Phenotypic diversity among Croatian common bean (Phaseolus vulgaris L.) landraces was assessed by analysing 12 qualitative and six quantitative traits in 338 accessions collected from all production areas in Croatia. Out of 12 qualitative morphological traits, seed coat colour was the most polymorphic among accessions having the highest Shannon’s information index and discriminating power. Out of six quantitative traits, the highest coefficient of variation was detected in duration of flowering. A weak correlation was found between distance matrices among accessions based on qualitative and quantitative traits

Boris Lazarević

Klaudija Carović-Stanko

Martina Grdiša

Monika Vidak

Sara Malešević

Zlatko Šatović

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133Agriculturae Conspectus Scientifi cus . Vol. 80 (2015) No. 3 (133-137)ORIGINAL SCIENTIFIC PAPERSummaryPhenotypic diversity among Croatian common bean (Phaseolus vulgaris L.) landraces was assessed by analysing 12 qualitative and six quantitative traits in 338 accessions collected from all production areas in Croatia. Out of 12 qualitative morphological traits, seed coat colour was the most polymorphic among accessions having the highest Shannon’s information index and discriminating power. Out of six quantitative traits, the highest coeffi  cient of variation was detected in duration of fl owering. A weak correlation was found between distance matrices among accessions based on qualitative and quantitative traits.Key wordscommon bean, landraces, morphological diversity, qualitative traits, quantitative traits, Phaseolus vulgaris L.Phenotypic Diversity among Croatian Common Bean (Phaseolus vulgaris L.) Landraces Monika VIDAK 1 ( )Sara MALEŠEVIĆ 1Martina GRDIŠA 1, 2Zlatko ŠATOVIĆ 1, 2Boris LAZAREVIĆ 3Klaudija CAROVIĆ-STANKO 1, 21 University of Zagreb, Faculty of Agriculture, Department of Seed Science and Technology, Svetošimunska cesta 25, 10000 Zagreb, Croatia e-mail: mvidak@agr.hr2 Centre of Excellence for Biodiversity and Molecular Plant Breeding, Svetošimunska cesta 25, 10000 Zagreb, Croatia3 University of Zagreb, Faculty of Agriculture, Department of Plant Nutrition, Svetošimunska cesta 25, 10000 Zagreb, CroatiaReceived: March 5, 2015 | Accepted: September 16, 2015ACKNOWLEDGEMENTS This work has been fully supported by Croatian Science Foundation under the project UIP-11-2013-3290.Agric. conspec. sci. Vol. 80 (2015) No. 3134 Monika VIDAK, Sara MALEŠEVIĆ, Martina GRDIŠA, Zlatko ŠATOVIĆ, Boris LAZAREVIĆ, Klaudija CAROVIĆ-STANKOIntroductionCommon bean (Phaseolus vulgaris L.) is a widely grown grain legume crop with global agricultural and dietary importance. It demonstrates a global adaptability and phenotypic diversity. It is believed that cultivated common bean originates from the wild species Phaseolus aborigineus Burk. Two independent centres of domestication were identifi ed: Mesoamerican (from northern Mexico to Colombia) and Andean (from southern Peru to north-western Argentina) (Gepts et al., 1986). Following the discovery of Americas, the common bean was transferred to Spain from where it was spread to Europe, Africa and Asia (Lešić et al., 2002).Common bean is a food crop of a high nutritive value, con-sumed mostly as a mature dry grain but also as immature pods. Common bean seed is a valuable source of high quality proteins, carbohydrates, vitamins, minerals, dietary fi bre, phytonutrients (fl avonoids, lignins, phytosterols) and antioxidants (Reynoso-Camacho et al., 2006; Cardador-Martínez et al., 2002). Many of those compounds have important benefi cial eff ects on human health (Duranti, 2006).In Croatia common bean is a traditional grain legume, grown by small farmers using traditional agricultural tech-niques in low input production systems. Croatian traditional landraces are adapted to the specifi c growing conditions and agro-environments showing a great morphological diversity. Th e common bean is primary characterized by a high variability in seed traits such as seed shape, size and colour. Th e aim of this research was to analyse morphological traits of the traditional common bean landraces.Materials