore, was more dependent on an adequate cover crop species than PL. The no-till method was not only dependent on an adequate cover crop species but also on its proper biomass production for sufficient weed control.

To evaluate the forage yield and quality of Berseem and Persian clover cultivars, a two-year experiment was conducted during two growing seasons (2017–2018 and 2018–2019). Irrigation was the main f...

The use of natural dyes in foods enhances the product quality and they are also associated with anti-inflammatory and anti-cancer effects. Therefore, they play significant role in human health. Chlorophyll is used as a natural pigment in a variety of foods. This study aims to prepare the extract and chlorophyll from Persian clover (Trifolium resupinatum L.), using enzymatic and ultrasound methods and evaluation of their antioxidant and physicochemical properties. In the present study, the chlorophyll stability was evaluated in different conditions including temperature (− 18, 4 and 10 °C), time (15, 30 and 45 days), pH (4.5 and 5.5) and NaCl concentration (50, 100 and 150 mM). The results showed the chlorophyll a concentration was approximately as twice as the chlorophyll b concentration. In all of the experiments, the enzymatic method has shown better efficiency than the ultrasound one. Also, with the increment of extract concentration, the DPPH free radical scavenging and ferric reducing antioxidant power (FRAP) as well as total phenolic compounds, anthocyanin, and flavonoids were increased dramatically. Due to the higher efficiency of enzymatic extraction, the stability assessments were performed based on this method. The result demonstrated that 50 mM of NaCl, temperature − 18 °C, pH 4.5 and time 15 days were associated with the highest chlorophyll a and b values. According to the results the enzymatic method to prepare clover extract and its chlorophyll can be used as an efficient method for indicator pigment production in food and pharmaceutical industries. However, further testing is needed to achieve more information.

The study was carried out between 2015 and 2016 in accordance with the split plots in randomized blocks research design with 3 replications, which aimed to determine the optimal mixture rate for the production design. The results of the research were made into agricultural implementation and cost analysis was performed. Relative profit margin was used in determining the most favorable mixture rate that will be included in the production design. According to the application results, the production threshold was exceeded in all mixtures. The highest gross production value (95.90 USD da-1) and production cost (59.05 USD da-1) was obtained from the sole Anatolian Clover among the forage crop mixtures. Whereas; the lowest gross production value (71.32 USD da-1) and production cost (58.60 USD) was detected in the sole Italian ryegrass application. In addition to this, it was found that the highest relative profit margin (1.62) had been achieved in sole Anatolian clover. 50% Anatolian clover + 50% Italian ryegrass (K2) (1.40), 25% Anatolian clover + 75% Italian ryegrass (K3) (1.38) were followed to sole Anatolian clover, in terms of mixture rates. Perdenpedicular row sowing (E3) had the highest relative profit margin (1.45) in this study according to the sowing methods. As a consequently, perpendicular rows (E3) and sole Anatolian clover (T), 25A.C + 75 I.R (K3) were the most suitable sowing method and mixture ratios under Anatolian conditions, respectively.

WOS: 000505981800017, This study was carried out at Experimental fields of Ege University Faculty of Agriculture Bornovailzmir/Turkey, during the years of 2012-2014. Three different harvest dates (Early Spring, Mid Spring and Late Spring) and mixture rates (100+0%, 80+20%, 60+40% 40+60%, 20+80% and 0+100% respectively) of Persian clover (Trifolium resupinatum) with annual ryegrass (Lolium multiflorum) were tested. the experiment was arranged in a split block design with three replications. As the average of 2 years results, highest yield were recorded in sole Persian clover sown as catch crop and harvested in late spring under Mediterrenean climatic conditions. the data also indicated that pure Persian cover sowing provided the highest green matter, dry matter and crude protein yields, being 36.04 t ha(-1), 5.61 t ha(-1) and 89 kg ha(-1), respectively. Legume ratio, NDF and ADF concentrations increased by advancing harvest dates.

Pollen morphology of 11 species of family Fabaceae that is, Trifolium alexandrinum, Trifolium resupinatum, Arachis hypogaea, Lathyrus aphaca, Medicago lupulina, Vicia sativa, Lathyrus odoratus, Pongamia pinnata, Melilotus indicus, Medicago polymorpha, Medicago sativa from Pakistan has been investigated by light and scanning electron microscopy. Pollen were generally tricolporate, radially symmetrical, isopolar, elliptic in equatorial view and triangular in polar view under LM. Results showed that pollens were per-prolate (T. alexandrinum), prolate (T. resupinatum, V. sativa, L. odoratus, Melilotus indicus, M. polymorpha, M. sativa) and sub-prolate (A. hypogaea, L. aphaca, M. lupulina, P. pinnata). The larger polar/equatorial (P/E) ratio was found in T. alexandrinum (2.26 μm) and the smallest was found in M. lupulina (1.21 μm). The exine of T. resupinatum was 3.00 μm in thickness while others posses smaller exine thickness. The larger pore diameter was found in P. pinnata (16.01 μm) while others have smaller. The length of colpi was larger in Arachis hypogaea (32.24) while others posses smaller. Eight types of surface ornamentation (Psilate, faintly rugulate). Perforate and rugulate to verrucate have been observed under SEM. The pollens were europalynous type. Pollen morphology proved to be useful for the specific delimitation and serve as a tool for the identification and classification of taxa at specific and generic levels and can also be used as a key for the taxonomic features. Diversity in exine sculpture is helpful indicative characters for the isolation of closely related species. Hence, it is clear that both qualitative and quantitative characters of pollen can be useful for differentiating between taxa at specific level.

Tomato plants, Solanum lycopersicum L., are often infested by the tomato fruit borer (TFB), Helicoverpa armigera (Hubner), in northern Iran. Persian clover, Trifolium resupinatum L., was selected as a companion plant in intercropping with tomato in our experiments, due to a key role in enhancing soil fertility and soil texture, attracting flower-visiting natural enemies by production of numerous flowers, and repelling insects by phenolic compounds secretion. Field experiments were conducted to evaluate the impact of intercropping tomato (T) and Persian clover (C) in the four patterns: 1 T: 2C, 2 T: 2C, 3 T: 2C and 4 T: 2C (row ratios) in comparison with sole tomato on the densities of TFB and natural enemies in tomato plantations during 2016 and 2017 growing seasons in Iran. Moreover, yields of both crops were measured to evaluate yield benefits of intercrops. Results indicated all intercropping patterns significantly reduced the density of TFB eggs and larvae compared with sole tomato in the two growing seasons. Lower densities of TFB eggs and larvae, among intercrops, were recorded in 1 T: 2C and 2 T: 2C in the two seasons. In 2016 and 2017 growing season, the average numbers of predatory bugs, coccinellids and Chrysoperla carnea (Stephans) were higher in intercrops than in sole tomato and their abundance, among intercrops, raised with increasing the proportion of clover/tomato rows. Moreover, the egg and larval parasitism rates were higher in intercrop plots than in the control plots. The highest value of land equivalent ratio (1.29 in 2016 and 1.30 in 2017) was found in 1 T: 2C among intercrop patterns in the two cropping seasons. Therefore, we could conclude that intercropping tomato and Persian clover, especially in 1 T:2C and 2 T:2C patterns are more profitable in TFB management programs.

