peer-reviewedThis work is part of the research project TR-31016, funded by the Ministry of Science and Technological Development, Republic of Serbia.A field trial with four red clover cultivars was established on acid soil in order to evaluate the effect of foliar boron application on seed yield. The crop received foliar boron treatment during the second growth of the second year at two applications. Although seed yield showed a significant increase in boron-treated plants in 2011 compared with control (26.0%), its relative increase was far higher in 2010 (43.2%), which had increased total rainfall amounts during flowering. Sufficient level of boron supply to red clover plants for seed production has a remarkably positive effect under conditions hampering pollination and fertilisation.Ministry of Science and Technological Development, Republic of Serbi

Bernie

Blevins

Dear

Dell

Dordas

Hanson

Hening

Matoh

Matsumoto

Rashid

Santos

Schon

Seguel

Yang

Ylstra

Irish Journal of Agricultural and Food Research

English

A field trial with four red clover cultivars was established on acid soil in order to evaluate the effect of foliar boron application on seed yield. The crop received foliar boron treatment during the second growth of the second year at two applications. Although seed yield showed a significant increase in boron-treated plants in 2011 compared with control (26.0%), its relative increase was far higher in 2010 (43.2%), which had increased total rainfall amounts during flowering. Sufficient level of boron supply to red clover plants for seed production has a remarkably positive effect under conditions hampering pollination and fertilisation

Tomić, Dalibor

Stevović, Vladeta

Durović, Dragan

Madić, Milomirka

Bokan, Nikola

Stanisavljević, Rade

 PlantaRum - Repository of the Institute for Plant Protection and Environment

Boron application in red clover (Trifolium pratense L.) seed production

Objective. To assess the efficacy of biologic drugs, beyond tumor necrosis factor- (TNF-) α inhibitors, in the management of noninfectious refractory scleritis, either idiopathic or associated with systemic immune-mediated disorders. Patients and Methods. This is a retrospective study assessing the efficacy of several biologic agents (rituximab, anakinra, tocilizumab, and abatacept) and the small molecule tofacitinib in the treatment of scleritis through assessment of scleral inflammation and relapses, as well as treatment impact on best-corrected visual acuity (BCVA) and safety profile. Results. Fourteen patients (19 eyes) were enrolled in the study. Scleritis inflammatory grading significantly improved from baseline to 3 months (p=0.002) and from baseline to the last follow-up visit (p=0.002). Scleritis relapses significantly decreased between the 12 months preceding and following biologic therapy (p=0.007). No differences regarding BCVA were observed (p=0.67). Regarding adverse events, only one patient developed pneumonia and septic shock under rituximab treatment. Conclusions. Our results, though limited to a low number of patients, highlight the effectiveness of different biologic therapies in the treatment of noninfectious refractory scleritis, showing to control scleral inflammation and allowing a significant reduction in the number of relapses

Claudia Fabiani

Jurgen Sota

Maite Sainz-de-la-Maza

Laura Pelegrín

Giacomo Emmi

Giuseppe Lopalco

Florenzo Iannone

Lorenzo Vannozzi

Silvana Guerriero

Maria Chiara Gelmi

Donato Rigante

Gian Marco Tosi

José Hernández-Rodríguez

Luca Cantarini

Directory of Open Access Journals

Mediators of Inflammation

New Potential Weapons for Refractory Scleritis in the Era of Targeted Therapy

PlantaRum - Repository of the Institute for Plant Protection and Environment, Belgrade

