Polimorfismo genetico delle caseine in razze caprine allevate in Lombardia. E\u2019 stato condotto uno studio della variabilit\ue0 genetica delle caseine in sei razze caprine allevate in Lombardia. La tipizzazione, condotta a livello proteico mediante isoelettrofocalizzazione, \ue8 preliminare ad un approccio molecolare dettagliato su un numero pi\uf9 limitato di soggetti, scelti in base a particolari fenotipi caseinici

A. Caroli

F. Chiatti

G. Pagnacco

P. Bolla

S. Chessa

Polimorfismo genetico delle caseine in razze caprine allevate in Lombardia. E’ stato condotto uno studio della variabilità genetica delle caseine in sei razze caprine allevate in Lombardia. La tipizzazione, condotta a livello proteico mediante isoelettrofocalizzazione, è preliminare ad un approccio molecolare dettagliato su un numero più limitato di soggetti, scelti in base a particolari fenotipi caseinici

AIR Universita degli studi di Milano

Full Terms & Conditions of access and use can be found athttp://www.tandfonline.com/action/journalInformation?journalCode=tjas20Download by: [Politecnico di Milano Bibl] Date: 28 November 2016, At: 02:36Italian Journal of Animal ScienceISSN: (Print) 1828-051X (Online) Journal homepage: http://www.tandfonline.com/loi/tjas20Casein genetic polymorphisms in goat breeds ofLombardyF. Chiatti, S. Chessa, P. Bolla, A. Caroli & G. PagnaccoTo cite this article: F. Chiatti, S. Chessa, P. Bolla, A. Caroli & G. Pagnacco (2005) Casein geneticpolymorphisms in goat breeds of Lombardy, Italian Journal of Animal Science, 4:sup2, 46-48,DOI: 10.4081/ijas.2005.2s.46To link to this article:  http://dx.doi.org/10.4081/ijas.2005.2s.46© 2005 Taylor & Francis Group LLCPublished online: 03 Mar 2016.Submit your article to this journal Article views: 14View related articles 46 ITAL.J.ANIM.SCI. VOL. 4 (SUPPL. 2), 46-48, 2005Casein genetic polymorphisms in goat breeds of LombardyF. Chiatti1, S. Chessa1, P. Bolla1, A. Caroli2, G. Pagnacco11 Dipartimento Scienze e Tecnologie Veterinarie per la Sicurezza Alimentare,Università di Milano, Italy2 Dipartimento Sanità e Benessere degli Animali, Università di Bari, ItalyCorresponding author: Francesca Chiatti. Dipartimento Scienze e Tecnologie Veterinarie per la Sicurezza Alimentare. ViaTrentacoste 2, 20134 Milano, Italy – Tel: +39 02 50315757 – Fax: +39 02 50315746 – Email: francesca.chiatti@unimi.itRIASSUNTO – Polimorfismo genetico delle caseine in razze caprine allevate in Lombardia. E’ stato condot-to uno studio della variabilità genetica delle caseine in sei razze caprine allevate in Lombardia. La tipizzazione,condotta a livello proteico mediante isoelettrofocalizzazione, è preliminare ad un approccio molecolare dettagliatosu un numero più limitato di soggetti, scelti in base a particolari fenotipi caseinici.KEY WORDS: casein, goat, genetics, polymorphisms.INTRODUCTION – In the last years, the genetic polymorphism of goat caseins has raised a considerableresearch interest due to relationships with milk quality and technological properties (Martin et al., 2002). Thefour caseins, αs1-casein (CSN1S1), β-casein (CSN2), αs2-casein (CSN1S2), and k-casein (CSN3), are coded by acluster of genes (Ferretti et al., 1990; Threadgill and Womack, 1990). Due to the tight association occurringamong casein genes, the estimation of the relationship between casein variants and milk traits can beimproved by considering the entire casein haplotype instead of single gene typing, as already shown in cattle(Boettcher et al., 2004). Caprine casein typing, however, requires great analytical effort due to the high poly-morphism shown, as well as to the presence of quantitative variations. Phenotype screening at the proteinlevel provides useful preliminary information on the genetic polymorphisms