CN110060739A - A kind of aquatic bird PCM Genetic diversity evaluation method - Google Patents
A kind of aquatic bird PCM Genetic diversity evaluation method Download PDFInfo
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Abstract
The invention discloses a kind of aquatic bird PCM Genetic diversity evaluation method, this method is by aquatic bird (duck/goose) birth weight (g), adult weight (g), 72 week old egg productions (a)；No. 1 chromosome (%), Z chromosome (%), W chromosome (%) relative length；And group's effective number of allele, heterozygosity, polymorphism information content are measured, and are further carried out multiple attribute synthetical evaluation to above-mentioned 9 indexs using Principal Component Analysis, are finally completed the diversity overall merit of aquatic bird genetic resources.Compared with the existing technology, the method of the present invention efficiently solves single index and is difficult to assess the technical problem of aquatic bird genetic diversity, the overall picture of aquatic bird genetic diversity is presented comprehensively, for aquatic bird (duck/goose), rationally conservation and scientific breeding provide effective technical support, have a good application prospect.
Description
Technical field
The present invention relates to a kind of aquatic bird PCM Genetic diversity evaluation methods, belong to aquatic bird PCM Genetic diversity evaluation technology
Field.
Background technique
Agricultural is the basis of human development, and animal husbandry is that the group of agricultural wants component part, and livestock and poultry breeds resources are animal husbandry
The foundation stone of development.China is aquatic bird cultivation and consumption big country, and in recent years, the meat duck amount of delivering for sale is about 3,000,000,000/year, and the goose amount of delivering for sale is
5.34 hundred million.China is also an aquatic bird resource big country, the goose heredity being included in " Animal Genetic Resources in China will-poultry will "
Resource has 31, duck genetic resources 34.These excellent local genetic resources appearance traits bright characteristics, in adaptability, disease-resistant
Power and flavor quality etc. have apparent advantage, contain huge genetic diversity and Selection potentiaD, be high-quality meat,
The inexhaustible source of egg matter varieties distribution and the basic guarantee of industry sustainable development.Aquatic bird germ plasm resource is protected, future is exactly protected
Aquatic bird industry.And how while development and utilization, aquatic bird resource is protected, is that waterfowls in China worker urgently solves in recent years
The problem of.Its key is a kind of method for developing aquatic bird Genetic diversity evaluation, this is the important of effective protection aquatic bird resource
Technological means.
Currently, waterfowls in China genetic resources Study on Diversity mostly uses detection microsatellite method to evaluate its genetic diversity,
Such as propitious essay woods and Xiao Tianfang step on (the seven family's duck population genetic diversity detections of national aquatic bird gene pool, propitious essay woods etc., middle country
Fowl, 2016,28 (24): 72-74；Using microsatellite marker research duck varieties genetic diversity, Xiao Tianfang etc., 2009,20
(1): 190-196 certain achievement), is achieved, the protection and developmental utilization for aquatic bird genetic resources provides theoretical foundation.But
It is only to be unable to the diversity of overall merit aquatic bird resource with single microsatellite index.
Summary of the invention
Goal of the invention: in view of the above technical problems, the present invention provides a kind of aquatic bird PCM Genetic diversity evaluation method,
It is simple reliable, provides scientific basis for aquatic bird Genetic diversity evaluation.