Research Article • DOI: 10.1515/ijafr-2015-0006  IJAFR • 54(1) • 2015 • 59–63Irish Journal of Agricultural and Food Research59Boron application in red clover (Trifolium pratense L.) seed production1University of Kragujevac, Faculty of Agronomy, Cara Dušana 34, 32000 Čačak, Serbia2Institute for Plant Protection and Environment, Teodora Drajzera 9, 11 000 Beograd, SerbiaDalibor Tomić1*, Vladeta Stevović1, Dragan Đurović1, Milomirka Madić1, Nikola Bokan1, Rade Stanisavljević2AbstractA field trial with four red clover cultivars was established on acid soil in order to evaluate the effect of foliar boron application on seed yield. The crop received foliar boron treatment during the second growth of the second year at two applications. Although seed yield showed a significant increase in boron-treated plants in 2011 compared with control (26.0%), its relative increase was far higher in 2010 (43.2%), which had increased total rainfall amounts during flowering. Sufficient level of boron supply to red clover plants for seed production has a remarkably positive effect under conditions hampering pollination and fertilisation.Keywordsboron • red clover • seed yield • yield componentsIntroductionIn Southeastern Europe, red clover (Trifolium pratense L.) seed crops are often grown on acid soils where certain macro- and micronutrients are less available to plants (Dear and Lipsett, 1987).As a micronutrient, boron (B) is vital to many cellular processes in plants (Hu and Brown, 1994; Matoh et al., 2000; Dos Santos et al., 2004). Sufficient B levels also enhance nitrogenase activity and nitrogen fixation by Rhizobium, thereby facilitating the growth of legume plants (Blevins and Lukaszewski, 1998).Specifically, insufficient B supply has an unfavourable effect on the growth and development of generative organs of many plants. B deficiency generally leads to precocious flowering (Hanson, Chaplin and Breen, 1985) and poor quality fruit (Gupta, 1993). Positive effects of sufficient B levels include an increased number of fertile flowers (Dear and Lipsett, 1987; Noppakoonwong et al., 1997), higher pollen viability (Ylstra et al., 1992) and improved seed and fruit growth (Rashid, Rafique and Bughio, 1994).B is uptaken from the soil mostly as undissociated boric acid (Hening and Patrick, 1997). B uptake in acid soils is adversely affected by a high concentration of aluminium ions (Yang and Zhang, 1998; Matsumoto, 2000; Seguel et al., 2012). Therefore, foliar B application in acid soils would be an efficient strategy to establish optimal B status in plants compared with its supplementation via soil. The objective of the present study was to evaluate seed yield and yield * Corresponding author: Dalibor Tomic E-mail: dalibort@kg.ac.rscomponents in red clover cultivars when supplied with foliar application of boron, a micronutrient not readily available to plants in acid soils.Materials and methodsSoil propertiesA field experiment was set up at a trial field of the Veterinary Extension Service in Čačak (43°54’39.06” N, 20°19’10.21” E, 246 m a.s.l.) in 2009–2011. The trial was established on a fluvisol (IUSS Working Group WRB, 2014), pHH2O 4.8, with 3.18% organic matter, 0% CaCO3, 22.08mg P 100g−1 soil and 30.0mg K 100g−1 soil (Gupta 2008). The preceeding crop was natural meadow. In autumn, prior to seeding, 45 kg ha−1 N, 45 kg ha−1 P2O5 and 45 kg ha−1 K2O were incorporated into the soil by primary tillage.Weather conditionsData on mean monthly temperatures and rainfall were recorded throughout the experiment at a weather station located near the experiment. The mean annual air temperature in 2010, 2011 and long-time period (1992–2002) was 12.57, 12.37 and 11.97ºC, respectively. Monthly rainfall (Figure 1) showed large variations during the growing season (April–September).60Irish Journal of Agricultural and Food ResearchExperimental designThe experiment, which included four diploid red clover cultivars and two treatments with boron (control, foliar B), was conducted as a randomised block design with four replications, with a plot size of 5 m2 (5 x 1 m). Red clover cvs. K-17, K-39 (Institute of Forage Crops, Kruševac, Serbia), Una (Institute of Field and Vegetable Crops Novi Sad, Serbia) and Viola (a Polish cultivar) were planted at 20 cm row spacing at a seed rate of 18 kg ha−1. The cultivars were sown on 3 April 2009 and 27 March 2010. The crop was subjected to foliar treatment with B (Bor-feed, Haifa, Izrael, with 11% of boron) at a concentration of 0.1% and water rate of 1000 l ha−1 (0.11 kg ha− B) at two applications, that is, the first during intensive growth (June 5), the second before the onset of flowering (June 25). Weeds were controlled mechanically. No irrigation was employed.MeasurementsSeed yield and yield components were determined from the second cut in the second year of cultivation (in 2010 and 2011 for establishments in 2009 and 2010, respectively). In the field, the following yield components were determined at flowering time: stem number m-2 and inflorescence number m−2 (by counting within an area of  0.2 m2 per plot), and inflorescence number per stem (by counting in a sample of 10 randomly selected middle-row stems per plot). A sample of 10 inflorescences was taken from each plot at the time of full maturity and the following traits were determined: flower number per inflorescence, seed number per inflorescence and thousand seed weight (based on the weight of 5 x 100 seeds extracted from the same sample). Fertility was calculated from the number of flowers and seeds per inflorescence and expressed in percentages. The actual seed yield was determined on the basis of yield components (inflorescences per unit area, seeds per inflorescence, thousand seed weight) and converted to seed yield in kg ha−1.The results were subjected to analysis of variance using SPSS (SPSS, 1995). Differences between means