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Grubisic Abstract: In this study was tested influences of three N fertilizers (urea = 46% N; calcium ammonium nitrate or CAN; ammonium sulphate) and their distribution (N-autumn; N- spring; N-50% autumn + 50% spring; N-33% autumn + 67% spring; N-100% spring based on N-min method test) on 1000-grain weight of two maize inbred lines (IL1 and IL2) under conditions of Zemun Polje calcaric chernozem for three growing seasons (2001, 2002 and 2003) with aim of N fertilization optimization for seed-maize growing. The use of the N-min method test (N ranging from 17 to 35 kg ha-1, in dependence on the soil mineral nitrogen content), especially in years with lower precipitation sums, resulted in the highest increase in 1000–grain weight (3.2%) of observed maize inbred lines in relation to the control. The application of different forms of nitrogen did not result in statistically significant differences in 1000–grain weight of observed maize inbred lines. Key words: Time of nitrogen application, Nitrogen form, Maize inbred lines, 1000-grain weight Introduction Maize ist the most spread arable crop in Serbia. For example, in the period 2000–2004 it was grown on 1,203 139 ha/annually with mean yield 4.12 t ha–1 (Statistical Yearbook of Serbia, 2001–2005). For this production is needed adequate quantities of seed, but growing of parents of maize hybrids (mainly inbred lines) is connected with more susceptibility to environmental stress including nutirients supplies, in conmparison with growing of maize hybrids. Forms and time of nitrogen (N) application are important factors of plants growth and development, especially for maize inbred lines, which are used as parents in maize seed production [1] [17]. The application of N fertilizers in the maize seed production represents an important cropping practice as it significantly affects the yield level and certain seed (grain) traits. Kling and Okoruwa [7] and Okoruwa [13] refer to several biological seed traits, while Miric and Brkic [11] describe several dozens of different seed traits affecting sowing and having the importance for seed drying, transport, storing, packing and conservation. However, Miric et al. [12] conclude that germination and 1000-grain weight are traits among a dozen of the most important sowing-technical and production-economical traits, because they specify both, quantity and quality. The variety, soil tillage, crop density and uniformity, irrigation regime and fertilisation have the greatest impact on 1000-grain weight, and then effects of weather conditions, except during pollination and maturity, are less important, while the fractioning has a crucial effect on 1000-grain weight. In this study was tested influences of three N fertilizers (urea = 46% N in amide form; calcium ammonium nitrate or CAN = 50% NH4–N + 50% NO3–N; ammonium sulphate = NH4–N) and their distribution (N–autumn; N–spring; N–50% autumn + 50% spring; N–33% autumn + 67% spring; N–100% spring based on N–min method test) under conditions of Zemun Polje calcaric chernozem with aim of N fertilization optimization for seed–maize growing. Materials and methods The field experiment The field experiment was conducted for three growing seasons (the factor A: 2001, 2002 and 2003) with two maize inbred lines (the factor B: L1 and L2), six fertilization (the factor C–Table 1) and three N–fertilizers applications (the factor D–Table 1) on the constant P and K fertilization (kg ha-1: 60 P2O5 + 60 K2O as superphosphate and KCl ) was conducted on chernozem soil (Maize Research Institute in Zemun Polje). Table 1. Forms and distributions of N by fertilization N Fertilizer (the factor C)Distribution of N in kg ha-1 (the factor D)PeriodD1D2D3D4D5D6**C1 = urea (46% N: 100% NH2-N)Autumn01005034C2 = CAN* (50% NH4-N + 