Technical solution: in order to achieve the above object of the invention, the technical solution adopted in the present invention is as follows:
A kind of aquatic bird PCM (P:phenotype；C:cytogenetics marker；M:molecular marker) heredity
Diversity method, comprising the following steps:
(1) by aquatic bird birth weight, adult weight, 72 week old egg productions, No. 1 chromosome, Z chromosome, W chromosome
Relative length and group's effective number of allele, heterozygosity, polymorphism information content be measured, using principal component analysis
Method carries out multiple attribute synthetical evaluation to above-mentioned 9 indexs and weight coefficient determines, establishes aquatic bird PCM genetic diversity Comprehensive Assessment
Model y (d)=azp1+bzp2+czp3+dzc1+ezc2+fzc3+gzm1+hzm2+izm3, in which: zp1, zp2, zp3, zc1, zc2,
Zc3, zm1, zm2, zm3 respectively represent standardized birth weight, adult weight, 72 week old egg productions, No. 1 chromosome, Z dyeing
Body, the relative length of W chromosome, group's effective number of allele, heterozygosity, polymorphism information content measured value；a,b,c,d,e,
F, g, h and i are respectively associated weight coefficient；
(2) different cultivars aquatic bird, raising to 72 week old are taken, measurement birth weight (p1), adult weight (p2), 72 week old are laid eggs
It measures (p3)；
(3) aquatic bird chromosome is prepared using Peripheral blood culture method, measures No. 1 chromosome (c1), Z chromosome (c2), W dye
Colour solid (c3) relative length；
(4) fluorescence-multiple PCR technique is used, carries out aquatic bird STR parting, respectively statistic mass's Gene frequency distribution, respectively
Effective number of allele (m1), heterozygosity (m2), the polymorphism information content (m3) of statistic mass；
(5) by said determination value or calculated value (p1, p2, p3；c1,c2,c3；P1, p2, p3) it pressesCarry out data
Standardization (wherein, X is the measured value of the character,For the average value of the character, S is the standard deviation of the character)；
(6) it is calculated according to step (5) each as a result, substituting into step (1) described PCM genetic diversity Comprehensive Assessment model
Y (d)=azp1+bzp2+czp3+dzc1+ezc2+fzc3+gzm1+hzm2+izm3, calculate aquatic bird (duck/goose) PCM genetic diversity
Comprehensive score, and be ranked up according to comprehensive score, determine different aquatic bird kind genetic diversity height.
As preferred:
The aquatic bird includes duck or goose.
The kind of the duck includes Gaoyou duck, Youxian County's sheldrake, Jingjiang shelduck, Liancheng white duck, Dual Gold, Linwu duck, Ji'an
Red duck with feather and Beijing duck.
The kind of the goose includes lion-headed goose, Southern Anhui, East Zhejiang province white goose, Sichuan White Goose, Taihu goose, slit eye goose and Wugang
Copper goose.
Determination condition is that ground is flat feeding in the step (2), and the body weight determination quantity of every kind of aquatic bird is 50-70, male and female
It is fifty-fifty；Egg production group 100-300 only, counts 72 week old group egg productions.
Aquatic bird is raised to more than 10 week old in the step (3), and wing venous acquires peripheral blood 10 or more, and separation lymph is thin
Born of the same parents prepare aquatic bird chromosome using periphery hemolymph cultivation, and every aquatic bird selection 5 is sprawled good, periphery and clearly dyed
Body division phases are observed and relative length are calculated by formula to 10 pairs of macrochromosomes before split coil method,The relative length of No. 1 chromosome, Z, W chromosome is included in carefully
Born of the same parents' science of heredity (cytogenetics marker) evaluation index (c1, c2 and c3).
Aquatic bird is raised to more than 10 week old in the step (4), and wing venous acquires peripheral blood 60 or more, and male and female is fifty-fifty,
It is flat with ABI 3730XL DNA mdk gene analyzer that poba gene group DNA, which is extracted, using fluorescence-multiple PCR technique
Platform carries out STR parting, counts respectively:
The effective number of allele (m1) of groupWherein, on the Pij: the i-th seat
The frequency of j allele；The number of alleles at M: j-th seat；Effective number of allele on Nei: the i-th seat；
N: the sum at measured seat；
Heterozygosity (m2)Wherein, k: number of alleles；The frequency of Pi: i-th allele；
Polymorphism information content (m3)Wherein,
K: allele number；The frequency of Pi: i-th allele；The frequency of Pj: j-th allele.
