Agronomic characteristics related to grain yield and crude protein content in common vetch (Vicia sativa) accessions of diverse geographic origin

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1 New Zealand Journal of Agricultural Research ISSN: (Print) (Online) Journal homepage: Agronomic characteristics related to grain yield and crude protein content in common vetch (Vicia sativa) accessions of diverse geographic origin A Mikić, V Mihailović, B Ćupina, S Vasiljević, B Milošević, S Katanski, R Matić, V Radojević & M Kraljević-Balalić To cite this article: A Mikić, V Mihailović, B Ćupina, S Vasiljević, B Milošević, S Katanski, R Matić, V Radojević & M Kraljević-Balalić (2013) Agronomic characteristics related to grain yield and crude protein content in common vetch (Vicia sativa) accessions of diverse geographic origin, New Zealand Journal of Agricultural Research, 56:4, , DOI: / To link to this article: Published online: 30 Oct Submit your article to this journal Article views: 226 View related articles Citing articles: 1 View citing articles Full Terms & Conditions of access and use can be found at

2 New Zealand Journal of Agricultural Research, 2013 Vol. 56, No. 4, , RESEARCH ARTICLE Agronomic characteristics related to grain yield and crude protein content in common vetch (Vicia sativa) accessions of diverse geographic origin A Mikić a *, V Mihailović a,bćupina b, S Vasiljević a, B Milošević a, S Katanski a, R Matić c, V Radojević b and M Kraljević-Balalić b a Department of Forage Crops, Institute of Field and Vegetable Crops, Novi Sad, Serbia; b Department of Field and Vegetable Crops, University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia; c South Australian Research and Development Institute, Adelaide, Australia (Received 4 September 2012; accepted 13 September 2013) We analysed the grain yield and quality in common vetch accessions to assess their breeding potential. A small-plot trial was carried out in 2005 and 2006 at the experimental field of the Institute of Field and Vegetable Crops at Rimski Šančevi. 14 accessions, of diverse geographic origin and status, from the Novi Sad Vicia collection were tested. The highest grain yield was in accession VIC 006 (2205 kg ha 1 ), while the highest grain dry matter crude protein was in accession Topaze (329 g kg 1 ). Significant positive correlations were found between the time from sowing to harvest and grain dry matter crude protein content (r gxy = 0.754). A cluster analysis of the tested common vetch accessions showed four main groups that are assumed to correlate with geographic and biological origin. The variability measured offers a basis for developing novel common vetch cultivars that could increase the use of this crop among the farmers in Serbia and the Balkans. Keywords: breeding; common vetch; cluster analysis; crude protein proportion; genetic correlations; grain yield; grain yield components; phenotypic correlations; Vicia sativa Introduction There are more than 160 vetch species with a prevailing Euro-Asiatic distribution (Maxted 1995). From a molecular taxonomic viewpoint, common vetch (Vicia sativa L.) is an aggregate species comprising several polymorphic annual autogamous taxa in a dynamic evolution that either are separate species or one species (V. sativa sensu lato) with subspecies and varieties (Potokina et al. 2000; Shiran & Raina 2001). Common vetch is a self-pollinating species, with 2n = 12chromosomes, namely two pairs of one metacentric, four subacrocentrics and one acrocentric, and is one of the vetch species with the smallest genome (Navrátilová et al. 2003; Kovářová et al. 2007). Common vetch originated in the Near Eastern centre of diversity along with many other annual legume species (Zeven & Zhukovsky 1975) and was a well-known crop to the ancestors of modern Eurasian nations (Mikić 2011; Mikić &Perić 2011). Available archaeological evidence suggests that common vetch initially was a weed that spread from the Mediterranean to Europe during the Neolithic agricultural revolution (van de Wouw et al. 2003b), particularly with Lens culinaris Medik., the cultivated lentil (Erskine et al. 1994), while it was domesticated in Asia Minor (Harlan 1971). Numerous gene banks worldwide deal with the genetic resources of common vetch. The State Scientific Centre N.I. Vavilov All-Russian Research Institute of Plant Industry of the Russian Academy of Agricultural Science (VIR) in St Petersburg, Russia, has about 700 accessions of landraces and local cultivars of common vetch (Potokina et al. *Corresponding author. aleksandar.mikic@nsseme.com; aleksandar.mikich@gmail.com 2013 The Royal Society of New Zealand

