Effect of <i>LCORL</i> gene polymorphism on body size traits in horse populations

The aim of this study was to determine polymorphism of LCORL gene in horse breeds and its association with body size. PCR-RFLP technique was performed using AluI for genotyping of 306 horses. Results showed that C is the rare allele in Iranian Breeds, because these horses have been used since ancient times as a courier and for war and archery, hence selection has done to benefit of spiky horses with medium body that need less food and are tireless. While, for foreign breeds; frequency of C allele was high that can be concluded these breeds used in fields, forests, and mines. A UPGMA dendrogram based on the Nei's standard genetic distance among studied breeds showed separate clusters for Iranian native and exotic breeds. Statistical association analysis of three observed genotypes with body size showed that there is an association between this polymorphism and body size criteria (p < 0.01). Overall, it can be concluded that studied mutation in LCORL gene can be used as candidate marker for improving body weight in horse.

Keywords:
Association; foreign breeds; Iranian Breeds; PCR-RFLP technique; SNP

Authorship

Ali Mostafavi

Animal Science Department, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, 7616914111, Iran. Shahid Bahonar University of KermanIranKerman, IranAnimal Science Department, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, 7616914111, Iran.

Masood Asadi Fozi

Ali Esmailizadeh Koshkooieh

Mohammadreza Mohammadabadi ^** Author for correspondence. E-mail: mrm@uk.ac.ir

https://orcid.org/0000-0002-1268-3043

Olena Ivanivna Babenko

Department of Animal Science, Bila Tserkva National Agrarian University, Bila Tserkva, Ukraine. Bila Tserkva National Agrarian UniversityUkraineBila Tserkva, UkraineDepartment of Animal Science, Bila Tserkva National Agrarian University, Bila Tserkva, Ukraine.

Nataliia Ihorivna Klopenko

* Author for correspondence. E-mail: mrm@uk.ac.ir

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables | Formulas

Figures (4)
Tables (5)
Formulas (1)

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Figure 1
Some samples of the extracted DNA from the studied animals on the 1% agarose gel.

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Figure 2
Ethidium bromide-stained agarose gel of amplified PCR products representing amplification of LCORL gene in some studied horses. Lines 1-9 are some samples, M is size marker and -C is negative control.

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Figure 3
PCR amplified products of LCORL gene digested with AluI in some studied horses. M is DNA size marker and other lanes are TT, TC and CC genotypes.

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Figure 4
UPGMA phylogenetic tree based on Nei genetic distance.

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Table 1
Sampling design of the studied animals.

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Table 2
Genotypic and allelic frequencies of LCORL gene in studied horses.

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Table 3
The values of the population genetics parameters in studied horses.

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Table 4
Least-square means and standard errors of the studied traits for the different genotypes of LCORL gene in all studied horse populations.

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Table 5
Least-square means and standard errors of the studied traits for the different genotypes of LCORL gene in the studied breeds.

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Figure 1 Some samples of the extracted DNA from the studied animals on the 1% agarose gel.

Figure 2 Ethidium bromide-stained agarose gel of amplified PCR products representing amplification of LCORL gene in some studied horses. Lines 1-9 are some samples, M is size marker and -C is negative control.

Figure 3 PCR amplified products of LCORL gene digested with AluI in some studied horses. M is DNA size marker and other lanes are TT, TC and CC genotypes.

Figure 4 UPGMA phylogenetic tree based on Nei genetic distance.

Table 1 Sampling design of the studied animals.

Breed type	Place of sampling within Iran	№ of samples
Pure-bred Arab horse (Persian Arab)	Kerman, Zarand, Rafsanjan	25
Turkmen (Akhal teke)	Kerman, Isfahan	42
Turkmen (Yamut)	Kerman	16
Dareshouri	Kerman, Sirjan, Rafsanjan, Rayen	28
Kurd	Kerman, Rafsanjan, Sirjan, Isfahan	25
Caspian	Rasht, Kerman, Karaj	42
Taleshi	Guilan (Rezvanshahr)	14
Egyptian Arabian	Karaj, Kerman, Sirjan, Zarand, Rafsanjan, Rayen	21
Hanoverian	Tehran, Karaj, Kerman	16
Holesteiner	Tehran, Karaj, Kerman	11
KWPN	Tehran, Karaj, Kerman	20
Thoroughbred	Tehran, Kerman	4
Oldenburg	Tehran, Kerman	4
Westphalian	Tehran, Karaj	2
Selle Francais	Kerman	2
Foreign Arab	Tehran, Kerman	32
Zangersheide	Kerman	2

The association between the LCORL gene polymorphism and the body size traits of the 306 horses were studied using the following statistical fixed-model:

Table 2 Genotypic and allelic frequencies of LCORL gene in studied horses.