and methodsPlant materialTh e plant material consisted of 338 common bean landrac-es collected from all the production areas in Croatia and in-cluded into the Grain legume collection of the University of Zagreb, Faculty of Agriculture, Department of Seed Science and Technology. Th e fi eld trial was set up in Maksimir, Zagreb, in 2014.Morphological analysisMorphological analysis was carried out using the descriptor list for common bean used in Phaseolus Database (http://www.genbank.at/en/ecpgr-phaseolus.html). Th e accessions of common bean landraces were described based on 12 qualitative (QL) and six quantitative (QN) traits: fl ower colour of standard (vexillum) (QL01), fl ower colour of wings (QL02), plant growth habit (QL03), pods position (QL04), seed coat colour (QL05), seed pattern colour (QL06), character of pattern (QL07), seed brilliance (QL08), seed shape - longitudinal (QL09), seed shape - cross (QL10), colour of hilum (QL11), colour of hilum ring (QL12), number of days from sowing to fl owering (QN1), duration of fl owering (QN2), seed length (mm) (QN3), seed width (mm) (QN4), seed height (mm) (QN5), and weight of 100 seeds (g) (QN6).Data analysisQualitative traitsShannon’s information index (H; Shannon, 1949) as a meas-ure of qualitative morphological trait diversity across common bean accessions was calculated for each trait as follows: ∑=−=IiiijppH12logwhere pi is the frequency of the ith state and I is a total number of states of the jth trait (Lewontin, 1972).To compare the effi  ciency of the morphological traits in ac-cession identifi cation, the confusion probability (C) and dis-criminating power (D) of each trait were estimated following the approach developed by Tessier et al. (1999) for molecular marker data. Confusion probability (C) of the jth morphologi-cal trait is thus calculated as follows:( )111−−=∑=NNppCiIiijwhere N is the sample size. Th e confusion probability of a given morphological trait provides an estimate of the probabil-ity that two randomly chosen accessions from the sample of N accessions share identical states for a given morphological trait.Th e number of indistinguishable pairs of accessions is equal to: ( )jjCNNP21−=Th us, discriminating power (Dj) of the jth morphological trait is equal to:( )11111−−−=−= ∑=NNppCDiIiijjdenoting the proportion of distinguishable pairs of accessions.Phenotypic dissimilarity between pairs of bean accessions was calculated using the proportion-of-shared-alleles distance (Bowcock et al., 1994) as implemented in MICROSAT (Minch, 1997). In this case, the results obtained by using the propor-tion-of-shared-alleles distance formula were identical to those obtained by 1 - simple matching coeffi  cient: DSM = 1 - SSM = 1 - (m / n), where m is the number of morphological traits shared between a pair of accessions and n is the total number of traits.Quantitative traitsPearson’s correlation coeffi  cients were calculated between the six quantitative traits using the SAS soft ware PROC CORR procedure (SAS Institute, 2004). Principal Component Analysis (PCA) was applied in order to examine the interrelationships between common bean landraces and their quantitative traits using command PROC PRINCOMP in SAS program. Th e biplot was constructed by two principal PCs showing accessions and quantitative traits (as vectors). Th e standardized scores of the fi rst three principal components (eigenvalue ˃1) were multi-plied by the square root of their eigenvalues, and the pairwise Euclidean distance between accessions based on quantitative traits was calculated using the SAS soft ware PROC DISTANCE. Distance matrices between accessions based on qualitative and Agric. conspec. sci. Vol. 80 (2015) No. 3135Phenotypic Diversity among Croatian Common Bean (Phaseolus vulgaris L.) Landracesquantitative traits were compared by calculating the correlation coeffi  cient and implementation Mantel’s test (Mantel, 1967). Mantel’s nonparametric test was conducted based on 1,000 per-mutations using NTSYS-pc (Rohlf, 2005).Results and discussionQualitative traitsAnalysis of 338 common bean landraces revealed high pheno-typic diversity. Th e