were tested by LSD test. When the interactions were significant, the main effects were not discussed. To satisfy the requirement of normality, the data for stem number m−2, inflorescence number m−2, flower number per inflorescence, seed number per inflorescence are transformed before analysis of variance using the form √x or √x + 1 for inflorescence number per stem.Results and discussionThe role of B in nitrogen fixation in legumes lies in its positive effect on nitrogenase activity (Blevins and Lukaszewski, 1998) and growth of Rhizobium bacteria (Loomis and Durst, 1992). In the present study, stem number m−2 after B treatment was significantly higher only in cvs. K-17 and Viola, compared with the control in both years (cultivar/foliar treatment interaction) (Table 1), which was due to their relatively higher leaf percentage during the treatment (the authors’ note). Regardless of foliar B application, significantly higher stem number m−2 in 2011 was produced in cv. K-17 compared with cvs. K-39 and Viola (year/cultivar interaction), which was likely indicative of its higher tolerance to soil moisture deficiency.B generally exhibits a markedly higher effect on the development of generative plant organs than on vegetative 020406080100120Apr May Jun Jul Aug Sep201020111992-2002MonthMonthly rainfall (mm)Figure 1. Monthly rainfall distribution for experimental years and 10-year (1992–2002) average61Irish Journal of Agricultural and Food Researchorgans (Bernie and Longbin, 1997). In the present study, foliar B application had no significant effect on inflorescence number m−2 and inflorescence number per stem in all cultivars in both years. Wilczek and Ćwintal (2008) found no significant effect of foliar B application in red clover during the bud formation stage on inflorescence number per stem. Schon and Blevins (1990) reported significantly more branches per plant and pods per branch in soybean as the result of B application at the onset of intensive growth. These results suggest that the effect of foliar-applied B on certain yield components is dependent on plant development stage during the treatment.B application significantly increased the number of flowers per inflorescence in cv. Una in both years (cultivar/foliar application interaction), although a positive response was observed in all cultivars except cvs. K-17. B deficiency inhibits or retards flower development, particularly in plants developing flowers in compact terminal inflorescences (Dell and Huang, 1997).As the result of foliar B application, seed number per inflorescence significantly increased in all cultivars in 2010. B enhances pollen viability and germination and ensures the increased presence of insect pollinators in red clover (Wilczek and Ćwintal, 2008). Regardless of foliar B application, seed Table 1. Seed yield components: SNM, stem number m−2;, INM, inflorescence number m−2;, INS, inflorescence number per stem; FNI, flower number per inflorescence; SNI, seed number per inflorescence; F (%), fertility; TSW (g), thousand seed weight and SY, seed yield (kg ha−1) in red clover cultivars as affected by foliar treatment with boron (control - 0).  SNM INM INS FNI SNI F TSW SYYear 2010 338.3 568b 1.66b 67.7b 34.2b 50.8 1.38 268b 2011 350.9 853a 2.43a 99.6a 52.4a 53.7 1.63 621aCultivar K-39 339.1ab 732ab 2.17 83.5ab 43.2b 51.0b 1.41 453aK-17 358.8a 761a 2.09 83.1ab 41.8b 51.0b 1.48 497aUna 357.5a 740ab 2.06 94.7a 52.8a 58.6a 1.39 561a Viola 321.1b 609b 1.87 73.2b 35.5c 48.4b 1.75 267bFoliar t. O 331.7b 675 2.02 76.9b 40.4b 51.4 1.62 385b B 357.5a 746 2.08 90.3a 46.2a 53.1 1.39 504a2010 K-39 347.5abc 657cde 1.89 68.2cd 33.0 48.1b 1.44 303cdK-17 338.1c 613de 1.77 69.8cd 31.1 47.8b 1.54 289cdUna 345.0bc 534e 1.55 72.4cd 42.9 58.9a 1.40 310cdViola 322.5c 467e 1.45 60.3d 29.8 48.3b 1.14 169d2011 K-39 330.6c 806abc 2.45 98.9ab 53.4 54.0ab 1.38 602bK-17 379.4a 910ab 2.41 96.4ab 52.5 54.1ab 1.42 704abUna 370.0ab 945a 2.56 117.0a 62.7 58.3ab 1.38 811a Viola 323.8bc 751bcd 2.30 86.1bc 40.9 48.5ab 2.35 364c2010 O 330.3b 550 1.64 61.1 30.5c 48.8c 1.35 220cB 346.2ab 586 1.69 74.2 37.9b 52.8b 1.41 315c2011 O 333.1b 799 2.39 92.8 50.2a 54.1a 1.89 549b B 368.8a 906 2.47 106.4 54.5a 53.8a 1.37 692aK-39 O 323.8cd 682 2.12 80.3bc 40.1 49.0 1.43 398B 354.4abc 780 2.21 86.8b 46.3 53.1 1.39 508K-17 O 340.6bc 710 2.04 83.9bc 39.5 45.9 1.45 424B 376.9a 812 2.15 82.4bc 44.1 56.1 1.52 570Una O 356.9ab 767 2.14 79.9bc 49.6 57.7 1.32 508B 358.1ab 712 1.97 109.4a 55.9 59.5 1.45 614Viola O 305.6d 539 1.76 63.7c 32.2 47.2 2.28 209 B 340.6bc 679 1.99 82.7bc 38.5 49.5 1.21 324Values followed by different small letters within columns are significantly different (P < 0.05) according to the LSD test.62Irish Journal of Agricultural and Food Researchnumber per inflorescence was significantly higher in cv. Una compared with the other cultivars in both years (42.9 and 62.7, respectively). Wilczek and Ćwintal (2008) reported a range of 61 to 74 for seed number per inflorescence in cv. Parada. In the present study, all cultivars had a relatively low number of seeds per inflorescence, which was attributable to high plant density and less favourable environmental conditions.A significant increase in fertility in all cultivars was obtained with B treatment in 2010. Foliar B application and declining rainfall during flowering (the beginning of July, Figure 1) induced a significant increase in flower fertility in red clover (Wilczek and Ćwintal, 2008). The results obtained suggest that the positive effect of foliar B application on pollen grain quality, pollination and fertilisation is remarkably high under increased rainfall during clover flowering. Lewis (1980), Dell and Huang (1997) and Dordas (2006) also indicated positive effects of B on pollination, fertilisation and seed and fruit growth.Foliar-applied B induced a significant increase in seed yield in all cultivars in 