Meat Science 97 (2014) 143–150

Contents lists available at ScienceDirect

Meat Science journal homepage: www.elsevier.com/locate/meatsci

Association of calpastatin gene polymorphisms and meat quality traits in pig K. Ropka-Molik a,⁎, A. Bereta b, M. Tyra b, M. Różycki b, K. Piórkowska a, M. Szyndler-Nędza b, T. Szmatoła a a b

Laboratory of Genomics, National Research Institute of Animal Production, Krakowska 1, 32-083 Balice, Poland Department of Animal Genetics and Breeding, National Research Institute of Animal Production, Krakowska 1, 32-083 Balice, Poland

a r t i c l e

i n f o

Article history: Received 30 October 2013 Received in revised form 30 December 2013 Accepted 30 January 2014 Available online 8 February 2014 Keywords: Calpastatin CAST gene Pork quality Meat texture traits Pig

a b s t r a c t Calpastatin is associated with the rate of post mortem degradation of structural proteins due to the regulation of calpain activity. In the present research, the associations between polymorphisms within 6th intron of porcine CAST gene and several meat quality traits were analyzed. The CAST gene polymorphisms affected meat colour, pH, water holding-capacity (WHC) and texture parameters (toughness, firmness, cohesiveness, chewiness, and resilience) measured in longissimus dorsi and semimembranosus muscles. The analysis performed on the most numerous breeds maintained in Poland, suggested that the most interesting polymorphisms were CAST/HpaII and CAST/RsaI, which had the greatest effect on WHC regardless of the breed analyzed and had an effect on meat pH, firmness and toughness for most breeds. Interestingly, for almost all breeds, the significant effect of both mutations on intramuscular fat content (IMF) was detected. The provided data confirmed the use of CAST gene as a genetic marker in breeding programmes which allows performing a selection focussed on improving the quality of pork. © 2014 Elsevier Ltd. All rights reserved.

1. Introduction Tenderness, meat colour and water holding-capacity are considered to be the most important factors of meat quality (Bredahl, Grunert, & Fertin, 1998; Grunert, Bredahl, & Brunsø, 2004). According to the consumers, the most desirable features of pork are its colour, external fatness and tenderness (Fortomaris et al., 2006). On the other hand, for the meat industry the most important are traits like meat and fat content in carcasses and those which determine meat quality after thermal treatment i.e. drip loss, water-holding capacity, pH and texture characteristics (Jaworska et al., 2009). These quality traits are mainly dependent on the muscle tissue structure and influence technological and culinary values of the meat. The activity of proteolytic enzymes – calpains and their inhibitor – calpastatin, corresponds to the meat (beef, lamb, pork) flavour and texture attributes: tenderness, juiciness, cooking loss, and cooked and fresh meat colour. Calpastatin, an endogenous inhibitor specific for the calpains, is also associated with the rate of degradation of structural proteins post mortem by controlling the activity of calpains (Wendt, Thompson, & Goll, 2004). Furthermore, the calpastatin protein is

⁎ Corresponding author at: Krakowska 1, 32-083 Balice, Poland. Tel.: +48 666081208; fax: +48 12 285 67. E-mail address: [email protected] (K. Ropka-Molik).

http://dx.doi.org/10.1016/j.meatsci.2014.01.021 0309-1740/© 2014 Elsevier Ltd. All rights reserved.

assumed to influence the expression levels of genes encoding structural or regulatory proteins. Melody et al. (2004) showed significant differences in calpastatin activity between three porcine muscles which correlate with various levels of desmin degradation. Thus, the changes of calpastatin activity in muscle cells can be partially explained by the differences in proteolysis rate and pork quality (Huff-Lonergan & Lonergan, 2005). To date, numerous research indicated that several QTLs related with pork quality traits were localized on SSC2 within the CAST gene region (Meyers & Beever, 2008; Meyers, Rodriguez-Zas, & Beever, 2007; Rohrer, Thallman, Shackelford, Wheeler, & Koohmaraie, 2006). For the first time, the CAST gene was proposed as a candidate gene associated with pork tenderness by Ernst, Robic, Yerle, Wang, and Rothschild (1998). In the recent years, intensive researches focussed on the identification of the functional CAST polymorphisms were performed. Ciobanu et al. (2004) indicated significant linkage disequilibrium between alleles of calpastatin and reported that both haplotypes and individual polymorphisms affected firmness, Instron force, and juiciness score of pork. The following studies confirmed the significant association of CAST SNPs with tenderness, cooking loss or favourable meat quality score (Lindholm-Perry et al., 2009; Nonneman et al., 2011). Rohrer et al. (2012) reported that CAST was one of the genes with the largest effects on pork quality and could be useful for breeding selection. Due to calpastatin function, the CAST gene is considered to be a candidate gene responsible for meat quality in many domestic animals

144

K. Ropka-Molik et al. / Meat Science 97 (2014) 143–150

including pigs. According to the fact that the significance of CAST markers differed in populations and analyzed mutations (Rohrer et al., 2007), the objective of the present study was to determine the effect of CAST gene polymorphisms on important pork quality traits in five pig breeds maintained in Poland.

positive force input during the first compression while chewiness was obtained by multiplying hardness, cohesiveness and springiness (Meullenet, Lyon, Carpenter, & Lyon, 1998). The data obtained was collected and calculated by Texture Expert, version 1.20 software.

2. Material and methods

2.4. Genotyping

2.1. Animals

The genomic DNA was isolated from whole blood using the Genomic Wizard Purification Kit (Promega) following the instruction provided in the protocol. All animals analyzed were free of C1843T mutation in the RYR1 gene. The three CAST polymorphisms, localized in intron 6, were genotyped by PCR-RFLP method using HpaII, HinfI and RsaI endonucleases according to Ernst et al. (1998).

The study was performed on 554 pigs represented by four purebreds used in Polish breeding programmes (Polish Landrace, Polish Large White, Pietrain and Duroc; 146, 158, 115, 76, respectively) and one conservative breed — Puławska pigs (n = 59). The Puławska pigs are kept locally and are characterized by high meat quality parameters, especially a high level of IMF content. All gilts were maintained in the Pig Test Station (SKURTCh) of the National Research Institute of Animal Production (in Pawłowice, Mełno, Chorzelów) under the same housing and feeding conditions. The animals were fed ad libitum from 30 kg up to 100 (±2.5) kg until they were slaughtered and dissected.

2.5. Statistical analysis The association analyses between meat quality traits and different CAST genotypes were performed using GLM procedure (SASv. 8.02). The most comprehensive model was:

