Abstract. Stephanie Witten*, Herwart Böhm*, and Karen Aulrich*

Transcription

1 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) DOI: /LBF Effet of variety and environment on the ontent of rude nutrient, lyine, methionine and yteine in organially produed field pea (Pium ativum L.) and field bean (Viia faba L.) Stephanie Witten*, Herwart Böhm*, and Karen Aulrih* Abtrat The ompoition of organially produed field pea and field bean a a oure of valuable protein i of peial interet for the planned 100 % organi feeding regulation in organi farming. For thi reaon, the influene of environment and variety on the ontent of rude nutrient and the amino aid lyine, methionine, and yteine were examined over three year. Laboratory analye were onduted uing NIRS. Pea ontained on average 21.9 g rude protein 100 g 1 dry matter with 8.0 g lyine 100 g -1, 1.0 g methionine 100 g -1, and 1.4 g yteine 100 g 1. In field bean 29.6 g rude protein 100 g -1 dry matter with 6.4 g lyine 100 g -1, 0.7 g methionine 100 g 1, and 1.1 g yteine 100 g 1 were found. Signifiant differene between varietie were found for rude protein and ether extrat in field pea, a well a for all omponent in field bean. Environmental fator and interation alo had influene on the ompoition of both legume peie. Furthermore, ignifiantly negative orrelation were found between the ontent of rude protein and tarh (r = -0.69), ugar (r = -0.47), lyine (r = 0.76), and methionine (r = -0.51) in field bean, a well a of rude protein and tarh (r = -0.79), ugar (r = -0.55), lyine (r = -0.78), methionine (r = -0.61), and yteine (r = -0.55) in field pea. The hift in ompoition were often undireted and for that reaon not preditable. Hene, it i reommended to analyze every bath before formulating a diet. Keyword: Nutrient ompoition, grain legume, fator of influene, amino aid Zuammenfaung Einflu von Sorte und Umwelt auf die Gehalte an Rohnährtoffen, Lyin, Methionin und Cytein in ökologih angebauten Futtererben (Pium ativum L.) und Akerbohnen (Viia faba L.) Die Gehalte an Rohnährtoffen und vor allem an limitierenden Aminoäuren von ökologih erzeugten Futtererben und Akerbohnen ind vor dem Hintergrund der 100 %-Biofütterung von Interee. Um den Einflu von Sorte und Umweltfaktoren auf die Rohnährtoff-, Lyin-, Cytein- und Methioningehalte zu unteruhen, wurden in drei Jahren Proben au Sortenveruhen in Deuthland genommen und mittel NIRS analyiert. Im Mittel enthielten Futtererben 21,9 g und Akerbohnen 29,6 g Rohprotein in 100 g Trokenubtanz. Da Rohprotein von Futtererben und Akerbohnen enthielt je 100 g im Mittel 8,0 g Lyin, 1,0 g Methionin und 1,4 g Cytein bzw. 6,4 g Lyin, 0,7 g Methionin und 1,1 g Cytein. Der Faktor Sorte beeinflute die Rohprotein- und Rohfettgehalte von Futtererben und deren Ernteerträge owie alle Inhalttoffe von Akerbohnen ignifikant. Umweltbedingte Faktoren und Interaktionen zwihen den Faktoren hatten ebenfall Effekte auf die Zuammenetzung beider Kulturarten. Der Rohproteingehalt war in Akerbohnen ignifikant negativ mit dem Stärke- (r = -0,69), Zuker- (r = -0,47), Lyin- (r = -0,76) und Methioningehalt (r = -0,51) und in Futtererben mit dem Stärke- (r = -0,79), Zuker- (r = -0,55), Lyin- (r = -0,78), Methionin- (r = -0,61) und Cyteingehalt (r = -0,55) korreliert (p < 0,01). Die Shwankungen der Inhalttoffgehalte ind oftmal ungerihtet und omit niht vorheragbar, oda eine laboranalytihe Unteruhung der Futtermittel vor jeder Rationberehnung empfohlen wird. Shlüelwörter: Nährtoffzuammenetzung, Körnerleguminoen, Einflufaktoren, Aminoäuren * Thünen Intitute of Organi Farming, Trenthort 32, D Weterau, Germany Contat: tephanie.witten@ti.bund.de, karen.aulrih@ti.bund.de

2 206 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) Introdution In animal prodution, a diet i targeted whih mathe the animal requirement and hene inreae the feed onverion ratio. Furthermore, the goal i the ue of a ombination of feedtuff with an amino aid (AA) profile equal to that needed in monogatri metabolim (Boien et al, 2000). In onluion, knowledge of the ompoition of the available feedtuff i required for preie diet formulation. The exat ontent of rude nutrient (CN) and eential AA in the available feedtuff are partiularly intereting. For eonomi reaon, table value are often ued to get information on the amount of valuable ingredient in feedtuff rather than reult from laboratory analye. Organi farming ha to deal with limitation in diet formulation. Extration meal annot be ued ine they are produed uing olvent. Oilake are ued intead. They ontain le rude protein (CP) and more ether extrat (EE) in omparion to extration meal, and an therefore not be fed in the ame amount. Some feed additive, like free AA, are not allowed in animal feed either. Furthermore, the organi prodution apaity of ome protein feedtuff, like protein onentrate, i not uffiient. Neverthele, it i intended to implement a 100 % organi feeding requirement in Until then, 5 % of peial protein feedtuff omponent of agriultural origin an be produed onventionally (EC, 2007; EC, 2008; EU, 2014). The main non-organi protein feedtuff ued are potato protein for piglet and maize gluten for poultry. Moreover, the ue of loally produed rop i intended. Due to retrition in field management, the yield (De Ponti et al., 2012) and the CP ontent (Janen and Seddig, 2007; Worthington, 2001) of organially produed rop an be lower than that of onventional farming. But they are main fator of the eonomi quality of rop. Depite their ontent of eondary plant metabolite (like viin/onviin, tannin, and trypin inhibitor), whih limit their appliability in nutrition (Krupa, 2008; Muzquiz, 2004), grain legume are important protein feedtuff, epeially in organi farming. Both the rop yield and the CN ontent of grain legume are influened by ite of ultivation, harvet year and variety (Wang and Daun, 2004). Even in onventional farming, the amount of the CN, mainly of arbohydrate and CP, an vary widely (Avola et al., 2009; Burtin et al., 2011; De Almeida Cota et al., 2006; Du et al., 1999). The retrition of organi farming ould be onidered to intenify thoe variation. Thee are reaon not to ue the ommon