50% NO3-N)Spring01005033N-minC3 = (NH4)2SO4 (100% NH4-N)Dressing0330D2-D6 (kg ha-1) : 60 P2O5 + 60 K2O* Calcium ammonium nitrate (26% N); ** N–min to 120 cm of depth + fertilization = = 100 kg N ha-1 (kg N ha-1 = 35, 31 and 17, for 2001, 2002 and 2003, respectively). The experiment was conducted in the randomized block design in four replicates. Gross of the basic plot was 28 m2. Maize was sown by pneumatic sowing maschine in the terms as follows: May 8, April 24 and April 15, for 2001, 2002 and 2003, respectively (crop rotation after winter wheat). Weed control was made by incorporation of Eradicane (6.0 l ha-1) by presowing soil tillage and pre–emergence application of Atrazin + Prometrin + Monosan (l ha-1 = 1.0 +1.5+1.5) as well. Plant number reduction to level of 60,000 plants ha-1 was made in early growth stage. The harvest was done on October 5, 2001; September 10, 2002; September 5, 2003 at the grain physiological maturity of maize inbred lines. Harvested ears were dried at the seed drying and processing plant at the Maize Research Institute, Zemun Polje. Statistical analyses (LSD–test) were performed according Mead [10]. Weather conditions Concerning precipitation quantities for the 6–month April–September period, the growing season 2002 was in range of 30–year means (LTM), while in 2001 and 2003 they were for 46% higher and for 30% lower than LTM, respectively. Temperature regime for maize growth was more favorable in 2001, compared to remaining two years (Table 2). In general, low yield of maize are in connection with drougt stress, especially during July and August [5] [8] [15] Table 2 . Weather characteristics (Zemun Polje Weather Bureau: 44o49’N, 20o27’E) Weather characteristics (LTM = long–term means: 1961-1990)MonthPrecipitation (mm)Mean air–temp. (oC)LTM200120022003200120022003mmoCApril (IV) May (V) June (VI) July (VII) August (VIII) Sept. (IX)148.8 46.2 168 41.8 35.0 70.854.8 29.4 65.0 34.8 105.2 55.414.6 36.4 19.0 105.4 26.4 41.211.0 17.6 14.1 22.4 23.6 15.911.6 19.5 22.0 23.4 21.6 16.511.2 20.5 24.0 22.5 24.3 17.248.3 61.2 79.4 63.5 52.3 44.711.2 16.9 19.3 20.3 22.1 17.2IV-IX: Total IV-IX: Mean510.6 344.6 243.0 17.4 19.1 19.9349.4 17.8 Results and discussion Thousand-grain weight significantly varied over investigation years and genotypes (Table 3). Fertilization based on the Nmin method resulted in a significantly greater 1000-grain weight in relation to remaining fertilization variants, on the average for three years and all three forms of nitrogen. Applied nitrogen in the form of fertilizer did not differ in 1000-grain weight. Table 3. 1000-grain weight (g ) of maize inbred lines FactorYear 2001 (A1)Year 2002 (A2)Year 2003 (A3)Genotype (B)MeanGenotype (B)MeanGenotype (B)MeanMeanIL-1IL-2IL-1IL-2IL-1IL-2Influences of A, B C and interactions AB, AC and ABCYear (A)264.0297.9242.1Genotype (B)3-year means:277.2258.7Interaction AB297.9230.1Y2001318.9276.4Y2002214.5269.6Y2003Interaction BC277.2258.2NH2-N275.9259.3CAN278.7258.6NH4-NFertilizer (C)ABCACABCACABCACCNH2-N299.1228.6263.9316.6277.8297.2215.8268.1242.0267.7CAN294.8232.9263.9322.8274.9298.9210.0270.2240.1267.6NH4-N299.6228.9264.3318.9276.5297.7217.5270.5244.0268.7Influences D and interactions AD, BD and ABDDistribution (D)ABDADABDADABDADDN-0297.6231.7264.7325.5276.2300.9210.4266.6238.5268.0N-100 autumn306.9211.9259.4327.8270.6299.2213.5267.0240.3266.3N-100 spring284.2241.5262.9312.2267.5289.9226.6257.0241.8397.3N 50a+50spr282.4234.6258.5316.0278.5297.3219.9268.1244.0266.6N 34a+66spr309.5229.2269.4312.9264.3288.6216.4261.0238.7265.6N-min spring306.6232.0269.3322.1301.2311.7200.0297.8248.9276.6Interactions ACD and CD (a= NH2-N; b=CAN; c= NH4-N)Year 2001 (A1)Year 2002 (A2)Year 2003 (A3)3-year meansabcabcabcabcACDACDACDCDN-0264.7264.7264.7300.9300.9300.9238.5238.5238.5268.0268.0268.0N-100 autumn262.4258.2257.6289.4313.1299.7224.1255.9240.8258.7275.7264.5N-100 