2011 (year/foliar application interaction). However, relative yield increase was far higher in 2010 (43.2%), although not significant (high variability over replications, CV = 26%), compared with 2011 (26.0%). This indicating that B supply in red clover comes to the forefront during conditions hampering pollination and fertilisation. Positive effects of foliar B application on seed yield in red clover were also reported by Lewis (1980), Wilczek and Ćwintal (2008) as well as by Schon and Blevins (1990) in soybean and Dordas (2006) in alfalfa. Favourable weather in the first part of the growing season and lower rainfall during flowering in 2011 contributed to increasing total seed yield compared with 2010, which is in agreement with the findings of Gorski and Bawolski (1989), and Wilczek and Ćwintal (2008).In this research, foliar B application had a positive effect on yield components and seed yield in all cultivars. A sufficient level of B supply to red clover plants for seed production has a remarkably positive effect, especially under conditions hampering pollination and fertilisation. The results suggest that foliar B application in red clover can increase seed production on acid soils, particularly when less favourable weather conditions occur during flowering.AcknowledgmentsThis work is part of the research project TR-31016, funded by the Ministry of Science and Technological Development, Republic of Serbia.Bernie, D. and Longbin, H. 1997. Physiological response of plants to low boron. Plant and Soil 193: 103-120.Blevins, G.D. and Lukaszewski, K.M. 1998. Boron in plant structure and function. Annual Review of Plant Physiology, Plant Molecular Biology 49: 481–500.Dear, S.B. and Lipsett, J. 1987. The effect of boron supply on the growth and seed production of subterranean clover (Trifolium subterraneum L.). Australian Journal of Agricultural Research 38: 537-546.Dell, B. and Huang, L. 1997. Physiological response of plants to low boron. Plant and soil 193: 103-120.Dordas, C. 2006. Foliar Boron Application Improves Seed Set, Seed Yield and Seed Quality of Alfalfa. Agronomy Journal 98: 907-913.Dos Santos, R.A., De Mattos, T., Da Silva Almeida, A., Monteiro, F.A., Correa, B.D. and Gupta, U.C. 2004. Boron nutrition and yield of alfalfa cultivar crioula in relation to boron supply. Scientia Agricola 61: 496-500.Górski, T. and Bawolski, S. 1989. Agroklimatyczne podstawy rejonizacji upraw koniczyny czerwonej na nasiona (Agro-climatic bases of cultivation regionalisation of red clover cultivated for seeds). Zeszyty Problemowe Postepow Nauk Rolniczych 224: 285-289.Gupta, U.C. 1993. Factors affecting boron uptake by plants. Boron and its role in crop production. Boca Raton: CRC Press Inc. 87-104.Gupta, P.K. 2008. Soil, Water, Plant and Fertilizer Analysis. Jodhpur, India: Agrobios, 265.Hanson, E.J., Chaplin, H.M. and Breen, J.P. 1985. Movement of foliar applied boron out of leaves and accumulation in flower buds and flower parts of “Italian“ prune. Horticultural Science 20: 747-748.Hening, H. and Patrick, B.H. 1997. Absorption of boron by plant roots. Plant and Soil 193: 49–58.Hu, H. and Brown, H. 1994. Localization of boron in cell walls of squash and tobacco and its association with pectin. Plant Physiology 105: 681-689.IUSS Working Group WRB. 2014. World Reference Base for Soil Resources 2014. International soil classification system for naming soils and creating legends for soil maps. World Soil Resources Reports No. 106. FAO, Rome, 7-8.Lewis, H.D. 1980. Are there inter-relations between the metabolic role of boron, synthesis of phenolic phytoalexins and the germination of pollen. New Phytology 84(2): 261-270.Loomis, W.D. and Durst, R.W. 1992. Chemistry and biology of boron. Bio Factors 4: 229-239.References63Irish Journal of Agricultural and Food ResearchMatoh, T., Tadasaki, M., Kobayashi, M. and Takabe, K. 2000. Boron nutrition of cultured tobacco BY-2 cells. III. Characterization of the boron-rhamnogalacturonan II complex in cells acclimated to low levels of boron. Plant Cell Physliology 41: 363-366.Matsumoto, H. 2000. Cell biology of aluminum toxicity and tolerance in higher plants. International Review of Cytology 200: 1-46.Noppakoonwong, R.N., Rerkasem, B., Bell, W.R., Dell, B. and Loneragan, F.J. 1997. Prognosis and diagnosis of boron deficiency in black gram (Vigna mungo L. Hepper) in the field by using plant analysis. In: Proceedings on Boron in Soils and Plants. Eds. R. W. Bell and B. Rerkasem. Kluwer Academic Publishers, Dordrecht, The Netherlands.Rashid, A., Rafique, E. and Bughio, N. 1994. Diagnosing boron deficiency in rapeseed and mustard by plant analysis and soil testing. Communications in Soil Science and Plant Analysis 25(17/18): 2883-2897.Schon, K.M. and Blevins, D.G. 1990. Foliar Boron Applications Increase The Final Number of Branches and Pods on Branches of Field-Grown Soyabeans. Plant Physiology 92: 602-607.Seguel, A., Medina, J., Rubio, R., Cornejo, P. and Borie, F. 2012. Effects of soil aluminum on early arbuscular mycorrhizal colonization of wheat and barley cultivars growing in an andisol. Chilean Journal of Agricultural Research 72(3): 449-455.SPSS. Inc. 1995. STATISTICA for Windows (Computer program manual). Tulsa. OK.Wilczek, M. and Ćwintal, M. 2008. Effect of the methods of additional feeding with microelements (B, Mo) on the yield structure and seed yield of red clover. Electronic Journal of Polish Agricultural Universities. Retrieved December 2012 from the World Wide Web: http://www.ejpau.media.pl. Yang, Y.H. and Zhang, Y.H. 1998. Boron amelioration of aluminium toxicity in mungbean seedlings. Journal of Plant Nutrition 21: 1045-1054.Ylstra, B., Toyraev, A., Moreno, B.M.R., Stoger, E., Tunen, J.A., Vicente, O., Mol, M.N.J. and Heberle-Bors, E. 1992. Flavonols stimulate development, germination, and tube growth of tobacco pollen. Plant Physiology 100: 902–907.