Intercropping and rotation systems involving legumes can be useful means to achieve a sustainable agricultural production. Biological nitrogen fixation (BNF) is widespread among leguminous species thanks to their ability to establish symbiosis with nitrogen (N) fixing microorganisms. Moreover, the application of intercropped legumes increases the agricultural biodiversity and helps farmers to reduce N leaching preventing soil erosion and weed spreading. Little information is available regarding the contribution of N2 (atmospheric nitrogen) from symbiotic fixation of pea and clover when involved in intercropping or in succession systems with other vegetable crops. This study, carried out in a mountain agroecosystem (Venosta Valley, South Tyrol, Italy), has been conceived as part of a more complex research trial on N optimization of cauliflower intercropped and in succession with other vegetables. We have applied the 15N natural abundance technique to field grown clover and dwarf pea, with or without external N supplied, to assess the potential contribution of N2 for intercropping and crop rotation. The technique exploits naturally occurring differences in 15N abundance between plant available N sources in the soil (more enriched in the 15N isotope) and that of N2 in the atmosphere (δ15N=0‰). When a legume crop fixes N2, its 15N abundance should be lower than that of a no-N fixing crop, exploiting only the soil N pool. Reference plant N derived from atmosphere (%Ndfa) was the white cauliflower (Brassica oleracea var. botrytis), one of the most important vegetables on a world scale belonging to the Brassicaceae family and the first vegetable crop cultivated for fresh consumption in South Tyrol. In our study, clover and pea crop residues contained around 74 and 43 kg N ha‑1, respectively. The %Ndfa for the two legumes was similar and ranged from 20% (dwarf pea) to 22% (clover). In presence of external N fertilizers, values of δ15N of the species resulted lower in comparison with the unfertilized control, making the BNF estimation difficult. The study indicated clover and dwarf pea as a potential tool to reduce inorganic fertilizers supply when included in a rotation system. Further trials are needed to confirm our results on BNF of the analysed species in different agroecosystems.

Cadmium(II) is well known for its toxicity and its removal is a prominent task in the field of environmental research. Trifolium resupinatum (TR) has been investigated for the biosorptive removal of Cd(II). The material was characterized by FTIR and SEM. These well established the nature of functional groups and the surface morphology. The effects of various process parameters including time of contact, concentration, pH and temperature were optimized. The maximum removal was observed at 20 min and a pH of7. The equilibrium was described by the Langmuir model showing the maximum biosorption of 31.95 mg/g. The binding of Cd(II) ions was found to be endothermic in nature. The biosorbent from T. resupinatum was found to adsorb potentially the toxic metal ions from its aqueous solutions.

The irrigation with sewage water can be useful if it has no negative effects on food crop yield, soil pollution, and health of humans. However, it includes various types of contaminants like heavy metals that pollute the soil and crops. In this regard, the aim of this study was to evaluate the possible health risks of heavy metals in forages. Forages both of summer and winter were grown with different water treatments (sewage water and tap water) in Department of Botany, University of Sargodha. The concentration of iron (Fe) in water, soil, and plant samples was determined. The Fe values in tap and sewage water were observed as 0.090 and 0.115 mg/L, respectively. The highest mean concentration of Fe was 9.608 mg/kg in the soil where Trifolium alexandrinum is grown, and the lowest mean concentration was 0.154 mg/kg which occurred in the soil where Trifolium resupinatum is grown in winter. The maximum mean concentration of Fe in the root samples of plants was observed as 2.483 mg/kg in Pennisetum typhoideum, and the minimum mean concentration occurred as 0.390 mg/kg in Zea mays grown in summer. The maximum bioconcentration factor value of Fe was observed for T. resupinatum (5.259) grown in winter. The maximum pollution load index value of Fe was observed for T. alexandrinum (0.1688). The maximum value of daily intake of metals was observed as 0.0731 in Medicago sativa, and the maximum health risk index value was determined as 0.1091 in P. typhoideum.

Clover which is an herbaceous, annual, and self-pollinated plant belongs to fabaceae family (legumes) and has become naturalized in Iran, Asia Minor and the Mediterranean eastern suburban countries. The aim of the present study is ITS molecular evaluation of the nuclear ribosomal genes of lines and landraces of Persian Clover. The sequences were aligned using ClustalW method and by MegAlign software and the dendrogram of different phylogenetic and matrix relationships between the sequences were drawn. The results showed little genetic diversity between the lines and the landrace. The conserved sequence of the analyzed gene in the Persian clover is 561 base. Totally, 740 loci (69 and 671 loci, respectively, with and without removal and addition), 9 Singletons, and 5 haplotypes were identified. The highest rate of transfer was observed in pyrimidine (%16.3). The numerical value of the ratio (dN/dS) was 0.86, and since it was less than 1, the pure selection on the studied gene happened. The lines and landraces were not separated based on their geographic locations. In general, the results indicated that the highest rate of the regional diversity belonged to the clover plants in Lorestan region. Moreover, ITS markers did not seem suitable enough for evaluating the intra- species genetic variation, but it was quite well- suited for inter-species or intergeneric evaluation. The nanotechnology is a relatively new technology that has recently entered the field of agriculture. Nanotechnology covers the integration or manipulation of individual atoms, molecules or molecular masses to a diverse array of structures allowing the production of new characteristics and traits of interest. The aim of this study was to evaluate the effects of foliar application of TiO2 nanoparticles on quantitative traits (plant height, number of branches, dry weight of shoots and roots) and the essential oil content of thyme under different levels of field capacity. Our results showed that the application of TiO2 nanoparticles had significant effects on thyme growth, while the essential oil concentrations not affected. These results imply that the application of TiO2 nanoparticles in plants increase agronomic value under reduced irrigation conditions but has not different significant on essential oil.

Seed production of four annual clover species, arrowleaf (Trifolium vesiculosum), balansa (Trifolium michelianum), gland (Trifolium glanduliferum) and Persian (Trifolium resupinatum) was quantified from four sowing dates in 2010. Following initial sowing rate at 4–6 kg/ha, these clovers produced seed yields up to ∼2340 kg/ha for balansa and gland, 914 kg/ha for arrowleaf and ∼814 kg/ha for Persian. After herbage was removed the clover seedlings that regenerated produced a maximum ground cover of 91% for balansa, 65% for gland, 17% for Persian and 6% for arrowleaf. The seeds of ‘Mihi’ Persian clover were all soft and none were recovered in the soil after three months of burial. In contrast, arrowleaf clover had the highest percentage of hardseededness (>97%) and therefore retained most of its viable seeds in the soil into the second year. The differences in hardseededness and regeneration potential mean different management strategies for their successful establishment into dryland farming systems.

The most suitable land for agricultural use has been gradually occupied by crops around the world. Large, uninterrupted croplands have been created, while disproportionate amounts of fertilizers, insecticides, fungicides and herbicides are applied on them. As a result, agricultural activity has a negative impact on biodiversity the ecological intensification of cultivated lands has become necessary. Multifunctional Margins (MFM), the establishment of native flora margins adjacent to croplands, provide a semi-natural habitat for food and wildlife refuge. Three different species mixtures sown in MFM were studied in this paper. The large capacity of six species used in MFM (Borago officinalis, Glebionis coronaria, Coriandrum sativum, Sinapis alba, Trifolium resupinatum and Vicia sativa) was determined. Reductions of up to 65% in the appearance of weeds and increases of 36% in pollinator biodiversity in sown MFM with respect to the MFM of spontaneous flora were observed. The biodiversity of the epigeal fauna increased by 15% in the MFM of spontaneous flora and by 32% in sown MFM, with respect to annual crops.