occurring at the DNA level.The aim of this work was to screen the casein variation of goat breeds reared in Lombardy by the use of theisoelectrofocusing (IEF). The investigation was preliminary to a more accurate typing, performed at the DNAlevel, on a selected sample for breeding of goat lines devoted to specific productions (cheese making or freshmilk with nutritional properties).MATERIAL AND METHODS – Milk samples were collected from the following goat breeds: Saanen (SA),Camosciata (CA), Verzaschese (VZ), Orobica (OR), Frisa (FR), and Bionda dell’Adamello (BA). Sampling wasperformed in different flocks of Lombardy, characterised for the presence of local breeds and for the officialrecording of goats’ productions. Casein phenotyping was carried out by isolectric focusing (IEF) according toCaroli et al. (2001).RESULTS AND CONCLUSIONS – The phenotype frequencies at CSN1S1, CSN1S2, and CSN3 areshown respectively in table 1, 2, and 3. At IEF level, typing of CSN1S1 allows to define three levels of expres-sion of this casein: 0 (including genotypes with null or weak alleles), 2 (including genotypes with two strongalleles), and 1 (other genotypes). A more detailed definition of the two variants carried by each individual needsa complex molecular approach analysing several polymorphisms, as described by Sacchi et al. (2005). However,the preliminary IEF typing provided useful information on the CSN1S1 variation in Lombardy goats. In par-ASPA_imp01_02_03  15-07-2005  9:54  Pagina 46ITAL.J.ANIM.SCI. VOL. 4 (SUPPL. 2), 46-48, 2005 47PROC. 16th NAT. CONGR. ASPA, TORINO, ITALYticular, animals with low CSN1S1 level still represents a high percentage of the population (68%), and aremainly concentrated in Orobica breed (84%). Cosmopolite breeds and Verzaschese show rather high frequencyof animals carrying both strong CSN1S1 alleles (SA: 27%; CA: 22%; VZ: 18%).Table 1. Distribution of animals per breed as a function of CSN1S1 level.CSN1S1 (level) BA SA CA FR VZ OR Total0 10 9 15 124 71 648 8771 5 10 34 103 65 99 3172 1 7 14 8 29 24 86Total 16 26 63 235 165 771 1280As far as CSN1S2 is concerned, IEF allows identifying four variants associated to a normal CSN1S2 content:CSN1S2*A, B, C, and G (table 2). The last allele was identified at the protein level by IEF (Erhardt et al., 2002).Recently, it’s most probably coincidence with CSN1S2*E allele, characterised at the DNA level (Lagonigro etal., 2001), has been widely discussed (Sacchi et al., 2005), and the importance of casein variant identificationtaking into account also the phenotypic expression has been focused. The G variant, except one heterozygousanimal (AG) in CA and OR respectively, occurred in Frisa breed. Frequencies of CSN1S2 variants per breedare shown in table 4. A clear predominance was found for CSN1S2*A, ranging from 0.81 to 0.93. The B vari-ant occurred in all breeds, but with noticeable differences in its incidence, ranging from 0.01 (VZ) to 0.19 (BA).The C variant was found with the highest frequency (0.09) in VZ.Table 2. Distribution of animals per breed as a function of CSN1S2 phenotype.CSN1S2 BA SA CA FR VZ OR TotalAA 10 22 55 157 133 638 1018AB 6 4 2 49 3 85 150AC 4 11 26 28 69 AG 1 11 1 13BB 1 2 11 14BC 3 3 6CC 1 1 2CG 1 1GG 2 2Total 16 26 63 236 163 767 1275Table 3. Distribution of animals per breed as a function of CSN3 IEF phenotype.CSN3 BA SA CA FR VZ OR TotalAA 12 22 54 199 135 441 867AB 3 4 8 35 27 266 343BB 1 58 59Total 16 26 62 234 162 765 1269Recently, extensive polymorphism was shown in goat CSN3 coding gene. A total of 13 polymorphic sites wereidentified in domestic goat (Jann et al., 2004) allowing the identification of 14 DNA alleles corresponding to 11protein variants. At the protein level, only two variants can be identified by IEF, because all the CSN3 vari-ants found in domesticated goat so far cluster in two groups on the basis of the isoelectric point (PrinzenbergASPA_imp01_02_03  15-07-2005  9:54  Pagina 47PROC. 16th NAT. CONGR. ASPA, TORINO, ITALY48 ITAL.J.ANIM.SCI. VOL. 4 (SUPPL. 