spring256.6264.9267.2292.6294.9282.1244.2245.1236.1264.4268.3261.8N 50a+50spr241.7269.6264.2293.2301.5297.1238.4249.5244.1257.8273.6268.5N 34a+66spr275.4264.7268.1296.5265.4304.0243.7218.4254.2271.9249.5275.4N-min spring282.7261.1264.1310.7317.6306.8263.2233.3250.3285.5270.7273.7Analyses of variance (LSD-test to levels 5% and 1%)ABCDABACADBCCDABCABDACDLSD 5% LSD 1%3.3 4.420.1 26.53.3 4.44.7 6.34.7 6.35.8 7.78.2 10.84.7 6.38.2 10.85.2 7.06.3 8.77.1 9.3 According to the fertilization date x nitrogen form interaction it can be concluded that significant differences occurred within certain fertilization variants. The inbred lines fertilization based on the Nmin method and distribution of N fertilizers in autumn and spring (D5: 34% + 66%) by the application of the amidic nitrogen (urea) resulted in a significantly higher grain weight in relation to the using of CAN. On the other hand, 1000-grain weight was very significantly higher in the fertilizing variants D2 (single application of N fertilizers in autumn) and D4 (application of N fertilizers in autumn and spring: 50% + 50%) with CAN in relation to the urea. Grain weights did not differ over D3 variant (single application of N fertilizers in spring). Observed maize inbred lines differently responded to nitrogen application dates. The inbred line IL1 had significantly lower seed weight in treatment with single application of fertilizers in autumn in relation to other treatments. The highest 1000-grain weight in the inbred IL2 was obtained by the application of the Nmin method. These data differ from results obtained by other study [6] which showed that the highest 1000-grain weight had been obtained by the application of the greatest fertilizer rate (N-150 kgha-1, P2O5 -120 kgha-1 and K2O-80 kgha-1). Similar investigations were made by other studies [2] [3] [4] [9][14] [16]. For example, by testing 1000-grain weight of maize inbred lines and hybrids by fertilization for 3-year period, increase of soil moisture and application of N fertilizers affected the increase of 1000-grain weight Using of N fertilizers had considerably influences on maize 1000-grain weight and resulted for 30-40% higher yield in comparison with the unfertilized plot. 1000-grain weight was depended on application N fertilizers and these effects were different for individual maize hybrids. CONCLUSIONS AND FUTURE WORK According to the statistical analysis of effects of fertilisation dates and nitrogen forms on the 1000-grain weight of maize inbred lines, very significant differences were obtained over investigation years and genotypes. It can be concluded that each inbred has its own potential for magnitude and range of variations of observed trait. Fertilizing on the basis of the Nmin method resulted in very significantly higher 1000-grain weight in relation to other fertilization variants, on the average for years, genotypes and all three nitrogen forms. The use of the N-min method test (N ranging from 17 to 35 kg ha-1, in dependence on the soil mineral nitrogen content), especially in years with lower precipitation sums, resulted in the highest increase in 1000–grain weight (3.2%) of observed maize inbred lines in relation to the control. The application of different forms of nitrogen did not result in statistically significant differences in 1000–grain weight of observed maize inbred lines. Thousand-grain weight is an important sowing-technical trait mainly conditioned by inheritance, but partially subjected to certain cropping practices. It is considered that 1000-grain weight close to a mean for a given hybrid is desirable. Namely, the overweight means a small number of grain per a unit of the harvested area, as it is a case with the inbred lines. This trial shows variations of this trait within a population, which is a result