2.2. Meat quality traits Several meat quality traits such as: pH, meat colour, water holdingcapacity and intramuscular fat content (IMF) were evaluated. The pH was measured using pH-Star Mathaus in the longissimus dorsi (at the last rib) and semimembranosus muscles, 45 min (pH45) and 24 h (pH24) after slaughter. Meat colour parameters (L* — lightness, a* — redness and b* — yellowness) were estimated using reflectance spectrophotometer Minolta CR-310 on loin samples 24 h after dissection. Intramuscular fat content was measured (IMF) on thawed longissimus dorsi homogenates by the Soxhlet method using Soxtherm SOX 406 — Gerhardt, whereas water holding-capacity was evaluated according to the Graua–Hamma method (Hamm, 1986). Because the data concerning meat quality traits were not available for all animals, the number of pigs in each group for the genotype association analysis was different. 2.3. Texture analysis The meat texture parameters were determined for longissimus dorsi and semimembranosus muscles of 262 animals belonging to four breeds: Polish Landrace, Polish Large White, Duroc and Puławska pigs (73, 93, 37, and 59; respectively). All muscle samples were collected after dissection and stored at − 20 °C. Texture analysis was performed at room temperature using the Texture Analyser TA-XTplus (Stable Micro Systems, Godalming, UK). The WBS analyses (Warner–Bratzler shear force) were performed for both raw and cooked meat. The muscle slices (3.5 cm wide, approximately 200 g) were placed in a polyethylene bag, cooked in a water bath until a core temperature reached 80 °C and chilled for 24 h at 4 °C. The 3 cores (15 mm diameter) were taken from the muscle slice parallel to the muscle fibre direction and sheared using a WB triangular blade at 4.5 mm/s on a Texture Analyser TA-XTplus. The peak force was recorded in N. The two cores for texture profile (TPA) determination were doubly compressed by a cylinder (SMS P/25, base diameter 50 mm) to 70% of their height at a rate of 2 mm/s with a 3 s break between the storage of compression. Selected texture parameters (texture profile analysis; TPA) such as hardness, cohesiveness, chewiness and resilience for both muscles (raw meat) were measured from force–deformation curves. Hardness was defined as the maximum force applied to the samples during the first compression cycle and expressed in N. Cohesiveness was calculated as the ratio of the area under the second curve to the area under the first curve. Springiness (mm) was determined as a ratio of time of contact with the sample during the second compression to the first compression. Resilience was defined as the ratio of the negative force input to

Yijklm ¼  þ si þ d j þ bk þ gl þ ðb  gÞkl þ eijklm

where, Yijklm μ si dj bk gl (b*g)kl eijklm

ijklm observation, overall average, the effect of pig station, the effect of date of slaughter (for meat quality traits: pH, colour, water holding-capacity), the fixed effect of breed, the fixed effect of different CAST genotypes, the interaction between CAST genotypes and breeds (included in the model when significant), the random error.

Additive and dominant effects were estimated using the REG procedure of SAS, where the additive effect was denoted as 1, 0 and −1 for genotypes AA/CC/EE (0), AB/CD/EF (1) and BB/DD/FF (2), respectively, and the dominance effects represented as 1, − 1 and 1 for AA/CC/EE (0), AB/CD/EF (1) and BB/DD/FF (2), respectively. The Hardy–Weinberg equilibrium for each genotype and breed was estimated by using Court Lab — HW calculator. The linkage disequilibrium between CAST polymorphisms was evaluated using PowerMarker V3.25 software.

3. Results 3.1. Genotype frequencies Concerning three CAST polymorphisms, frequencies of genotypes did not differ significantly within all analyzed breeds. In each breed the most numerous were BB CAST/HinfI, DD CAST/HpaII, and FF CAST/ RsaI genotypes (frequencies about 67%, 58%, 61%, respectively) while less numerous were pigs with opposite genotypes: AA CAST/HinfI, CC CAST/HpaII and EE CAST/RsaI (about 6%, 12%, and 6%, respectively). Only in Polish Large White pigs the highest number of CC homozygotes and the lowest DD genotypes compared to other breeds were observed (28% and 42%, respectively). Analyzed polymorphisms were in linkage disequilibrium, the r2 values of CAST/HinfI and CAST/HpaII, CAST/HinfI and CAST/RsaI, and CAST/HpaII and CAST/RsaI were 0.813, 0.663, and 0.671, respectively. All populations were consistent with the Hardy– Weinberg equilibrium (Table S1).

K. Ropka-Molik et al. / Meat Science 97 (2014) 143–150

145

Table 1 Effect of breed, day of slaughter and CAST polymorphisms on pork quality traits. Meat quality traits

WHC a* L* b* PH45 (LD) PH24 (LD) PH45(SEMI) PH24(SEMI) IMF

N

Mean

357 304 304 304 554 554 119 119 368

34.84 54.23 15.76 3.19 6.31 5.60 6.31 5.61 1.79

Standard deviation

Coefficient of variation

GLM procedure Breed

Day of slaughter

6.67 2.92 1.34 1.67 0.33 0.13 0.26 0.19 0.77

19.14 5.38 8.51 52.44 5.27 2.34 4.24 3.49 43.01

** * * *

*** *** * ***

*

***

*

**

* *

** ***

***

*** *

CAST HpaII

CAST HinfI

*

CAST RasI

*

** *

WHC — water holding-capacity, meat colour: L* — luminosity, a* — redness, b* — yellowness; PH45 and PH24 — pH measured 45 min and 24 h after slaughter in longissimus dorsi (LD) or semimembranosus (SEMI) muscles, IMF — intramuscular fat content. N — the number of animals analyzed in each group; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.

3.2. Basic statistical characteristics for analyzed meat traits

3.3. Association of CAST polymorphisms and meat quality traits

The preliminary statistical analyses for meat quality traits and texture parameters with regard to the effects of breed, day of slaughter and investigated CAST polymorphisms were presented in Tables 1 and 2. The obtained results showed statistically significant effects of breed factor and day of slaughter on majority of the investigated meat quality traits excluding pH in longissimus dorsi and semimembranosus muscles measured 45 min after slaughter. Concerning texture parameters, breed factor influenced only firmness and toughness in m. semimembranosus and in m. longissimus dorsi hardness and chewiness. The GLM procedure showed that all three CAST polymorphisms (HinfI, HpaII and RsaI) had a highly significant (p ≤ 0.001; p ≤ 0.01) effect on pH in semimembranosus muscle measured 24 h after slaughter. Furthermore, the CAST/HpaII and CAST/RsaI affected pH in loin 24 h after slaughter. Both the CAST/HpaII and CAST/HinfI polymorphisms influenced meat colour — redness (p ≤ 0.01), while the CAST/HinfI and CAST/RsaI affected intramuscular fat content (Table 1). For texture parameters only the CAST/HpaII and CAST/RsaI polymorphisms had a significant (p ≤ 0.01) effect on firmness (m. longissimus dorsi) and hardness (m. semimembranosus) measured in raw muscles. The breed factor affected mainly WBS texture parameters (Table 2).

In all investigated breeds a significant association between the CAST/ HpaII and CAST/RsaI mutations and water holding-capacity was observed. The meat of homozygous pigs: DD (CAST/HpaII) and EE (CAST/ RsaI) was characterized by the lowest water holding-capacity (WHC) in each breed, except Pietrain pigs, where the opposite trend was observed (Tables 3, 4). The meat colour was also affected by different CAST polymorphisms. For DD homozygotes (CAST/HpaII) the highest values of redness (a*) and the lowest of luminosity (L*) were observed for all breeds. A similar trend was obtained for CAST/RsaI mutation: the meat from FF pigs was characterized by the lowest red colour intensities and the highest luminosity, however without statistical significance (Tables 3, 4). Furthermore, the analysis performed in total for all breeds showed that the meat of pigs with AB genotype (CAST/HinfI) had the highest level of yellowness (b*) and luminosity (L*) (p ≤ 0.01; p ≤ 0.001) (Table 5). In turn, the significant (p ≤ 0.05; p ≤ 0.01) additive effect of all the three polymorphisms on a* parameter was obtained (Table 6). The CAST genotypes had an effect on meat pH, which was the most obviously observed for pH measured 24 h post mortem. Generally, the significantly lower pH24 in semimembranosus muscle was obtained for

Table 2 Basic statistical characteristics for meat texture parameters; the effect of breed and CAST polymorphisms on pork quality traits. Texture parameters

N

Mean

Standard deviation

Coefficient of variation

m. longissimus dorsi Firmness(R) Toughness(R) Firmness(C) Toughness (C) Hardness Springiness Cohesiveness Chewiness Resilience

260 260 262 262 262 262 262 262 262

25.88 73.78 75.34 186.52 7.54 0.69 0.62 3.41 0.26

7.49 22.02 19.45 58.28 3.39 0.06 0.05 1.72 0.03

28.96 29.85 25.82 31.24 44.96 8.94 9.33 50.54 13.38

m. semimembranosus Firmness(R) Toughness(R) Firmness(C) Toughness (C) Hardness Springiness Cohesiveness Chewiness Resilience

262 262 260 260 260 260 260 260 260

25.82 77.66 83.59 202.11 18.38 0.71 0.63 4.86 0.26

6.31 19.47 20.67 52.03 89.95 0.08 0.05 2.18 0.03

24.44 25.07 24.73 25.74 489.39 12.65 9.17 44.99 13.35

GLM procedure Breed

CAST HpaII

CAST HinfI

CAST RsaI

*** *** * ** *

*

*

*

*

*

* *** *** **

*

N — the number of animals analyzed in each group; R — raw meat, C — cooked meat; TPA parameters were measured only on cooked meat; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.