2), 46-48, 2005et al., 2005). The phenotype distribution shown in table 3 is, therefore, relative to these two IEF variants, whichcould also be named as AIEF and BIEF to avoid confusions with CSN3 alleles detectable at the DNA level, as pro-posed by Prinzenberg et al. (2005). The predominant IEF variant was CSN3*AIEF in all breeds (table 4), withfrequency ranging from 0.75 (OR) to 0.94 (CA). The rather high frequency of CSN3*BIEF in OR (0.25) is remark-able, and could allow to assess relationships with productive traits. An interesting difference has been alreadysuggested by Chianese et al. (2000) between the two CSN3 IEF variants, as BIEF seems to be associated withhigher milk casein content than AIEF. If this result will be confirmed by the future development of the presentinvestigation, local breeds like Orobica, as well as Bionda dell’Adamello, could be valorised exploiting BIEF vari-ant effects.Table 4. Frequencies of CSN1S2 and CSN3 IEF variants in the different breeds.IEF Variant BA SA CA FR VZ ORCSN1S2*A 0.81 0.92 0.93 0.82 0.90 0.91CSN1S2*B 0.19 0.08 0.03 0.12 0.01 0.07CSN1S2*C 0.00 0.00 0.03 0.03 0.09 0.02CSN1S2*G 0.00 0.00 0.01 0.03 0.00 0.00CSN3*AIEF 0.84 0.92 0.94 0.93 0.92 0.75CSN3*BIEF 0.16 0.08 0.06 0.07 0.08 0.25ACKNOWLEDGMENTS – Research supported by CARIPLO foundation. The authors wish to thankARAL (Crema) for the help in collecting milk samples.REFERENCES – Boettcher, P. J., Caroli, A., Chessa, S., Budelli, E., Stella, A., Canavesi, F., Ghiroldi, S.,Pagnacco, G., 2004. Effects of casein haplotypes on production traits in Italian Holstein and Brown Cattle. J.Dairy Sci. 87: 4311–4317. Caroli, A., Jann, O., Budelli, E., Bolla, P., Jäger, S., Erhardt, G., 2001. Genetic poly-morphism of goat _-casein (CSN3) in different breeds and characterization at DNA level. Anim. Genet. 32: 226-230. Chianese, L., Portolano, B., Troncone, E., Pizzolongo, F., Ferranti, P., Addeo, F., Alicata, M.L., Pilla, F.,Calcagna, G. 2000. The quality of Girgentana goat milk. 7th International Conference on Goats, Tours, France:946-949. Erhardt, G., Jäger, S., Budelli, E., Caroli, A., 2002. Genetic polymorphism of goat αS2-casein(CSN1S2) and evidence for a further allele. Milchwissenschaft, 57, 137-140. Ferretti, L., Leone, P., Sgaramella,V., 1990. Long range restriction analysis of the bovine casein genes. Nucleic Acids Res. 18: 6829-6833. Jann,O., Prinzenberg, E.M., Luikart, G., Caroli, A., Erhardt, G. 2004. High polymorphism in the k-casein (CSN3)gene from wild and domestic caprine species revealed by DNA sequencing. J. Dairy Res. 71: 188-195.Lagonigro, R., Pietrola, E., D’Andrea, M., Veltri, C., Pilla, F., 2001. Molecular genetic characterization of thegoat s2-casein E allele. Anim. Genet. 32: 391-393. Martin, P., Szymanowska, M., Zwierzchowski, L., Leroux, C.,2002. The impact of genetic polymorphisms on the protein composition of ruminants milks. Reprod. Nutr. Dev.42: 433-459. Prinzenberg, E.M., Gutscher, K., Chessa, S., Caroli, A., Erhardt, G., 2005. Caprine κ -casein(CSN3) polymorphism: new developments of the molecular knowledge. J. Dairy Sci. 88:1490-1498. Sacchi, P.,Chessa, S., Budelli, E., Bolla, P., Ceriotti, G., Soglia, D., Rasero, R., Cauvin, E., Caroli, A., 2005. Casein haplo-type structure in five Italian goat breeds. J. Dairy Sci. 88:1561-1568. Threadgill, D.W., Womack, J.E., 1990.Genomic analysis of the major bovine milk proteins genes. Nucleic Acids Res. 18: 6935-6942.ASPA_imp01_02_03  15-07-2005  9:54  Pagina 48