Objective. To assess the efficacy of biologic drugs, beyond tumor necrosis factor- (TNF-) α inhibitors, in the management of noninfectious refractory scleritis, either idiopathic or associated with systemic immune-mediated disorders. Patients and Methods. This is a retrospective study assessing the efficacy of several biologic agents (rituximab, anakinra, tocilizumab, and abatacept) and the small molecule tofacitinib in the treatment of scleritis through assessment of scleral inflammation and relapses, as well as treatment impact on best-corrected visual acuity (BCVA) and safety profile. Results. Fourteen patients (19 eyes) were enrolled in the study. Scleritis inflammatory grading significantly improved from baseline to 3 months () and from baseline to the last follow-up visit (). Scleritis relapses significantly decreased between the 12 months preceding and following biologic therapy (). No differences regarding BCVA were observed (). Regarding adverse events, only one patient developed pneumonia and septic shock under rituximab treatment. Conclusions. Our results, though limited to a low number of patients, highlight the effectiveness of different biologic therapies in the treatment of noninfectious refractory scleritis, showing to control scleral inflammation and allowing a significant reduction in the number of relapses

Rigante D (ORCID:0000-0001-7032-7779)

PubliCatt

New potential weapons for refractory scleritis in the era of targeted therapy

Tomic, Dalibor

Stevovic, Vladeta

Durovic, Dragan

Madic, Milomirka

Stanisavljevic, Rade

T-Stór

Research Article • DOI: 10.1515/ijafr-2015-0006  IJAFR • 54(1) • 2015 • 59–63Irish Journal of Agricultural and Food Research59Boron application in red clover (Trifolium pratense L.) seed production1University of Kragujevac, Faculty of Agronomy, Cara Dušana 34, 32000 Čačak, Serbia2Institute for Plant Protection and Environment, Teodora Drajzera 9, 11 000 Beograd, SerbiaDalibor Tomić1*, Vladeta Stevović1, Dragan Đurović1, Milomirka Madić1, Nikola Bokan1, Rade Stanisavljević2AbstractA field trial with four red clover cultivars was established on acid soil in order to evaluate the effect of foliar boron application on seed yield. The crop received foliar boron treatment during the second growth of the second year at two applications. Although seed yield showed a significant increase in boron-treated plants in 2011 compared with control (26.0%), its relative increase was far higher in 2010 (43.2%), which had increased total rainfall amounts during flowering. Sufficient level of boron supply to red clover plants for seed production has a remarkably positive effect under conditions hampering pollination and fertilisation.Keywordsboron • red clover • seed yield • yield componentsIntroductionIn Southeastern Europe, red clover (Trifolium pratense L.) seed crops are often grown on acid soils where certain macro- and micronutrients are less available to plants (Dear and Lipsett, 1987).As a micronutrient, boron (B) is vital to many cellular processes in plants (Hu and Brown, 1994; Matoh et al., 2000; Dos Santos et al., 2004). Sufficient B levels also enhance nitrogenase activity and nitrogen fixation by Rhizobium, thereby facilitating the growth of legume plants (Blevins and Lukaszewski, 1998).Specifically, insufficient B supply has an unfavourable effect on the growth and development of generative organs of many plants. B deficiency generally leads to precocious flowering (Hanson, Chaplin and Breen, 1985) and poor quality fruit (Gupta, 1993). Positive effects of sufficient B levels include an increased number of fertile flowers (Dear and Lipsett, 1987; Noppakoonwong et al., 1997), higher pollen viability (Ylstra et al., 1992) and improved seed and fruit growth (Rashid, Rafique and Bughio, 1994).B is uptaken from the soil mostly as undissociated boric acid (Hening and Patrick, 1997). B uptake in acid soils is adversely affected by a high concentration of aluminium ions (Yang and Zhang, 1998; Matsumoto, 2000; Seguel et al., 2012). Therefore, foliar B application in acid soils would be an efficient strategy to establish optimal B status in plants compared with its supplementation via soil. The objective of the present study was to evaluate seed yield and yield * Corresponding author: Dalibor Tomic E-mail: dalibort@kg.ac.rscomponents in red clover cultivars when supplied with foliar application of boron, a micronutrient not readily available to plants in acid soils.Materials and methodsSoil propertiesA field experiment was set up at a trial field of the Veterinary Extension Service in Čačak (43°54’39.06” N, 20°19’10.21” E, 246 m a.s.l.) in 2009–2011. The trial was established on a fluvisol (IUSS Working Group WRB, 2014), pHH2O 4.8, with 3.18% organic matter, 0% CaCO3, 22.08mg P 100g−1 soil and 30.0mg K 100g−1 soil (Gupta 2008). The preceeding crop was natural meadow. In autumn, prior to seeding, 45 kg ha−1 N, 45 kg ha−1 P2O5 and 45 kg ha−1 K2O were incorporated into the soil by primary tillage.Weather conditionsData on mean