For overall merit aquatic bird genetic diversity, aquatic bird (duck/goose) the Genetic diversity evaluation side PCM of the present invention
Method is both from phenotype, cell and molecular level by producing to aquatic bird (duck/goose) birth weight (g), adult weight (g), 72 week old
Egg amount (a)；No. 1 chromosome (%), Z chromosome (%), W chromosome (%) relative length；And group's effective number of alleles
Number, heterozygosity, polymorphism information content are measured, and further carry out multi objective to above-mentioned 9 indexs using Principal Component Analysis
Overall merit is finally completed the diversity overall merit of aquatic bird resource, provides for the protection and utilization of waterfowls in China genetic resources
Theoretical basis.
Technical effect: compared with the existing technology, the method for the present invention efficiently solves single index and is difficult to assess aquatic bird heredity
Multifarious technical problem presents the overall picture of aquatic bird genetic diversity comprehensively for the first time, is aquatic bird (duck/goose) rationally conservation and section
It learns breeding and effective technical support is provided, have a good application prospect.
Specific embodiment
Technical solutions according to the invention are further described in detail below by specific embodiment.
Embodiment 1
Front-end process is established:
Determination condition is ground online raising, and the measurement quantity of every kind of duck is 100-300, by normal in current production
Duck varieties (Gaoyou duck, Youxian County's sheldrake, Jingjiang shelduck, Liancheng white duck, Dual Gold, Linwu duck, the red duck with feather in Ji'an and Beijing
Duck) birth weight (g), adult weight (g), 72 week old egg productions (a)；No. 1 chromosome (%), Z chromosome (%), W chromosome
The relative length of (%)；And 9 indexs such as group's effective number of allele, heterozygosity, polymorphism information content are measured, and are adopted
Multiple attribute synthetical evaluation is carried out to above-mentioned 9 indexs with Principal Component Analysis and weight coefficient determines, establishes duck PCM genetic diversity
Property Comprehensive Assessment model y=0.054zp1+0.094zp2-0.045czp3 +0.024zc1+0.003zc2+0.024zc3+
0.021zm1+0.075zm2+0.062zm3, in formula: zp1, zp2, zp3, zc1, zc2, zc3, zm1, zm2, zm3 respectively represent mark
The birth weight of standardization, adult weight, 72 week old egg productions, No. 1 chromosome, Z chromosome, the relative length of W chromosome, group have
Imitate number of alleles, heterozygosity, polymorphism information content measured value.
On the basis of obtaining the above duck PCM genetic diversity Comprehensive Assessment model, carry out Comprehensive Assessment by the following method
Duck passes diversity:
(2) each 300 of Gaoyou duck, 1 age in days duckling of Dual Gold are taken, raising to 72 week old randomly selects 30 public affairs, and 30 is female, electricity
Sub- balance accurately measures duck measurement birth weight (g), 43 week old weight (g), its male and female birth weight average value are respectively as follows: 47.5g,
47.8g；43 week old Weight averages are respectively 2653.8g, 1796.0g；And 72 week old group egg productions are counted, respectively 210
It is a, 288.
(3) Gaoyou duck, Dual Gold were raised to 70 days, and every kind randomly selects 10 public affairs, and 10 is female, wing venous blood sampling, gradient
Be centrifugated lymphocyte, using periphery hemolymph cultivation prepare duck or goose chromosome, RMPI1640 culture medium culture 72h,
And cell is harvested, Gimsa dyeing, good, periphery is sprawled in every duck selection 5, and clearly the progress of chromosome division phases is micro-
Simultaneously relative length is calculated by formula to 10 pairs of macrochromosomes before split coil method in sem observationGaoyou duck, No. 1 chromosome of Dual Gold, Z, W chromosome it is flat
Equal relative length is respectively 24.3%, 9.66%, 4.87%；25.98%, 10.06% and 4.81%.
