Sugar beet response to N fertilization as assessed by late season chlorophyll and leaf area index measurements in a semi-arid environment

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1 Interntionl Journl of Plnt Prodution 2 (1), Jnury 2008 ISSN: (Print), (Online) This is refereed journl nd ll rtiles re professionlly sreened nd reviewed. GUASNR Sugr eet response to N fertiliztion s ssessed y lte seson hlorophyll nd lef re index mesurements in semi-rid environment J.T. Tsilts, *, N. Mslris Helleni Sugr Industry SA, Lriss ftory, Deprtment of Experimenttion, 4 10 Lriss, Greee Agronomi Reserh Servie, Sindos, Greee * Corresponding uthor. E-mil: tsilts01@hotmil.om Aepted 7 Otoer 2007; Pulished online 25 Novemer Astrt In 3-yer experiment (2002-), five N rtes (0, 60, 0, 1, 240 kg N h -1 ) were pplied to sugr eets rrnged in Rndomized Complete Blok design with six replitions. Experimenttion took ple under the semi-rid, irrigted onditions of entrl Greee. The im of this work ws to study the lte seson response of sugr eet rop to N fertiliztion s ssessed y non-destrutive mesurements of hlorophyll (SPAD) nd Lef Are Index (). Physiologil mesurements were onduted from erly August to mid-septemer, every two weeks. Sugr yield response to N fertiliztion ws evident only in. The highest root nd sugr yields were found in 2002 s result of the unusully high, for Mediterrnen onditions, rinfll during summer. The highest petiole NO 3 -N onentrtions nd the lowest surose ontent in root fresh weight were lso reorded in In 2003, sugr eets were wter stressed t lte August nd thus ws minimum. Thus, the lowest root nd sugr yields were found in In, SPAD nd vlues were relted with root nd sugr yields. SPAD redings of were the optiml for mximum yield. Optiml vlues for root nd sugr yields were 3.99 nd 3.88, respetively. SPAD ws, lso, relted with petiole NO 3 -N onentrtion nd α-mino N in roots. A helthy nd green nopy t the lte stges of the growing seson n ontriute to high sugr eet yield. Optiml N rte for mximum root nd sugr yield ws lose to or higher thn 200 kg h -1 N. Chlorophyll (SPAD) nd determintion y instruments ould e sensitive tool in reveling sugr eet response to N fertiliztion nd nondestrutively ssessing sugr eet N nutrition sttus. Keywords: Drought; Photosynthesis; Plnt nutrition; SPAD; Sugr yield Introdution Sugr eet produtivity worldwide is hevily ffeted y the intertion of wether vriility with nutrient vilility (Frekleton et l., 1999; Märländer et l., 2003). Thus, optimizing nutrient nd espeilly N input, whih is the most limiting nutrient, is provoking gol for sugr eet rop (de Koeijer et l., 2003). This effort is more hllenging