We report two populations of Trifolium resupinatum (Reversed Clover, trèfle résupiné) from southern Ontario, confirming it as established in Canada. This Eurasian and north African species was reported in the late 1800s in New Brunswick and Quebec, where it apparently did not persist. Its distribution across the United States is sporadic.

In this study, effect of the 4-day germination on the chemical composition, antioxidant activity and fatty acids profile of Persian clover seeds ( Trifolium resupinatum ) was investigated. In addition, clover sprouts was immersed in 0.5, 1 and 2% thyme ( Thymus vulgaris ) essential oil (EO) solutions, packaged in polyethylene containers and stored at refrigerated temperature (4 °C) for 10 days. As seedling progressed, the moisture, protein, crude fibre and ash content increased, whilst fat and carbohydrate reduced. After 4-day germination, ascorbic acid, total phenolic content and antioxidant activity indicated 1.3, 2 and 1.5 fold increase, respectively. Sprouting caused significantly (p 10 unit reduction, just after treatment. During storage, the population of natural microflora increased significantly (p

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

Plants from the genus Trifolium have been utilized in the treatment of chronic diseases by many cultures. The aim of this study was to investigate the antioxidant activity of flavonoids isolated from Trifolium resupinatum L. using an in vitro radical scavenging assay and a computational study of the structure-antioxidant activity relationship (SAR). A phytochemical fractionation of the ethanolic extract of T. resupinatum resulted in the isolation of one new flavonoid along with six known flavonoids. The structure of the isolated compounds was elucidated by data obtained from UV, IR, MS, 1D NMR and 2D NMR spectra. The order of antioxidant efficacies of the isolated flavonoids, obtained by DPPH assay, was correlated to that predicted by the computational analysis. To verify the antioxidant potential, the effect of T. resupinatum on oxidative stress, and the expression of the antioxidant enzymes and the redox sensitive nuclear factor erythroid 2-related factor 2 (Nrf2), in a cyclophosphamide-induced hepatotoxicity rat model was investigated. T. resupinatum protected against cyclophosphamide-induced liver injury through attenuation of oxidative stress and inflammation, and up-regulation of Nrf2. Therefore, the present study provides the first clarification of the detailed antioxidant SARs of T. resupinatum flavonoids and points to the involvement of Nrf2 in their hepatoprotective activity.

Leaf appearance rate and time to canopy expansion of four annual clover species (arrowleaf, balansa, gland and Persian) were quantified in field and controlled environment studies. Crops sown in autumn, which experienced shortening daylengths at emergence, had a slower rate of leaf production and consequently took a longer time to initiate branching, than spring-sown crops. When autumn-sown ‘Bolta’ balansa clover emerged on the shortest day in winter (21 June), the rate of leaf appearance was lengthened by 4 °C d/leaf/h. When the same species emerged after the shortest day, into an increasing photoperiod, the phyllochron was shortened by 5 °C d/leaf/h. This influence of photoperiod on the phyllochron consequently altered the time to axillary leaf production (branching). Throughout all sowing dates, phyllochron was the fastest for ‘Prima’ gland (33–91 °C d/leaf) and slowest for ‘Cefalu’ arrowleaf (53–116 °C d/leaf) clovers. ‘Bolta’ balansa was 44–82 °C d/leaf and ‘Mihi’ Persian 61–93 °C d/leaf. The response of phyllochron to photoperiod suggests these annual clovers should be sown in late summer or early autumn to initiate axillary leaf production as soon as possible to ensure maximize dry matter for early spring.

Cover crop (CC) benefits and adoption in crop rotations may be limited by the short planting window after the cash crop. Interseeding or relay intercropping of CC is a technique that could overcome short planting windows constraints and allow CC establishment and growth. However, in Mediterranean regions this technique has been poorly implemented. Field experiments were established in Central Spain during two consecutive growing seasons to assess the performance of different species as interseeded crops into irrigated maize (Zea mays L.). The treatments included annual ryegrass (Lolium multiflorum L.) and eight legumes: burr medic (Medicago polymorpha L.), barrel medic (Medicago truncatula L.), yellow sweetclover (Melilotus officinalis L.), berseem clover (Trifolium alexandrinum L.), balansa clover (Trifolium michelanium L.), red clover (Trifolium pratense L.), Persian clover (Trifolium resupinatum L.), and common vetch (Vicia sativa L.). A bare soil was included as control treatment. After the maize harvest, the four species with the best performance were planted as autumn cover crops in adjacent plots. The ground cover was studied throughout the experimental period, and biomass and N content for each species were determined in the autumn and in the following spring prior to the cover crop termination. The soil inorganic N content was determined in spring. Barrel medic, yellow sweetclover, common vetch and annual ryegrass were the species with the best performance in terms of soil coverage during the summer. Red clover and Persian clover had good establishment in one out of two years. None of the interseeded species impaired maize grain yield or quality. Interseeded species achieved better soil coverage and increased the biomass accrued at autumn compared to cover crops sown in October. Therefore, interseeded species ensured longer ground coverage, increasing the potential for soil quality improvement or weed suppression. Moreover, the higher biomass achieved by interseeded species ensured the survival of yellow sweetclover, which was winter killed when planted in the autumn. Interseeded cover crops enhanced the potential of N leaching reduction compared to autumn cover crops. This experiment confirms the suitability of different species interseeded in summer into irrigated maize in Mediterranean regions, and highlights the extra benefits of this practice in annual rotations.

Background The various extracts of Trifolium resupinatum (Persian clover) sprout was obtained by using different solvents and microwave assisted extraction in the present study. Then, the bifunctionalized nanoliposomes were prepared and added to soybean oil for evaluating their effect on deferring the oxidation process. Methods The total phenol and antioxidant activity of the extracts was determined by using the free radical scavenging assay. Then, various nanoliposomal structures of the methanolic extract of Persian clover sprout (PCSE) were prepared by using six several formulations containing different ratios of soybean oil, lecithin and the extract. Afterward, the most stable nanoliposome was bifunctionalized by using WPC and pectin (PCSEN-W and PCSEN-WP, respectively). The size and zeta potential of nanoparticles were measured. Furthermore, in order to evaluate the effects of PCSE, PCSEN, PCSEN-W and PCSEN-WP at 100–300 ppm concentrations in deferring the oxidation process of soybean oil, the heat treatment tests were applied (PV and TBA) at 63 °C within a 20-day period. Results The methanolic extract had the highest level of total phenol and antioxidant activity. The results of creaming index and microencapsulation efficiency were exhibited that formulation containing 30% oil, 5% lecithin and 2% the extract was led to the production of the most stable nanoliposomal structure (PCSEN). The size of nanoparticles was in the range of 282.5–491.2 nm. Zeta potential of the samples was obtained in the range between − 56.9 and − 36.3 mV. Polydispersity index of them was ranged from 0.424 to 0.541. The results were confirmed the existence of stable nanoliposomal systems. The results of the PV and TBA values of the extracts in free and nanoliposomal forms were shown that the nanoliposomal forms had very good antioxidant activity against the oxidation process in soybean oil.