(4) Gaoyou duck, Dual Gold were raised to 70 days, and every kind randomly selects 10 public affairs, and 10 is female, and wing venous blood sampling is extracted
Poba gene group DNA, using fluorescence-multiple PCR technique (primer is shown in Table 1), with the analysis of ABI 3730XL DNA mdk gene
Instrument is platform, carries out STR parting, respectively the effective number of allele (m1) of statistic mass
(wherein, on the Pij: the i-th seat j-th of allele frequency；M: the number of alleles at jth seat；On Nei: the i-th seat
Allele effective number；N: the sum at measured seat.)；Heterozygosity (m2)(wherein, k: equipotential
Gene number；The frequency of Pi: i-th allele.) polymorphism information content (m3)(wherein, k: allele number；Pi: i-th etc.
The frequency of position gene；The frequency of Pj: j-th allele).Through counting, Gaoyou duck, Dual Gold group effective number of allele,
Heterozygosity and polymorphism information content are respectively 5.92,0.475,0.534；7.67,0.416,0.560.
The combination of 1 duck STR parting fluorescence primer of table and optimizing reaction conditions
(5) said determination value or calculated value are pressedProgress data normalization (wherein, X is the measured value of the character,For the average value of the character, S is the standard deviation of the character)；
(6) it is calculated according to step (5) each as a result, pressing duck PCM genetic diversity Comprehensive Assessment model y=
0.054zp1+0.094zp2-0.045czp3+0.024zc1+0.003zc2+0.024zc3+0.021zm1+0.075zm2+
0.062zm3, in formula: zp1, zp2, zp3, zc1, zc2, zc3, zm1, zm2, zm3 respectively represent standardized birth weight, adult
Weight, 72 week old egg productions, No. 1 chromosome, Z chromosome, the relative length group effective number of allele of W chromosome, heterozygosis
Degree, polymorphism information content measured value, calculating Gaoyou duck, Dual Gold PCM genetic diversity comprehensive score are respectively 0.201,
0.086, from the point of view of comprehensive score, Gaoyou duck PCM genetic diversity is better than Dual Gold.
Embodiment 2
Front-end process is established:
Determination condition is ground online raising, and the measurement quantity of every kind of goose is 100-300, by normal in current production
Measurement is just for goose kind (lion-headed goose, Southern Anhui, East Zhejiang province white goose, Sichuan White Goose, Taihu goose, slit eye goose and Wugang copper goose)
Raw weight (g), adult weight (g), 72 week old egg productions (a)；No. 1 chromosome (%), Z chromosome (%), W chromosome (%)
Relative length；Group's effective number of allele, heterozygosity, polymorphism information content, using Principal Component Analysis to above-mentioned 9 indexs
It carries out multiple attribute synthetical evaluation and weight coefficient determines, establish goose PCM genetic diversity Comprehensive Assessment model y=0.041zp1+
0.036zp2-0.035czp3-0.055zc1+0.048zc2+0.027zc3+0.074zm1+ 0.100zm2+0.099zm3, in formula:
Zp1, zp2, zp3, zc1, zc2, zc3, zm1, zm2, zm3 respectively represent standardized birth weight, adult weight, 72 week old and lay eggs
Amount, No. 1 chromosome, Z chromosome, the relative length of W chromosome, group's effective number of allele, heterozygosity, polymorphism information content
Measured value.
On the basis of obtaining the above goose PCM genetic diversity Comprehensive Assessment model, carry out Comprehensive Assessment by the following method
Goose passes diversity:
(1) each 300 of Southern Anhui, 1 age in days duckling of East Zhejiang province white goose are taken, raising to 72 week old randomly selects 30 public affairs, and 30
Mother, electronic balance accurately measures goose measurement birth weight (g), 43 week old weight (g), its male and female birth weight average value are respectively as follows:
103.5g,109.0g；43 week old Weight averages are respectively 6100.0g, 4750.3g；And 72 week old group egg productions are counted,
Respectively 34,36.
