CN103404483A

CN103404483A - Breeding method for Gansu meat sheep multiple-birth strain

Info

Publication number: CN103404483A
Application number: CN2013103355186A
Authority: CN
Inventors: 刘建斌; 孙晓萍; 郭健; 王凡; 张万龙; 冯瑞林; 岳耀敬; 郎侠; 杨博辉; 郭婷婷; 曾玉峰; 王宏博
Original assignee: Lanzhou Institute of Animal Husbandry and Veterinary Medicine CAAS
Current assignee: Lanzhou Institute of Animal Husbandry and Veterinary Medicine CAAS
Priority date: 2013-08-05
Filing date: 2013-08-05
Publication date: 2013-11-27
Anticipated expiration: 2033-08-05
Also published as: CN103404483B

Abstract

The invention discloses a breeding method for a Gansu meat sheep multiple-birth strain. Poll Dorset and Borderdale which serve as a male parent are hybridized with small tail han sheep carrying mutant coincidental genotype (BB) serving as a female parent; through the hybridization and crossbreed and fixation, the Gansu meat sheep multiple-birth strain is obtained. The Gansu meat sheep multiple-birth strain has the advantages of high lambing rate, good meat producing performance, fast growth, tall build, high adaptation and the like, is adaptable to feeding and captive fattening in farming and pastoral areas of the Gansu Hexi Corridor, and accords with the development trend of Gansu sheep industry.

Description

The breeding method of Gansu Mutton Sheep polyembryony strain

Technical field

The present invention relates to the animal breeding technology field, particularly relate to and utilize the cold sheep polyembryony kind of domestic little tail and introduction Mutton Sheep Poll Dorset Saite and baud for variety source, carry out the breeding method of Gansu Mutton Sheep multiparous new strain.

Background technology

It is that the important economical trait of Mutton Sheep, particularly litter size and growth rate are the key factors that determines Hexi Corridor In Gansu farming-pastoral region sheep husbandry benefit that Growth and Reproduction is grown.The cold sheep of little tail is one of world-renowned rare local polyembryony sheep variety of China, its prlificacy causes many Sheep Breeding scientists' concern very early, its endocrine mechanism and phenotype characteristics have been carried out deeply to research widely, controlled the major gene resistance (Fec of the cold sheep polyembryony of little tail ^BGene) essence is sheep bone morphogenetic protein receptor IB gene (Bone morphogenetic protein receptor IB, BMPR-IB) on 746 of code areas, A → G single base mutation having occurred, makes recipient cell intracellular kinase district encoding proteins by the glutamine of wild type, be sported the arginine (Q249R) of the cold sheep polyembryony of little tail.Therefore, the BMPR-IB gene is one of major gene resistance of the cold sheep polyembryony litter size of little tail.Take full advantage of and introduce Mutton Sheep Poll Dorset Saite and baud generation fast, the advantage such as the price of deed is high, meat yield is many and performance is good of growing, comprehensively performance is as the Poll Dorset Saite of male parent and baud generation and as the good characteristic of the cold sheep polyembryony kind of maternal little tail, cultivates that lambing percentage is high, meat performance good, grow fast, physique is tall and big and adaptable Gansu Mutton Sheep multiparous new strain.

Summary of the invention

The invention discloses the breeding method of a kind of Gansu Mutton Sheep multiparous new strain, by the Poll Dorset Saite as male parent and baud generation respectively with as maternal and carry the cold sheep of little tail that sudden change overlaps genotype (BB) and hybridize, the generation of selecting and remain is outstanding and carry the hybridization ewe (the cold generation of ripple and road tremble with fear a generation) that sudden change overlaps genotype (BB) or sudden change heterozygous genes type (B+); With the cold generation of the cold generation of the first cross ewe ripple selected and remain and road, with Poll Dorset Saite and baud, hybridize for kind of a sheep respectively, in two generations of selecting and remain, are outstanding and carry hybridization ram (cold two generations of road ripple and radio frequency channel cold two generations) and the ewe (cold two generations of road ripple and radio frequency channel were trembled with fear for two generations) that sudden change overlaps genotype (BB) or sudden change heterozygous genes type (B+); By select and remain first cross ewe and two generations hybridization ewe, with two generations of selecting and remain, hybridize the ram mating respectively, produce Gansu Mutton Sheep multiparous new strain zero sheep from generation to generation; From ram and the ewe outstanding and that carry sudden change coincidence genotype (BB) or sudden change heterozygous genes type (B+) of selecting and remain zero sheep from generation to generation; Adopt colony's subculture selection-breeding method and molecular marker assisted selection method to carry out the traversed by of 4 generations and fix, obtain Gansu Mutton Sheep multiparous new strain.The Gansu Mutton Sheep polyembryony strain of this breeding method seed selection has that lambing percentage is high, meat performance good, it is fast to grow, physique is tall and big and the advantage such as strong adaptability, be applicable to the drylot feeding of Hexi Corridor In Gansu farming-pastoral region and stable breeding and fatten, meet Gansu sheep husbandry development trend.