2 58 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: under Mediterrnen onditions euse supplementl irrigtion omplites the intertion. Nitrogen ffets oth root nd sugr yields (Dryott nd Christenson, 2003). Adequte N supply supports Lef Are Index () of 3 or more mximizing sun-light intereption, surose synthesis in leves nd sugr umultion in roots. Exessive N ffets negtively the prtitioning of photosyntheti produt inresing the proportion of petiole nd lef mss (Sott nd Jggrd, 1993; Dryott nd Christenson, 2003). Exept from dense nopy, effetive rdition turn over into surose requires N investment in photo-ssimiltory mhinery nd thus in hlorophyll, the min ompound prtiipting in photosynthesis. Nitrogen vilility ffets the totl mount of surose produed y sugr eets nd it hs detrimentl effets on white sugr reovering, during industril proessing, vi the inrese of non-sugr molssigeni ftors (K, N, α-mino N) in roots (Hrvey nd Dutton, 1993). Reently, there is trend to sustitute lorious nd time-onsuming physiologil mesurements with instrumentl, non-destrutive methods. SPAD-502 hs een developed s non-destrutive lef hlorophyll meter nd lose reltionships etween SPAD redings nd destrutively determined lef hlorophyll onentrtion in mny speies were estlished (Ymmoto et l., 2002; Wng et l., ). Sine hlorophyll is nitrogenous ompound, lose reltionships etween SPAD redings nd lef N onentrtion were found (Duru, 2002; Wng et l., ) nd thus SPAD is used s rop N inditor (Singh et l., 2002; Wiesler et l., 2002) or yield preditor (Rmesh et l., 2002; Singh et l., 2002). Non-destrutive methods hve, lso, een evolved for ssessing nopy hrteristis suh s the perentge of rop over nd. In sugr eet, Röver nd Koh (19) found strong reltionship etween destrutive nd instrumentl determintions. Although muh work hs een onduted to understnd physiology t the erly stges of sugr eet, rop responses t the lter stges hve not een studied in depth. Mintenne of helthy nd green nopy during utumn ould ontriute signifintly to hrvested yield (Oer et l., 2003). Thus, instrumentl mesurements of physiologil trits like hlorophyll nd during the lte stges ould provide useful informtion out sugr eet physiology nd yield. The im of this work ws to study the effet of N fertiliztion on lte seson, nondestrutively determined hlorophyll ontent nd nd to relte these trits with sugr eet N nutrition nd yield. Mterils nd methods Site nd experimentl set up A 3-yer (2002-) experiment ws onduted in Niki, Thessly Plin, entrl Greee (39º 33 N, 22º 27 E, 98m sl). Five N rtes (N 0 =ontrol, N 60 = 60, N 0 = 0, N 1 = 1 nd N 240 = 240 kg N h -1 ) were pplied in Rndomized Complete Blok design with six replitions. Seeds of v Rizor (SESVANDERHAVE NV/SA, Tienen, Belgium) were mehnilly sown in eight rows (8 m long) per plot, t 50 m prt nd t m sping in the row. The 2:3 of N were pplied s sl (mmonium sulphte) nd the rest s top-dressing (mmonium nitrte) given during the first hlf of My. Before sowing, 200 kg P 2 O 5 h -1 (hyper-phosphte) nd 320 kg K 2 O h -1 (potssium sulphte) were inorported

3 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: into soil. Tle 1 presents gronomi informtion nd Tle 2 some soil hrteristis determined in soil smples tken from the enter of ontrol plots efore fertilizer pplition. The limte of the region is typil Mediterrnen with July nd August eing the hottest months (25ºC nd 26.5ºC men monthly temperture, respetively) nd with negtive wter udget (preipittion + irrigtion evpotrnspirtion) during growing seson. Wter udget ws more defiit in 2003 (-54 mm), followed y (-38 mm) nd 2002 (-22.5 mm). Supplementl irrigtion ws provided ording to the reommendtions of Agronomi Servie of Lriss ftory nd the vilility of irrigtion wter. Full protetion ginst erospor, powdery mildew, insets nd weeds were tken y spryings. On erly July, petioles per plot were rndomly olleted, oven-dried (t o C for 48 h) nd nlyzed for NO 3 -N (Ulrih, 10). Chlorophyll ontent nd were mesured four times eginning t erly August nd every dys. Lef hlorophyll ontent ws ssessed y SPAD-502 (Minolt Co Ltd, Osk, Jpn) on ten full-expnded, intt nd full sun-lit leves per plot. ws determined using SunSn Cnopy Anlysis System (Delt-T Devies Ltd, Cmridge, UK). Two mesurements were tken etween the 4th nd 5th rows in eh plot. The three internl rows (3 rd, 4 th, 5 th ) were hrvested y hnd in length of 7 m (10.5 m 2 ) per plot. Sugr eets were topped y hnd, root numer ws ounted nd root nd top weights were mesured. A rndomly seleted root smple (25-30 roots), from eh plot, ws trnsferred to ftory s tre house for qulittive determintions (% surose ontent in fresh root, K, N, α-mino N ontent). Root qulity ws determined using Venem utomti eet lortory system (Venem utomtion.v., Groningen, Hollnd) onneted with BETALYSER nlyzing system (Dr Wolfgng Kernhen GmH, Seelze, Germny). Moisture ontent of roots nd tops ws estimted fter drying smple of ~200 g t o C till onstnt weight. Hrvest Index ws lulted s the perentge of root dry weight to the totl dry mss (roots + tops). On hrvest dte, soil profile (0-30 m nd m) smples were tken from the enter of eh plot using ylindril proe nd soil NO 3 -N ws determined y olorimetri method (Bremner, 1965). Dt nlysis Soil dt efore experiment estlishment were sujeted to one ftor (depths) Anlysis of Vrine (ANOVA) omined over yers. Dt of CEC were sujeted to one ftor (yers) ANOVA. Produtive trits (root numer, root nd sugr yields, Hrvest Index, surose ontent in fresh or dry weight, α-mino N) nd NO 3 -N onentrtion in petioles were nlyzed s one ftor (N rtes) Rndomized Complete Blok design omined over yers. SPAD nd were nlyzed s two ftor (N rtes nd dtes) Rndomized Complete Blok design with split plot omined over yers. NO 3 -N in soil t hrvest ws nlyzed s two ftor (N rtes nd depths) Rndomized Complete Blok design with split plot omined over yers. Mens were ompred y LSD test using M- STAT sttistil pkge (MSTAT-C, version 1.41, Crop nd Soil Sienes Deprtment, Mihign Stte University, USA).