data olleted in onventional agriulture for diet formulation without quetioning it. Information on the ompoition of organially produed rop and the impat of different influening fator i not omprehenive. Aordingly, expanding the data bai an poibly improve the preditability of the information needed for formulating a ration. In regarding an oberved negative orrelation between the ontent of CP and the eential AA (Igbaan et al., 1996), it ha to be onidered that the eletion of high CP ontent an have a detrimental effet on the protein quality. The AA profile of the protein an hange negatively due to higher inreae of noneential AA, like proline and arginine, than of eential one (Monti and Grillo, 1983; Partanen et al., 2001; Wang and Daun, 2004). Thi phenomenon an alo our under drought tre ondition, when even abolutely higher ontent of nitrogen and ulfur an be deteted in rop (Shumaher et al., 2011). The organi prodution of oybean, whih are, due to their high protein ontent and -quality, the main protein feedtuff ued in European ountrie, i not yet uffiient. Furthermore, it i deired to upport loal organi prodution without import of protein feedtuff originating from Third World ountrie. Therefore, the importane of loal grain legume, like field pea and field bean, a valuable oure of protein i further inreaed. High lyine (Ly) ontent, in omparion to grain ereal, make their AA profile benefiial, epeially for wine. In ontrat, the ontent of ulfur-ontaining AA (SAA), namely methionine (Met) and yteine (Cy), are relatively low in field pea and field bean. In total, the SAA an often be limiting in diet for poultry and, beide Ly, even in diet for wine (Boien et al., 2000). A mentioned above, they are, in addition to tryptophan, alo limiting in field pea and field bean (Emmert and Baker, 1997; Wang and Daun, 2004; Zdunzyk et al., 1997). Aordingly, it i favorable to harvet large quantitie of grain legume with high CP and total SAA ontent. When ompared with field bean, field pea ontain lower amount of CP and relatively higher amount of SAA (Shumaher et al., 2011). In addition to a variety of environmental fator, multiple geneti fator an alo aue differene in the ompoition between varietie. Therefore, the meaured ontent of CN and of AA in field pea and field bean alo range widely (Kotlarz et al., 2011; Makkar et al., 1997; Nikolopoulou et al., 2007; Urbatzka et al., 2011). Of partiular benefit would be the extenion of the data bai and further knowledge on influene of the variety and the environment (ite and harvet year), a well a on relation between CN and AA, whih eventually would allow predition. The preditability of thoe variation in the ompoition of rop would be of great interet for diet formulation. The aim of the tudy wa to examine whether the amount of CN, Ly, Met, and Cy differ markedly in field pea and field bean varietie whih are urrently ued under pratial ondition on organi farm. Furthermore, the preditability of the trength of their reation to geneti and environmental differene wa quetioned. Another iue wa whether the maximization of CP ontent an be an adequate breeding objetive for thoe grain legume with regard to their protein quality for ue a protein feedtuff for monogatri animal. 2 Material and method 2.1 Sampling Field pea (Pium ativum L.) and field bean (Viia faba L.) ample from field variety trial of variou organially managed experimental ite in Germany were olleted in the year 2011, 2012, and In total, 113 ample of 12

3 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) white-flowering pring field pea varietie and 105 pring field bean ample of 12 varietie were available for further analye (Table 1). Table 1 Charateriti of the field pea (Pium ativum) and field bean (Viia faba) varietie of the ample et Leaftype Pium ativum L. Flowertype Viin/ Conviin Tannin Viia faba L. Abarth Alexia Alveta Bioro Aukland Divine - Caablana Epreo La Manha KWS Fabelle - Mythi Fanfare Navarro Fuego Natura n Iabell Proteta Julia Rebel Pyramid Repet Taifun w Salamana Tangenta w - Organi agriultural prodution area (a above) n = normal-leafed, = emi-leafle, w = white-flowering, = olored-flowering 1 Sandy area in the north-eat 2 Sandy area in the north-wet Sample of field pea and field bean varietie originated from 13 trial per peie. The trial took plae in different part of Germany and therefore in different organi agriultural prodution area with homogenou limati ondition (aording to JKI, 2014; Figure 1). Table 2 Number of varietie, trial, and ultivation ite of pring field pea (Pium ativum L.) and pring field bean (Viia faba L.) ample in three harvet year and in total Pium ativum L. Viia faba L. Varietie Trial Site Region Total Total Total Field pea varietie were ultivated at ten ite of the Area 1, 2, 3, 4, and 6. Field bean varietie grew at eight ite of the Area 2, 3, and 6 (Table 2). In the field bean trial, 5 to 11 varietie, and in the field pea trial, 7 to 10 varietie were teted, repetively. 3 Loamy area in the wet 4 Area with loe in the eat 6 Agriultural area in the outh and altitude in the outh-wet Figure 1 Organi agriultural prodution area with homogenou limati ondition in Germany (aording to JKI, 2014) from whih the ample originated 2.2 Laboratory analye The ample were dried for 48 hour at 60 C and then tored in a ooling hamber at 8 C. A part of eah ample wa either ground to pa through a 1.0 mm ieve for CN analye or through a 0.5 mm ieve for AA analyze. CP (Kjeldahl N*6.25), ah, EE, rude fiber (CF), tarh, and ugar a well a the AA Ly, Met, and Cy were analyzed with near infrared refletane petroopy (NIRS). Spetral data for NIRS analye were reorded uing the Fourier-Tranform NIR petrometer (NIRLab N-200, Fa. Bühi, Een) in the petral range from 1000 to 2500 nm and then exported to the NIRCal hemometri oftware (Fa. Bühi, Een). Subequently, the ontent of CN, Ly, Met, and Cy were predited uing exiting alibration. The alibration for eah ingredient were baed on referene analye of 430 pea and 315 bean ample a deribed by Aulrih et al. (2016). 2.3 Statiti When available, rop yield were extrated from open ae field trial reult and attributed to the ample.