Population	Genotype	N	Genotypic freq.	Allele	Allelic freq.
Pure-bred Arab horse (Persian Arab)	TT	23	0.92	T	0.94
	TC	1	0.04	C	0.06
	CC	1	0.04	C	0.06
Turkmen (Akhal teke)	TT	42	1.00	T	1.00
Turkmen (Akhal teke)				C	0.00
Turkmen (Yamut)	TT	15	0.94	T	0.97
Turkmen (Yamut)	TC	1	0.06	C	0.03
Dareshouri	TT	26	0.93	T	0.97
Dareshouri	TC	2	0.07	C	0.03
Kurd	TT	25	1.00	T	1.00
Kurd				C	0.00
Caspian	TT	42	1.00	T	1.00
Caspian				C	0.00
Taleshi	TT	14	1.00	T	1.00
Taleshi				C	0.00
Egyptian Arabian	TT	21	1.00	T	1.00
Egyptian Arabian				C	0.00
Hanoverian	TT	4	0.25	T	0.56
	TC	10	0.62	C	0.44
	CC	2	0.13	C	0.44
Holesteiner	TT	5	0.46	T	0.60
	TC	3	0.27	C	0.40
	CC	3	0.27	C	0.40
KWPN	TT	5	0.25	T	0.52
	TC	11	0.55	C	0.48
	CC	4	0.20	C	0.48
Thoroughbred	TT	1	0.25	T	0.50
	TC	2	0.50	C	0.50
	CC	1	0.25	C	0.50
Oldenburg	TT	2	0.50	T	0.62
	TC	1	0.25	C	0.38
	CC	1	0.25	C	0.38
Westphalian	TC	1	0.50	T	0.25
Westphalian	CC	1	0.50	C	0.75
Selle Francais	TC	1	0.50	T	0.25
Selle Francais	CC	1	0.50	C	0.75
Foreign Arab	TT	32	100	T	1.00
Foreign Arab			0.00	C	0.00
Zangersheide	TC	1	0.50	T	0.25
Zangersheide	CC	1	0.50	C	0.75
All	TT	257	0.84	T	0.90
	TC	34	0.11	C	0.10
	CC	15	0.05	C	0.10

Table 3 The values of the population genetics parameters in studied horses.

Population	№ of observed alleles	№ of effective alleles	Observed heterozygosity	Expected heterozygosity	Shanon’s Index
Pure-bred Arab horse (Persian Arab)	2	1.12	0.04	0.12	0.23
Turkmen (Akhal teke)	1	1	0	0	0
Turkmen (Yamut)	2	1.06	0.06	0.06	0.14
Dareshouri	2	1.07	0.07	0.07	0.16
Kurd	1	1	0	0	0
Caspian	1	1	0	0	0
Taleshi	1	1	0	0	0
Egyptian Arabian	1	1	0	0	0
Hanoverian	2	1.96	0.62	0.50	0.68
Holesteiner	2	1.93	0.27	0.50	0.67
KWPN	2	1.99	0.55	0.51	0.69
Thoroughbred	2	2	0.50	0.57	0.69
Oldenburg	2	1.88	0.25	0.54	0.66
Westphalian	2	1.06	0.50	0.50	0.56
Selle Francais	2	1.06	0.50	0.50	0.56
Foreign Arab	1	1	0	0	0
Zangersheide	2	1.06	0.50	0.50	0.56
All	2	1.23	0.11	0.18	0.40

Table 4 Least-square means and standard errors of the studied traits for the different genotypes of LCORL gene in all studied horse populations.

Genotype (№)	WHT	GRC	BLG	EBW	HDL	HWD	OBI
Genotype (№)	***	***	***	***	***	**	N.S.
CC (15)	162.6 ±1.37^a	185.9 ±2.23^a	166.7 ±2.18^a	509.2 ±13.15^a	55.6 ±0.59^a	19.5 ±0.29^a	1.14 ±0.012^a
TC (34)	157.4 ±1.07^b	178.2 ±1.74^b	158.4 ±1.7^b	437.2 ±10.27^b	53.4 ±0.46^b	18.9 ±0.22^ab	1.13 ±0.009^a
TT (257)	151.4 ±0.64^c	172.6 ±1.04^c	156.0 ±1.01^c	403.2 ±6.12^c	52.4 ±0.28^b	18.4 ±0.13^b	1.14 ±0.006^a

EBW: estimated body weight (kg), OBI: obesity index, WHT: withers height (cm), BLG: body length (cm), GRC: girth chest (cm), HDL: head length (cm), HWD: head width. N.S. = not Significant, **: significant effect of genotype on studied traits in 1% (p < 0.01), ***: significant effect of genotype on studied traits in 0.1% (p < 0.001).

Table 5 Least-square means and standard errors of the studied traits for the different genotypes of LCORL gene in the studied breeds.