Prangos ferulacea is one of the widely used, nutritional and popular fodders in livestock industry. This species is also considered as an important option in rangeland restoration and management. In this study, the comparative phytotoxic activity of aqueous and hydroalcoholic extracts obtained from different organs (flower, shoot and leaf) of P. ferulacea on proline content, seed germination and seedling growth of Trifolium resupinatum has been investigated. According to the results, the hydroalcoholic extract of P. ferulaceae flower possesses the highest total phenolic and flavonoid content and the uppermost phytotoxic effect on T. resupinatum. The extracts significantly decreased seed germination and seedling growth of T. resupinatum and increased the proline content. Our findings indicate that hydroalcoholic extract induced a stronger oxidative stress in T. resupinatum. Finally, based on the results, aqueous allelochemicals that originated from P. ferulacea played a significant role in the successful propagation and development of T. resupinatum in rehabilitated pastures. According to our results, the phytotoxicity effect of the hydroalcoholic extract was significantly higher than that of the aqueous extract. Since in nature, the allelopathic interaction between plants is closer to the aqueous method, primary evaluations of rangeland restoration using this method is suggested.

There is an ever-increasing demand for high quality forage legumes, essential to feed livestock. Micropropagation may be useful to preserve elite allelic compositions in allogamous and auto-incompatible forage legume species, as is the case of Trifolium resupinatum L. (Persian clover). Etiolation-like phenotypes in in vitro cultured plants, with long weak stems and sparse leaves, together with flowering, compromises the long term establishment and maintenance of explants and the adequate development of plants when transferred to ex vitro. Aiming to develop a solution to limit the impact of in vitro etiolation-like effects, flowering and plant viability, we used a very prone to etiolate elite T. resupinatum genotype maintained through in vitro propagation for over 1 year. Stem segments were subcultured in Murashige and Skoog (MS) supplemented with 2.22, 4.56 or 9.12 µM of zeatin; with 4.44 or 8.88 µM of benzyladenine or with 2.89 or 14.44 µM of gibberellic acid. Gibberellic acid supplementation was detrimental and ultimately led to the death of the explants. Supplementation with 2.22 µM zeatin produce no significant differences from control; however higher concentrations of zeatin, as well as of benzyladenine, diminished the longer internodes and increased the development of shoots with more stout stems. Although rooting was prevented by cytokinins, it was successfully induced when explants were transferred to MS media without plant growth regulators. The 9.12 µM zeatin condition was the most favorable since it reversed the etiolation-like phenotype, inhibited in vitro flowering and improved acclimation from 28.7 to 90 % in the FTTr07.13 T. resupinatum genotype.

The development process from floral bud formation to seed maturity of four top flowering annual clovers was quantified from a field experiment across 10 sowing dates at Lincoln University, Canterbury, New Zealand. For each species, a numeric reproductive scale was created as a field guide to document morphological changes as the bud progresses through reproductive development. The duration from bud visible to open flower was 341 °C days for 'Cefalu' arrowleaf, 215 °C days for 'Bolta' balansa, 196 °C days for 'Prima' gland and 186 °C days for 'Mihi' Persian clover. The inflorescence then required a further 274-689 °C days, 185 °C days , 256 °C days and 425 °C days for each respective species to reach physiological maturity. This was indicated when 50% of seeds had turned red/brown for 'Cefalu' arrowleaf, 100% pods turned yellow for 'Bolta' balansa, 100% of seeds were yellow and hard for 'Prima' gland, and pods turned brown with the first sign of colour change in seeds for 'Mihi' Persian clover. These results can be used to facilitate on farm decision making in relation to grazing management or seed set for subsequent regeneration. Keywords: floral development chart, harvest maturity, peak flowering, pollination, seed filling, thermal time, Trifolium vesiculosum, T. michelianum, T. glanduliferum, T. resupinatum

Trifolium resupinatum L. (Fabaceae) is native in Southern Europe and Southwesten Asia. This species is known as a naturalized plant, which is widely distributed in the worl d. We first found it in the Eushincheon river bank of Jindo Isl. , Jeollanam-do, South Korea. T. resupinatum can be easily distinguished from the other species of the same genus by its resupinate flower. The Korean name “Geo-kkul-kkot-to-kki- pul” indicates its distinctive characteristic of flower. Key words - Trifolium resupinatum , Naturalized plant, Resupinate flower * 교신저자(E-mail): megalamen@hanmail.net ⓒ 본 학회지의 저작권은 (사)한국자원식물학회지에 있으며, 이의 무단전재나 복제를 금합니다.This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. 서 언 콩과(Fabaceae)는 전 세계적으로 광범위하게 분포하는 식물군으로 약 650속, 18,000분류군이 알려져 있으며(Choi, 2007; Xu et al., 2010), 우리나라에는 38속 101분류군이 분포한다(Choi, 2007). 이 중 토끼풀속(Trifolium L.)은 약 250분류군이 전 세계 온대와 아열대에 광범위하게 분포한다 (Choi, 2007; Xu et al., 2010).우리나라 토끼풀속에 대한 연구는 Nakai (1911)가 토끼풀(T. repens L.)과 붉은토끼풀(T. pratense L.) 2분류군을 보고한 이래, Chung (1956) 은 달구지풀(T. lupinaster L.) 등 3분류군을 기록한 바 있고, 여러 학자들이 3~4분류군(Lee, 1996; Lee, 2003; Lee, 2006) 을 기록하고 있다 . 최근에 Park (2009) 은 토끼풀속 귀화식물로 노랑토끼풀 (T. campestre Schreb.), 애기노랑토끼풀(T. dubium Sibth.), 선토끼풀(T. hydridum L.), 붉은토끼풀(T. pratense L.), 토끼풀(T. repens L.) 등 5분류군을 정리하여 기록하였다. 그리고 제주도와 전남 보성에서 진홍토끼풀(T. incarnatum L.)의 분포가 확인된 바 있다 (Lee et al., 2008). 한편, 달구지풀에 비해 전체가 소형인 분류군을 제주달구지풀(T. lupinaster L. for. alpinium (Nakai) M. Park) 로 구분하기도 하는데(Lee, 1996), Choi (2007) 는 제주달구지풀의 기본종인 달구지풀의 분포만 보고하였다 . 따라서 제주달구지풀을 기본종인 달구지풀에 포함할 경우 우리나라에 분포하는 토끼풀속 식물은 7분류군이며, 본 연구에서 새롭게 분포가 확인된 분류군을 포함하면 8분류군이다. 본 연구에서는 우리나라 자생식물로 보고되지 않은 T. resupinatum을 전라남도 진도에서 발견하여, 외부형태를 도해하고 기재하여 보고한다(Fig. 1, 2). 국명은 같은 속 내에서 유일하게 꽃이 뒤집혀 달리는 특징을 반영하여 '거꿀꽃토끼풀'이라 새로 명명하였다.

Clovers were chosen on the basis of traditional medicine recommendations, agricultural value, or available information on their promising chemical profiles.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.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.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.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.