4 208 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) The normal ditribution of the variable and the reidual were proved with hitogram and the Shapiro-Wilk-Tet. No tranformation of the data wa required. Statitial analye for unbalaned data were onduted uing pro MIXED with the REML etimation method and the Kenward-Roger- Etimation in SAS 9.4 (SAS Intitute In.) to oberve differene in the leat quare mean of the ontent of CN and AA between the field pea and field bean varietie. Furthermore, the ovariane of the fator ite and harvet year hould be oberved. Therefore, variety wa et a fixed effet, while ite and year a well a the two-ided interation between the three fator (ite*year, variety*ite, variety*year) were initially et a random effet. A model optimization wa done for eah analyzed ingredient on the bai of the Bayeian information riterion (BIC) a a meaure of the relative quality of the model. If the model howed a tatitially ignifiant influene of the fixed effet, a Tukey-tet wa added a potho-tet (Moll and Piepho, 2001). The maro %mult.a wa ued to deribe ignifiant differene (p < 0.05) with letter (Piepho, 2012). Furthermore, Pearon orrelation analye (pro CORR, SAS 9.4) were performed to examine relation between the ontent of the oberved CN and AA in field bean and field pea. In the following text, ignifiant orrelation are marked with ** (p < 0.01). 3 Reult and diuion 3.1 Field pea (Pium ativum L.) High variation of the ompoition of field pea were oberved. In addition, the relation of the SAA to Ly varied (Table 3). Table 3 Range and mean with tandard deviation (SD) of the ontent of rude nutrient (g 100 g -1 dry matter), lyine, methionine and yteine in organially produed field pea (Pium ativum L.) (g 100 g -1 rude protein) Mean SD Min Max Dry Matter Protein Lyine Methionine Cyteine MethionineCyteine (SAA) SAA:Lyine Ether Extrat Ah Fiber Starh Sugar In pig nutrition a ratio of about 0.6 between SAA and Ly i reommended a optimal (Boien, 1997). Poultry need even more SAA in their metabolim, for example for feather growth. Thu, the SAA are limiting in field pea. Neverthele, ine the ratio of SAA:Ly i notably higher in ereal, the limiting AA in ereal-baed diet i Ly. Therefore, field pea an ontribute to a balaned AA profile in the diet. Shumaher et al. (2011) reported even higher variation in the CP ontent of their organially produed field pea ample (18 39 g CP 100 g -1 DM) of variou origin. In 50 varietie of forage, marrowfat, ath rop and winter pea available at the Norddeuthe Pflanzenzuht KG, the CP ontent were higher, the Ly ontent lower and the SAA ontent idential to the preent one. Although the data wa derived from analye of onventionally produed rop, Evonik Indutrie (2010) publihed lightly lower value (AMINODat 4.0) in omparion to the preent data. They found lower mean Ly and Met ontent in 18 field pea ample harveted in Germany. The ondition of ultivation and the analyzed varietie are unknown. DLG (DLG, ) ummarized higher mean ontent of CP (n = 75), Met and Cy, but lower Ly ontent (n = 46) in organially produed field pea. Wang and Daun (2004) found lightly lower Ly and higher SAA ontent in onventionally produed field pea. Table value for CN derived from onventionally produed field pea (Staudaher and Potthat, 2014) differed partially from the reult. For EE and tarh they were lower than the oberved value, wherea for ugar and ah they were higher. Other author oberved diverting reult epeially for CP and tarh, a well a omparable reult for EE, ah, and CF. High variation of the ompoition of field pea were found (Jezierny et al., 2011; Ravindran et al., 2010; Shumaher et al., 2009; Batianelli et al., 1998) with regard to high differene in the eletion of varietie. The preent reult were generally in agreement with thoe outome. Flutuation ould be additionally due to varying environmental influene. Several tudie onfirmed that the variety of field pea ha an influene on it CP ontent a well a on many omponent part like other CN, AA, mineral or trypin inhibitor (Guillamón et al., 2008; Kotlarz et al., 2011; Wang et al., 2008; Canbolat et al., 2007). In the preent tudy, ignifiant differene of the CP and EE ontent were oberved between field pea varietie (Table 4). Caablana had on average the highet CP ontent, whih wa ignifiantly different from Navarro, Natura, Aukland, Repet, Alveta, and Abarth. Proteta had ignifiantly higher CP ontent than Repet, Alveta, and Abarth. In addition, KWS La Manha differed ignifiantly from Aukland. The EE ontent wa highet in Abarth and imilarly high in Mythi, Navarro, Proteta, and Rebel. It wa ignifiantly lower in Alveta, La Manha KWS, and Salamana. Salamana had the lowet EE ontent, whih wa alo ignifiantly lower than in Aukland. The only normal-leafed variety Natura had relatively low ah and CF ontent, whih did not differ ignifiantly from any of the emi leafle varietie, and performed on a medium level in all other parameter. There eem to be an invere relationhip between the CP ontent and the ontent of tarh, Ly, and Met between the varietie (Table 4). Thi might be due to geneti fator and/or to differene in the proe of maturation (Flinn and Pate, 1968; Geervani and Devi, 1988).