146

K. Ropka-Molik et al. / Meat Science 97 (2014) 143–150

Table 3 The association of CAST/HpaII genotypes and selected WBS parameters measured in raw longissimus dorsi and semimembranosus muscles. Texture traits

Firmness m. longissimus dorsi Toughness m. longissimus dorsi Firmness m. semimembranosus Toughness m. semimembranosus

Genotypes LMS ± S.E.

PLW (93)

CC CD DD CC CD DD CC CD DD CC CD DD

26.29 28.20 26.60 81.17 81.23 74.70 28.50 25.56 28.08 84.21 80.53 85.59

± ± ± ± ± ± ± ± ± ± ± ±

1.87 1.09 1.63 5.70 3.04 5.34 1.59 0.91 0.98 4.99 3.26 3.16

PL (73)

21.01 26.12 28.44 68.30 77.33 81.76 21.01 26.12 28.44 68.30 77.33 81.76

± ± ± ± ± ± ± ± ± ± ± ±

Puławska (59)

2.78b 1.51ab 1.02a 7.23 4.16 2.84 0.64b 1.61ab 0.95a 4.31 4.83 2.98

19.44 21.67 23.47 51.78 58.32 65.24 23.33 26.55 23.83 71.08 75.86 66.96

± ± ± ± ± ± ± ± ± ± ± ±

0.96b 1.54ab 1.06a 3.11b 3.86ab 3.03a 1.03ab 1.15a 0.88b 1.90ab 2.67a 2.65b

Duroc (37)

27.41 27.27 26.17 67.25 82.13 77.15 27.27 30.33 30.70 83.53 92.67 82.44

± ± ± ± ± ± ± ± ± ± ± ±

1.18 1.01 0.71 5.16 2.07 3.05 3.13 5.51 2.11 8.27 6.54 7.30

Total (262)

22.48 25.33 27.40 67.08 72.29 73.90 25.27 25.87 26.01 75.54 78.34 76.95

± ± ± ± ± ± ± ± ± ± ± ±

1.13B 0.79A 1.10A 3.56 2.22 2.12 0.91 0.65 0.58 2.75 2.06 1.87

Effect Additive

Dominance

1.131 (0.665)

−0.474 (0.502)

2.373 (1.963)

−1.441 (1.482)

−0.161 (0.546)

0.073 (0.417)

−0.882 (1.721)

−0.844 (1.313)

PLW — Polish Large White, PL — Polish Landrace; values (LSM ± S.E.) with different letters show significant differences between genotypes (A, B… = p ≤ 0.01. a, b… = p ≤ 0.05). Effects are means (±SE).

all heterozygotes AB, CD and EF compared to other genotypes. In turn, for these genotypes intermediate values of pH24 in m. longissimus dorsi were observed. For IMF content, ambiguous results were obtained, because of the different distributions of IMF values between CAST genotypes, depending on breed. In Pietrain and Polish Landrace pigs, the DD and BB genotypes were characterized by the lowest level of intramuscular fat, while in Duroc the converse trend was observed. According to CAST/RsaI polymorphism, the opposite direction of CAST mutation effect was showed in PL pigs when compared to Pietrain and Duroc.

3.4. Association of CAST polymorphisms and texture parameters The results obtained showed that CAST gene polymorphisms were associated with WBS parameters: firmness and toughness in both analyzed muscles, but only in raw meat (Tables 7–9). The pigs with CC, AA and FF genotypes had the lowest value of these two parameters in the longissimus dorsi muscle. The parallel trend was observed for most of the analyzed breeds. On the other hand, in semimembranosus muscle, heterozygotes AB and EF were characterized by the lowest firmness

Table 4 The association of CAST/RsaI genotypes and selected WBS parameters measured in raw longissimus dorsi and semimembranosus muscles. Texture traits

Firmness m. longissimus dorsi Toughness m. longissimus dorsi Firmness m. semimembranosus Toughness m. semimembranosus

Genotypes LMS ± S.E

PLW (93)

EE EF FF EE EF FF EE EF FF EE EF FF

26.33 28.74 26.03 75.07 82.75 77.76 28.2 25.12 28.90 86.45 79.73 84.48

± ± ± ± ± ± ± ± ± ± ± ±

PL (73)

1.58 1.16 1.69 5.12 3.31 5.27 0.9a 0.90b 1.56a 3.15 3.29 4.91

28.36 25.94 21.14 82.09 76.95 65.08 28.36 25.94 21.14 82.09 76.95 67.08

± ± ± ± ± ± ± ± ± ± ± ±

Puławska (59)

1.03a 1.47ab 3.40b 2.86 4.01 7.93 0.94a 1.60ab 0.68b 2.94 4.87 4.53

23.18 22.12 19.44 64.40 59.70 51.78 23.26 27.26 23.33 66.11 76.91 71.08

± ± ± ± ± ± ± ± ± ± ± ±

0.95 1.66 0.96 2.86a 4.14ab 3.11b 0.88B 1.06A 1.03B 2.66B 2.47A 1.90AB

Duroc (37)

27.40 26.35 27.87 77.20 68.23 74.20 30.70 22.95 32.19 82.00 76.99 93.98

± ± ± ± ± ± ± ± ± ± ± ±

1.51 1.93 0.83 4.01 15.96 2.34 1.33a 1.87b 1.88a 4.20 9.13 3.72

Total (262)

25.96 25.60 22.20 73.85 73.13 64.87 25.84 26.00 25.50 76.86 78.35 76.29

± ± ± ± ± ± ± ± ± ± ± ±

0.69a 0.82a 1.04b 2.10a 2.32ab 3.29b 0.58 0.64 0.95 1.88 2.04 2.81

Effect Additive

Dominance

−1.293 (0.676)*

−0.763 (0.502)

−3.362 (1.993)

−2.165 (1.481)

0.566 (0.543)

0.268 (0.418)

1.585 (1.715)

−0.268 (1.318)

PLW — Polish Large White, PL — Polish Landrace; values (LSM ± S.E.) with different letters show significant differences between genotypes (A, B… = p ≤ 0.01. a, b… = p ≤ 0.05). Effects are means (±SE).

Table 5 The association of CAST/HinfI genotypes and selected WBS parameters measured in raw longissimus dorsi and semimembranosus muscles. Texture traits

Genotypes LMS ± S.E.