The accompanying drawing explanation

Fig. 1: Gansu Mutton Sheep polyembryony strain development scheme.

Embodiment

1, the selection of the cold sheep ewe of little tail

(1) select the cold sheep of little tail of physical health, to the cold sheep blood sample collection 5mL of every little tail of preliminary election, and carry out the ear tag record;

(2) to the cold sheep blood sample of the little tail gathered, utilize high-resolution melting curve analysis technology (HRM) to carry out the BMPR-IB genetic test, determine that the sheep individuality of Q249R sudden change has occurred at BMPR-IB gene coded sequence the 746th bit base place, namely carry the female parent (1500) of the cold sheep individuality of little tail of sudden change coincidence genotype (BB) as Gansu Mutton Sheep multiparous new strain.

2, utilize the HRM technology to detect the BMPR-IB gene

(1) utilize LightScanner Primer Design Software (Idaho company, the U.S.) to be designed for the primer that HRM method SNP loci gene type detects.

Upstream: 5 '-AGATTGGAAAAGGTCGCTATG-3 '

Downstream: 5 '-ACCCTGAACATCGCTAATACA-3 '

(2) HRM-PCR amplification system (11 μ L)

The genomic DNA 1 μ L of 20ng μ L-1, each 0.2 μ L of the upstream and downstream primer of 10pmol, 2 * Taq PCR MasterMix (includes TaqDNA polymerase, Mg ²⁺, dNTPs etc.) (Beijing, day root) 5 μ L, sterilizing ultra-pure water 3.6 μ L, 10 * LC Green saturable dye (U.S., Idaho) 1 μ L.

(3) HRM-PCR response procedures

95 ℃ of denaturation 5min; 94 ℃ of sex change 20s, Tm renaturation 20s, 72 ℃ are extended 20s, 35 circulations; 72 ℃ are extended 53min, 15 ℃ of preservations.

(4) mark dilution process in high low temperature

Mark strand+400 μ L water in 1OD.

(5) mark annealing system in high low temperature

Saturated sodium-chloride 1 μ L, complementary double-stranded each the 1 μ L of interior mark, moisturizing to 10 μ L, 95 ℃ of water-bath 3min, then slowly be down to room temperature.

(6) the HRM fluorescence signal gathers

Each 1 μ L of mark in the high low temperature diluted is added in HRM-PCR product single hole to 95 ℃ of water-bath 30s; 25 ℃ of water-bath 30s; Double-click the Lightscanner icon, open file.Click " RUN ", scanning imaging system is set.96 orifice plates after reaction are put into to LS96 to be scanned.Waiting temperature is raised to the standby temperature, and 96 orifice plates are inserted in machine, notes having an end of corner cut first to insert, and treats that door shuts.Click " Start Run ", eject a window, be used for store files.Software is collected data.Lightscanner melts setting, 75 ℃ of initial temperatures, and 94 ℃ of end temps, keep 72 ℃ of temperature, and exposure is selected automatically.

(7) interpretation of result

Open LightScanner software, click " the Small Amplicon " of " Genotyping " below, open the data that will analyze.After data are divided into groups to have named, amplified fragments is carried out to genotyping: row's option of operation appears in " Genotyping " software left side central portion of clicking LightScanner software top, clicks and just can complete the analysis to sample successively in order.That at first occur is " Negative Filter ", is used for selecting feminine gender or positive findings, chooses rear click " Apply Change "; Click again Calibration and proofread and correct, if adopted inner correction, set by inner correction parameter, if do not proofread and correct, directly carry out next step.Then click " Normalize ", curve is carried out to normalization; Click " Group ", " Auto Group " and " Use Internal Standards " two options are arranged in the drop-down menu on Standards mono-hurdle, after selecting suitable option, click " Compute Groups " and just genotype can be divided into to different curves.