number of categories (i), Shannon’s informa-tion index (H) and discriminating power (D) of each qualitative trait are shown in Table 1. All the traits were found polymorphic except QL12 Colour of hilum ring. Th e highest number of catego-ries (i = 19) as well as the highest Shannon’s information index (H = 1.945) and discriminating power (D = 0.815) were detect-ed for QL05 Seed coat colour. A high discriminating power (D > 0.600) was also detected for traits QL06 Seed pattern colour, QL04 Pods position and QL07 Character of pattern. Th ese re-sults coincide with those reported by Piergiovanni et al. (2000), Pereira et al. (2009) and Scarano et al. (2014).Th e average distance between 338 common bean accessions based on 11 qualitative traits was DSM = 0.527, ranging from 0.000 (accessions matching in all traits) to 1.000 (accessions diff ering in all traits). From the total of 56,953 pairs of acces-sions, the number of indistinguishable ones was 520 (0.913%). Th e number of pairs of accessions that diff ered in all the traits was 307 (0.539%). Quantitative traitsTable 2. shows the descriptive statistical parameters of six quantitative traits. Th e highest coeffi  cient of variation was de-tected for the trait QN2 Duration of fl owering (28.89), while the lowest f was detected for or the trait QN1 Number of days from sowing to fl owering (12.36).A few of the six quantitative traits appeared to be highly intercorrelated (Table 3). A strong positive correlation was re-vealed between QN4 Seed width and QN6 Weight of 100 seeds (r = 0.769) and a moderate positive correlation between QN3 Seed length and QN4 Seed width (r = 0.605) as well as between QN3 Seed length and QN6 Weight of 100 seeds (r = 0.672).Principal component analysis (PCA) based on six quanti-tative traits revealed that fi rst three components accounted for 82.16% of the total variance. Th e fi rst and second principal com-ponents had eigenvalues greater than one, while the largest dif-ference was determined between the fi rst and second eigenvalues. Th e correlations between the fi rst three principal components No. Trait Abbr. i H D 1 Flower colour of  standard (vexillum) QL01 5 1.174 0.504 2 Flower colour of wings QL02 5 1.167 0.500 3 Plant growth habit (type) QL03 2 0.996 0.499 4 Pods position QL04 5 1.926 0.706 5 Seed coat colour QL05 19 2.945 0.815 6 Seed pattern colour  QL06 17 2.504 0.730 7 Character of pattern QL07 10 1.960 0.662 8 Seed brilliance QL08 3 0.831 0.300 9 Seed shape - longitudinal QL09 2 0.589 0.244 10 Seed shape - cross QL10 5 0.873 0.295 11 Colour of hilum QL11 9 1.716 0.543 12 Colour of hilum ring QL12 1 –* –* i - number of categories of trait; H - Shannon's information index; D - discriminating power; *- monomorphic trait No. Trait Abbr. Average SD CV Min Max 1 No. of days to flowering QN1 58.81 7.27 12.36 49.00 76.00 2 Duration of flowering QN2 22.64 6.54 28.89 7.00 37.00 3 Seed length (mm) QN3 14.46 2.16 14.95 7.75 20.51 4 Seed width (mm) QN4 8.20 1.05 12.81 5.51 12.96 5 Seed height (mm) QN5 6.19 0.86 13.90 4.26 8.62 6 Weight of 100 seeds (g) QN6 48.51 13.15 27.10 13.18 110.10 SD - standard deviation; CV - coefficient of variation Table 1. Diversity of 12 qualitative morphological traits in 338 Croatian common bean accessionsTable 2. Diversity of six quantitative traits in 338 Croatian common bean accessionsAgric. conspec. sci. Vol. 80 (2015) No. 3136 Monika VIDAK, Sara MALEŠEVIĆ, Martina GRDIŠA, Zlatko ŠATOVIĆ, Boris LAZAREVIĆ, Klaudija CAROVIĆ-STANKOand six quantitative traits are shown in Table 4. Th e biplot was constructed by the fi rst two PCs showing common bean landraces and six quantitative traits (as vectors) (Fig. 1).Th ree out of six traits showed a strong pos-itive correlation with the fi rst principal com-ponent which explained 45.36% of the total variance (QN6 Weight of 100 seeds, QN4 Seed width, and QN3 Seed length). Th e second prin-cipal component accounting for 22.39% of the total variance showed a moderate positive corre-lation with QN2 Duration of fl owering and QN1 No. of days to fl owering and a moderate nega-tive correlation with QN5 Seed height (Table 4).Th e average Euclidean