PLW (93)

PL (73)

Puławska (59)

Duroc (37)

Total (262)

Firmness m. longissimus dorsi

AA AB BB AA AB BB AA AB BB AA AB BB

23.45 28.54 27.67 69.34 84.18 76.88 31.28 25.27 28.14 95.34 78.69 84.93

16.61– 26.38 ± 27.50 ± 56.67– 78.86 ± 79.50 ± 16.61– 26.38 ± 27.50 ± 56.67– 78.86 ± 79.50 ±

23.18 22.12 19.44 64.40 59.70 51.78 22.33 27.18 23.44 71.08 76.86 66.48

26.18 27.46 27.96 61.73 80.14 77.45 25.30 24.82 33.53 77.32 86.13 97.58

19.83 25.49 26.24 59.26 73.76 73.89 26.18 26.06 25.79 79.58 78.03 76.31

Toughness m. longissimus dorsi Firmness m. semimembranosus Toughness m. semimembranosus

± ± ± ± ± ± ± ± ± ± ± ±

1.62b 1.17a 1.44ab 4.03b 3.49a 4.48ab 2.59A 0.83B 0.86A 7.72a 3.00b 2.73ab

1.54 0.99 3.97 2.79 1.50 0.87 4.52 2.75

± ± ± ± ± ± ± ± ± ± ± ±

0.96 1.63 0.99 3.11a 4.04ab 2.91b 1.03B 1.11A 0.87B 1.90AB 2.57A 2.61B

± ± ± ± ± ± ± ± ± ± ± ±

1.76 1.13 0.84 9.46 4.11 3.09 4.22 2.01 2.31 9.46 4.37 1.63

± ± ± ± ± ± ± ± ± ± ± ±

0.93b 0.85a 0.64a 2.73b 2.46a 1.91a 1.37 0.61 0.54 3.94 1.96 1.73

Effect Additive

Dominance

2.443 (0.744)⁎⁎

−1.166 (0.524)⁎

7.157 (2.179)⁎⁎

−4.599 (1.536)⁎⁎

−0.481 (0.643)

0.237 (0.4444)

−2.516 (2.022)

0.385 (1.397)

PLW — Polish Large White, PL — Polish Landrace; values (LSM ± S.E.) with different letters show significant differences between genotypes (A, B… = p ≤ 0.01. a, b… = p ≤ 0.05). Effects are means (±SE).

K. Ropka-Molik et al. / Meat Science 97 (2014) 143–150

147

Table 6 The additive and dominance effects obtained for three CAST polymorphisms and analyzed pork quality traits. Traits

IMF (%) L* a* b* PH45(LD) PH24(LD) PH45(SEMI) PH24(SEMI) WHC (%)

Effect (mean ± SE) CAST/HpaII

Effect (mean ± SE) CAST/HinfI

Effect (mean ± SE) CAST/RsaI

Additive

Dominance

Additive

Dominance

Additive

Dominance

−0.057 (0.058) −0.172 (0.247) 0.339 (0.115)** −0.148 (0.130) −0.056 (0.021)** −0.011 (0.007) −0.047 (0.041) −0.023 (0.031) −0.941 (0.580)

−0.036 (0.043) −0.362 (0.192) 0.117 (0.089) −0.108 (0.101) 0.012 (0.015) 0.009 (0.005) 0.023 (0.029) 0.053 (0.022) −0.232 (0.431)

−0.110 (0.063) 0.131 (0.269) 0.337 (0.124)** −0.105 (0.137) −0.061 (0.023)* −0.022 (0.008)** −0.066 (0.051) −0.024 (0.039) −0.281 (0.646)

−0.008 (0.047) −0.373 (0.211) 0.061 (0.098) −0.284 (0.108)** −0.008 (0.016) 0.006 (0.005) 0.046 (0.035) 0.025 (0.026) −0.300 (0.483)

0.005 (0.059) 0.106 (0.284) −0.373 (0.126)** 0.126 (0.146) 0.027 (0.021) 0.002 (0.007) 0.050 (0.039) −0.003 (0.031) 1.503 (0.602)*

−0.046 (0.044) −0.139 (0.207) 0.014 (0.092) 0.010 (0.106) −0.009 (0.015) 0.003 (0.005) 0.026 (0.028) 0.037 (0.022) −0.381 (0.446)

WHC — water holding-capacity, meat colour: L* — luminosity, a* — redness, b* — yellowness; PH45 and PH24 — pH measured 45 min and 24 h after slaughter in longissimus dorsi (LD) or semimembranosus (SEMI) muscles, IMF — intramuscular fat content; *p ≤ 0.05, **p ≤ 0.01.

and toughness traits, and statistically significant traits (p ≤ 0.01) were obtained mainly for Polish Landrace and Polish Large White pigs. Interestingly in the ham muscle of Puławska pigs, the opposite distributions of values depending on the genotype were obtained i.e. the semimembranosus muscle of animals with AB, CD, and EF genotypes had the highest firmness and toughness parameters (p ≤ 0.001; p ≤ 0.01)(Tables 7–9). For CAST/HinfI and CAST/HpaII mutations significant additive and dominance effects on firmness and toughness traits measured in raw longissimus dorsi muscle were observed (Tables 7, 8). Moreover, all three polymorphisms influenced the cohesiveness in semimembranosus muscle. The chewiness and resilience were affected by CAST/RsaI and CAST/HpaII, respectively (Table S2). Because of the lack of effect of breed factor on TPA parameters the results obtained were shown in total for all breeds (Table S2). 4. Discussion Presently, consumers are looking for high quality pork and pork products. Unfortunately, the long-term selection aimed at improving leanness and reducing carcass fatness led to a significant decline of meat

quality — mainly its tenderness, succulence and intramuscular fat content. This resulted in a deterioration of the flavour of pork, which after heat treatment became dry and hard. The increasing demands of consumers have prompted farmers to produce high quality meat, while maintaining the ability of animals to improve the characteristics of fattening and slaughter. One of the most important meat quality traits is tenderness. In pork, the tenderisation processes, which have the fastest rate when compared to those of beef or lamb, are related to the lowest ratio of calpastatin to calpain in the porcine muscle (Ilian et al., 2001; Koohmaraie, 1992). Via controlling of calpain activity, calpastatin plays a key role in post mortem proteolysis. In addition to changes in water holding-capacity and drip loss, calpain/calpastatin system affects a majority of the processes of post mortem tenderisation and conversion of muscle to meat (Huff-Lonergan & Lonergan, 1999; Kemp, Sensky, Bardsley, Buttery, & Parr, 2010). Thus, the CAST gene is considered to be a candidate gene responsible for meat quality in many domestic animals including pigs. The CAST gene is intensively studied in cattle due to the fact that the majority of beef cuts are characterized by both high shear force and high toughness compared to lamb or pork. Polymorphisms in bovine CAST

Table 7 The association between CAST/HinfI polymorphism and several meat quality traits in different pig breeds. Meat quality traits WHC (%)

a*

L*

b*

PH45(LD)

PH24(LD)

PH45(SEMI)

PH24(SEMI)

IMF (%)

AA AB BB AA AB BB AA AB BB AA AB BB AA AB BB AA AB BB AA AB BB AA AB BB AA AB BB

PLW (146)

PL (158)

38.81 34.79 35.95 14.92 14.98 15.03 55.51 53.45 54.49 3.79 3.72 3.50 6.34 6.36 6.22 5.44 5.46 5.48 6.20 6.36 6.19 5.82 5.29 5.81 1.50 1.51 1.53

34.14 36.84 34.45 15.93 16.12 16.45 51.26 55.26 54.35 2.22 2.24 2.14 6.65 6.29 6.27 5.56 5.54 5.56 6.52 6.29 3.32 5.49 5.63 5.63 1.71 1.39 1.30