Breed	Genotypes	WHT	GRC	BLG	EBW	HDL	HWD	OBI
Thoroughbred	CC	168.2±4.63	197.7±7.45	183.7±7.19	602.5±43.2	58.4±1.92	18.1±0.93	1.2±0.04
	TC	164.3±3.28	183.8±5.28	168.8±5.09	479.6±30.6	54.1±1.26	18.2±0.66	1.1±0.03
	TT	158.2±4.63	183.7±7.45	186.7±7.19	528.9±43.2	55.9±1.92	20.6±0.93	1.2±0.04
Selle Francais	CC	172.2±4.63	207.7±7.45	192.7±7.19	697.5±43.2	61.9±1.92	22.1±0.093	1.2±0.04
Selle Francais	TC	168.2±4.63	194.7±7.45	183.7±7.19	584.4±43.2	61.4±1.92	21.1±0.93	1.2±0.04
Zangersheide	CC	170.8±4.63	198.3±7.45	166.3±7.19	548.5±43.2	57.6±1.92	20.9±0.93	1.2±0.04
Zangersheide	TC	166.8±4.63	199.3±7.45	166.3±7.19	554±1.92	58.6±1.92	19.9±0.93	1.2±0.04
Oldenburg	CC	173.8±4.63	196.3±7.45	164.3±7.19	531±43.2	56.6±1.92	19.4±0.93	1.1±0.04
	TC	167.8±4.63	190.3±7.45	172.3±7.19	523.1±43.2	57.6±1.92	19.9±0.93	1.1±0.04
	TT	162±3.27	190.5±5.25	171.3±5.07	521.8±30.5	53.5±1.35	19.3±0.66	1.2±0.02
Westphalian	CC	173.8±4.63	195.3±7.45	169.3±7.19	541.5±43.2	58.6±1.92	19.4±0.93	1.1±0.04
Westphalian	TC	171.8±4.63	207.3±7.45	169.3±7.19	610.3±43.2	58.6±1.92	19.9±0.93	1.2±0.04
Holesteiner	CC	174.8±2.67	202.4±4.29	173.4±4.14	600.6±24.9	63.2±1.11	21±0.54	1.2±0.02
	TC	169.5±2.69	189.6±4.32	161±4.17	486±25.1	57.8±1.11	19.4±0.54	1.1±0.02
	TT	166.8±2.07	186.1±3.32	161.7±3.21	470.7±19.3	56.9±0.86	19.1±0.42	1.1±0.02
Hanoverian	CC	177.3±3.28	205.8±5.28	178.8±5.09	635.9±30.6	59.3±1.36	20.7±0.66	1.2±0.02
	TC	171.5±1.47	195.4±2.26	169.9±2.28	546.3±13.7	57.2±0.61	20.1±0.29	1.1±0.01
	TT	163.4±2.32	187.9±3.72	166.6±3.59	494.4±21.6	54.5±0.96	18.4±0.47	1.2±0.02
KWPN	CC	176±2.32	202.6±3.72	177.9±3.59	614.2±21.6	59.7±0.96	20.7±0.47)	1.2±0.02
	TC	168.8±1.4	194±2.24	168±2.17	530.5±13	58±0.58	19.9±0.28	1.2±0.01
	TT	162.5±2.08	190.7±3.34	161.6±3.22	493.6±19.4	57.6±0.86	19.6±0.42	1.2±0.02
Egyptian Arab	TT	149.8±4.63	163.3±7.45	146.3±7.19	326.9±43.2	50.6±1.92	16.9±0.93	1.1±0.04
Pure-bred Arab horse (Persian Arab)	CC	155.2±4.63	177.7±7.45	170.7±7.19	452.5±43.2	51.4±1.92	18.1±0.93	1.1±0.04
	TC	148.8±4.63	174.3±7.45	157.8±7.19	401.8±43.2	48.6±1.92	19.4±0.93	1.2±0.04
	TT	145.3±0.97	166.1±1.55	154.6±1.5	358.9±9	50.1±0.4	17.9±0.19	1.1±0.01
Foreign Arab	TT	149.9±0.83	171.3±1.33	155.1±1.29	382.6±7.7	49.9±0.34	17.9±0.17	1.1±0.01
Turkmen (Akhal teke)	TT	154.2±0.84	174.3±1.35	163.6±1.3	418.4±7.8	53.2±0.35	18.4±0.17	1.1±0.01
Turkmen (Yamut)	TC	156.8±4.63	180.3±7.45	165.3±7.19	450.4±43.2	52.6±1.92	18.9±0.93	1.2±0.04
Turkmen (Yamut)	TT	149.1±1.2	168.9±1.93	161.9±1.86	389.4±11.2	52.5±0.51	18.4±0.25	1.1±0.01
Dareshouri	TC	156±3.27	175.5±5.25	156.5±5.07	404.2±30.5	51.5±1.35	17.3±0.66	1.1±0.03
Dareshouri	TT	148.9±0.93	168.8±1.5	154.7±1.45	371.7±8.7	51.1±0.41	17.6±0.2	1.1±0.01
Kurd	TT	146±0.95	169.9±1.53	157.3±1.47	382.3±8.9	51.8±0.39	18.4±0.19	1.2±0.01
Caspian	TT	117±0.77	125.8±1.24	120.5±1.2	162±7.2	43.5±0.32	16.2±0.15	1.1±0.01
Taleshi	TT	125.9±2.08	137.3±3.34	127.3±0.3.22	201.1±19.4	46.5±1.11	17.5±0.54	1.1±0.02

y_{i j k l} = µ + B_{i} + S_{j} + {G (B)}_{k (i)} + e_{i j k l}

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Effect of <i>LCORL</i> gene polymorphism on body size traits in horse populations

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[1] * Author for correspondence. E-mail: mrm@uk.ac.ir