3 298 A Mikić et al. 2002), while the International Center for Agricultural Research in Dry Areas (ICARDA) in Aleppo, Syria, uses the common vetch genetic resources in a sustainable way in developing novel cultivars suitable for cultivation in the Eastern Mediterranean (Al-Doss et al. 1996). Common vetch is basically a multi-purpose crop (Mikić et al. 2006) and is used in a similar way to pea (Pisum sativum L.) (Akopian et al. 2010), faba bean (Vicia faba L.) or grass pea (Lathyrus sativus L.). The grain of common vetch is rich in globulins and albumins (Ribeiro et al. 2004) and is used as a valuable source of plant protein for feeding animals (Saki et al. 2008), especially in Australia (Chowdhury et al. 2004). Among the major constraints to the use of common vetch grain as feed are its limited amino acid availability (Farran et al. 2001a) and the presence of the neurotoxin, γ-glutamyl-β-cyanoalanine, which affects the conversion of methionine to cysteine, and has indirect effects on glutathione metabolism (Enneking et al. 1993). Later advances in common vetch breeding resulted in the development of the cultivars with non-shattering pods (Abd El-Moneim 1993b) and decreased neurotoxin content, such as the Australian Morava. This enabled its application in feeding pigs (Collins et al. 2002), broilers and laying hens (Farran et al. 2001b) and lambs (Gül et al. 2005). The wild and agricultural flora in Serbia comprises 35 vetch species (Mihailović et al. 2011). Common vetch in Serbia today is cultivated on about 7500 ha (Mihailović et al. 2005). The only institution in Serbia dealing with breeding common vetch is the Institute of Field and Vegetable Crops in Novi Sad, with 10 registered autumnand spring-sown cultivars to date in Serbia and abroad (Mikić et al. 2011a). It collaborates with the Faculty of Agriculture of the University of Novi Sad, which curates the Novi Sad Vicia collection, with 21 vetch species and about 1500 accessions of diverse geographic origin and status (Mihailović et al. 2010). This collection is enriched each year with newly collected local landraces and wild populations from various regions of Serbia (Mikić et al. 2009). This study aimed to analyse the grain yield components in common vetch accessions in order to assess their breeding potential and suitability for developing novel common vetch lines with improved agronomic characteristics related to grain production and quality. Materials and methods A small-plot trial was carried out in 2005 and 2006 at the experimental field of the Institute of Field and Vegetable Crops at Rimski Šančevi, N and E and 84 m asl, in the vicinity of Novi Sad, Serbia, in rainfed conditions and on an aridic Kastanozem (FAO 1974). The chemical composition of the sowing layer of the slightly carbonated chernozem soil (Table 1) showed a slightly alkaline soil reaction (ph 7.90 in water and 7.41 in KCl), as well as the presence of all essential elements, such as nitrogen, phosphorus, potassium and calcium, and organic matter. It included 14 common vetch accessions from the Novi Sad Vicia collection of diverse geographic origin and status (Table 2). These accessions formed the entire core collection before the recent germplasm collection expanded the current collection to several hundred accessions. The 14 accessions used here were the first to be characterized and evaluated in a detailed way. The trial was set up as a randomized block design with three replicates. Plot size measured 1 5 m, consisting of five rows spaced 20 cm apart. In both years, all the accessions were sown by hand in the first week of March, at a sowing depth of 4 cm and at a sowing density of 150 viable seeds m 2, roughly Table 1 Chemical composition of the sowing layer of an aridic Kastanozem during the growing season of common vetch accessions at Rimski Šančevi in 2005 and ph (H 2 O) ph (KCl) N (%) P 2 O 5 (mg 100 g 1 ) K 2 O (mg 100 g 1 ) CaCO 3 (%) Humus (%)