Casein genetic polymorphisms in goat breeds of Lombardy

Chessa, S.

Institutional Research Information System University of Turin

46 ITAL.J.ANIM.SCI. VOL. 4 (SUPPL. 2), 46-48, 2005Casein genetic polymorphisms in goat breeds of LombardyF. Chiatti1, S. Chessa1, P. Bolla1, A. Caroli2, G. Pagnacco11 Dipartimento Scienze e Tecnologie Veterinarie per la Sicurezza Alimentare,Università di Milano, Italy2 Dipartimento Sanità e Benessere degli Animali, Università di Bari, ItalyCorresponding author: Francesca Chiatti. Dipartimento Scienze e Tecnologie Veterinarie per la Sicurezza Alimentare. ViaTrentacoste 2, 20134 Milano, Italy – Tel: +39 02 50315757 – Fax: +39 02 50315746 – Email: francesca.chiatti@unimi.itRIASSUNTO – Polimorfismo genetico delle caseine in razze caprine allevate in Lombardia. E’ stato condot-to uno studio della variabilità genetica delle caseine in sei razze caprine allevate in Lombardia. La tipizzazione,condotta a livello proteico mediante isoelettrofocalizzazione, è preliminare ad un approccio molecolare dettagliatosu un numero più limitato di soggetti, scelti in base a particolari fenotipi caseinici.KEY WORDS: casein, goat, genetics, polymorphisms.INTRODUCTION – In the last years, the genetic polymorphism of goat caseins has raised a considerableresearch interest due to relationships with milk quality and technological properties (Martin et al., 2002). Thefour caseins, αs1-casein (CSN1S1), β-casein (CSN2), αs2-casein (CSN1S2), and k-casein (CSN3), are coded by acluster of genes (Ferretti et al., 1990; Threadgill and Womack, 1990). Due to the tight association occurringamong casein genes, the estimation of the relationship between casein variants and milk traits can beimproved by considering the entire casein haplotype instead of single gene typing, as already shown in cattle(Boettcher et al., 2004). Caprine casein typing, however, requires great analytical effort due to the high poly-morphism shown, as well as to the presence of quantitative variations. Phenotype screening at the proteinlevel provides useful preliminary information on the genetic polymorphisms occurring at the DNA level.The aim of this work was to screen the casein variation of goat breeds reared in Lombardy by the use of theisoelectrofocusing (IEF). The investigation was preliminary to a more accurate typing, performed at the DNAlevel, on a selected sample for breeding of goat lines devoted to specific productions (cheese making or freshmilk with nutritional properties).MATERIAL AND METHODS – Milk samples were collected from the following goat breeds: Saanen (SA),Camosciata (CA), Verzaschese (VZ), Orobica (OR), Frisa (FR), and Bionda dell’Adamello (BA). Sampling wasperformed in different flocks of Lombardy, characterised for the presence of local breeds and for the officialrecording of goats’ productions. Casein phenotyping was carried out by isolectric focusing (IEF) according toCaroli et al. (2001).RESULTS AND CONCLUSIONS – The phenotype frequencies at CSN1S1, CSN1S2, and CSN3 areshown respectively in table 1, 2, and 3. At IEF level, typing of CSN1S1 allows to define three levels of expres-sion of this casein: 0 (including genotypes with null or weak alleles), 2 (including genotypes with two strongalleles), and 1 (other genotypes). A more detailed definition of the two variants carried by each individual needsa complex molecular approach analysing several polymorphisms, as described by Sacchi et al. (2005). However,the preliminary IEF typing provided useful information on the CSN1S1 variation in Lombardy goats. In par-ASPA_imp01_02_03  15-07-2005  9:54  Pagina 46ITAL.J.ANIM.SCI. VOL. 4 (SUPPL. 