The influence of sowing method and plant density on the biomass of spring wheat and Persian clover was evaluated. In a pot experiment conducted in three series during the years 2010–2012, plants were cultivated as mixed and pure crop at higher (consistent with agronomic recommendations) and lower density, decreased by 20% compared to it. Dry mass accumulation tests for both species were conducted during the following wheat growth stages (BBCH): leaf development (12–14), tillering (21–23), stem elongation (31–32), inflorescence development (54–56), and ripening (87–89). Based on the results obtained, the biomass growth rate for both species in question was also determined. It was shown that the mass of shoots of spring wheat cultivated as mixed crop was lower than that of wheat shoots grown as pure crop during the stem elongation and ripening stages. Mixed sowing limited aboveground accumulation in the heads and grain the most and in the stems the least. During the leaf development and stem elongation stages, the wheat presented a more pronounced response to the presence of clover expressed by biomass decrease in case of the treatment with lower plant density and during ripening in the treatment with recommended plant density. In case of both sowing methods and plant densities, the mass of wheat roots was similar. Biomass accumulation in Persian clover shoots and roots in mixed sowing was lower than in pure crop during the entire growing period. The wheat limited biomass accumulation of Persian clover in inflorescences the strongest and in the roots the least. The spring wheat growth rate in both sowing methods was similar as opposed to Persian clover in the case of which a decrease in the growth rate was observed in the mixed crop during the generative development period.

Natural products are being searched for the potential source of medicines and drugs. Trifolium resupinatum is a significant hay crop in cold regions of Iran, Iraq, and Middle East Asia. In this study, assessment of the property of elicitors and precursors on the antioxidant ability of T. resupinatum and on the capitulate improvement of metabolites were done aiming to advance the metabolite and antioxidant production. Explants and containerized plants of T. resupinatum were cultivated before treatment with elicitors and precursors. Plants grown from different explants and culture conditions such as somatic embryos, shade-grown, dark-induced callus, and light-induced callus were treated with different concentrations of elicitors and precursors. The variations in the biosynthesis of secondary metabolites were tissue and environment-specific. Though there was similarity in the relation of content with the concentration of elicitors and precursors shows tissue specificity. The results showed that Methyl Jasmonate (MJ), Salicylic acid (SA), and Glutathione (Glu) were effective on treatments on the bio-production of phenolic and flavonoid compounds. The overall results indicate significant effects of treatments in improving the shade-grown plant activity. The correlation between phenolic and flavonoid content was statistically evaluated with antioxidant activity. Enzymatic activity was influenced by all three effectors and precursors; it was quite more under the influence of SA. The current study displayed the enhancement of metabolites (phenolic and flavonoid) production and increase of antioxidant and enzymatic activities, particularly in the presence of glutamine as precursors and elicitors.

The genotype of nodule occupants in four annual clovers – balansa (Trifolium michelianum), Persian (Trifolium resupinatum), gland (Trifolium glanduliferum) and arrowleaf (Trifolium vesiculosum) – was investigated. The clovers were inoculated with the ALOSCA® group C granule preparation of strain WSM1325. A total of 224 strains were recovered from root nodules with between 55 and 58 strains for each clover species. Genotyping showed that no strains had fingerprints identical to strain WSM1325. The nodule occupants were diverse with 26, 35, 31 and 32 genotypes identified on arrowleaf, balansa, gland and Persian clovers, respectively. Arrowleaf clover had some specificity for genotype A with 43% of nodules occupied by this strain. The most dominant strain for the other three clovers ranged between 13%–18% occupancy. This work demonstrates a high diversity of naturalized rhizobia strains in New Zealand soils that had the ability to nodulate these top flowering annual clover species.

The present study provides information about the binding of Pb(II) ions on an eco-friendly and easily available biodegradable biomass Trifolium resupinatum. The powdered biomass was characterized by FTIR, potentiometric titration and surface area analyses. The FTIR spectrum showed the presence of hydroxyl, carbonyl and amino functional groups and Pb(II) ions bound with the oxygen- and nitrogen-containing sites (hydroxyl and amino groups). The acidic groups were also confirmed by titrations. Effects of various environmental parameters (time, pH and concentration) have been studied. The biosorption process achieved equilibrium in a very short period of time (25 min). Non-linear approach for Langmuir and Freundlich models was used to study equilibrium process and root mean-square error was used as an indicator to decide the fitness of the mathematical model. The biosorption process was found to follow pseudo-second-order kinetics and was very fast. Thus, the biomass can be cost-effectively used for the binding of Pb(II) ions from aqueous solutions.

In recent years, biosynthesis and the utilisation of silver nanoparticles (AgNPs) has become an interesting subject. In this study, the authors investigated the biosynthesis of AgNPs using Trifolium resupinatum (Persian clover) seed exudates. The characterisation of AgNPs were analysed using ultraviolet–visible spectroscopy, X‐ray diffraction (XRD), transmission electron microscopy (TEM) and Fourier transform infra‐red spectroscopy. Also, antifungal efficacy of biogenic AgNPs against two important plant‐pathogenic fungi (Rhizoctonia solani and Neofusicoccum Parvum) in vitro condition was evaluated. The XRD analysis showed that the AgNPs are crystalline in nature and have face‐centred cubic geometry. TEM images revealed the spherical shape of the AgNPs with an average size of 17 nm. The synthesised AgNPs were formed at room temperature and kept stable for 4 months. The maximum distributions of the synthesised AgNPs were seen to range in size from 5 to 10 nm. The highest inhibition effect was observed against R. solani at 40 ppm concentration of AgNPs (94.1%) followed by N. parvum (84%). The results showed that the antifungal activity of AgNPs was dependent on the amounts of AgNPs. In conclusion, the AgNPs obtained from T. resupinatum seed exudate exhibit good antifungal activity against the pathogenic fungi R. solani and N. Parvum.

Foram estudados os efeitos de três frequências e duas intensidades de desfolhação nas características morfogênicas e estruturais, na produção de forragem e na relação folha/caule de duas variedades de trevo-persa (Trifolium resupinatum L. var. resupinatum Gib & Belli. cv. Kyambro e var. majus Boiss cv. BRS Resteveiro). O delineamento utilizado foi de blocos completos ao acaso, em esquema fatorial 3 × 2 × 2, com cinco repetições. Foram avaliadas as seguintes variáveis: taxa de aparecimento de folhas, filocrono, taxa de alongamento e alargamento de folíolos, largura e comprimento de folíolos, número de ramificações, altura de planta, número de folhas vivas abertas, produção de matéria seca total e relação folha/caule. Apesar de a pastagem apresentar maior número de folhas vivas em intervalos de desfolhação maiores e a maior produção de forragem ter sido obtida na altura residual de 5 cm e no intervalo de 6 folhas surgidas, cortes mais frequentes proporcionam melhora da relação folha/caule, maior número de ramificações, maior taxa de surgimento de folhas e maior tamanho de folíolos. The effects of three frequencies and two intensities of defoliation on the morphogenic and structural characteristics, in the herbage production and leaf/stem ratio of two varieties of persian clover (Trifolium resupinatum L. var. resupinatum Gib & Belli. cv. Kyambro and Trifolium resupinatum var. majus Boiss cv. BRS Resteveiro) were studied. The experimental design consisted of complete randomized blocks in a 2 × 3 × 2 factorial arrangement, with five replications. The following variables were evaluated: leaf appearance rate, phyllochron, leaflet extension and elongation rate, leaflet width and length, number of ramifications, plant height, number of open live leaves, total dry matter production and leaf/stem ratio. Although the forage presented higher number of live leaves in bigger defoliation intervals and the higher forage production was obtained in the residual height of 5 cm and in the interval of 6 leaves appearing, more frequent cuttings provide improvement of the leaf/stem ratio, higher number of ramifications, higher leaf appearance rate and bigger leaflet size.