monthly temperatures and rainfall were recorded throughout the experiment at a weather station located near the experiment. The mean annual air temperature in 2010, 2011 and long-time period (1992–2002) was 12.57, 12.37 and 11.97ºC, respectively. Monthly rainfall (Figure 1) showed large variations during the growing season (April–September). - 10.1515/ijafr-2015-0006Downloaded from PubFactory at 08/09/2016 04:26:17PMvia free access60Irish Journal of Agricultural and Food ResearchExperimental designThe experiment, which included four diploid red clover cultivars and two treatments with boron (control, foliar B), was conducted as a randomised block design with four replications, with a plot size of 5 m2 (5 x 1 m). Red clover cvs. K-17, K-39 (Institute of Forage Crops, Kruševac, Serbia), Una (Institute of Field and Vegetable Crops Novi Sad, Serbia) and Viola (a Polish cultivar) were planted at 20 cm row spacing at a seed rate of 18 kg ha−1. The cultivars were sown on 3 April 2009 and 27 March 2010. The crop was subjected to foliar treatment with B (Bor-feed, Haifa, Izrael, with 11% of boron) at a concentration of 0.1% and water rate of 1000 l ha−1 (0.11 kg ha− B) at two applications, that is, the first during intensive growth (June 5), the second before the onset of flowering (June 25). Weeds were controlled mechanically. No irrigation was employed.MeasurementsSeed yield and yield components were determined from the second cut in the second year of cultivation (in 2010 and 2011 for establishments in 2009 and 2010, respectively). In the field, the following yield components were determined at flowering time: stem number m-2 and inflorescence number m−2 (by counting within an area of  0.2 m2 per plot), and inflorescence number per stem (by counting in a sample of 10 randomly selected middle-row stems per plot). A sample of 10 inflorescences was taken from each plot at the time of full maturity and the following traits were determined: flower number per inflorescence, seed number per inflorescence and thousand seed weight (based on the weight of 5 x 100 seeds extracted from the same sample). Fertility was calculated from the number of flowers and seeds per inflorescence and expressed in percentages. The actual seed yield was determined on the basis of yield components (inflorescences per unit area, seeds per inflorescence, thousand seed weight) and converted to seed yield in kg ha−1.The results were subjected to analysis of variance using SPSS (SPSS, 1995). Differences between means were tested by LSD test. When the interactions were significant, the main effects were not discussed. To satisfy the requirement of normality, the data for stem number m−2, inflorescence number m−2, flower number per inflorescence, seed number per inflorescence are transformed before analysis of variance using the form √x or √x + 1 for inflorescence number per stem.Results and discussionThe role of B in nitrogen fixation in legumes lies in its positive effect on nitrogenase activity (Blevins and Lukaszewski, 1998) and growth of Rhizobium bacteria (Loomis and Durst, 1992). In the present study, stem number m−2 after B treatment was significantly higher only in cvs. K-17 and Viola, compared with the control in both years (cultivar/foliar treatment interaction) (Table 1), which was due to their relatively higher leaf percentage during the treatment (the authors’ note). Regardless of foliar B application, significantly higher stem number m−2 in 2011 was produced in cv. K-17 compared with cvs. K-39 and Viola (year/cultivar interaction), which was likely indicative of its higher tolerance to soil moisture deficiency.B generally exhibits a markedly higher effect on the development of generative plant organs than on vegetative 020406080100120Apr May Jun Jul Aug Sep201020111992-2002MonthMonthly rainfall (mm)Figure 1. Monthly rainfall distribution for experimental years and 10-year (1992–2002) average - 10.1515/ijafr-2015-0006Downloaded from PubFactory at 08/09/2016 04:26:17PMvia free access61Irish Journal of Agricultural and Food Researchorgans (Bernie and Longbin, 1997). In the present study, foliar B application had no significant effect on inflorescence number m−2 and inflorescence number per stem in all cultivars in both years. Wilczek and Ćwintal (2008) found no significant effect of foliar B application in red clover during the bud formation stage on inflorescence number per stem. Schon and Blevins (1990) reported significantly more branches per plant and pods per branch in soybean as the result of B application at the onset of intensive growth. These results suggest that the effect of foliar-applied B on certain yield components is dependent on plant development stage during the treatment.B application significantly increased the number of flowers per inflorescence in cv. Una in both years (cultivar/foliar application interaction), although a positive response was observed in all cultivars except cvs. K-17. B deficiency inhibits or retards flower development, particularly in plants developing flowers in compact terminal inflorescences (Dell and Huang, 1997).As the result of foliar B application, seed number per inflorescence significantly increased in all cultivars in 2010. B enhances pollen viability and germination and ensures the increased presence of insect pollinators in red clover (Wilczek and Ćwintal, 2008). Regardless of foliar B application, seed Table 1. Seed yield components: SNM, stem number m−2;, INM, inflorescence number m−2;, INS, inflorescence number per stem; FNI, flower number per inflorescence; SNI, seed number per inflorescence; F (%), fertility; TSW (g), thousand seed weight and SY, seed yield (kg ha−1) in red clover cultivars as affected by foliar treatment with boron (control - 0).  