4 60 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: Tle 1. Agronomi informtion of the experiments. Yer Previous rop Sowing dte Hrvest dte * Irrigtion numer Wter (mm) 2002 Cotton 19 Mrh 3 Otoer (198) Whet 18 Mrh 1 Otoer (196) Cotton 23 Mrh 24 Septemer (184) In prenthesis is given Dys After Sowing (DAS) Tle 2. Soil hrteristis, t two depths (0-30 m nd m), of the experimentl sites. In the sme olumn, mens leled with the sme letter did not differ signifintly t P<0.05. Soil nlysis Exhngele tions Snd Silt Cly ph CCO 3 Org. mtter Totl N NO 3 -N P-Olsen K N CEC (g kg -1 ) (1:1) (g kg -1 ) (mg kg -1 ) (mg kg -1 ) (mol kg -1 ) Depth m d m m d 1.00 d d m 0-30 m m Results NO 3 -N in petioles nd soil t hrvest The highest NO 3 -N onentrtion ws mesured in 2002 (8223 mg kg -1 ) nd the lowest in 2003 nd (4309 nd 2461 mg kg -1, respetively). Higher N rtes (N 0, N 1, N 240 ) inresed signifintly NO 3 -N in petioles (Figure 1). Soil NO 3 -N onentrtion t hrvest ws higher in the upper lyer (0-30 m). The highest onentrtion ws found in 2003 when n inrese of the residul NO 3 -N in the soil with inresing N rte ws evident. In 2002 nd, no signifint differenes were found etween yers or N tretments (Figure 1). SPAD nd mesurements Yers nd dtes signifintly ffeted oth SPAD nd. Nitrogen rtes hd no signifint effet on SPAD ut ws strongly ffeted ut the simple intertions (yers N rtes, yers dtes, N rtes dtes) were signifint for oth trits (Figure 2). ws highest in 2002 nd lowest in 2003 ut in oth yers, no signifint effets of N rtes were evident. In, positive retion of to N fertiliztion ws found sine the higher N tretments (N 0, N 1, N 240 ) hd higher vlues thn the lower rtes (N 0, N 60 ). In ll yers, showed deresing trend with the progress of seson. At erly August, did not differ signifintly etween yers. In 2003, ollpse of plnt nopy ourred etween mid-august nd erly Septemer when ws redued to hlf (Figure 2).