Prangos ferulacea is a member of Apiaceae family, is a popular fodder for livestock production and an important species in rangeland restoration. The objectives of the research are the comparative phytotoxic activity of aqueous and hydroalcoholic extracts obtained from different organs (flower, shoot and leaf) of P. ferulacea on antioxidant response of Trifolium resupinatum was investigated in a laboratory bioassay. Antioxidant enzyme activities including catalase (CAT), peroxidase (POD), ascorbic peroxidase (APX), and superoxide dismutase (SOD), in line with content of malondialdehyde (MDA) and hydrogen peroxide were measured. Results indicated that hydroalcoholic extract of P. ferulaceae flower possess the highest total phenolic content as well as highest phytotoxic effect on T. resupinatum . The highest antioxidant enzyme activity belonged to hydroalcoholic treatment. The treated T. resupinatum seedlings experienced lipid peroxidation at high extract concentrations (12% of hydroalcoholic and 100% of aqueous extract) as evidenced by increased concentration of MDA. In response to this, the activities of SOD, CAT, POD and APX increased at lower extract concentrations but significantly dropped as concentrations increased. According to results of this study, rehabilitation of T. resupinatum sites through the use of P. ferulaceae will probably not be successful.

The small-seeded legumes are important forage crops for grazing animals and contribute nitrogen to succeeding crops in crop rotation systems. However, the susceptibility of several of the forage legumes to the specialist pea aphid Acyrthosiphon pisum (Harris) has never been investigated. The present study on aphid probing behaviour using the Electrical Penetration Graph technique revealed that the forage legumes studied were (i) highly acceptable (common vetch Vicia sativa L.), (ii) acceptable (wooly vetch Vicia villosa Roth), (iii) moderately acceptable (fodder galega Galega orientalis Lam., crimson clover Trifolium incarnatum L., Persian clover Trifolium resupinatum L., white clover Trifolium repens L.), (iv) barely acceptable (common bird’s-foot-trefoil Lotus corniculatus L., yellow lucerne Medicago falcata L., alfalfa Medicago sativa L., sand lucerne Medicago × varia Martyn, common bird’s-foot Ornithopus sativus Brot., alsike clover Trifolium hybridum L., red clover Trifolium pratense L., common sainfoin Onobrychis viciifolia Scop.), and (v) unacceptable (white melilot Melilotus albus Medik.) to the pea aphid. On (i) plants, probing occupied 85% of experimental time, all aphids (100%) succeeded in feeding on phloem sap, phloem phase occupied 50% of probing time, sap ingestion periods were long (mean duration: 100.8 ± 28.2 min.) and engaged 97% of the phloem phase. On (ii) plants, probing occupied 73% of exp. time, feeding activity occurred in 66.7% of aphids, phloem phase occupied 30% of probing time, sap ingestion periods were long (mean duration: 115.5 ± 46.7 min) and engaged 80% of the phloem phase. On (iii) plants, probing ranged from 53% of exp. time on T. repens to 70% on T. incarnatum and T. resuspinatum, feeding occurred in 35.3% of aphids on T. resuspinatum up to 54.5% on T. incarnatum, phloem phase occupied 10% of exp. time on G. orientalis, T. incarnatum, and T. resuspinatum and 20% on T. repens, sap ingestion periods were from 9.8 ± 1.8 min. on G. orientalis to 51.9 ± 20.7 min. long on T. resuspinatum and engaged from 30% of phloem phase on G. orientalis to 80% on T. incarnatum. On (iv) plants, probing occupied 25% of exp. time on O. viciifolia up to 38% on O. sativus and T. hybridum, feeding occurred in 6.7% of aphids on T. hybridum to 28% on O. sativus, phloem phase occupied less than 1% of probing time on all plants except O. viciifolia (4%) and O. sativus (5%) and it consisted mainly of salivation. On M. albus (v), probing occupied 22% of experimental time, the probes were short (1.8 ± 0.3 min), and no aphid on M. albus showed feeding on phloem sap. M. albus can be recommended for intercropping, ‘push-pull’ strategies, or as a barrier crop against A. pisum in sustainable agricultural practices.

Summary Sustainable agriculture uses nature as the model in designing agricultural systems. Because nature consistently integrates her plants and animals into a diverse landscape, a major tenet of sustainable agriculture is to create and maintain diversity. The effects of earthworm inoculation (Pheretima sp., Ew), arbuscular mycorrhizal fungi (Glomus mosseae, AMF) and mixed cropping systems on forage yield, mycorrhizal colonization rate, nitrogenase activity (NA) of free-living rhizosphere bacteria, soil microbial biomass (SMB) carbon, and the growth of clovers were studied at various mixed cropping ratios of berseem clover (Trifolium alexandrinum L., B) to Persian clover (Trifolium resupinatum L., P) (B:P=1:0, 3:1, 1:1, and 1:3). The effects of AMF and Ew on plant yield were positive. Mixed cropping gave more stable yields than monoculture. The greatest mycorrhizal colonization rate was observed at a B:P ratio of 1:1 with a combined AM+Ew inoculant. With the AMF+Ew inoculant, the greatest NA of free-living rhizosphere bacteria was observed with B:P=3:1. Although Ew had no significant effect on SMB, AMF increased SMB from 256 to 444 mg carbon kg−1. The greatest SMB (379 mg kg−1) was observed at B:P=1:1. The greatest nitrogen accumulation in the aboveground biomass, 31.5 mg g−1 forage dry matter, was obtained with mixed cropping (B:P=3:1) in the presence of the AMF+Ew inoculant. A combination of AMF, Ew, and mixed cropping increased the yield, mycorrhizal colonization rate, SMB, and nitrogen uptake of the clover plants, confirming the positive effects of diversity on an agriecosystem.

This experiment was conducted to compare the agronomic characteristics, flowering condition and productivity of introduced annual legumes at paddy field of Seonghwan(Cheonan, Chungnam, Korea) from September 2006 to May 2007. Annual legumes used in this study were hairy vetch(Vicia villsa, Rosa), crimson clover(Trifolium incarnatum, Contea), berseem clover(Trifolium alexandrinum, Alexandria), persian clover(Trifolium resupinatum, Prolific), balansa clover(Trifolium michelianum, Paradona), sweet clover(Melilotus officinalis, Yellow) and forage pea(Pisum sativum, Austrian). Emergency rate after seeding were 90% or more in crimson clover, hairy vetch, forage pea and berseem clover. Wintering of hairy vetch and crimson clover were excellent as 98% and 95%, respectively. Flowering rate of harvesting date(May 10) was 100% in crimson clover, 98% in balansa clover, 5% in persian clover and others were not flowering. Fresh and dry matter yield of crimson clover were highest as 72,556 kg/ha and 16,062 kg/ha, respectively. Crude protein yield of hairy vetch was highest as 2,929 kg/ha but not significant with crimson clover(2,169 kg/ha). TDN yield of crimson clover was highest as 9,007 kg/ha but not significant with hairy vetch(7,366 kg/ha). According to the results from this study, it is suggested that crimson clover would be recommendable for autumn-sowing annual legume at paddy field of Central Provinces.