4 Agronomic characteristics related and crude protein content in common vetch 299 Table 2 Common vetch accessions from the Novi Sad Vicia collection at Rimski Šančevi in 2005 and Accession number Accession name Country of origin Status VIC 002 Beograd Serbia Cultivar VIC 003 Novi Serbia Cultivar Beograd VIC 004 L-10 Serbia Line VIC 005 L-15 Serbia Line VIC 006 VIC 006 Bulgaria Landrace VIC 007 VIC 007 Bulgaria Landrace VIC 008 Skopje Former Yugoslav Cultivar Republic of Macedonia VIC 009 Slavej Former Yugoslav Cultivar Republic of Macedonia VIC 010 Serva-174 Spain Cultivar VIC 011 Armantes Spain Cultivar VIC 012 Topaze France Cultivar VIC 013 Verdor Spain Cultivar VIC 014 Labari Spain Cultivar VIC 015 1/2000 Serbia Cultivar corresponding to kg ha 1. This is the optimum method for the conditions in the Balkans (Aydoğdu & Açikgöz 1995) and differs from the recommended sowing density of 40 plants m 2 (about kg ha 1 ) for southwest Australia (Seymour et al. 2002). Each common vetch accession was harvested at full grain maturity, which varied by accession. For each accession, number of days from sowing to harvest was counted, and 30 plants were taken from the middle part of the third row of each replicate plot directly before the harvest. These plant samples were used to measure the number of pods (plant 1 ), number of grains (plant 1 ) and thousand grains mass (g). Grain yield was measured after the harvest, at a grain moisture content of 14% and recalculated from g plot 1 to kg ha 1. Harvest index was determined as the ratio of grain mass to total above-ground biomass at the moment of harvest. Crude protein content in grain dry matter (g kg 1 ) was determined on the basis of nitrogen (N) content in grain dry matter multiplied by 6.25 (Carouée & Gatel 1995). Data were processed using MSTAT-C software with calculations of mean (x), mean standard error (s x ), standard deviation (δ) and coefficient of variation (CV) determined. Analysis of variance (ANOVA) with a least significant test (LSD) applied at the levels of 0.05 and 0.01 was also performed. The same software was used to calculate the coefficients of genetic (r gxy ) and phenotypic (r pxy ) correlations among the monitored characteristics on the basis of ANOVA and an analysis of covariance with t-test applied. A cluster analysis of the tested common vetch accessions was performed using SYSTAT 10 software. Results Weather and growing season The growing season in 2005 (Table 3) had the same average seasonal temperature as the long-term Table 3 Weather data during the growing season of common vetch accessions at Rimski Šančevi in 2005 and 2006 compared with a long-term average. Month March April May June July Average/sum Year Average monthly temperature ( C) Long-term average Year Precipitation sum (mm) Long-term average