5 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) Table 4 Leat quare mean of rude nutrient ontent (g 100 g -1 dry matter) and ontent of the amino aid lyine, methionine, and yteine (g 100 g -1 rude protein) with tandard error (SE) and with letter to mark ignifiant differene (p < 0.05) in variou field pea (Pium ativum L.) varietie Protein Ether Extrat Ah Fiber Starh Sugar Lyine Methionine Cyteine SAA:Lyine Abarth d 2.00 a 3.00 d SE Alveta d 1.91 d 3.04 bd SE Aukland d 1.96 a 3.10 b SE Caablana a 1.93 ad 3.09 abd SE KWS La Manha ab 1.90 d 3.10 b SE Mythi ad 2.00 ab 3.10 ab SE Natura bd 1.91 bd 3d SE Navarro bd 1.99 ab 3.10 b SE Proteta a 1.98 ab 3.19 a SE Rebel ad 1.99 ab 3.07 bd SE Repet d 1.96 ad 3.05 bd SE Salamana ad 1.89 d 3.09 b SE SAA = MethionineCyteine Neither the other CN nor the AA in the CP of field pea differed ignifiantly between the varietie of the data et. Even Ravindran et al. (2010) did not find ignifiant differene in the CN and AA ontent between five pea ultivar ultivated in New Zealand. Canbolat et al. (2007) examined only light but ignifiant differene of the ontent of CP, CA, and CF between white- and purple-flowered pea from two oneutive harvet year in Turkey. Furthermore, the teta olor had an effet on the CN ontent a brown olored pea did ontain higher amount of CP, CA, and CF but lower amount of N-free extrat (arbohydrate). Moreover, Gronle et al. (2015) found a higher CP and CF ontent, but a lower EE, tarh, and ugar ontent a well a a lower energeti value in a olored-flowering winter pea ompared to a Table 5 Etimated ovariane of random fator on rude nutrient, ome amino aid, and rop yield of field pea (Pium ativum L.) Parameter Protein Ether Extrat Ah Fiber Starh Sugar Lyine Methionine Cyteine SAA:Ly Yield ite 1.869* * year year*ite 2.53** 0.007* 0.009* 0.045* * * 0.003* ** year*variety 0.37* 0.293* * * Site*variety Reidual 0.57*** 0.006*** 0.008*** 0.04*** 0.436*** 0.038*** 0.043*** 0.001*** 0.002*** *** *** Cell without a value how parameter removed during model eletion by BIC; * p < 0.05, ** p < 0.01, *** p < 0.001

6 210 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) white-flowering winter pea. The ize of a pea eed an impat the ompoition, partiularly the CF ontent (Batianelli et al., 1998). However, in the preent tudy only yellow, whiteflowering pea were oberved. Some further tudie with variou experimental et-up did not reveal remarkable differene between varietie although ingle varietie howed ignifiantly differing ontent of ome CN (Kotlarz et al., 2011; Wang et al., 2008). Neverthele, the variability of thoe ingredient due to varietal harateriti influene diet formulation and hould therefore be regarded. Environmental fator are known to affet the nutrient ompoition of rop (Wang et al., 2010). For intane, the year an affet the amount of CN (Kotlarz et al., 2011). Borreani et al. (2007) did not find any ignifiant differene in the ompoition of mature rop of onventionally produed field pea between the oberved harvet year and varietie. However, different environmental fator of year and ite an interat and therefore have unpreditable effet on the ompoition of field pea (Nikolopoulou et al., 2007). In the preent tudy, the random fator year*ite had an effet in the eleted model of all analyzed parameter (Table 5). Hene, it an be onluded that there i an interation between the environmental effet, whih lead to differene in the ompoition. However, thee interating effet are a unpreditable a the environmental alteration (like limati ondition or poible alteration of the ite). Additionally, year*variety affeted the ontent of CP, tarh, and the AA. Thi interation wa rather low and ould not be found by other (Borreani et al., 2007; Kotlarz et al., 2011). The rop yield of the field pea wa on average 4.4 ± 1.2 t ha -1 and differed ignifiantly between the examined varietie. Mythi had the highet (4.8 ± 3.7 t ha -1 ) and Rebel the lowet (4.1 ± 3.5 t ha -1 ) rop yield. Due to a high CP ontent, Mythi alo had the highet CP yield per hetare. In onequene of low CP ontent, Abarth howed the lowet CP yield per hetare. Additionally, there were differene between the varietie regarding the yield of AA per hetare. The interation between ite and harvet year influened the yield (Table 5). Hene, there i apparently an effet by the ultivation ite, whih might mainly be due to ite-peifi harateriti of the oil (plu oil fertility management, tillage, time of eeding and harveting, row paing, eed rate, pet and dieae infetation, and further fator) ombined with weather ondition and differene of the ultivation management onneted to the harvet year. The preent reult indiate that individual varietie an have an improved AA profile and/or eential AA yield. For diet formulation, the ompoition of the protein of harveted rop determine part of it quality. High amount of limiting AA an inreae the value of a feedtuff. With regard to the amino aid gap it ould be of inreaed interet to reward thee harateriti intead of the CP yield in rop for ue in monogatri nutrition. Thu, it i of