Flowering time plasticity is a commonly occurring adaptive characteristic of fodder crops, including legumes, in arid and semiarid environments of the Mediterranean regions. Time of flowering is mainly influenced by genotype, temperature and photoperiod. Field experiments were carried out at Foggia (southern Italy) during successive growing seasons (from 8 to 16 growing cycles according to species) to study the relation among air temperature, photoperiod and duration of the morphological development of flowering in eight forage legume species: sulla (Hedysarum coronarium L.), sainfoin (Onobrychis viciifolia Scop.), pea (Pisum sativun L.), berseem clover (Trifolium alexandrinum L.), Persian clover (Trifolium resupinatum L.), faba bean (Vicia faba L.), common vetch (Vicia sativa L.) and hairy vetch (Vicia villosa Roth). Time to reach 10% flowering (EF) and 100% flowering (FF) were recorded. Rate of progress to flowering, defined as the inverse of time from sowing to EF and FF, was related to mean daily temperature, or to both mean daily temperature and mean photoperiod. Using the linear equations, the thermal time requirements (Tt) and the base temperature (Tb) expressed as heat units were determined by the x-intercept method for both EF and FF stages. Evaluation of flowering time was also based on days after planting (DAP), day of year (DOY) and on a photothermal index (PTI). For all species, a significant negative correlation (P ≥ 0.01) was found between planting date (PD) and DAP whereas PTI showed a significant negative relationship (P ≥ 0.05) only for faba bean, pea, berseem clover and common vetch. In sainfoin, sulla and berseem clover, the rate of progress to flowering was affected significantly (P ≥ 0.05) by both mean temperature and photoperiod. The Tt requirements to reach the EF and the FF stage ranged from 871 to 1665 °C day and from 1043 to 1616 °C day, respectively, for the studied species. Both phenological stages considered depended upon accumulated thermal time above a species-specific base temperature. Furthermore, in all legumes the onset of flowering only occurred when dual thresholds of a minimum Tt and a minimum photoperiod were reached, which were specific to each species.

The aim of this experiment was to investigate the growth and residual-nitrogen (-N) effects of different catch-crop species on a low–N fertility coarse sandy soil. Six legumes (white clover [Trifolium repens L.], red clover [Trifolium pratense L.], Persian clover [Trifolium resupinatum L.], black medic [Medi-cago lupulina L.], kidney vetch [Anthyllis vulneraria L.], and lupin [Lupinus angustifolius L.]), four non legumes (rye grass [Lolium perenne L.], chicory [Cichorium intybus L.], fodder radish [Raphanus sativus L.],and sorrel [Rumex Acetosa L.]), and one mixture (rye/hairy vetch [Secale cereale L./Vicia villosa L.])were tested in a field experiment with three repli-cates in a randomized block design. Four reference treat-ments without catch crops and with N application (0, 40, 80, and 120 kg Nha–1) to a succeeding spring barley were included in the design. Due to their ability to fix N2, the legume catch crops had a significantly larger abovegrounddry-matter production and N content in the autumn than the non legumes. The autumn N uptake of the nonlegumes was 10–13 kg Nha–1 in shoots and approx. 9 kg ha–1 in the roots. The shoot N content of white clover, black medic, red clover, Persian clover, and kidney vetch was 55–67 kg ha–1, and the root N content in white clover and kidney vetch was approx. 25 kg ha–1. The legume catch crops, especially white and red clover, seemed to be valuable N sources for grain production on this soil type and their N fertilizer-replacement values in a following unfertilized spring barley corresponded to 120 and 103 kg Nha–1, respectively. The N fertilizer–replacement values exceeded the N content of shoots and roots.

Fifteen years ago subterranean clover (Trifolium subterraneum) and annual medics (Medicago spp.) dominated annual pasture legume sowings in southern Australia, while limited pasture legume options existed for cropping areas of subtropical Australia. Since then a number of sustainability and economic challenges to existing farming systems have emerged, exposing shortcomings in these species and the lack of legume biodiversity. Public breeding institutions have responded to these challenges by developing 58 new annual and short-lived perennial pasture legumes with adaptation to both existing and new farming systems. This has involved commercialisation of new species and overcoming deficiencies in traditional species. Traits incorporated in legumes of Mediterranean Basin origin for the Mediterranean, temperate and southern subtropical climates of Australia include deeper root systems, protection from false breaks (germination-inducing rainfall events followed by death from drought), a range of hardseed levels, acid-soil tolerant root nodule symbioses, tolerance to pests and diseases and provision of lower cost seed through ease of seed harvesting and processing. Ten new species, French serradella (Ornithopus sativus), biserrula (Biserrula pelecinus), sulla (Hedysarum coronarium), gland (Trifolium glanduliferum), arrowleaf (Trifolium vesiculosum), eastern star (Trifolium dasyurum) and crimson (Trifolium incarnatum) clovers and sphere (Medicago sphaerocarpos), button (Medicago orbicularis) and hybrid disc (Medicago tornata × Medicago littoralis) medics have been commercialised. Improved cultivars have also been developed of subterranean (T. subterraneum), balansa (Trifolium michelianum), rose (Trifolium hirtum), Persian (Trifolium resupinatum) and purple (Trifolium purpureum) clovers, burr (Medicago polymorpha), strand (M. littoralis), snail (Medicago scutellata) and barrel (Medicago truncatula) medics and yellow serradella (Ornithopus compressus). New tropical legumes for pasture phases in subtropical cropping areas include butterfly pea (Clitoria ternatea), burgundy bean (Macroptilium bracteatum) and perennial lablab (Lablab purpureus). Other species and cultivars of Mediterranean species are likely to be released soon. The contributions of genetic resources, rhizobiology, pasture ecology and agronomy, plant pathology, entomology, plant chemistry and animal science have been paramount to this success. A farmer survey in Western Australia has shown widespread adoption of the new pasture legumes, while adoption of new tropical legumes has also been high in cropping areas of the subtropics. This trend is likely to increase due to the increasing cost of inorganic nitrogen, the need to combat herbicide-resistant crop weeds and improved livestock prices. Mixtures of these legumes allows for more robust pastures buffered against variable seasons, soils, pests, diseases and management decisions. This paper discusses development of the new pasture legumes, their potential use and deficiencies in the current suite.

This experiment was conducted in order to determine if Persian clover (Trifolium resupinatum) and Berseem clover (Trifolium alexandrium) contained water-soluble and methanol-soluble constituents that could affect the seedling growth of four test species; namely, Amaranthus retroflexus, Convolvulus arvensis (field bindweed), Secale cereale, and Sinapis arvensis (wild mustard). The above-ground tissues of the Persian and Berseem clover plants were collected during the vegetative growth stage and oven-dried. Three concentrations of aqueous and methanolic extracts were used at full-strength (33.3 g L−1), half-strength (16.7 g L−1), and quarter-strength (8.3 g L−1). Distilled water was used as the control. The weed seeds were placed in Petri dishes containing the legume extract or distilled water (control). The seedling growth of the weed species declined with the increasing concentration of the clover extracts. Wild mustard exhibited the highest sensitivity to both the legume extracts. Compared to the aqueous extract, the methanolic extract caused a greater decline in the seedling growth of the weeds. Berseem clover was the stronger inhibitor of the weed seedling growth, as compared to Persian clover. Therefore, the amounts of allelochemicals present might differ in these two clover species. Field bindweed showed the least sensitivity to both the legume extracts. Field bindweed showed more tolerance to the allelochemicals produced by the clover species.