5 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: LSD= NO 3-N in petioles (mg kg -1 ) NO3-N in soil (mg kg -1 ) Nrtes(kgNh -1 ) LSD=.29 N rtes (kg N h -1 ) Figure 1. Conentrtion of NO 3 -N in petioles nd soil for the five N rtes. The highest SPAD vlues were found in 2003, the lowest in while SPAD vlues in 2002 were moderte. In 2002 nd 2003, SPAD ws unffeted y N rtes ut in, the higher N tretments (N 1, N 240 ) showed higher SPAD redings. In, SPAD grdully inresed towrd mid-septemer (Figure 2). Produtive trits Fertiliztion ffeted signifintly sugr yield, surose ontent in dry weight nd α- mino N in roots. The yers N rtes intertion ws signifint only for sugr yield. Root numer ws ffeted y yer eing higher in 2002 nd 2003 ( nd plnts h -1, respetively) ompred with (84 0 plnts h -1 ). Root yield ws highest in 2002 (8.4 t h -1 ), lowest in 2003 (73.7 t h -1 ) nd moderte in (87.4 t h -1 ). Surose ontent in fresh weight ws signifintly ffeted y yers, eing highest in nd 2003 (.9 nd.5%, respetively) nd lowest in 2002 (.4%). The highest sugr yield ws found in 2002 nd (.7 t h -1 nd.8 t h -1, respetively) nd the lowest in 2003 (.4 t h -1 ). When dt omined over yers, higher N rtes hd positive effet on sugr yield (N 0, N 1, N 240 ) ompred with the lower ones (N 0, N 60 ). Also, signifint effets of N rtes on sugr yield were evident in (Tle 3).

6 62 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: Surose ontent in dry weight did not differ etween yers lthough surose onentrtion in 2002 (68.1%) ws 2% lower thn tht found in 2003 nd (70.2 nd 70.0%, respetively). Signifintly higher surose ontent in dry weight ws found in N 60 nd N 0 (72.3% nd 69.9%, respetively). Hrvest Index ws highest in 2003 (82.35 %), lowest in 2002 (79.56 %) nd moderte in (.63 %) ut it ws not ffeted y N rtes. Hrmful N in roots inresed y the highest N doses (N 1, N 240 ). The lowest α-mino N ws mesured in (199.1 mg N g -1 sugr) nd the highest in 2002 nd 2003 (408.7 nd mg N g -1 sugr, respetively). Reltionships etween yield nd fertiliztion or physiologil trits Figure 3 presents the reltionships etween root nd sugr yields with N rtes for dt omined over yers nd in. From the qudrti funtions est fitted to dt, the optimum N doses were lulted. For oth omined nd dt, root yield ws mximum t higher N rtes ( nd kg N h -1, respetively) ompred with sugr yield ( nd kg N h -1, respetively). We lso plotted root nd sugr yields ginst nd SPAD vlues in order to determine the optimum of physiologil trits mximizing yields in (Figure 4). Optimum for mximum root yield ws lulted t 3.99 nd for sugr yield t Optimum SPAD were nd 38.26, respetively. Figure 5 presents the reltionship etween SPAD nd NO 3 -N onentrtion in petioles or α-mino N in roots during. Qudrti funtions reveled tht NO 3 -N in petioles ws mximized t SPAD reding nd α-mino N t SPAD. No signifint reltionship etween SPAD or vlues nd yields, NO 3 -N in petioles or α-mino N in roots ws found in 2002 nd 2003 dt. Tle 3. Comprison of mens for yers, N rtes nd their intertion of the trits (root nd sugr yield, surose ontent in fresh weight, surose ontent in dry weight, Hrvest Index) determined. Mens leled with the sme letter did not differ signifintly t P<0.05. N rte (kg h -1 ) Root yield (t h -1 ) Sugr yield (t h -1 ) Hrvest Index (%) f d-f e f e f ef f 81.4-d f N rte (kg h -1 ) Surose ontent in fresh weight (%) Surose ontent in dry weight (%) d 69.6-d d d 68.4-d d 69.1-d d d d

7 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: LSD = 0.79 N rtes (kg N h -1 ) LSD = de d d e e de f f erly Aug. mid-aug. erly Sept. mid-sept. Dte LSD = 2.41 SPAD e de de d N rtes (kg N h -1 ) LSD = 1.06 SPAD e e d e d 40 f fg g fg erly Aug. mid-aug. erly Sept. mid-sept. Dte Figure 2. SPAD nd vlues of the N rtes nd dtes.