interet to oberve the hange in the AA profile with varying CP ontent in the rop. The ontent of the firt-limiting AA for wine and poultry, namely Ly, Met and Cy, were ignifiantly negatively orrelated with the CP ontent (Figure 2). -1 Lyine [g 100g CP] 10,00 9,00 8,00 7,00 6,00 5,00 4,00 3,00 2,00 1,00 Lyine Methionine Cyteine 0,00 16,00 18,00 20,00 22,00 24,00 26,00-1 Protein [g 100 g DM] r = 0.78** r = 0.55** r = 0.61** 2,50 2,00 1,50 1,00 0,50 0,00 Figure 2 Correlation of lyine and the ulfur-ontaining amino aid (SAA), methionine and yteine, with rude protein in field pea (Pium ativum L.) with the oeffiient of orrelation (r, **= p < 0.01) Wang and Daun (2004) alo found a ignifiant negative orrelation between the ontent of Ly and CP (r = -0.52**) in field pea. However, their analye of 24 ample did not reveal a relationhip between the amount of the SAA and CP. In ontrat, a more or le trong negative orrelation between the CP ontent and the amount of SAA in the CP of field pea i deribed by other author, who ued more than 190 ample (Igbaan et al., 1996; Reihert and MaKenzie, 1982). Shumaher et al. (2011) found Ly and CP orrelated with r = -0.50**, Cy and CP with r = -0.70** a well a SAA with r = -0.67** in 50 field pea ample from different varietie grown organially at two ite in the year Thee reult indiate a negative relationhip between the protein quality, more preiely the AA profile of the CP, and the CP quantity. Further invetigation regarding the whole AA profile are required to gain omprehenive knowledge. The AA in the profile, whih aount for thoe dereaing eential AA, mut be tudied. Moreover, there were orrelation between different CN (Table 6). A trong negative orrelation of CP and tarh wa found with r = -0.79**. Atually, it i tated that tarh aount for variation in the CP ontent of field pea in the firt plae (Batianelli et al., 1998; Holl and Voe, 1980; Reihert and MaKenzie, 1982; Wang et al., 2008). A reaon for thi negative orrelation between tarh and CP ontent an be the higher inreae of tarh in omparion to CP in the maturation proe of the eed (Borreani et al., 2007). Thi hange in the relationhip between the ingredient during maturation ould alo be a poible reaon for deribed orrelation between other nutrient like CP and ugar. Correlation with partly varying trength have been found by different reearher. Reihert and MaKenzie (1982) alo found negative orrelation of tarh, EE, and CF -1 SAA [g 100g CP]

7 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) with CP. Nikolopoulou et al. (2007) publihed higher orrelation between CP and ah and further orrelation between ah and EE, ah and tarh, a well a EE and tarh than within the preent data. They found CP and EE poitively orrelated in field pea. Batianelli et al. (1998) found EE with CP, tarh, and CF negatively orrelated. Moreover, thi invere orrelation between CP and EE wa oberved when drought tre ourred during the maturation of oybean (Dornbo and Mullen, 1992). The orrelation oeffiient eem trongly dependent on the data bai. Hene, orrelation are not uitable for prediting the ompoition of harveted rop due to a high variability of geneti and environmental influene on the maturation proe (Gallardo et al., 2008; Gutierrez et al., 2007; Weber et al., 2005). Table 6 Pearon orrelation oeffiient (n = 113, r > 0.30, p < 0.01) between rude nutrient (g 100 g -1 DM) of field pea (Pium ativum L.) Protein Ether Extrat Ah Fiber Starh Sugar Protein Ether Extrat 1 Ah Fiber Starh 1 Sugar 1 The rop yield of the pea wa poitively orrelated with the CP ontent (r = 0.50*) and the ah ontent (r = 0.38*) a well a negatively orrelated with the tarh ontent (r = -0.44*) and the CF ontent (r = -0.39*). However, it wa oberved to be negatively orrelated with the CP ontent and poitively orrelated with the tarh ontent (Al-Karaki and Ereifej, 1999; Tar an et al., 2004). Thoe negative orrelation between CP and yield are alo deribed for other legume (Baudoin and Maquet, 1999; Dahiya et al., 1977; Erkine et al., 1985; Malik et al., 2006; Malik et al., 2007). Neverthele, Pandey and Gritton et al. (1975) found variable but motly poitive orrelation between CP ontent and the yield of pea. Tar an et al. (2004) oberved the variation of rop yield and CP ontent aounted for by two different trait loi. They onluded that it i poible to breed field pea varietie ombining high rop yield and high CP ontent. Thu, it i poible that orrelation between rop yield and ompoition are influened by the oberved varietie. Another poible explanation i that the rop yield of field pea i highly variable depending on fator of the ultivation ite and harvet year. A rop hortfall due to pet or weather phenomena doe not have to affet the CP ontent equally and an therefore alter the outome of orrelation analye. 