5 300 A Mikić et al. average (15 C), while the growing season during 2006 was, on average, one degree warmer (16 C) than the long-term average. The growing season during 2006 also had a higher average monthly temperature in July (23 C) compared with 21 C in 2005 and the long-term average for July (21 C). The growing seasons in both 2005 and 2006 had 16% 20% more precipitation (360 and 347 mm, respectively) than the long-term ( ) average precipitation of 299 mm. The monthly distribution of precipitation over the two growing seasons varied markedly with the majority of the precipitation occurring in the second half of 2005 growing season and in the beginning of the 2006 growing season. The overall mean time from sowing to harvest, for both years, was 125 days with differences at the levels of both 0.05 and 0.01 for the tested common vetch accessions (Table 4). The longest growing season was in accession 1/2000 (136 days), followed by Beograd, L-10 and Topaze (all 133 days) and the shortest growing seasons were in Serva-174 and Armantes (both 116 days) and Verdor and Labari (both 117 days). Grain yield and components of yield The accessions Labari and VIC 006 had the greatest two-year average number of both pods plant 1 (36.7 and 34.7, respectively) and grains plant 1 (128.4 and 124.9, respectively), while the accessions L-10 and Armantes had the smallest twoyear average number of both pods plant 1 (20.6 and 20.8, respectively) and grains plant 1 (63.9 and 62.4, respectively), with significant differences at the levels of both 0.05 and The twoyear average values for thousand grain mass in the tested common vetch accessions varied from 51 g in Novi Beograd to 76 g in Verdor, with an overall mean of 61 g. There were differences at the levels of both 0.05 and 0.01 in two-year average values of grain yield among the tested common vetch accessions (Table 5). The highest two-year average grain yield was in accession VIC 006 (2205 kg ha 1 ), while the lowest two-year average grain yield was in accession 1/2000 (1084 kg ha 1 ), with significant differences among many of the tested common vetch accessions (Table 5). It may be noted that many of the tested common vetch accessions with high grain yields also had a high harvest index, such as Verdor and Labari (both 0.41) and VIC 006, VIC 007, Skopje and Serva-174 (0.39). Conversely, the lowest harvest index was in 1/ 2000 (0.21) which also had the lowest grain yield. Crude protein content The overall mean crude protein content was 313 g kg 1 and varied significantly among the tested common vetch accessions (Table 5). The two-year average values of grain dry matter crude protein content ranged between 299 g kg 1 in Slavej to 329 g kg 1 in Topaze. Of all the monitored agronomic characteristics related to grain yield and crude protein content, the widest range of coefficient of variation was found in the number of pods plant 1 (Table 4), with 7.17% in the line L-15 and 61.58% in the cultivar Labari. On the other hand, very small values for coefficient of variation were determined for the time period from sowing to harvest, mostly <1%. Correlations Analysis of the genetic correlations among the monitored agronomic characteristics related to grain yield and quality in the tested common vetch accessions revealed the existence of several significant positive coefficients (Table 6), namely between time from sowing to harvest and grain dry matter crude protein content (r gxy = 0.754; P < 0.01), grain yield and harvest index (r gxy = 0.696; P < 0.01), number of grains and grain yield (r gxy = 0.687; P < 0.01) and thousand grains mass and grain yield (r gxy = 0.607; P < 0.01) and between number of pods and number of grains (r gxy = 0.613; P < 0.05) and thousand grains mass and harvest index (r gxy = 0.611; P < 0.05). On the other hand, three significant negative correlations were also found, such as between thousand grains mass and grain dry matter crude protein content (r gxy = 0.716; P < 0.01), time from sowing to harvest and thousand grains mass (r gxy = 0.699;

6 Table 4 Two-year average values of time from sowing to harvest and grain yield components in common vetch accessions at Rimski Šančevi in 2005 and Time from sowing to harvest (days) Number of pods (plant 1 ) Number of grains (plant 1 ) Thousand grains mass (g) Accession x s x δ CV (%) x s x δ CV (%) x s x δ CV (%) x s x δ CV (%) Beograd 133 ± ± ± ± Novi Beograd 131 ± ± ± ± L ± ± ± ± L ± ± ± ± VIC ± ± ± ± VIC ± ± ± ± Skopje 124 ± ± ± ± Slavej 125 ± ± ± ± Serva ± ± ± ± Armantes 116 ± ± ± ± Topaze 133 ± ± ± ± Verdor 117 ± ± ± ± Labari 117 ± ± ± ± / ± ± ± ± Mean LSD LSD Agronomic characteristics related and crude protein content in common vetch 301

7 Table 5 Two-year average values of grain yield, harvest index and grain dry matter crude protein content in common vetch accessions at Rimski Šančevi in 2005 and A Mikić et al. Grain yield (kg ha 1 ) Harvest index Grain dry matter crude protein content (g kg 1 ) Accession x s x δ CV (%) x s x δ CV (%) x s x δ CV (%) Beograd 1347 ± ± ± Novi Beograd 1393 ± ± ± L ± ± ± L ± ± ± VIC ± ± ± VIC ± ± ± Skopje 1944 ± ± ± Slavej 1888 ± ± ± Serva ± ± ± Armantes 1616 ± ± ± Topaze 1984 ± ± ± Verdor 1737 ± ± ± Labari 2015 ± ± ± / ± ± ± Average LSD LSD