2), 46-48, 2005 47PROC. 16th NAT. CONGR. ASPA, TORINO, ITALYticular, animals with low CSN1S1 level still represents a high percentage of the population (68%), and aremainly concentrated in Orobica breed (84%). Cosmopolite breeds and Verzaschese show rather high frequencyof animals carrying both strong CSN1S1 alleles (SA: 27%; CA: 22%; VZ: 18%).Table 1. Distribution of animals per breed as a function of CSN1S1 level.CSN1S1 (level) BA SA CA FR VZ OR Total0 10 9 15 124 71 648 8771 5 10 34 103 65 99 3172 1 7 14 8 29 24 86Total 16 26 63 235 165 771 1280As far as CSN1S2 is concerned, IEF allows identifying four variants associated to a normal CSN1S2 content:CSN1S2*A, B, C, and G (table 2). The last allele was identified at the protein level by IEF (Erhardt et al., 2002).Recently, it’s most probably coincidence with CSN1S2*E allele, characterised at the DNA level (Lagonigro etal., 2001), has been widely discussed (Sacchi et al., 2005), and the importance of casein variant identificationtaking into account also the phenotypic expression has been focused. The G variant, except one heterozygousanimal (AG) in CA and OR respectively, occurred in Frisa breed. Frequencies of CSN1S2 variants per breedare shown in table 4. A clear predominance was found for CSN1S2*A, ranging from 0.81 to 0.93. The B vari-ant occurred in all breeds, but with noticeable differences in its incidence, ranging from 0.01 (VZ) to 0.19 (BA).The C variant was found with the highest frequency (0.09) in VZ.Table 2. Distribution of animals per breed as a function of CSN1S2 phenotype.CSN1S2 BA SA CA FR VZ OR TotalAA 10 22 55 157 133 638 1018AB 6 4 2 49 3 85 150AC 4 11 26 28 69 AG 1 11 1 13BB 1 2 11 14BC 3 3 6CC 1 1 2CG 1 1GG 2 2Total 16 26 63 236 163 767 1275Table 3. Distribution of animals per breed as a function of CSN3 IEF phenotype.CSN3 BA SA CA FR VZ OR TotalAA 12 22 54 199 135 441 867AB 3 4 8 35 27 266 343BB 1 58 59Total 16 26 62 234 162 765 1269Recently, extensive polymorphism was shown in goat CSN3 coding gene. A total of 13 polymorphic sites wereidentified in domestic goat (Jann et al., 2004) allowing the identification of 14 DNA alleles corresponding to 11protein variants. At the protein level, only two variants can be identified by IEF, because all the CSN3 vari-ants found in domesticated goat so far cluster in two groups on the basis of the isoelectric point (PrinzenbergASPA_imp01_02_03  15-07-2005  9:54  Pagina 47PROC. 16th NAT. CONGR. ASPA, TORINO, ITALY48 ITAL.J.ANIM.SCI. VOL. 4 (SUPPL. 