8 64 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers Root yield (t h -1 ) All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers Root yield (t h -1 ) All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers All yers x x , r 2 = x x , r 2 = 0.92 All yers S All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers Sugr yield (t h -1 ) All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers Nrtes (kg N h -1 ) All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers S All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers Sugr yield (t h -1 ) All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers Nrtes (kg N h -1 ) All yers All yers All yers x x +.406, r 2 = x x +.563, r 2 = 0.91 All yers Figure 3. Qudrti funtions etween root or sugr yields nd N rtes. Disussion Experiments took ple on inorgni, lyey soils, typil of Thessly Plins, entrl Greee. In those soils, seed-ed ggregtes redued plnt emergene in (Dürr nd Auertot, 2000) ut N fertiliztion or rop sequene hd no effet on plnt popultion on ontrry to other reports (Dryott nd Christenson, 2003; Moyer et l., ). However, in ll ses, plnt numer exeeded 000 plnts h -1, the lower limit for mximum yield (Sott nd Jggrd, 1993). Sine sugr eet response to N is highly site- nd yer-speifi (de Koeijer et l., 2003; Märländer et l., 2003), positive effet of N fertiliztion on yield ws found only in. Optiml N rte, for mximum root nd sugr yields, ws lose to or exeeded 200 kg N h -1, in ordne with previous reports in Greee (Tsilts nd Mslris, 2005).

9 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r Root yield (t h -1 ) x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r 2 = x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r Root yield (t h -1 ) x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r x x , r 2 = x x 88.2, r 2 = x x 88.2, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = 0.97 Sugr yield (t h -1 ) x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = 0.97 Sugr yield (t h -1 ) x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x 2 +.7x.62, r 2 = x x.1, r 2 = SPAD x x.1, r 2 = SPAD Figure 4. Qudrti funtions etween root or sugr yields nd or SPAD in. Root yield (t h -1 ) Sugr yield (t h -1 ) SPAD

10 66 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: NO3-N in petioles (mg kg -1 ) x x , r 2 = SPAD α-mino N (mg N g -1 sugr) x x , r 2 = SPAD Figure 5. Reltionships etween or SPAD nd root or sugr yields in. Totl N need of sugr eet is 250 kg N h -1, hlf of whih is provided y residul nd minerlized N (Märländer et l., 2003). Under Greek onditions, high optiml N rtes re proly sried to the low levels of soil minerlizle N sine the two trits re inversely relted (Allison et l., 1996). The highest root yield hrvested in 2002 is the result of the unusul, for Mediterrnen onditions, rinfll ourred from July onwrd. In semi-rid res, soil N vilility depends on wter vilility (Tsilts et l., 2001) nd thus, in 2002, sugr eets took up more N s it ws lso indited y the high α-mino N in roots (Pook et l., 19). Nitrogen uptke ws trnslted to higher NO 3 -N onentrtion in petioles nd higher folige nd totl iomss prodution (Sott nd Jggrd, 1993). In 2002, soil resoures (wter, nitrogen) vilility fvored prtitioning of photo-ssimiltes to the oveground iomss deresing Hrvest Index (Werker et l., 1999). Moreover, even not signifint, surose ontent in dry weight ws lowest in 2002; n indition of prtitioning to oveground prts. The lowest surose ontent in fresh weight found in 2002 is lso