of inheritance. References [1] Binder, D.L., Sander, D.H. Walters, D.T. 2000. Maize response to time of nitrogen application as affected by level of nitrogen deficiency. Agron. J., 92, 1228-1236. [2] Cirilo. A.G., Andrade, E.H. 1994. Sowing date and maize productivity: II Kernel number detrmination. Crop Sci. 34, 1044-1046. [3] Cirovic, M. 1985. Prou avanje bioloakih osobina samooplodnih linija kukuruza pri razli itim gustinama i optimalnim uslovima gajenja sa i bez navodnjavanja. Doktorska disertacija. Poljoprivredni fakultet, Novi Sad. [4] Gotlin, J., Pucaric, A., Varga, B. 1981. Utjecaj gnojidbe duaikom na prinos i komponente prinosa hibrida kukuruza raznih vegetacijskih grupa. Zbornik radova sa nau nog skupa: Ekosistemi i mogunosti njihovog racionalnog koriaenja , Matica Srpska, Novi Sad, 389-400. [5] Josipovic M., Kovacevic V., Petosic D., Sostaric Jasna 2005. Wheat and maize yield variations in the Brod-Posavina area. Cereal Research Communications 33 (1):229-233. [6]Jovin, P., Veskovi, M. (1997): Uticaj gustine setve i doza mineralnih ubriva na prinos i broj zrna u semenskom kukuruzu. Selekcija i semenarstvo, Vol. 4, 3-4, 93-97, Novi Sad. [7] Kling, J.G., A.E. Okoruwa 1994. Influence of variety and environment on maize grain quality for food uses in Africa. p. 119. In Agronomy abstracts. ASA, Madison, W.I. 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Badu-Apraku et al. 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Contributing to food self-suficiency: maize resaearch and development in West and Central Africa. Proc. Regional Maize Workshop. IITA.Cotonou, Benin Republic. 29 May – 2 June 1995. WECAMAN/IITA, Nigeria. [14] Purcino, A.A.C., Silva, M.R., Andrade, S.R.M., Belele, C.I., Parentoni, S.N., dos Santos, M.X. 2000. Grain filling in maize: The effect of nitrogen nutrition on the activities of nitrogen assimilating enzymes in the Pedicel-placento-chalaza region. Maydica 45, 95-103. [15] Shaw R. H. 1988. Climatic requirement. pp. 609-638. In: G. F. Sprague (Ed.). Corn and corn improvement, Agronomy Monograph No 18 ASA-CSSA-SSSA, Madison, Wisconsin, USA. [16] Uhart, S.A., Andrade F.H. 1995. Nitriogen deficiency in maize:I. Effects on crop growth, development, dry matter partitioning and kernel set. Crop Sci., 35,1376-1383. [17] Vetsch, J.A., Randall, G.W. 2004. Corn production as affected by nitrogen application timing and tillage. Agronomy J., 96, 2, 502-509. ABOUT THE AUTHORS Dr Zdravko Hojka, dr Predrag Jovin Maize Research Institute „Zemun Polje”, Slobodana Bajica 1, Belgrade–Zemun, Serbia; E–mail HYPERLINK "mailto:zhojka@mrizp.co.yu" zhojka@mrizp.co.yu Ivica Djalovic, University in Kragujevac, Faculty of Agronomy–Cacak, Cara Dusana 34, 32000 Cacak, Serbia, E–mail: HYPERLINK "mailto:sanjas@telekom.yu" sanjas@telekom.yu Prof. Dr. Vlado Kovacevic, University J. J. Strossmayer in Osijek, Faculty of Agriculture, Trg Sv. Trojstva 3, HR-31000 Osijek, Croatia; E-mail: HYPERLINK "mailto:vladok@pfos.hr" vladok@pfos.hr Mirko Grubisic, ITNMS, Belgrade, Serbia Research People and Actual Tasks on Multidisciplinary Sciences 6 – 8 June 2007, Lozenec, Bulgaria PAGE ÑÙÙÙåÙçÙèÙúÙSÚfÚ„Ú”Ú–Ú—Ú™Ú¤Ú·Ú¸Ú'Û/Û@ÛBÛCÛfÛ¯Û¸ÛñäÚÐÚÃµÃ¦˜Š˜yk˜yYyÚÐÚL>hbŒh±Y6OJQJ^Jh%.h±YOJQJ^J#hbŒh8ß6OJQJ^JmHsHhbŒOJQJ^JmHsH h%.h8ßOJQJ^JmHsHhÙl3OJQJ^JmH sH hbŒOJQJ^JmH sH h%.h[×OJQJ^JaJhbŒh[×6OJQJ^Jh%.h[×OJQJ^JhÙl3OJQJ^JhbŒOJQJ^Jh%.hVjOJQJ^JhbŒhVj6OJQJ^J¸ÛºÛÎÛÏÛáÛÜ Ü ÜÜÜÜÜÜ#Ü$Ü)Ü,Ü5Ü6Ü<Ü@ÜHÜIÜNÜPÜVÜXÜYÜZÜ^Ü`ÜaÜbÜôçØÆ¸©–Š–Š–zŠ–Š–Š–Š–Š–Š–Š–g–g–T–%jhÞhß2‹OJQJU^JaJ$hÞhß2‹OJQJ^JaJmHsHhß2‹OJQJ^JaJmHsHhß2‹OJQJ^JaJ$hÀhß2‹OJQJ^JaJmHsHhÞhß2‹OJQJ^JaJhß2‹OJQJ^JmHsH#hxf(hß2‹5OJQJ^JmHsHhÙE³5OJQJ^JmHsHh%.h±YOJQJ^JhbŒ6OJQJ^J bÜkÜmÜsÜtÜzÜ{Ü€ÜÜƒÜ„Ü†ÜˆÜ‰ÜŠÜÜ‘Ü–Ü—Ü™ÜšÜœÜÜžÜ£Ü¤Ü¬Ü®Ü¸Ü¹Ü»Ü¼ÜÆÜÈÜÏÜÐÜÒÜÓÜÛÜÜÜíÚíÚíÚíÚíÚíÚÄ±œ‡œ‡œ‡œ±Úveveveveveveveve hÀhß2‹OJQJ^JmHsH hÀhß2‹OJQJ^JmH sH 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