3.2 Field bean (Viia faba L.) The analyzed field bean ontained on average 29.6 g CP 100 g -1 DM with 6.4 g Ly 100 g -1 CP, 0.7 g 100 g -1 Met, and 1.1 g 100 g -1 Cy. The relation of the SAA to Ly varied between 0.2 and 0.3 (Table 7), even more widely than in field pea, and wa motly le favorable than the ratio found in field pea. Shumaher et al. (2011) found lower ontent of CP and Ly in organially produed field bean. However, the ontent of Met and Cy did not differ from the reult in Table 7. Although their field bean ample alo originated from different varietie (not all of them atually in ue), ite and harvet year, they reported lower variation of the CP ontent. The data of AMINODat 4.0 derived from analye of 18 onventionally produed field bean ample from Germany (Evonik Indutrie, 2010) with unknown ultivation ondition and varietie, publihed lower CP and Ly ontent a well a higher Cy ontent. The mean ontent of CP in 59 organially produed field bean ample wa higher in the online table of the DLG ( ). The ample ontained le Ly while the average amount of Met and Cy (43 ample) were imilar to the preent finding. Conventionally derived table value (Staudaher and Potthat, 2014) were lower for CP but higher for Ly, Met, and Cy. The table value for EE, tarh, and CF were alo lower, while ugar ontent were higher. Taken a a whole, the oberved ompoition agree with the widely varying literature reult for both onventionally and organially ultivated field bean. Table 7 Range and mean with tandard deviation (SD) of the ontent of rude nutrient (g 100 g -1 dry matter), lyine, methionine and yteine in organially produed field bean (Viia faba L.) (g 100 g -1 rude protein) Mean SD Min Max Dry Matter Protein Lyine Cyteine Methionine MethionineCyteine (SAA) SAA:Lyine Ether Extrat Ah Fiber Starh Sugar Jut a for field pea, the geneti influene of the variety an alo affet the ompoition of field bean (Ghaouti, 2007). The field bean variety had a ignifiant effet on the ontent of all CN and AA (p < 0.05) (Table 8). The varietie low in CP (Alexia, Epreo, Pyramid, and Fuego), howed ignifiantly higher amount of CF, Ly, and Met than the varietie high in CP (Bioro, Divine, Julia, and

8 212 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) Table 8 Leat quare mean of rude nutrient ontent (g 100 g -1 dry matter) and ontent of the amino aid lyine, methionine, and yteine (g 100 g 1 rude protein) with tandard error (SE) and with letter to mark ignifiant differene (p < 0.05) in variou field bean (Viia faba L.) varietie Protein Ether Extrat Ah Fiber Starh Sugar Lyine Methionine Cyteine SAA:Lyine Alexia d 1.70 df 3.77 d 9.78 d a 2.97 f 6.8 a 0.73 ab 1.09 b 0.27 bd SE Bioro ab 1.66 f 3.96 b 9.48 ef bd 3.07 de 6.61 e 0.69 fgh 1.04 def 0.26 de SE Divine ab 1.65 f 3.98 b 9.48 ef d 3.06 de 6.65 de 0.68 h 1.04 e 0.26 e SE Epreo d 1.74 ef 3.84 d ab a 3.15 b 6.76 ad 0.73 ab 1.08 be 0.27 b SE Fabelle a 1.69ef 4.06 ab 9.24 g d 3.06 de 6.64 de 0.69 gh 1.05 ef 0.26 e SE Fanfare ad 1.74 de 3.98 b 9.86 b bd 3.08 bd 6.66 bde 0.71 e 1.07 bf 0.27 b SE Fuego d 1.86 b 4.05 ab a d 3.11 bd 6.64 de 0.71 d 1.07 b 0.27 b SE Iabell a 1.74 de 4.04 a 9.63 de bd 3.07 d 6.61 e 0.70 def 1.06 ef 0.27 b SE Julia ab 1.57 g 3.89 d 9.39 fg b 3.01 ef 6.6 e 0.69 eg 1.04 ef 0.26 de SE Pyramid d 1.77 d 4.20 a a bd 3.14 b 6.78 ab 0.73 b 1.07 bd 0.27 b SE Taifun bd 1.95 a 4.15 ab 9.69 e a 3.11 bd 6.75 ad 0.75 a 1.14 a 0.28 a SE Tangenta d 1.83 b 4.20 a 9.78 d ab 3.27 a 6.77 a 0.72 b 1.07 be 0.26 be SE SAA = MethionineCyteine Fabelle). Even the Cy ontent were on average higher in varietie with low CP ontent. Furthermore, varietie with average ontent of CP, like Taifun or Tangenta, had ignifiantly the highet EE, ah, tarh, and ugar ontent a well a high amount of Ly, Met, and Cy. Jezierny et al. (2011) alo examined the ompoition of Divine, Epreo, and Fuego. They found different amount of ome CN in eah variety. Divine wa harveted in 2005 and had equal CP, lower EE and ah but higher tarh and ugar ontent than the preent finding. Epreo and Fuego were harveted in 2004 and ontained more CP but le tarh and ugar. Even the ontent of ah and EE were lower in Fuego harveted in Thee differene are probably dependent on the ultivation ondition. Du et al. (1999) found no influene of the viin-gene on the ompoition of field bean. However, the tannin-gene eemed to lead to higher CP- and lower CFontent. In the preent ample, the tannin-free variety Tangenta performed on a medium level and the viin-free varietie Divine and Fabelle had high CP ontent a well a low CF ontent. Thee finding upport the uggetion that fator other than varietal effet might be oniderably high. Furthermore, it i poible that twelve year of breeding ontributed to the altered ompoition with regard to thi omparion. Taken a a whole, the reult indiate that the hoie of the field bean variety an mot likely be of benefit for diet formulation for monogatri. Further breeding trategie might be able to ue thoe geneti differene to their advantage. In addition to geneti influene, the environment an affet the ompoition of field bean, too (Römer, 1998). Jut a in field pea, the fator year*ite wa found to affet all ingredient in the individual tatitial model (Table 9). The ombination of the environmental fator eem to be important with regard to the ompoition of rop. Furthermore, variety*ite influened the ontent of CP and tarh. Remarkable i the fat that field bean, whih were harveted in 2011 on Loation 56_6, had lowet average ontent of CP a well a very low Cy-ontent, medium Met-, and the highet Ly-ontent. Hene, the relationhip between Cy:Met a well a SAA:Ly were partiularly low. Even the ontent of CF, ah, ugar, and tarh were high in omparion and the ontent of EE wa low. No aue for thee effet ould be determined for ertain. However, the generative development of thoe field bean wa quikened due