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.97 1.44 1.29 0.43 0.42 0.37 1.02 1.21 0.93 0.42 0.54 0.41 0.03a 0.01a 0.08b 0.02 0.01 0.01 0.23 0.02 0.18 0.06 0.33 0.14 0.09 0.08 0.05

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.03 1.28 1.03 1.69 0.22 0.10 3.98b 0.34a 0.27a 0.16 0.13 0.08 0.05a 0.05ab 0.01b 0.04 0.01 0.01 0.03 0.04 0.03 0.09 0.06 0.02 0.25A 0.06B 0.02B

Pietrain (115)

Duroc (76)

Total (554)

29.47 ± 5.80 32.92 ± 1.62 33.61 ± 0.86 16.68 ± 0.89ab 15.92 ± 0.33b 16.86 ± 0.21a 54.33 ± 0.37 54.98 ± 1.22 53.26 ± 0.35 3.63 ± 0.14 3.63 ± 0.51 3.21 ± 0.26 – 6.20 ± 0.11 6.30 ± 0.06 – 5.57 ± 0.04 5.55 ± 0.01 – 6.22 ± 0.16 6.28 ± 0.05 – 5.66 ± 0.07 5.60 ± 0.01 – 1.88 ± 0.13 1.72 ± 0.08

32.10 33.70 31.41 15.67 15.82 15.73 53.55 53.41 53.23 3.02 3.95 3.13 6.40 6.46 6.31 5.74 5.62 5.59 6.60 6.38 6.57 5.89 5.55 5.69 2.22 2.36 2.67

34.19 34.48 33.19 15.80 15.81 16.10 54.31 54.68 53.73 3.09 3.60 3.15 6.34 6.33 6.27 5.62 5.60 5.61 6.46 6.32 6.37 5.73 5.58 5.61 1.70 1.76 1.72

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.59 0.94 2.13 0.31 0.23 0.34 0.51 0.46 0.63 0.35 0.36 0.39 0.09 0.08 0.09 0.06a 0.04ab 0.02b 0.21 0.11 0.10 0.08a 0.05b 0.04ab 0.18 0.18 0.30

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.15 0.67 0.57 0.25 0.15 0.10 0.49ab 0.39a 0.21b 0.23B 0.20A 0.11B 0.03 0.02 0.02 0.02 0.01 0.01 0.14 0.04 0.03 0.09 0.04 0.02 0.11 0.07 0.05

PLW — Polish Large White, PL — Polish Landrace, WHC — water holding-capacity, meat colour: L* — luminosity, a* — redness, b* — yellowness; PH45 and PH24 — pH measured 45 min and 24 h after slaughter in longissimus dorsi (LD) or semimembranosus (SEMI) muscles, IMF — intramuscular fat content. Values (LSM ± S.E.) with different letters show significant differences between genotypes (A, B… = p ≤ 0.01. a, b… = p ≤ 0.05).

148

K. Ropka-Molik et al. / Meat Science 97 (2014) 143–150

Table 8 The association between CAST/HpaII polymorphism and several meat quality traits in different pig breeds. Meat quality traits WHC (%)

a*

L*

b*

PH45(LD)

PH24(LD)

PH45(SEMI)

PH24(SEMI)

IMF (%)

CC CD DD CC CD DD CC CD DD CC CD DD CC CD DD CC CD DD CC CD DD CC CD DD CC CD DD

PLW (146)

PL (158)

39.79 33.52 36.27 15.12 14.87 14.95 55.07 53.95 53.70 3.93 3.55 3.46 6.36 6.33 6.21 5.45 5.46 5.48 6.24 6.36 5.93 5.44 5.53 5.59 1.51 1.52 1.53

37.66 36.67 36.67 16.06 16.06 16.58 54.37 55.47 53.94 2.12 2.29 2.11 6.32 6.30 6.27 5.57 5.54 5.57 6.37 6.32 6.30 5.82 5.56 5.64 1.46 1.38 1.30

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.80a 1.16b 1.48 0.42 0.34 0.48 1.11 0.98 1.09 0.52 0.43 0.51 0.03a 0.02ab 0.10b 0.02 0.01 0.01 0.17 0.12 0.35 0.14 0.16 0.25 0.08 0.07 0.06

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.33a 1.06ab 1.23b 0.38ab 0.17b 0.11a 1.08ab 0.38a 0.27b 0.06 0.14 0.09 0.07 0.04 0.02 0.02a 0.01b 0.01a 0.09 0.05 0.03 0.14a 0.03b 0.02b 0.13a 0.04ab 0.02b

Pietrain (115)

Duroc (76)

Total (554)

27.42 ± 3.31b 33.69 ± 1.18a 33.61 ± 1.02 16.73 ± 0.89 16.38 ± 0.32 16.73 ± 0.23 54.25 ± 0.37 54.16 ± 1.02 53.49 ± 0.34 3.68 ± 0.14 3.57 ± 0.43 3.19 ± 0.27 – 6.19 ± 0.08 6.32 ± 0.06 – 5.55 ± 0.02 5.56 ± 0.01 – 6.22 ± 0.16 6.28 ± 0.04 – 5.66 ± 0.07 5.60 ± 0.01 2.23 ± 0.45a 1.92 ± 0.12a 1.64 ± 0.09b

31.78 34.02 28.41 15.72 15.72 15.93 53.32 53.69 52.26 3.03 3.68 3.20 6.42 6.40 6.46 5.74 5.62 5.65 6.60 6.39 6.47 5.90 5.50 5.75 2.13 2.36 2.82

35.32 34.24 32.72 15.87 15.78 16.14 54.71 54.62 53.46 3.22 3.43 3.15 6.35 6.31 6.29 5.61 5.62 5.61 6.44 6.36 6.35 5.73 5.55 5.62 1.71 1.78 1.69

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.40ab 0.80a 2.34 0.33 0.20 0.49 0.45 0.45 0.68 0.36 0.30 0.67 0.08 0.07 0.11 0.05 0.03 0.03 0.21 0.12 0.10 0.08a 0.05b 0.03ab 0.18b 0.16ab 0.31a

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.00a 0.54ab 0.68 0.22ab 0.13b 0.12 0.45a 0.33b 0.21 0.23 0.16 0.13 0.02 0.01 0.02 0.01 0.01 0.01 0.09 0.04 0.03 0.10a 0.03b 0.50ab 0.09 0.06 0.06

PLW — Polish Large White, PL — Polish Landrace, WHC — water holding-capacity, meat colour: L* — luminosity, a* — redness, b* — yellowness; PH45 and PH24 — pH measured 45 min and 24 h after slaughter in longissimus dorsi (LD) or semimembranosus (SEMI) muscles, IMF — intramuscular fat content. Values (LSM ± S.E.) with different letters show significant differences between genotypes (a, b… = p ≤ 0.05).

gene have been extensively studied and are significantly associated with meat tenderness (Corva et al., 2007; Pinto et al., 2010; Schenkel et al., 2006), cooking loss, meat colour: brightness, yellowness, and redness (Juszczuk-Kubiak, Rosochacki, Wicińska, Szreder, & Sakowski, 2004; Li et al., 2013). In sheep, Palmer, Robert, Hickford, and Bickerstaff (1998) and Palmer, Morton, Roberts, Ilian, and Bickerstaffe

(1999) showed that several polymorphic sites in the CAST gene were associated with live weight gain, age-corrected carcass weight and longissimus dorsi shear force. Furthermore, the calpastatin gene interacts with the callipyge gene and affects muscling — paternal polar overdominance accounted for CLPG genotypic effects on calpastatin protein content (Freking et al., 1999). In turn, Hu, Zhang, and Zhu (2011) confirmed