2), 46-48, 2005et al., 2005). The phenotype distribution shown in table 3 is, therefore, relative to these two IEF variants, whichcould also be named as AIEF and BIEF to avoid confusions with CSN3 alleles detectable at the DNA level, as pro-posed by Prinzenberg et al. (2005). The predominant IEF variant was CSN3*AIEF in all breeds (table 4), withfrequency ranging from 0.75 (OR) to 0.94 (CA). The rather high frequency of CSN3*BIEF in OR (0.25) is remark-able, and could allow to assess relationships with productive traits. An interesting difference has been alreadysuggested by Chianese et al. (2000) between the two CSN3 IEF variants, as BIEF seems to be associated withhigher milk casein content than AIEF. If this result will be confirmed by the future development of the presentinvestigation, local breeds like Orobica, as well as Bionda dell’Adamello, could be valorised exploiting BIEF vari-ant effects.Table 4. Frequencies of CSN1S2 and CSN3 IEF variants in the different breeds.IEF Variant BA SA CA FR VZ ORCSN1S2*A 0.81 0.92 0.93 0.82 0.90 0.91CSN1S2*B 0.19 0.08 0.03 0.12 0.01 0.07CSN1S2*C 0.00 0.00 0.03 0.03 0.09 0.02CSN1S2*G 0.00 0.00 0.01 0.03 0.00 0.00CSN3*AIEF 0.84 0.92 0.94 0.93 0.92 0.75CSN3*BIEF 0.16 0.08 0.06 0.07 0.08 0.25ACKNOWLEDGMENTS – Research supported by CARIPLO foundation. The authors wish to thankARAL (Crema) for the help in collecting milk samples.REFERENCES – Boettcher, P. J., Caroli, A., Chessa, S., Budelli, E., Stella, A., Canavesi, F., Ghiroldi, S.,Pagnacco, G., 2004. Effects of casein haplotypes on production traits in Italian Holstein and Brown Cattle. J.Dairy Sci. 87: 4311–4317. Caroli, A., Jann, O., Budelli, E., Bolla, P., Jäger, S., Erhardt, G., 2001. Genetic poly-morphism of goat _-casein (CSN3) in different breeds and characterization at DNA level. Anim. Genet. 32: 226-230. Chianese, L., Portolano, B., Troncone, E., Pizzolongo, F., Ferranti, P., Addeo, F., Alicata, M.L., Pilla, F.,Calcagna, G. 2000. The quality of Girgentana goat milk. 7th International Conference on Goats, Tours, France:946-949. Erhardt, G., Jäger, S., Budelli, E., Caroli, A., 2002. Genetic polymorphism of goat αS2-casein(CSN1S2) and evidence for a further allele. Milchwissenschaft, 57, 137-140. Ferretti, L., Leone, P., Sgaramella,V., 1990. Long range restriction analysis of the bovine casein genes. Nucleic Acids Res. 18: 6829-6833. Jann,O., Prinzenberg, E.M., Luikart, G., Caroli, A., Erhardt, G. 2004. High polymorphism in the k-casein (CSN3)gene from wild and domestic caprine species revealed by DNA sequencing. J. Dairy Res. 71: 188-195.Lagonigro, R., Pietrola, E., D’Andrea, M., Veltri, C., Pilla, F., 2001. Molecular genetic characterization of thegoat s2-casein E allele. Anim. Genet. 32: 391-393. Martin, P., Szymanowska, M., Zwierzchowski, L., Leroux, C.,2002. The impact of genetic polymorphisms on the protein composition of ruminants milks. Reprod. Nutr. Dev.42: 433-459. Prinzenberg, E.M., Gutscher, K., Chessa, S., Caroli, A., Erhardt, G., 2005. Caprine κ -casein(CSN3) polymorphism: new developments of the molecular knowledge. J. Dairy Sci. 88:1490-1498. Sacchi, P.,Chessa, S., Budelli, E., Bolla, P., Ceriotti, G., Soglia, D., Rasero, R., Cauvin, E., Caroli, A., 2005. Casein haplo-type structure in five Italian goat breeds. J. Dairy Sci. 88:1561-1568. Threadgill, D.W., Womack, J.E., 1990.Genomic analysis of the major bovine milk proteins genes. Nucleic Acids Res. 18: 6935-6942.ASPA_imp01_02_03  15-07-2005  9:54  Pagina 48

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Casein genetic polymorphisms in goat breeds of Lombardy

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