9 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) Table 9 Etimated ovariane of random fator on rude nutrient, ome amino aid, and rop yield of field bean (Viia faba L.) Parameter Protein Ether Extrat Ah Fiber Starh Sugar Lyine Methionine Cyteine SAA:Ly Yield ite year year*ite 1.40* 0.02** 0.15** 0.22* 3.19* 0.01** 0.04* * * * 89.12* variety*ite 0.23* 0.23** 14.95* year*variety Reidual *** 0.03*** 0.04*** 0.22** 0.005*** 0.01*** *** *** *** 12.37*** Cell without a value how parameter removed during model eletion by BIC; * p < 0.05, ** p < 0.01, *** p < to a dry period until May. Neverthele, rop yield of about 5 t ha -1 were reahed after a low maturation. It i poible that drought and heat had detrimental effet on the nodulation and ymbioi in the early tage of development, repetively on N-fixation, and therefore on the CP ontent (Hungria and Varga, 2000) but there were no tudie in the temperate zone. No ignifiant differene were oberved for the rop yield of field bean varietie. It ranged between 3.9 ± 0.6 t ha -1 (Tangenta) and 5.3 ± 0.7 t ha -1 (Fanfare). The random fator year*ite affeted the rop yield. Furthermore, effet of ite*variety and year led to the model with the lowet BIC (Table 9). The CP yield did not differ ignifiantly between varietie. In onluion, the um of environmental influene trongly affeted the yield of the field bean in the data et, independent of the variety. In field bean, Ly and Met howed high negative orrelation with CP. However, the Cy ontent wa not orrelated with the CP ontent of thi peie (Figure 3). -1 Lyine [g 100g CP] 8,00 7,00 6,00 5,00 4,00 3,00 2,00 2,00 1,80 1,60 1,40 1,20 1,00 0,80 0,60 0,40 1,00 0,20 Lyine Methionine Cyteine 0,00 0,00 24,00 26,00 28,00 30,00 32,00 34,00-1 Protein [g 100 g DM] r = -0.76** r = -0.16** r = -0.51** Figure 3 Correlation of lyine and the ulfur-ontaining amino aid (SAA), methionine and yteine, with rude protein in field bean (Viia faba L.) with the oeffiient of orrelation (r, **= p < 0.01) -1 SAA [g 100g CP] A negative orrelation between the CP ontent and the amount of SAA in the CP of field bean ha already been deribed (Gatel, 1994; Monti and Grillo, 1983). Shumaher et al. (2011), for example, found Met and CP orrelated with r = -0.83**, Cy and CP orrelated with r = n a well a SAA and CP orrelated with r = -0.61**. In addition, Ly and CP had a trong negative orrelation. Thu, alo in field bean, a negative relationhip between the quality and the quantity of the protein wa found. Further orrelation were even found in field bean. The ontent of tarh and ugar were, for example, negatively orrelated with the CP ontent (Table 10). Du et al. (1999) found an equal relationhip of tarh and CP within 74 genotype of faba bean (r = -0.67**). The maturation proe might aount for thi orrelation in field bean jut a it doe in field pea (Borreani et al., 2007; Holl and Voe, 1980). EE, ah and CF dereaed with inreaing tarh ontent and were poitively orrelated with eah other. Furthermore, light orrelation were found between the ugar ontent and the ontent of ah and CF (Table 10). Thoe orrelation are not deribed onitently in the literature. Therefore, influene of parameter within the trial, like the varietie, the harvet year with their limati fator and the loation with different oil harateriti, tillage, time of eeding and harveting, row paing, eed rate, and further fator, might be very high. Table 10 Pearon orrelation oeffiient (n = 105, r > 0.30, p < 0.01) between rude nutrient (g 100 g -1 dry matter) of field bean (Viia faba L.) Protein Ether Extrat Ah Fiber Starh Sugar Protein ,47 Ether Extrat 1 0,57 0, Ah 1 0,56-0,43 0,41 Fiber 1-0,37 0,39 Starh 1 Sugar 1

10 214 S. Witten, H. Böhm, K. Aulrih Landbauforh Appl Agri Foretry Re 3/ (65) A orrelation of r = 0.55** between rop yield and tarh ontent ould be oberved in pring faba bean of the preent tudy. Furthermore, the rop yield wa alo lightly negatively orrelated with the amount of SAA (Cy: r = -0.40**, Met: r = -0.36**). Al-Karaki and Ereifej (1999) found the rop yield of field pea to be poitively orrelated with the tarh ontent, whih equal the preent finding in field bean. Neverthele, rop yield are trongly influened by environmental fator and an therefore vary depending on harvet year and ultivation ite. Thee influene an affet the orrelation oeffiient, epeially in an unbalaned dataet. In the preent tudy, a well a in earlier tudie (Shumaher et al., 2011; Wang and Daun, 2004; Wang et al., 2008), both grain legume peie howed variation in their ompoition. Regarding