Weeds often form a major problem in weakly competitive vegetable crops, particularly in low input systems. Undersown cover crops can be used to suppress weeds, but often put too high a competitive pressure on the main crop. Cover crop selection is one of the potential means that can be used to design or optimize these intercropping systems. The objective of the current research was to investigate the variability among a range of clover species in morphological and physiological traits that are considered relevant for interplant competition. To this purpose, field experiments with pure stands of eight clover species (2001) and a selection of three clover species (2002) were conducted, in which regular observations and periodic harvests were taken. Clear differences in the time in which full soil cover was obtained, total accumulated biomass, growth duration, height development and N-accumulation were observed. Persian clover (Trifolium resupinatum L.) and subterranean clover (T. subterraneum L.) were the two most contrasting species in this study, particularly differing in the period in which full soil cover was obtained. Persian clover's faster soil cover could not be attributed to a single trait, but resulted from a number of intrinsic characteristics, like light extinction coefficient, light use efficiency and specific leaf area that together determine the relative growth rate. The study also demonstrated the importance of differences in relative starting position, caused by, for instance, seed size, seeding rate and fraction establishment, for the analysis of early growth characteristics. Alsike clover (T. hybridum L.), berseem clover (T. alexandrinum L) and crimson clover (T. incarnatum L.) developed slower than Persian clover, but all produced a higher amount of accumulated dry matter, due to a longer growing period. Clear differences in height and height development between species were observed. These differences were not associated with dry matter accumulation, as the tallest (red clover; 80 cm) and the shortest species (subterranean clover; 12 cm) produced similar amounts of dry matter. A strong positive correlation between early soil cover development and N-accumulation was observed. The large variability among clover species indicates that species selection is a very important aspect of the development of cropping systems that include clover as a cover crop.

The nutritive value of 4 straws, obtained after thrashing of seeds from fodder crops, was assessed as complete feed for ruminants. Sixteen male Murrah buffaloes (liveweight 365.8±19.5 kg), were divided into 4 equal groups and offered ad lib. straw of either Trifolium resupinatum, Trifolium alexandrium, Medicago saliva or Lolium perenne, supplemented with minerals and vitamin A, for 40 days in a completely randomized design. Simultaneously, each straw was offered to 3 rumen fistulated male buffaloes in order to assess the biochemical changes in the rumen. Compared to other straws M. sativa straw had higher (p

An experiment on the effect of row distance and weed control on the herbage yield and some characteristics of Trifolium resupinatum was conducted in 1998–1999 in Bornova, Izmir, Turkey. Row distances were 20, 30 and 40 cm and the herbicides were imazethapyr, imazamox and bentazone. The dry matter yields of clover were highest in 20 cm row distance in both years. The highest crude protein rate was found in 30 cm row distance and bentazone application in second year. The highest crude protein yields were taken from the 20 and 30 cm row distances in both years. Imazethapyr was unsuitable for weed control in Persian clover. In case of a problem originated from the broad-leaved weeds such as Anagallis caerula, Fumaria parviflora, Matricaria chamomilla, Polygonum aviculare, Sinapis arvensis and Stellaria media, bentazone was a suitable herbicide for Persian clover, whereas in the case of presence of broad-leaved weeds and wild oat (Avena sterilis) imazamox was a suitable herbicide for controlling these weeds. Although the herbicide application did not have an important effect on the green herbage yield of Persian clover, it is thought that it is possible to achieve a satisfactory herbage yield by using imazamox. The most suitable row distance was found to be 20 cm in the trial.

In vitro regeneration protocol was developed in Trifolium resupinatum, an important fodder legume in Northwestern India. Out of 6 explants (leaf, petiole, hypocotyl, cotyledon, collar and root) tried for callus induction in 2 different media, good response was observed from hypocotyl and root explants in ‘A’ medium. Successful regeneration was obtained from callus induced from hypocotyl, cotyledon and root in ‘A’ followed with subculture in ‘E’ medium. One of the regenerated plantlet was characterized and compared with mother plant for isozyme-banding pattern. Wide variation for isozyme banding pattern for SOD, Peroxidase and GOT enzyme indicates the regenerant to be a somaclonal variant. Thus, in vitro regeneration will help in widening the genetic base of the species.

This is the first report on the development of interspecific hybrids between T. alexandrinum and T. resupinatum, using embryo rescue. T. resupinatum is an important donor species for traits such as resistance to root and stem rot diseases, and tolerance to alkalinity for the potential improvement of T. alexandrinum. T. alexandrinum, as female parent, was crossed with T. resupinatum and embryos were excised 10–12 days after pollination and cultured in vitro on EC3 medium. Of the recovered embryos, 23.08% showed plumule emergence. For subculturing, shoot-induction medium LSP3 was used. Roots were induced on RL1 medium. Regenerants were hardened and inoculated with Rhizobium before transfer to the field. Morphological appearance of the hybrids was intermediate to both the parents. The hybrids were erect, leafy with short internodes and possessed distinct dwarf bushy compact appearance and were profusely branched. The plants were late flowering with pollen fertility more than 90%. Three out of five hybrid plants set seed following selfing. Hybridity of the plants was also confirmed through Est, PRX, SOD, GOT isozymes and protein banding patterns.

The course of N-NO 3 concentrations in soil, N transfer from the catch crops Vicia sativa L. and Trifolium resupinatum L. to the companion grass Lolium multiflorum ssp. gaudinii (Parl.) Schinz et Keller, and the preceding crop effect on Lolium multiflorum ssp. italicum (A. Br.) Volkart were studied in a field experiment. Catch crops were sown in pure stands and grass/legume swards (= two species in alternating rows) at two sowing dates and harvested at different dates from August to November. Vicia sativa was more effective concerning N 2 fixation than Trifolium resupinatum, this was also evident from above ground N yield and N-NO 3 amount of the soil. The factor sowing date had the greatest influence on N yield. N transfer to the companion grass was generally low. Early sown legumes in pure stands and in mixture temporarily caused slightly increased N-NO 3 amounts in soil compared with unfertilized grass in winter and following spring, whereas late sown legumes apparently had no effect on N-NO 3 amounts. Both legumes had a considerable preceding crop effect in the mixture and in the pure stand for both sowing dates.

An efficient and reliable micropropagation system for Persian clover (Trifolium resupinatum L.) was developed using different explants and media. Node, hypocotyl and cotyledonary node explants were cultured on Murashige and Skoog (MS) medium supplemented with combinations of either 6-benzyladenine (BA) and indole-3-butyric acid (IBA) or BA, Kinetin (KIN) and IBA. Direct multiple shoots developed within 6 weeks in all explants in most media tested. The best shoot multiplication capacity was obtained from cotyledonary node explants on MS medium containing 7.1 μM BA and 1 μM IBA or 14.1 μM BA and 1 μM IBA. Elongated shoots were rooted on either MS medium alone or combination with different concentrations of indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and α-naphthaleneacetic acid (NAA). High rooting was achieved in half strength MS medium containing 8 μM IBA.