8 Agronomic characteristics related and crude protein content in common vetch 303 Table 6 Coefficients of genetic correlations (r gxy ) among the monitored agronomic characteristics related to grain yield and quality in common vetch accessions at Rimski Šančevi in 2005 and Number of pods Number of grains Thousand grains mass Grain yield Harvest index Grain dry matter crude protein content Time from sowing ** 0.549* ** to harvest Number of pods 0.613* Number of grains ** Thousand 0.607** 0.611* 0.716** grains mass Grain yield 0.696** Harvest index *Significant at **Significant at P < 0.01) and time from sowing to harvest and grain yield (r gxy = 0.549; P < 0.05). In contrast, there was only one significant coefficient in the analysis of the phenotypic correlations among the monitored agronomic characteristics related to grain yield and quality in the tested common vetch accessions (Table 7), namely between thousand grains mass and harvest index (r pxy = 0.581; P < 0.05). Cluster analysis Based on the branching relationships, there are four clusters and two quite distinct accessions. One related cluster includes Labari, VIC006, and L15, a second cluster consists of VIC 007 and Topaze, a third very closely related cluster comprises of Verdor, Skopje and Armantes, and a fourth related cluster is made of Novi Beograd, Slavej, 1/2000 and Beograd (Fig. 1). L-10 and Serva-174 are genetically quite distant from the other accessions. Discussion Growing season The growing seasons in both trial years were characterized by favourable weather conditions. The variations in the average monthly temperatures, compared with the long-term average were minimal, providing optimal conditions for the growth and development of the common vetch accessions. Similarly, there was sufficient precipitation during Table 7 Coefficients of phenotypic correlations (r pxy ) among the monitored agronomic characteristics related to grain yield and quality in common vetch accessions at Rimski Šančevi in 2005 and Number of pods Number of grains Thousand grains mass Grain yield Harvest index Grain dry matter crude protein content Time from sowing to harvest Number of pods Number of grains Thousand * grains mass Grain yield Harvest index *Significant at 0.05.

9 304 A Mikić et al. Figure 1 A cluster analysis of the tested common vetch cultivars at Rimski Šančevi in 2005 and 2006 on the basis of the monitored agronomic characteristics related to grain yield and quality. the sensitive developmental stages, such as flowering, with no significant deviation in precipitation from the long-term average. In general, the conditions for comparing the common vetch accessions, in both trial years, were within the defined limits of the desirable precipitation range of between 200 and 500 mm (Papastylianou 1995). Likewise, soil fertility was sufficient, with phosphorus levels adequate for good expression of the economically important characteristics in common vetch such as grain yield, biological yield, pod length, plant height and harvest index (Turk & Tawaha 2001). Number of pods and grains Number of pods is generally considered one of the most important yield components for many grain legumes. With an average value of 27.4 plant 1, the tested common vetch accessions had significantly greater numbers of pods compared with those in field pea, with a range between 5.3 and 11.3 plant 1 (Mihailović & Mikić 2004), and grass pea, varying from 11.1 to 18.3 plant 1 (Mikić et al. 2010), in the trials carried out in the same agroecological conditions. Number of grains plant 1 is another significant grain yield component. In a trial conducted in the same environment with three annual vetch species, common vetch achieved an average of 80 grains plant 1 (Karagić et al. 2011). In the current study, the common vetch accessions had greater grain numbers compared with the results for field pea in Pleven, Bulgaria, that ranged from 17.9 to 69.1 plant 1 (Kosev et al. 2012). The tested common vetch accessions had a similar thousand grains mass as the local landraces of common vetch from the region of Tokat, Turkey, with an average of 59 g (Taser et al. 2005). Larger seeds in common vetch are regarded as having a positive effect on seedling vigour and thus may be one of the goals in common vetch breeding programmes (Acikgoz & Rumbaugh 1979). Grain yield The large variability in the average length of the growing season among the tested cultivars is potentially very significant when selecting breeding lines to develop new common vetch cultivars for the conditions of Serbia and neighbouring regions. All tested accessions are spring-sown, meaning that they finish their growing season in early summer, when the droughts usually begin and which often severely affect later sown crops, such as soybean (Glycine max (L.) Merr.). Although these droughts, in most seasons, do not have a negative impact on common vetch, it is generally desirable that the cultivars intended for grain production are characterised by a relatively short growing season in order to produce high and stable grain yields (Mihailović & Mikić 2010). This also allows for the possibility of cultivating a second short-season crop, such as forage brassicas, during the summer and in the conditions of irrigation. The two-year mean yield of the 14 vetch accessions (1744 kg ha 1 ) was considerably higher than in previous trials with common vetch cultivars carried out in Catalonia, where an average grain yield of 900 kg ha 1 was achieved (Lloveras et al. 2004). However, grain yields in the current study are lower than those of the traditional grain legumes in Serbia, such as field pea, varying between 3348 and 5252 kg ha 1 (Mikić et al. 2011c). This still