11 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: sried to the high wter vilility, whih inresed root moisture ontent nd diluted surose (Follett, 1991). In 2003, no response to N ws ompnied with moderte NO 3 -N onentrtion in petioles nd high prtitioning to roots (high HI). These findings ould e sried to limited exploittion of soil NO 3 -N y sugr eets proly due to soil wter defiit, whih resulted to inresed residul NO 3 -N. Low levels of residul NO 3 -N nd no signifint differenes etween N rtes were evident in 2002 nd, in ordne with Allison et l. (1996). In, positive response to N fertiliztion ws found for rtes higher thn 0 kg N h -1 with n optimum t 200 kg N h -1. NO 3 -N in petioles estimted suessfully the indequy of N nutrition sine N 0 nd N 60 tretments hd NO 3 - N onentrtion lower or lose to the ritil level of 0 mg NO 3 kg -1 (Ulrih nd Hill, 19). Crop showed typil response to N y inresing top nd root weights nd onsequently sugr yield without negtive effet of N on surose ontent in fresh weight. Hrmful N in roots inresed y N fertiliztion nd in ll ses, exeeded the eptle limit (0 mg N g -1 sugr) for inorgni soils (Plmer nd Csurn, 19; Tsilts nd Mslris, 2005). Reently, sustitution of lorious, destrutive physiologil mesurements with nondestrutive methods gins interest, espeilly under field onditions. is n importnt preditor of rop produtivity nd is used in growth models (Yin et l., 2000). Critil for mximum light intereption y sugr eets rnges etween 3.5 nd 4.0 (Sott nd Jggrd, 1993) nd uptke of kg N h -1 is neessry in order to e hieved (Mlnou et l., 2006). Hoffmnn nd Blomerg () found no reltionship etween determined in June or Septemer nd yield. In our work, root nd sugr yields in, were mximized t vlues (3.99 nd 3.88, respetively) within the optimum rnge for mximum light sorption. After mximum rehed, mintenne over 2.5 is neessry for yield mximiztion (Milford et l., 19). This ws evident in the highyielding tretments (N 0, N 1, N 240 ) in while in 2003, ollpsed etween mid- August nd erly Septemer, typil indition of rop sujetion to wter or nutrient stress. SPAD is non-destrutive ssessment of lef hlorophyll nd N ontent (Duru, 2002; Wiesler et l., 2002). As n indiret hlorophyll meter filed to distinguish high-yielding genotypes (Pulkráek et l., 2001) ut it is useful inditor of in seson sugr eet N demnd (Tugnoli nd Bettini, 2000; Wiesler et l., 2002). In, root nd sugr yields were mximized t SPAD vlues (38.30 nd 38.26, respetively) slightly higher thn the threshold vlue of set s the lower limit of N effiieny in sugr eet (Tugnoli nd Bettini, 2000). In Greee, sugr eets re left without irrigtion for out month in order surose onentrtion in fresh mss to e inresed. In tht se, lef SPAD drops elow nd thus yield is negtively ffeted. Consequently, higher N rtes ould mintin green folige for longer period during irrigtion withholding nd thus yield losses ould e restrited. In 2003, high SPAD vlues ould e sried to wter stress onditions (Grí-Vlenzuel et l., 2005) ut the drk green folige ws not dvntgeous for CO 2 ssimiltion nd surose umultion in root (Pulkráek et l., 2001; Dryott nd Christenson, 2003). In our work, SPAD ws linerly relted to NO 3 -N in petioles up to 3000 mg NO 3 kg -1 nd SPAD vlue of ws the optiml for mximizing NO 3 -N in petioles. Sexton nd Crroll (2002) found liner reltionship etween the two trits up to