(8) according to curve map, can determine the different genotype of BMPR-IB gene.

3, the Poll Dorset Saite of selection physical health and baud are for the male parent (Poll Dorset Saite 15 and baud for 10) of ram as Gansu Mutton Sheep multiparous new strain;

4, as the Poll Dorset Saite of male parent and baud generation respectively with as maternal and carry the cold sheep of little tail that sudden change overlaps genotype (BB) and carry out hybridization in continuous 2 years, by method 2, determine and carrying the hybridization ewe (the cold generation 890Zhi He of ripple tremble with fear in road a generation 950) that sudden change overlaps genotype (BB) or sudden change heterozygous genes type (B+);

5, with the cold generation of the cold generation of the first cross ewe ripple selected and remain and road, with Poll Dorset Saite and baud, hybridized for kind of a sheep respectively in continuous 2 years, determine that by method 2 carrying sudden change overlaps the hybridization ram (20 of road ripple cold two generations and radio frequency channel cold two generation 25) of genotype (BB) or sudden change heterozygous genes type (B+) and ewe (600 of road ripple cold two generations and radio frequency channel trembled with fear two generations 750);

6, by the first cross ewe of selecting and remain (1840) and two generations hybridization ewes (1350), with two generations of selecting and remain, hybridize ram (45) mating respectively, produce Gansu Mutton Sheep multiparous new strain zero sheep (1450) from generation to generation; From ram (10) and the ewe (1440) outstanding and that carry sudden change coincidence genotype (BB) or sudden change heterozygous genes type (B+) of selecting and remain zero sheep from generation to generation; Adopt colony's subculture selection-breeding method and molecular marker assisted selection method to carry out the traversed by of 4 generations and fix, obtain Gansu Mutton Sheep multiparous new strain, this new lines core group population scale reaches 2100, and the average lambing percentage of the every tire of multiparity ewe reaches more than 160%; The public lamb of 0～90 age in days average daily gain and ewe lamb have improved respectively 31.25% and 30.77% than the public lamb of the cold sheep of little tail and ewe lamb; During 90 age in days, public lamb and ewe lamb body weight than the public lamb of the cold sheep of little tail and the high 5.07kg of ewe lamb and 3.91kg, improve respectively 27.78% and 25.54% respectively; 4 monthly ages public lamb carcass weight 16.59 ± 1.49kg, dressing percentage is 51.6 ± 0.96%, eye muscle area is 11.18cm ².

The above, be only the specific embodiment of the present invention, but protection scope of the present invention is not limited to this, and any variation or replacement of expecting without creative work, within all should being encompassed in protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain that claims were limited.

Claims

1. the breeding method of a Gansu Mutton Sheep polyembryony strain, by the Poll Dorset Saite as male parent and baud generation respectively with as maternal and carry the cold sheep of little tail that sudden change overlaps genotype (BB) and hybridize, the generation of selecting and remain is healthy and carry hybridization ewe that sudden change overlaps genotype (BB) or sudden change heterozygous genes type (B+) and obtain the cold generation of ripple and the road generation of trembling with fear; With the cold generation of the cold generation of the first cross ewe ripple selected and remain and road, with Poll Dorset Saite and baud, hybridize for kind of a sheep respectively, two generations of selecting and remain healthy and carry cold two generations of road ripple that sudden change overlaps genotype (BB) or sudden change heterozygous genes type (B+) and radio frequency channel cold two generations hybridization ram and cold two generations of Dao Bo and the cold two godmother sheep of radio frequency channel; By select and remain first cross ewe and two generations hybridization ewe, with two generations of selecting and remain, hybridize the ram mating respectively, produce Gansu Mutton Sheep polyembryony strain zero sheep from generation to generation; From ram and the ewe of selecting and remain physical health and carrying sudden change coincidence genotype (BB) or sudden change heterozygous genes type (B+) zero sheep from generation to generation; Adopt colony's subculture selection-breeding method and molecular marker assisted selection method to carry out the traversed by of at least 4 generations and fix, obtain Gansu Mutton Sheep polyembryony strain.

2. breeding method according to claim 1, it is characterized in that utilizing high-resolution melting curve analysis technology (HRM) to carry out the BMPR-IB genetic test for offspring maternal or hybridization, determine and carry the sheep individuality that sudden change overlaps genotype (BB) or sudden change heterozygous genes type (B+).