10 Agronomic characteristics related and crude protein content in common vetch 305 represents a significant barrier to higher adoption of common vetch by Serbian farmers. Grain yield in common vetch, as in other annual legumes, is heavily susceptible to lodging and grain losses during mechanical harvest, but it could be significantly improved in the intercrops with faba bean or white lupin (Lupinus albus L.), grain legumes with a prominent standing ability (Ćupina et al. 2011). On average, the tested common vetch accessions had a similar mean harvest index to other annual legumes forming abundant aboveground biomass, such as the field pea and grass pea forage crops, where a harvest index of 0.35 has been recorded (Mikić et al. 2011b). The average grain dry matter crude protein content of the common vetch cultivar Morava in arid areas of South Australia was 284 g kg 1 (King & Eason 2001), confirming that the majority of the common vetch accessions tested in the current study had significant potential for grain protein production. As in many other European countries, Serbia has been suffering from a serious lack of quality plant protein for decades. By developing novel common vetch cultivars with high protein yield, the animal husbandry in the country and other similar environments could be provided with a good and reliable complementary feed for both imported and European soybean meal, especially in those seasons heavily affected by summer droughts (Mihailović et al. 2005). Correlations In a study with common vetch cultivars in the Mediterranean coastal region of Turkey, the length of the growing season was also found to correlate negatively with grain yield, unlike thousand grains mass and harvest index which both correlated positively (Çakmakci et al. 2006). Similar results were obtained in the trials with common vetch in Syria, where high grain yields were strongly correlated to high harvest indexes, indicating the need for early harvest accessions (Abd El-Moneim 1993a). In addition, in numerous trials in various regions of Turkey, it has also been found that grain yield positively correlates to both number of pods per plant and number of grains per plant (Çakmakci & Açikgöz 1994; Cakmakci et al. 2003; Fırıncıoğlu et al. 2009). This again highlights the significance of the role that these two yield components can play in developing common vetch cultivars with high and stable grain (Blum & Lehrer 1972). Cluster analysis Due to the fact that there are few breeding programmes on common vetch in Europe the exact pedigree of the tested common vetch cultivars are not known. However, based on the results of the cluster analysis, the French cultivar Topaze could have derived from the Bulgarian landrace VIC 007, L-15 appears to be very closely related or derived from the other Bulgarian landrace VIC 006, and 1/2000 looks to be highly related to Slavej. A possible explanation for the clusters could be that the groups comprise accessions with close geographic origins, especially local landraces or the cultivars they were used to develop (Potokina et al. 2002). This could be the case for the cluster containing Beograd and Novi Beograd. On the other hand, the clusters may be the result of the accessions belonging to similar latitudes and climates (van de Wouw et al. 2003a), as in the case of Labari, Verdor and Serva-174 or Skopje with Slavej and VIC 006 and VIC 007. Conclusions The 2-year study on 14 common vetch accessions of diverse geographic origin showed significant variability in their agronomic characteristics related to grain yield and crude protein. Despite the fact that common vetch is a traditional forage and grain crop in Serbia and other countries of the Balkans, it is cultivated on a small area, facing the same challenges as other grain legumes in Europe. However, we have demonstrated that common vetch represents a stable and reliable source of proteinrich grain in temperate regions such as Southeast Europe, and thus deserves a role in various crop rotations and farming systems. The existing genetic variability among the tested accessions offers a solid basis for selecting to improve yield and

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