12 68 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: mg NO 3 kg -1 in petioles nd supported tht NO 3 -N in petioles is more urte preditor of sugr eet N nutrition. Finlly, sugr eet N uptke, s ssessed y α-mino N in roots (Pook et l., 19) ws mximized t SPAD reding of Conlusions Yield response to N fertiliztion ws found only in. Optiml N rte for mximum root nd sugr yields pprohed or exeeded 200 kg N h -1 for or omined dt. In, root nd sugr yields were relted to SPAD nd. Optiml SPAD for mximum yields ws while optiml rnged from 3.88 for sugr yield to 3.99 for root yield. Also, in, signifint qudrti funtions etween SPAD nd NO 3 -N in petioles or α-mino N in roots were evident. Aknowledgments We thnk N. Athnsiou, P. Akrivosoulis, A. Zuhero Zhros, G. Goussniotis, K. Goulikou nd A. Kostrellou for their help during the ourse of experimenttion nd the personnel of Soil Siene L, Agronomi Reserh Servie, Helleni Sugr Industry SA, Sindos, for the nlyses. We re grteful to Mr E. Kourelos who kindly offered his lnd to undergo this experimenttion. Referenes Allison, M.F., Armstrong, M.J., Jggrd, K.W., Todd, A.D., Milford, G.F.J., An nlysis of the gronomi, eonomi nd environmentl effets of pplying N fertilizer to sugr eet (Bet vulgris). J Agri. Si. 7, Bremner, J.M., Inorgni forms of nitrogen. In: C.A. Blk et l. (Eds.), Methods of Soil Anlysis, Prt 2, Agronomy 9. ASA, Mdison, pp de Koeijer, T.J., de Buk, A.J., Wossink, G.A.A., Oenem, J., Renkem, J.A., Struik, P.C., Annul vrition in wether: its implitions for sustinility in the se of optimizing nitrogen input in sugr eet. Eur. J. Agron. 19, Dryott, A.P., Christenson, D.R., Nutrients for Sugr Beet Prodution: Soil-Plnt Reltionships. CABI Pulishing, Wllingford. Dürr, C., Auertot, J.-N., Emergene of seedlings of sugr eet (Bet vulgris L.) s ffeted y the size, roughness nd position of ggregtes in the seeded. Plnt Soil 219, Duru, M., Evlution of hlorophyll meter to ssess nitrogen sttus of oksfoot swrd. J. Plnt Nutr. 25, Follett, R.F., Sesonl surose, dry mtter, nd tion onentrtions of sugreet s influened y vriety nd N-fertiliztion. Commun. Soil Si. Plnt Anl. 22, Frekleton, R.P., Wtkinson, A.R., We, D.J., Thoms, T.H., Yield of sugr eet in reltion to wether nd nutrients. Agri. For. Meteorol. 93, Grí-Vlenzuel, X., Grí-Moy, E., Rsón-Cruz, Q., Herrer-Estrell, L., Agudo-Sntruz, G.A., Chlorophyll umultion is enhned y osmoti stress in grmineous hlorophylli ells. J. Plnt Physiol. 2, Hrvey, C.W., Dutton, J.V., Root qulity nd proessing. In: Cooke, D.A., Sott, R.K. (Eds.) The Sugr Beet Crop: Siene into Prtie. Chpmn & Hll, London, pp Hoffmnn, C.M., Blomerg, M.,. Estimtion of Lef Are Index of Bet vulgris L. sed on optil remote sensing dt. J. Agron. Crop. Si. 1,