Table 9 The association between CAST/RsaI polymorphism and several meat quality traits in different pig breeds. Meat quality traits WHC (%)

a*

L*

b*

PH45(LD)

PH24(LD)

PH45(SEMI)

PH24(SEMI)

IMF (%)

EE EF FF EE EF FF EE EF FF EE EF FF EE EF FF EE EF FF EE EF FF EE EF FF EE EF FF

PLW (146)

PL (158)

36.27 33.52 39.79 14.95 14.87 15.12 53.70 53.95 55.07 3.46 3.55 3.93 6.21 6.33 6.36 5.48 5.46 5.44 5.93 6.36 6.24 5.59 5.53 5.44 1.53 1.51 1.51

33.79 36.44 37.44 16.53 16.21 15.72 54.03 55.26 54.23 2.12 2.27 2.04 6.27 6.30 6.31 5.56 5.54 5.57 6.31 6.30 6.46 5.64 5.59 5.72 1.29 1.38 1.50

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.48b 1.16b 1.80a 0.48 0.34 0.42 1.09 0.98 1.11 0.51 0.43 0.52 0.09b 0.02a 0.02a 0.01a 0.01ab 0.02b 0.35 0.12 0.17 0.25 0.16 0.14 0.06 0.07 0.08

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

1.26b 1.05ab 1.66a 0.11a 0.18ab 0.40b 0.27b 0.38a 1.46ab 0.09 0.13 0.05 0.02 0.04 0.09 0.08 0.01 0.02 0.03 0.04 0.03 0.02 0.04 0.14 0.02b 0.04ab 0.20a

Pietrain (115)

Duroc (76)

Total (554)

32.07 36.08 36.63 16.61 16.72 16.40 54.06 52.35 55.22 3.45 2.82 3.87 6.30 6.25 6.85 5.58 5.53 5.55 6.28 6.20 6.21 5.68 5.53 5.64 1.76 1.87 1.29

28.46 33.91 31.92 15.91 15.77 15.54 52.35 53.52 53.77 3.29 3.52 3.27 6.45 6.44 6.33 5.62 5.66 5.66 6.47 6.39 6.60 5.75 5.50 5.90 2.84 2.33 2.16

32.00 34.60 36.12 16.10 15.93 15.82 53.66 54.19 54.60 3.23 3.23 3.33 6.28 6.32 6.31 5.62 5.60 5.60 6.34 6.35 6.45 5.63 5.56 5.67 1.74 1.75 1.67

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

0.91B 1.25A 1.78AB 0.23 0.30 0.58 0.58 1.09 0.20 0.31 0.39 0.19 0.03a 0.10a 0.20b 0.01 0.02 0.04 0.06 0.14 0.14 0.03a 0.04b 0.0ab 0.09a 0.14a 0.13b

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

2.32b 0.90a 1.59ab 0.58 0.19 0.47 0.71 0.40 0.61 0.78 0.27 0.54 0.11 0.07 0.08 0.03 0.05 0.03 0.10 0.06 0.21 0.01a 0.05b 0.08a 0.37a 0.15b 0.25b

± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

0.66B 0.56A 1.06A 0.12 0.13 0.26 0.26 0.31 0.56 0.15 0.14 0.29 0.02 0.02 0.02 0.01 0.01 0.01 0.03 0.04 0.08 0.02 0.03 0.08 0.06 0.06 0.09

PLW — Polish Large White, PL — Polish Landrace, WHC — water holding-capacity, meat colour: L* — luminosity, a* — redness, b* — yellowness; PH45 and PH24 — pH measured 45 min and 24 h after slaughter in longissimus dorsi (LD) or semimembranosus (SEMI) muscles, IMF — intramuscular fat content. Values (LSM ± S.E.) with different letters show significant differences between genotypes (A, B… = p ≤ 0.01. a, b… = p ≤ 0.05).

K. Ropka-Molik et al. / Meat Science 97 (2014) 143–150

that SNPs within chicken CAST gene influenced some carcass traits such as carcass weight, weight of breast and leg muscles. In pigs, during the last few years an intensive research focussed on the identification of causative mutations associated with pork quality was performed. The numerous authors showed that different CAST polymorphisms affected pork tenderness, firmness, juiciness, meat colour and pH (Ciobanu et al., 2004; Gandolfi et al., 2011; Lindholm-Perry et al., 2009; Meyers & Beever, 2008). Furthermore, CAST markers have been validated in the U.S. commercial swine populations (Nonneman et al., 2011; Rohrer, Nonneman, Miller, Zerby, & Moeller, 2012). Nonneman et al. (2011) identified and analyzed four SNPs which were consistently related with tenderness in several pig populations. To select the most functional mutation, the EMSA method (electrophoretic mobility shift assays) was performed and authors selected four of sixteen polymorphic sites, which may independently affect the expression of calpastatin. The proposed SNPs within the CAST regulatory region were more significantly associated with pork tenderness than three missense mutations investigated by Ciobanu et al. (2004) and could be useful in marker assisted selection. Rohrer et al. (2012) confirmed the significant impact of CAST polymorphisms described previously by Ciobanu et al. (2004) and Lindholm-Perry et al. (2009) and proposed calpastatin gene as a gene with the largest effects on pork quality traits. In this study, significant association between different CAST polymorphisms and meat colour, water holding-capacity and pH was observed. The meat of pigs with AB (CAST/HinfI) and CD (CAST/HpaII) genotypes had the higher intensiveness of yellowness (b*) and luminosity (L*) and lower redness (a*) compared to that with BB and DD homozygotes. On the other hand, the meat of EE homozygotes (CAST/RsaI) were characterized by the lowest values of L*, the highest of a* parameters and the lowest water holding-capacity. The results obtained were consistent with Van Laack et al. (1994) and Joo, Kauffman, Kim, and Park's (1999) findings, who indicated that meat colour is strongly correlated with denaturation of myofibrillar proteins as well as with water holding-capacity and meat pH. In Polish pigs, Rybarczyk, Kmieć, Terman, and Szaruga (2012) confirmed that CAST/RsaI mutation affected drip loss and water holdingcapacity. According to the authors, the meat of EE homozygotes had higher drip loss when compared to that of EF pigs, which was in accordance with our findings where meat of EE homozygotes was characterized by significantly lowest water holding-capacity (p ≤ 0.01). The authors presented that pigs with EE genotypes had higher yellowness (b*) and lower pH24 in longissimus dorsi compared to EF genotypes, while in our study the significant association between CAST/RsaI polymorphism and these traits was not observed. The obtained discrepancies could be due to the fact that in the present research different pig breeds were analyzed and FF genotype was not detected. Contrary to our results Gandolfi et al. (2011), who investigated CAST EU137105:g76.872G > A polymorphisms within the 6th intron, did not remark correlation between analyzed genotypes and meat colour (L*, a*, b*). The different CAST genotypes influenced autolyzed μ-calpain activity 24 and 72 h post mortem, as well as drip loss. The higher level of calpain activation corresponded to the lower rate of pH decline from 1 to 72 h post mortem and lower drip loss. In our results, only concerning CAST/HpaII polymorphisms, pigs with CC genotypes had meat (loin muscle) that maintained high water holding-capacity and high value of pH45 and pH24, which was observed for most analyzed breeds. This is consistent with the fact that the increase of pH in muscles is related with the increase of the ability of the post mortem muscles to retain water (WHC) and decrease drip loss. These changes are strongly associated with the proteolysis rate via modification of calpain and calpastatin activities (Huff-Lonergan & Lonergan, 2005). Consequently, the raw longissimus dorsi muscle of CC pigs (CAST/HpaII) was characterized by the lowest values of firmness and toughness (p ≤ 0.05) parameters observed for the analysis performed on all breeds (p ≤ 0.05). The CAST/HinfI and CAST/RsaI polymorphisms also influenced the WBS