SNM INM INS FNI SNI F TSW SYYear 2010 338.3 568b 1.66b 67.7b 34.2b 50.8 1.38 268b 2011 350.9 853a 2.43a 99.6a 52.4a 53.7 1.63 621aCultivar K-39 339.1ab 732ab 2.17 83.5ab 43.2b 51.0b 1.41 453aK-17 358.8a 761a 2.09 83.1ab 41.8b 51.0b 1.48 497aUna 357.5a 740ab 2.06 94.7a 52.8a 58.6a 1.39 561a Viola 321.1b 609b 1.87 73.2b 35.5c 48.4b 1.75 267bFoliar t. O 331.7b 675 2.02 76.9b 40.4b 51.4 1.62 385b B 357.5a 746 2.08 90.3a 46.2a 53.1 1.39 504a2010 K-39 347.5abc 657cde 1.89 68.2cd 33.0 48.1b 1.44 303cdK-17 338.1c 613de 1.77 69.8cd 31.1 47.8b 1.54 289cdUna 345.0bc 534e 1.55 72.4cd 42.9 58.9a 1.40 310cdViola 322.5c 467e 1.45 60.3d 29.8 48.3b 1.14 169d2011 K-39 330.6c 806abc 2.45 98.9ab 53.4 54.0ab 1.38 602bK-17 379.4a 910ab 2.41 96.4ab 52.5 54.1ab 1.42 704abUna 370.0ab 945a 2.56 117.0a 62.7 58.3ab 1.38 811a Viola 323.8bc 751bcd 2.30 86.1bc 40.9 48.5ab 2.35 364c2010 O 330.3b 550 1.64 61.1 30.5c 48.8c 1.35 220cB 346.2ab 586 1.69 74.2 37.9b 52.8b 1.41 315c2011 O 333.1b 799 2.39 92.8 50.2a 54.1a 1.89 549b B 368.8a 906 2.47 106.4 54.5a 53.8a 1.37 692aK-39 O 323.8cd 682 2.12 80.3bc 40.1 49.0 1.43 398B 354.4abc 780 2.21 86.8b 46.3 53.1 1.39 508K-17 O 340.6bc 710 2.04 83.9bc 39.5 45.9 1.45 424B 376.9a 812 2.15 82.4bc 44.1 56.1 1.52 570Una O 356.9ab 767 2.14 79.9bc 49.6 57.7 1.32 508B 358.1ab 712 1.97 109.4a 55.9 59.5 1.45 614Viola O 305.6d 539 1.76 63.7c 32.2 47.2 2.28 209 B 340.6bc 679 1.99 82.7bc 38.5 49.5 1.21 324Values followed by different small letters within columns are significantly different (P < 0.05) according to the LSD test. - 10.1515/ijafr-2015-0006Downloaded from PubFactory at 08/09/2016 04:26:17PMvia free access62Irish Journal of Agricultural and Food Researchnumber per inflorescence was significantly higher in cv. Una compared with the other cultivars in both years (42.9 and 62.7, respectively). Wilczek and Ćwintal (2008) reported a range of 61 to 74 for seed number per inflorescence in cv. Parada. In the present study, all cultivars had a relatively low number of seeds per inflorescence, which was attributable to high plant density and less favourable environmental conditions.A significant increase in fertility in all cultivars was obtained with B treatment in 2010. Foliar B application and declining rainfall during flowering (the beginning of July, Figure 1) induced a significant increase in flower fertility in red clover (Wilczek and Ćwintal, 2008). The results obtained suggest that the positive effect of foliar B application on pollen grain quality, pollination and fertilisation is remarkably high under increased rainfall during clover flowering. Lewis (1980), Dell and Huang (1997) and Dordas (2006) also indicated positive effects of B on pollination, fertilisation and seed and fruit growth.Foliar-applied B induced a significant increase in seed yield in all cultivars in 2011 (year/foliar application interaction). However, relative yield increase was far higher in 2010 (43.2%), although not significant (high variability over replications, CV = 26%), compared with 2011 (26.0%). This indicating that B supply in red clover comes to the forefront during conditions hampering pollination and fertilisation. Positive effects of foliar B application on seed yield in red clover were also reported by Lewis (1980), Wilczek and Ćwintal (2008) as well as by Schon and Blevins (1990) in soybean and Dordas (2006) in alfalfa. Favourable weather in the first part of the growing season and lower rainfall during flowering in 2011 contributed to increasing total seed yield compared with 2010, which is in agreement with the findings of Gorski and Bawolski (1989), and Wilczek and Ćwintal (2008).In this research, foliar B application had a positive effect on yield components and seed yield in all cultivars. A sufficient level of B supply to red clover plants for seed production has a remarkably positive effect, especially under conditions hampering pollination and fertilisation. The results suggest that foliar B application in red clover can increase seed production on acid soils, particularly when less favourable weather conditions occur during flowering.AcknowledgmentsThis work is part of the research project TR-31016, funded by the Ministry of Science and Technological Development, Republic of Serbia.Bernie, D. and Longbin, H. 1997. Physiological