13 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: Mlnou, C.S., Jggrd, K.W., Sprkes, D.L., A nopy pproh to nitrogen fertilizer reommendtions for the sugr eet rop. Eur. J. Agron. 25, Märländer, B., Hoffmnn, C., Koh, H.-J., Ldewig, E., Merkes, R., Petersen, J., Stokfish, N., Environmentl sitution nd yield performne of the sugr eet rop in Germny: heding for sustinle development. J. Agron. Crop Si. 189, Milford, G.F.J., Pook, T.O., Jggrd, K.W., Bisoe, P.V., Armstrong, M.J., Lst, P.J., Goodmn, P.J. 19. An nlysis of lef growth in sugr eet. IV. The expnsion of the lef nopy in reltion to temperture nd nitrogen. Ann. Appl. Biol. 107, Moyer, J.R., Nitshelm, J., Regitnig, P., Blkshw, R.E., Hung, H.C., Chng, C.,. Effet of tillge system nd rop sequene on irrigted sugreet prodution. Cn. J. Plnt Si. 84, Oer, E.S., Clrk, C.J.A., Jggrd, K.W., Ftors ontriuting to improved effiieny of sugreet rop during the utumn in the UK. Zukerind. 8, 1-4. Plmer, M., Csurn, C., 19. Amino nitrogen nlyses-ftory experienes. Brit. Sugr Beet Rev. 53, Pook, T.O., Milford, G.F.J., Armstrong, M.J., 19. Storge root qulity in sugreet in reltion to nitrogen uptke. J. Agri. Si. 1, Pulkráek, J., Jozefyová, L., Fměr, O., Šroller, J., Štěpánek, P., Differenes in hlorophyll ontent in leves of sugr eet (Bet vulgris L.). Rostlin Výro 47, Rmesh, K., Chndrsekrn, B., Blsurmnin, T.N., Bngrusmy, U., Sivsmy, R., Snkrn, N., Chlorophyll dynmis in rie (Oryz stiv) efore nd fter flowering sed on SPAD (hlorophyll) meter monitoring nd its reltion with grin yield. J. Agron. Crop Si. 188, Röver, A., Koh, H.J., 19. Indiret determintion of Lef Are Index of sugr eet nopies in omprison to diret mesurement. J. Agron. Crop Si. 174, Sott, R.K., Jggrd, K.W., Crop physiology nd gronomy. In: Cooke, D.A., Sott, R.K. (Eds.) The Sugr Beet Crop: Siene into Prtie. Chpmn & Hll, London, pp Sexton, P., Croll, J., Comprison of SPAD hlorophyll meter redings vs. petiole nitrte onentrtion in sugreet. J. Plnt Nutr. 25, Singh, B., Singh, Y., Ldh, J.K., Bronson, K.F., Blsurmmin, V., Singh, J., Khind, C.S., Chlorophyll meter- nd lef olor hrt-sed nitrogen mngement for rie nd whet in Northwestern Indi. Agron. J. 94, Tsilts, J.T., Mslris, N., Effet of N fertiliztion rte on sugr yield nd non-sugr impurities of sugr eets (Bet vulgris) grown under Mediterrnen onditions. J. Agron. Crop Si. 191, Tsilts, J.T., Kssioumi, M.T., Veresoglou, D.S., N-nihe sptiotemporl differentition, N uptke rtes nd ommunity struture in Mediterrnen grsslnd. J. Medit. Eol. 2, Tugnoli, V., Bettini, G., Nitrogen fertilizers in sugreet spring sowing: use of the SPAD optil instrument. Proeed. IIRB Congr. 65, Ulrih, A., 10. Critil nitrte levels of sugr eets estimted from nlysis of petioles nd ldes with speil referene to yields nd surose onentrtions. Soil Si. 69, Ulrih, A., Hill, F.J., 19. Plnt nlysis s n id in fertilizing sugr eet. In: Westermn, R.L. (Ed.), Soil Testing nd Plnt Anlysis. SSSA, Mdison, pp Wng, Q., Chen, J., Li, Y.,. Nondestrutive nd rpid estimtion of lef hlorophyll nd nitrogen sttus of pee lily using hlorophyll meter. J. Plnt Nutr. 27, Werker, A.R., Jggrd, K.W., Allison, M.F., Modelling prtitioning etween struture nd storge root in sugr eet: effets of drought nd soil nitrogen. Plnt Soil 207, Wiesler, F., Buer, M., Kmh, M., Engels, T., Reush, S., The rop s inditor for sidedress nitrogen demnd in sugr eet prodution-limittions nd perspetives. J. Plnt Nutr. Soil Si. 7, Ymmoto, A., Nkmur, T., Adu-Gymfi, J.J., Sigus, M., Reltionship etween hlorophyll ontent in leves of sorghum nd pigeonpe determined y extrtion method nd y hlorophyll meter (SPAD-502). J. Plnt Nutr. 25, Yin, X., Shpendonk, A.H.C.M., Kropff, M.J., vn Oijen, M., Bindrn, P.S., A generi eqution for nitrogen-limited Lef Are Index nd its pplition in rop growth models for prediting lef senesene. Ann. Bot.,

14 70 J.T. Tsilts, N. Mslris / Interntionl Journl of Plnt Prodution (2008) 2: 57-70

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