149

parameters particularly evident in loin muscle: the lowest were obtained for AA and FF homozygotes (p ≤ 0.05). On the other hand, in semimembranosus muscle, the CC, AA and FF homozygotes (in PL and Duroc pigs) had the meat, which is characterized by the higher toughness and firmness parameters. Interestingly, for Puławska breed — the highest values of analyzed WBS traits were observed for heterozygotes (AB, CD, EF; p ≤ 0.01, p ≤ 0.05, p ≤ 0.01; respectively), while the frequency of each genotypes did not differ compared to that of other analyzed breeds. The lack of differences in CAST genotype distribution between pig breeds could be due to the fact that meat quality traits have not been taken into account during the selection process, so far. The different directions of impact of genotypes on meat texture parameters in analyzed breeds may be a result of variation in the meat quality observed among breeds. The native Puławska pigs, which are not under selection, are characterized by very good meat quality traits, especially by high intramuscular fat content. On the other hand, results obtained by different authors indicated that some markers within the CAST locus were more significantly associated with pork quality than others and incomplete linkage disequilibrium of analyzed markers with the functional SNP or multiple QTN (quantitative trait nucleotide) in CAST could give conflicting results across populations (Rohrer et al., 2007, 2012). Lindholm-Perry et al. (2009) indicated a significant relationship between 8 SNPs in the CAST gene and share force (SSF) measured at days 7 and 14 post mortem. The authors proposed 41658_290i polymorphism in intron 19 as a marker with the largest effect on SSF in longissimus dorsi. Similar to our results, polymorphisms in calpastatin gene display additive effect on shear force. Moreover, Lindholm-Perry et al. (2009) confirmed the high linkage disequilibrium of CAST SNP and detected the two haplotype blocks with strong LD. However, according to the authors, the haplotype association analysis was less informative when compared to the single marker association. In turn, in four commercial lines of pigs, Ciobanu et al. (2004) indicated that both haplotypes and individual polymorphisms affected firmness (p < 0.01), Instron force (p < 0.01), and juiciness score (p < 0.05), and for chewiness the trend was obtained. The effect on meat tenderness was not significant. In the present study in semimembranosus muscle, all the three investigated polymorphisms influenced cohesiveness, the CAST/RsaI affected chewiness and CAST/HinfI — resilience. In beef muscles, Monteiro (2012) indicated that tenderness correlated negatively with chewiness and also showed that a trend for a negative correlation with resilience was obtained. In the present research, a similar observation was made for CAST/HpaII and CAST/RsaI polymorphisms, because the meat of pigs with CC and FF genotypes was characterized by the lowest toughness of ham muscle, as well as the significantly lower values of resilience (CC genotype) and chewiness (FF genotype) (p ≤ 0.05). In Polish breeds, similar observations were made for longissimus lumborum muscle by Kapelański et al. (2004), who presented a significant effect of CAST/HpaII and CAST/RsaI polymorphisms on firmness and springiness. 5. Conclusion It seems important to find a genetic marker associated with meat quality, and pork tenderness, especially, is a highly heritable trait (h2 = 0.45) (Lo, McLaren, McKeith, Fernando, & Novakofski, 1992). In the presented research, numerous meat quality features were taken into account and this comprehensive analysis confirmed that polymorphisms within the 6th intron of the CAST gene were associated with holding-capacity and texture parameters (toughness, firmness, cohesiveness, chewiness and resilience). The analysis performed on the most numerous breeds maintained in Poland, suggested that the most interesting polymorphisms were CAST/HpaII and CAST/RsaI which had the greatest effect on WHC despite the analyzed pig breed and an effect on meat pH, firmness and toughness for most breeds. Interestingly, for almost all breeds, a significant effect of both mutations on intramuscular fat content (IMF) was detected. The data provided indicates that the use

150

K. Ropka-Molik et al. / Meat Science 97 (2014) 143–150

of CAST gene as a genetic marker in breeding programmes could allow performing the selection focussed on improving the quality of pork. On the other hand, further research should be conducted to show whether such selection can be performed while maintaining a satisfactory level of leanness. Acknowledgements The study was supported by the Polish Ministry of Science and Higher Education (project no NN311 349139). Katarzyna Piórkowska is a recipient of the fellowship of the Foundation for Polish Science. References Bredahl, L., Grunert, K. G., & Fertin, C. (1998). Relating consumer perceptions of pork quality to physical product characteristics. Food Quality and Preference, 9, 273–281. Ciobanu, D. C., Bastiaansen, J. W. M., Lonergan, S. M., Thomsen, H., Dekkers, J. C. M., Plastow, G. S., & Rothschild, M. F. (2004). New alleles in calpastatin gene are associated with meat quality traits in pigs. Journal of Animal Science, 82, 2829–2839. Corva, P., Soria, L., Schor, A., Villarreal, E., Pérez Cenci, M., Motter, M., Mezzadra, C., Melucci, L., Miquel, C., Paván, E., Depetris, G., Santini, F., & Naón, J. G. (2007). Association of CAPN1 and CAST gene polymorphisms with meat tenderness in Bos taurus beef cattle from Argentina. Genetics and Molecular Biology, 30(4), 1064–1069. Ernst, C. W., Robic, A., Yerle, M., Wang, L., & Rothschild, M. F. (1998). Mapping of calpastatin and three microsatellites to porcine chromosome 2q2.1 ± q2.4. Animal Genetics, 29, 212–215. Fortomaris, P., Arsenos, G., Georgiadis, M., Banos, G., Stamataris, C., & Zygoyiannis, D. (2006). Effect of meat appearance on consumer preferences for pork chops in Greece and Cyprus. Meat Science, 72, 688–696. Freking, B. A., Keele, J. W., Shackelford, S. D., Wheeler, T. L., Koohmaraie, M., Nielsen, M. K., & Leymaster, K. A. (1999). Evaluation of the Ovine Callipyge locus: III. Genotypic effects on meat quality traits. Journal of Animal Science, 77, 2336–2344. Gandolfi, G., Pomponio, L., Ertbjerg, P., Karlsson, A. H., Nanni Costa, L., Lametsch, R., Russo, V., & Davoli, R. (2011). Investigation on CAST, CAPN1 and CAPN3 porcine gene polymorphisms and expression in relation to post-mortem calpain activity in muscle and meat quality. Meat Science, 88(4), 694–700. Grunert, K. G., Bredahl, L., & Brunsø, K. (2004). Consumer perception of meat quality and implications for product development in the meat sector: A review. Meat Science, 66, 259–272. Hamm, R. (1986). Functional properties of the myofibrilar system and their measurement. In P. J. Bechtel (Ed.), Muscle as food. London: Academic Press Inc. Hu, Y. D., Zhang, Z. R., & Zhu, Q. (2011). Identification and association of the single nucleotide polymorphisms in calpastatin (CAST) gene with carcass traits in chicken. Journal of Animal and Veterinary Advances, 22, 2968–2974. Huff-Lonergan, E., & Lonergan, S. M. (1999). Postmortem mechanisms of meat tenderization: The roles of the structural proteins and the calpain system. In Y. L. Xiong, C. -T. Ho, & F. Shahidi (Eds.), Quality attributes of muscle foods (pp. 229–251). New York: Kluwer Academic/Plenum Publishers. Huff-Lonergan, E., & Lonergan, S. M. (2005). Mechanisms of water-holding capacity of meat: The role of postmortem biochemical and structural changes. Meat Science, 71, 194–204. Ilian, M. A., Morton, J. D., Kent, M. P., Le Couteur, C. E., Hickford, J., Cowley, R., & Bickerstaffe, R. (2001). Intermuscular variation in tenderness: Association with the ubiquitous and muscle-specific calpains. Journal of Animal Science, 79, 122–132. Jaworska, D., Przybylski, W., Kajak-Siemaszko, K., Olczak, E., Skierka, E., & Niemyski, S. (2009). Evaluation of technological value and sensory quality of pork meat, originated from two paternal PenArLan lines. Roczniki Naukowe Polskiego Towarzystwa Zootechnicznego, 3, 105–113. Joo, S. T., Kauffman, R. G., Kim, B. C., & Park, G. B. (1999). The relationship of sarcoplasmic and myofibrillar protein solubility to colour and water-holding capacity in porcine longissimus muscle. Meat Science, 52(3), 291–297.