(2) Southern Anhui, East Zhejiang province white goose were raised to 70 days, and every kind randomly selects 10 public affairs, and 10 is female, wing venous blood sampling,
Gradient centrifugation separates lymphocyte, prepares duck or goose chromosome, RMPI1640 culture medium culture using periphery hemolymph cultivation
72h, and cell is harvested, Gimsa dyeing, every duck selects 5 to sprawl the clearly chromosome division phases progress of good, periphery
Simultaneously relative length is calculated by formula to 10 pairs of macrochromosomes before split coil method in micro- sem observationSouthern Anhui, No. 1 chromosome of East Zhejiang province white goose, Z, W chromosome
Average relative length be respectively 20.29%, 11.48%, 6.97%, 22.68%, 10.68%, 6.23%.
(3) Southern Anhui, East Zhejiang province white goose were raised to 70 days, and every kind randomly selects 10 public affairs, and 10 is female, wing venous blood sampling,
Poba gene group DNA is extracted, using fluorescence-multiple PCR technique (primer is shown in Table 2), with ABI 3730XL DNA mdk gene point
Analyzer is platform, carries out STR parting, respectively the effective number of allele (m1) of statistic mass
(wherein, on the Pij: the i-th seat j-th of allele frequency；M: the number of alleles at jth seat；On Nei: the i-th seat
Allele effective number；N: the sum at measured seat.)；Heterozygosity (m2)(wherein, k: equipotential
Gene number；The frequency of Pi: i-th allele.) polymorphism information content (m3)(wherein, k: allele number；Pi: i-th etc.
The frequency of position gene；The frequency of Pj: j-th allele).Through counting, Gaoyou duck, Dual Gold group effective number of allele,
Heterozygosity and polymorphism information content are respectively 6.90,0.622,0.563；4.80, 0.590,0.522.
The combination of 2 goose STR parting fluorescence primer of table and optimizing reaction conditions
(4) said determination value or calculated value are pressedCarrying out data normalization, (wherein, X is the measurement of the character
Value,For the average value of the character, S is the standard deviation of the character)；
(5) it is calculated according to step (5) each as a result, pressing goose PCM genetic diversity Comprehensive Assessment model y=
0.041zp1+0.036zp2-0.035czp3-0.055zc1+0.048zc2+0.027zc3+0.074zm1+0.100zm2+
0.099zm3, in formula: zp1, zp2, zp3, zc1, zc2, zc3, zm1, zm2, zm3 respectively represent standardized birth weight, adult
Weight, 72 week old egg productions, No. 1 chromosome, Z chromosome, the relative length of W chromosome, group's effective number of allele, heterozygosis
Degree, polymorphism information content measured value, calculate Southern Anhui, East Zhejiang province white goose PCM genetic diversity comprehensive score be respectively 0.666 ,-
0.103, from the point of view of comprehensive score, Southern Anhui PCM genetic diversity is better than East Zhejiang province white goose.