the preent reult a well a the reult of Shumaher et al. (2011), the reation of the amount of Met in the CP eem to be more table and dependent on the CP ontent than the reation of Cy in the eed of field bean. The range of the Met ontent i maller than the range of the Cy ontent. Statitially ignifiant orrelation between Met and CP ontent ould be found repeatedly, while there were no orrelation between Cy and CP ontent in the tudie. Reult of variety trial on ertain ultivation ite in one year indiate that varietie of both oberved peie reat within their geneti poibilitie depending on the environmental fator like oil, limate, and ultivation. The interation of both geneti and environmental fator at a ingle ite in one year i of interet. In multiannual trial, genetially indiated varietal differene an be derived from repetitive obervation under different environmental ondition (ite, weather ondition, ultivation management). However, varietal differene are more obviou when the variane of environmental differene i redued. Therefore, homogenou limati area, and ubequently ultivation area for different rop peie, were identified a a bai to improve reommendation for varietie of rop peie (Graf et al., 2009). Baed on the knowledge that varietie an how differene in their development, rop yield, and even quality parameter depending on the ultivation ytem, there i alo a eparation into organi and onventional variety trial. Every time a rop i ultivated, there are many different expreion of environmental effet a well a variou interation between thoe effet (preipitation, tillage, fertilization ), whih an affet further plant development. Thoe influene annot be overlooked. Therefore, a preie etimation of the influene of environmental fator on variation of rop yield, CP ontent or other parameter i not poible. Neverthele, in the preent tudy it wa poible to how that varietal differene in the nutrient ompoition of field pea and field bean our regardle of environmental effet related to ite and year. However, ine there i an undireted interferene between different environmental ondition and varietal harateriti, the reult of the plant development under different ondition i not yet preditable. Hene, it i reommended to analyze every bath before formulating a diet. Sine a negative orrelation between the CP ontent and the limiting AA, Ly, Met, and Cy (Monti and Grillo, 1983; Partanen et al., 2001; Wang and Daun, 2004) wa onfirmed, the ue of varietie with high CP ontent or a ultivation management that lead to high CP ontent might alo be diadvantageou for diet formulation. Therefore it i aimed to improve the SAA ontent in field bean and field pea. The fertilization with ulfur an have a poitive impat on the quantitie of SAA in the rop (Sherer, 2001). Beide ulfur-fertilization, the ue of breeding trategie to improve both CP ontent and protein quality i another relevant poibility in the prodution of grain legume a feedtuff. Regarding uee in oybean (Imande, 2001), the breeding of Met- or SAAenrihed varietie of native grain legume eem promiing (Shumaher et al., 2009). Thee finding are upported by the identifiation of gene, whih ontrol the amount of CP and SAA in legume eed (Gallardo et al., 2008). 4 Conluion Both geneti and environmental fator an have an influene on the ontent of CN, Ly, Cy, and Met in the eed of field pea and field bean and alo on the rop yield. The poibility to make predition for the nutrient ompoition of thee rop i very limited due to unforeeeable ombinatory effet of variety, ite (ultivation), and limate (year). It i yet not poible to make predition with regard to the influene of thoe ombined environmental fator. The effet of the variety i the mot ertain fator of influene, and i more or le affeted by other parameter. Mot of the urrently ued varietie, whih were part of the tudy, did not differ ignifiantly, but the hoie of the variety an be eential with regard to the amount of quality-determining ontituent. Some orrelation ould be onfirmed by the preent data. In general, the negative orrelation of the Ly-, Met-, and Cy ontent with the CP ontent wa proved. However, the trength of the orrelation wa not equal in omparion with other tudie and eemed to be dependent on the databae and it influening fator. The ultivation of varietie with higher CP ontent might dereae the amount of eential AA in the CP. Thi i not deired for grain legume, whih are ued to feed monogatri animal. Therefore, breeding for higher protein ontent alone i not adequate. An improvement in the protein quality to meet the requirement of monogatri animal mut be the fou of attention rather than protein yield. Sine the diretion of deribed variation i not preditable, it i learly reommended to analyze every bath before diet formulation to math the requirement of monogatri animal. Epeially in organi farming, the ue of thi information ha the potential to improve the feeding ytem. Further reearh i required regarding the protein quality of rop and the influening fator.

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