Juszczuk-Kubiak, E., Rosochacki, S. J., Wicińska, K., Szreder, T., & Sakowski, T. (2004). A novel RFLP/AluI polymorphism of the bovine calpastatin (CAST) gene and its association with selected traits of beef. Animal Science Papers and Reports, 22(2), 195–204. Kapelański, W., Grajewska, S., Kurył, J., Bocian, M., Jankowiak, H., & Wiśniewska, J. (2004). Calpastatin (CAST) gene polymorphism and selected meat quality traits in pig. Animal Science Papers and Reports, 4, 435–441. Kemp, C. M., Sensky, P. L., Bardsley, R. G., Buttery, P. J., & Parr, T. (2010). Tenderness—An enzymatic view. Meat Science, 84(22), 248–256. Koohmaraie, M. (1992). Effect of pH, temperature, and inhibitors on autolysis and catalytic activity of bovine skeletal muscle calpain. Journal of Animal Science, 70, 3071–3080. Li, Y. X., Jin, H. G., Yan, C. G., Seo, K. S., Zhang, L. C., Ren, C. Y., & Jin, X. (2013). Association of CAST gene polymorphisms with carcass and meat quality traits in Yanbian cattle of China. Molecular Biology Reports, 40, 1875–1881. Lindholm-Perry, A. K., Rohrer, G. A., Holl, J. W., Shackelford, S. D., Wheeler, T. L., Kooohmaraie, M., & Nonneman, D. (2009). Relationship among calpastatin single nucleotide polymorphisms, calpastatin expression and tenderness in pork longissimus. Animal Genetics, 40, 713–721. Lo, L. L., McLaren, D. G., McKeith, F. K., Fernando, R. L., & Novakofski, J. (1992). Genetic analyses of growth, real-time ultrasound, carcass, and pork quality traits in Duroc and Landrace pigs: II. Heritabilities and correlations. Journal of Animal Science, 70, 2387–2396. Melody, J. L., Lonergan, S. M., Rowe, L. J., Huiatt, T. W., Mayes, M. S., & Huff-Lonergan, E. (2004). Early postmortem biochemical factors influence tenderness and waterholding capacity of three porcine muscles. Journal of Animal Science, 82, 1195–1205. Meullenet, J. F., Lyon, B. G., Carpenter, J. A., & Lyon, C. E. (1998). Relationship between sensory and instrumental texture profile attributes. Journal of Sensory Studies, 13, 77–93. Meyers, S. N., & Beever, J. E. (2008). Investigating the genetic basis of pork tenderness: Genomic analysis of porcine CAST. Animal Genetics, 39, 531–543. Meyers, S. N., Rodriguez-Zas, S. L., & Beever, J. E. (2007). Fine-mapping of a QTL influencing pork tenderness on porcine chromosome 2. BMC Genetics, 8, 69. Monteiro, A. C. S. M. G. (2012). Relationship between texture analysis and sensory attributes of the three main beef types marketed in Portugal, Lisboa. (Chapter 6). Nonneman, D., Lindholm-Perry, A. K., Shackelford, S. D., King, D. A., Wheeler, T. L., Rohrer, G. A., Bierman, C. D., Schneider, J. F., Miller, R. K., Zerby, H., & Moeller, S. J. (2011). Predictive markers in calpastatin for tenderness in commercial pig populations. Journal of Animal Science, 89(9), 2663–2672. Palmer, B. R., Morton, J. D., Roberts, N., Ilian, M. A., & Bickerstaffe, R. (1999). Markerassisted selection for meat quality and the ovine calpastatin gene. Proceedings of the New Zealand Society of Animal Production, 59, 266–268. Palmer, B. R., Robert, N., Hickford, J. G. H., & Bickerstaff, G. (1998). Rapid communication: PCR-RFLP for MspI and NcoI in the ovine calpastatin gene. Journal of Animal Science, 76, 1499–1500. Pinto, L. F., Ferraz, J. B., Meirelles, F. V., Eler, J. P., Rezende, F. M., Carvalho, M. E., Almeida, H. B., & Silva, R. C. (2010). Association of SNPs on CAPN1 and CAST genes with tenderness in Nellore cattle. Genetics and Molecular Research, 9(3), 1431–1442. Rohrer, G. A., Holl, J., Perry, A., Nonneman, D., Shackelford, S. D., Wheeler, T., & Koohmaraie, M. (2007). Update on genetic markers for pork quality. Proc. Natl. Swine Improve. Fed. Annu. Conf., Kansas City, MO (Accessed Mar. 21, 2011. http:// www.nsif.com/conferences/2007/pdf/GeneticMarkers.pdf). Rohrer, G. A., Nonneman, D. J., Miller, R. K., Zerby, H., & Moeller, S. J. (2012). Association of single nucleotide polymorphism (SNP) markers in candidate genes and QTL regions with pork quality in commercial pigs. Meat Science, 92, 511–518. Rohrer, G. A., Thallman, R. M., Shackelford, S., Wheeler, T., & Koohmaraie, M. (2006). A genome scan for loci affecting pork quality in a Duroc–Landrace F2 population. Animal Genetics, 37, 17–27. Rybarczyk, A., Kmieć, M., Terman, A., & Szaruga, R. (2012). The effect of CAST and RYR1 polymorphisms on carcass and meat quality traits in Pietrain crossbred pigs. Animal Science Papers and Reports, 30(3), 241–248. Schenkel, F. S., Miller, S. P., Jiang, Z., Mandell, I. B., Ye, X., Li, H., & Wilton, J. W. (2006). Association of a single nucleotide polymorphism in the calpastatin gene with carcass and meat quality traits of beef cattle. Journal of Animal Science, 84, 291–299. Van Laack, R. L. J. M., Kauffman, R. G., Sybesma, W., Smulders, F. J. M., Eikelenboom, G., & Pinheiro, J. C. (1994). Is colour brightness (L-value) a reliable indicator of water-holding capacity in porcine muscle? Meat Science, 38(2), 193–201. Wendt, A., Thompson, V. F., & Goll, D. E. (2004). Interaction of calpastatin with calpain: A review. The Journal of Biological Chemistry, 385(6), 465–472.

Association of calpastatin gene polymorphisms and meat quality traits in pig.

Calpastatin is associated with the rate of post mortem degradation of structural proteins due to the regulation of calpain activity. In the present re...
282KB Sizes 2 Downloads 3 Views