Claims (7)
1. a kind of aquatic bird PCM Genetic diversity evaluation method, which comprises the following steps:
(1) by aquatic bird birth weight, adult weight, 72 week old egg productions, No. 1 chromosome, Z chromosome, W chromosome it is opposite
Length and group's effective number of allele, heterozygosity, polymorphism information content are measured, using Principal Component Analysis to upper
It states 9 indexs progress multiple attribute synthetical evaluations and weight coefficient determines, establish aquatic bird PCM genetic diversity Comprehensive Assessment model y
(d)=azp1+bzp2+czp3+dzc1+ezc2+fzc3+gzm1+hzm2+izm3, in which: zp1, zp2, zp3, zc1, zc2, zc3,
Zm1, zm2, zm3 respectively represent standardized birth weight, adult weight, 72 week old egg productions, No. 1 chromosome, Z chromosome, W dye
The relative length of colour solid, group's effective number of allele, heterozygosity, polymorphism information content measured value；A, b, c, d, e, f, g, h and
I is respectively associated weight coefficient；
(2) different cultivars aquatic bird is taken, raising to 72 week old measures birth weight (p1), adult weight (p2), 72 week old egg productions
(p3)；
(3) aquatic bird chromosome is prepared using Peripheral blood culture method, measures No. 1 chromosome (c1), Z chromosome (c2), W chromosome
(c3) relative length；
(4) fluorescence-multiple PCR technique is used, carries out aquatic bird STR parting, statistic mass's Gene frequency distribution, counts respectively respectively
Effective number of allele (m1), heterozygosity (m2), the polymorphism information content (m3) of group；
(5) by said determination value or calculated value (p1, p2, p3；c1,c2,c3；P1, p2, p3) it pressesCarry out data normalization
(wherein, X is the measured value of the character,For the average value of the character, S is the standard deviation of the character)；
(6) it is calculated according to step (5) each as a result, substituting into step (1) described PCM genetic diversity Comprehensive Assessment model y (d)
=azp1+bzp2+czp3+dzc1+ezc2+fzc3+gzm1+hzm2+izm3, it is comprehensive to calculate aquatic bird (duck/goose) PCM genetic diversity
Score, and be ranked up according to comprehensive score, determine different aquatic bird kind genetic diversity height.
2. aquatic bird PCM Genetic diversity evaluation method according to claim 1, which is characterized in that the aquatic bird includes duck
Or goose.
3. aquatic bird PCM Genetic diversity evaluation method according to claim 2, which is characterized in that the kind packet of the duck
Include Gaoyou duck, Youxian County's sheldrake, Jingjiang shelduck, Liancheng white duck, Dual Gold, Linwu duck, the red duck with feather in Ji'an and Beijing duck.
4. aquatic bird PCM Genetic diversity evaluation method according to claim 2, which is characterized in that the kind packet of the goose
Include lion-headed goose, Southern Anhui, East Zhejiang province white goose, Sichuan White Goose, Taihu goose, slit eye goose and Wugang copper goose.
5. aquatic bird PCM Genetic diversity evaluation method according to claim 1, which is characterized in that surveyed in the step (2)
Fixed condition, which is that ground is flat, supports, and the body weight determination quantity of every kind of aquatic bird is 50-70, and male and female is fifty-fifty；Egg production group 100-300
Only, 72 week old group egg productions are counted.
6. aquatic bird PCM Genetic diversity evaluation method according to claim 1, which is characterized in that water in the step (3)
More than poultry raising to 10 week old, wing venous acquires peripheral blood 10 or more, lymphocyte is separated, using periphery hemolymph cultivation
Prepare aquatic bird chromosome, every aquatic bird select 5 to sprawl good, periphery clearly chromosome division phases are observed and right
Relative length is calculated by formula in 10 pairs of macrochromosomes before split coil method,
The relative length of No. 1 chromosome, Z, W chromosome is included in cytogenetics (cytogenetics marker) evaluation index
(c1, c2 and c3).
7. aquatic bird PCM Genetic diversity evaluation method according to claim 1, which is characterized in that water in the step (4)
More than poultry raising to 10 week old, wing venous acquires peripheral blood 60 or more, and male and female is fifty-fifty, extracts poba gene group DNA, and use is glimmering
Light-multiple PCR technique carries out STR parting using ABI 3730XL DNA mdk gene analyzer as platform, counts respectively:
The effective number of allele (m1) of groupWherein, j-th on the Pij: the i-th seat
The frequency of allele；The number of alleles at M: j-th seat；Effective number of allele on Nei: the i-th seat；N: institute
Measure the sum at seat；
Heterozygosity (m2)Wherein, k: number of alleles；The frequency of Pi: i-th allele；
Polymorphism information content (m3)Wherein, k: equipotential
Number of genes；The frequency of Pi: i-th allele；The frequency of Pj: j-th allele.
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