distance between 338 analysed accessions based on six qualita-tive traits ranged from 0.009 to 12.279, with an average value of DEUC = 2.486. Th e correlation coeffi  cient between distance matrices among accessions based on qualitative and quantita-tive traits was r = 0.393 (PMantel < 0.0001), aft er 10.000 permutations of Mantel’s test.No. Trait Abbr. Trait QN1 QN2 QN3 QN4 QN5 QN6 1 No. of days to flowering QN1  *** * *** ns *** 2 Duration of flowering QN2 0.332  *** *** * *** 3 Seed length (mm) QN3 0.139 0.250  *** ns *** 4 Seed width (mm) QN4 0.259 0.243 0.605  *** *** 5 Seed height (mm) QN5 –0.035 –0.111 0.027 0.460  *** 6 Weight of 100 seeds (g) QN6 0.213 0.182 0.672 0.769 0.445  *** P < 0.001; ** 0.001< P < 0.01; * 0.01 < P < 0.05; ns P > 0.05 No. Trait Abbr. Principal component PC1  PC2  PC3  1 No. of days to flowering QN1 0.374 *** 0.589 *** 0.562 *** 2 Duration of flowering QN2 0.378 *** 0.695 *** 0.080 ns 3 Seed length (mm) QN3 0.765 *** 0.127 * -0.543 *** 4 Seed width (mm) QN4 0.906 *** -0.128 * 0.033 ns 5 Seed height (mm) QN5 0.457 *** -0.671 *** 0.489 *** 6 Weight of 100 seeds (g) QN6 0.908 *** -0.174 ** -0.086 ns  Eigenvalue  2.722  1.343  0.864   % Total variance  45.36  22.39  14.41  *** P < 0.001; ** 0.001< P < 0.01; * 0.01 < P < 0.05; ns P > 0.05 Table 3. Correlation coeffi  cients (r; below diagonal) and their signifi cance (P; above the diagonal) between six quantitative traits in 338 Croatian common bean accessionsTable 4. Pearson’s correlation coeffi  cient between six quantitative traits and values of the fi rst three main componentsQN1QN2QN3QN4QN5QN6-2.00-1.000.001.002.00-2.00 -1.00 0.00 1.00 2.00PC1 (45.36%)PC2 (22.39%)PC2 (22.39%) Figure 1. Biplot obtained by principal component analysis on the basis of six quantitative traits in 338 Croatian common bean accessionsAgric. conspec. sci. Vol. 80 (2015) No. 3137Phenotypic Diversity among Croatian Common Bean (Phaseolus vulgaris L.) LandracesConclusionIn Croatia common bean is traditional but neglected grain legume, grown by small farmers using traditional agricultural practices in low input production systems. Croatian landraces are adapted to specifi c growing conditions showing a great mor-phological variability.Phenotypic diversity among Croatian common bean (Phaseolus vulgaris L.) landraces was assessed by analysing 12 qualitative and six quantitative traits in 338 accessions collected from all production areas in Croatia. Out of 12 qualitative morphological traits, the trait QL5 Seed coat colour was the most polymorphic among accessions having the highest number of categories as well as the highest Shannon’s information index and discriminating power, while the trait QL12 Colour of hilum ring was found monomorphic.Out of six quantitative traits, the highest coeffi  cient of vari-ation was detected for the trait QN2 Duration of fl owering. A strong positive correlation was revealed between the trait QN4 Seed width and QN6 Weight of 100 seeds.Principal component analysis based on six quantitative traits revealed that the fi rst two principal components had eighenvalues greater than one, jointly explaining 67.75% of the total variance. Traits related to the fi rst principal component were QN6 Weight of 100 seeds, QN4 Seed width and QN3 Seed length, while QN2 Duration of fl owering, QN1 Number of days to fl owering and QN5 Seed height were related to the second principal component.Th e correlation coeffi  cient between distance matrices among accessions based on qualitative and quantitative traits was highly signifi cant, but weak.ReferencesBowcock A. M., Ruiz-Linares A., Tomfohrde J., Minch E., Kidd J. R. and Cavalli-Sforza L. L. (1994). High resolution of human evo-lutionary trees with polymorphic microsatellites. Nature 368: 455-457Cardador-Martínez A., Loarca-Piña G., Oomah B. D. (2002). 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Phenotypic Diversity among Croatian Common Bean (Phaseolus vulgaris L.) Landraces

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Phenotypic Diversity among Croatian Common Bean (Phaseolus vulgaris L.) Landraces

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