response of plants to low boron. Plant and Soil 193: 103-120.Blevins, G.D. and Lukaszewski, K.M. 1998. Boron in plant structure and function. Annual Review of Plant Physiology, Plant Molecular Biology 49: 481–500.Dear, S.B. and Lipsett, J. 1987. The effect of boron supply on the growth and seed production of subterranean clover (Trifolium subterraneum L.). Australian Journal of Agricultural Research 38: 537-546.Dell, B. and Huang, L. 1997. Physiological response of plants to low boron. Plant and soil 193: 103-120.Dordas, C. 2006. Foliar Boron Application Improves Seed Set, Seed Yield and Seed Quality of Alfalfa. Agronomy Journal 98: 907-913.Dos Santos, R.A., De Mattos, T., Da Silva Almeida, A., Monteiro, F.A., Correa, B.D. and Gupta, U.C. 2004. Boron nutrition and yield of alfalfa cultivar crioula in relation to boron supply. Scientia Agricola 61: 496-500.Górski, T. and Bawolski, S. 1989. Agroklimatyczne podstawy rejonizacji upraw koniczyny czerwonej na nasiona (Agro-climatic bases of cultivation regionalisation of red clover cultivated for seeds). Zeszyty Problemowe Postepow Nauk Rolniczych 224: 285-289.Gupta, U.C. 1993. Factors affecting boron uptake by plants. Boron and its role in crop production. Boca Raton: CRC Press Inc. 87-104.Gupta, P.K. 2008. Soil, Water, Plant and Fertilizer Analysis. Jodhpur, India: Agrobios, 265.Hanson, E.J., Chaplin, H.M. and Breen, J.P. 1985. Movement of foliar applied boron out of leaves and accumulation in flower buds and flower parts of “Italian“ prune. Horticultural Science 20: 747-748.Hening, H. and Patrick, B.H. 1997. Absorption of boron by plant roots. Plant and Soil 193: 49–58.Hu, H. and Brown, H. 1994. Localization of boron in cell walls of squash and tobacco and its association with pectin. Plant Physiology 105: 681-689.IUSS Working Group WRB. 2014. World Reference Base for Soil Resources 2014. International soil classification system for naming soils and creating legends for soil maps. World Soil Resources Reports No. 106. FAO, Rome, 7-8.Lewis, H.D. 1980. Are there inter-relations between the metabolic role of boron, synthesis of phenolic phytoalexins and the germination of pollen. New Phytology 84(2): 261-270.Loomis, W.D. and Durst, R.W. 1992. Chemistry and biology of boron. Bio Factors 4: 229-239.References - 10.1515/ijafr-2015-0006Downloaded from PubFactory at 08/09/2016 04:26:17PMvia free access63Irish Journal of Agricultural and Food ResearchMatoh, T., Tadasaki, M., Kobayashi, M. and Takabe, K. 2000. Boron nutrition of cultured tobacco BY-2 cells. III. Characterization of the boron-rhamnogalacturonan II complex in cells acclimated to low levels of boron. Plant Cell Physliology 41: 363-366.Matsumoto, H. 2000. Cell biology of aluminum toxicity and tolerance in higher plants. 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Chilean Journal of Agricultural Research 72(3): 449-455.SPSS. Inc. 1995. STATISTICA for Windows (Computer program manual). Tulsa. OK.Wilczek, M. and Ćwintal, M. 2008. Effect of the methods of additional feeding with microelements (B, Mo) on the yield structure and seed yield of red clover. Electronic Journal of Polish Agricultural Universities. Retrieved December 2012 from the World Wide Web: http://www.ejpau.media.pl. Yang, Y.H. and Zhang, Y.H. 1998. Boron amelioration of aluminium toxicity in mungbean seedlings. Journal of Plant Nutrition 21: 1045-1054.Ylstra, B., Toyraev, A., Moreno, B.M.R., Stoger, E., Tunen, J.A., Vicente, O., Mol, M.N.J. and Heberle-Bors, E. 1992. Flavonols stimulate development, germination, and tube growth of tobacco pollen. Plant Physiology 100: 902–907. - 10.1515/ijafr-2015-0006Downloaded from PubFactory at 08/09/2016 04:26:17PMvia free access

Fabiani, Claudia

Sota, Jurgen

Sainz de la Maza Serra, María Teresa

Pelegrin, Laura

Emmi, Giacomo

Lopalco, Giuseppe

Iannone, Florenzo

Vannozzi, Lorenzo

Guerriero, Silvana

Gelmi, Maria Chiara

Rigante, Donato

Tosi, Gian Marco

Hernández Rodríguez, José

Cantarini, Luca

Diposit Digital de la Universitat de Barcelona

AbstractA field trial with four red clover cultivars was established on acid soil in order to evaluate the effect of foliar boron application on seed yield. The crop received foliar boron treatment during the second growth of the second year at two applications. Although seed yield showed a significant increase in boron-treated plants in 2011 compared with control (26.0%), its relative increase was far higher in 2010 (43.2%), which had increased total rainfall amounts during flowering. Sufficient level of boron supply to red clover plants for seed production has a remarkably positive effect under conditions hampering pollination and fertilisation.</jats:p

Dalibor Tomić

Vladeta Stevović

Dragan Đurović

Milomirka Madić

Nikola Bokan

Rade Stanisavljević

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http://plantarum.izbis.bg.ac.rs/bitstream/123456789/369/1/367.pdf

Boron application in red clover (Trifolium pratense L.) seed production

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