| China have rich local goose breeds, and body conformation, performance and genetic traits were quite different among different varieties. To fully understand the genetic resources, genetic diversity and genetic relationships of geese in our country, the study collected 11 China local varieties which come from different locations of China,7 varieties of Jiangsu province,2 European domestic goose breeds, swan geese and gray geese. we obtained 10 pairs of microsatellite primers goose by using dual-suppression-PCR technique to analysis 437 Goose individual genomic DNA. Analysis of allele frequencies, heterozygosity (H), effective number of alleles, polymorphism information content, Nei’s standard genetic distance (DS) among populations, and the DA genetic distance, we analyzed 11 China local varieties and 7 local varieties in Jiangsu province by using UPGMA method, principal component analysis (FCA) and population genetic structure analysis method. Based on 79 individuals’ 681bp fragment of mitochondrial DNA COI gene sequences of 13 varieties and related sequences in GenBank to discussion taxonomic status of goose breeds in China. The results were as follows:1.12 pairs of microsatellite primers isolated from geese by dual-suppression PCR, of which only a pair of primer not completely amplified microsatellite fragments, the rest can do it, and the annealing temperature varied greatly, so it is easy for PCR amplification. Only G11 primer amplified fragments in all species were tested on a single state, the remaining 10 primers were polymorphic. we selected relatively small and clear alleles A and B from the amplified fragments by microsatellite primers, A allele was 11 (C A/TG)n, B was 14, The results suggest that there were a lot of (TG) n repeats in the genome of the geese.2.10 pairs of microsatellite primers with good polymorphism were screened from the obtained primers, which used to amplified genomic DNA of 13 breeds and to determined genotypes. The results calculated by Fstat software showed that 61 alleles were detected in 13 varieties; number of alleles ranged from 4-11 in each locus.12 alleles exist in all 11 local goose populations of China, the number of alleles accout for 19.56% in the total number of alleles; only seven alleles appeared in 11 chinese local goose populations and two exotic species. However, some specific alleles only occured in certain groups, such as in Xupu goose, Shitou goose, taihu goose, Huoyan goose. The results show that in addition to 143 bp fragment of G08 appeared in Huoyan goose has high allele frequency (0.868), while the rest rare alleles only appeared lower frequencies. Also we found that some breeds lack some alleles, such as the 143 bp fragment of G01 weren’t detected in Yili goose.3. The average heterozygosity of 10 microsatellites in 13 breeds was 0.545, in addition to average heterozygosity of G04, G09 and G12 are lower than 0.5, the remaining seven were more than 0.5. The heterozygosity of Changle geese was highest (0.605), and Zi Geese was lowest (0.488) in 11 local goose breeds of China. The heterozygosity of two exotic species were lower, so the heterozygosity of Rhine and Landes goose were 0.470 and 0.421, respectively, indicated that China local goose breeds have large genetic variation. Local breeds of China have a high genetic diversity, showed a high heterozygosity, the heterozygosity of Rhin and Landes goose are lower than our local varieties, indicated that the two exotic species were lower. The average of all loci polymorphic information content (PIC) were 0.485 in 13 goose breeds. Local breeds of China have a high PIC, two exotic species were lower PIC. The average of observed number alleles in 10 microsatellite loci were 3.53; the number of effective alleles were 2.539, that of Local breeds of China were high than the number of the two exotic species. Effective alleles of Sichuan white goose was 2.8270. FST (population differentiation factor) in the all groups, the average genetic differentiation was 21.39%(p<0.001), all sites were significant contribution to this result (p<0.001). FIT (total inbreeding coefficient) of 13 goose breeds was 23.73%, reached a significant level (p<0.001); and FIT value of G04, G06, G08, G09, G10 and G12 loci deviated from Hardy-Weinberg expectations, reached extremely remarkable level. Nm (gene flow value) range of 0.3863 of G04 to 1.7854 of G10, the average Nm was 0.8890. From the known Fst and Nm, Fst was larger, Nm was smaller, the degree of species differentiation was greater, so Nm was smaller. FST values of Yili geese and other 11 geese species were remarkable (P<0.05) by analysis of molecular variance. FST between two exotic species and 10 different varieties of China reached a extremely remarkable level (P<0.01). FST between large breeds and small breeds in China local Goose varieties were remarkable (P<0.05). From the results of Nm, Nm between Wanxi White goose and wild goose was maximum (9.554); Nm between Sichuan white goose and wild geese was followed (7.947); Nm between Landes and Zhedong Goose was minimum (0.325). FST between Rhine and Landes geese was smaller (FST=0.072), showed that two populations have weak genetic differentiation. In local varieties, the degree of genetic differentiation between wild geese and Wanxi White was smallest (FST=0.026). But overall, gene differentiation coefficient among populations and gene flow were at a low level in various varieties of Chinese local goose, indicated that the origin of China local goose were affected by the same ancestor.4. In general,13 goose populations can be divided into two categories by UPGMA and NJ method:first, wild geese, Sichuan White geese, Wanxi White geese, Xupu geese, Shitou geese, Taihu geese, Huoyan geese, Zi Geese, Zhedong geese and Changle geese clustered together, second, Rhin and Landes geese firstly clustered together, and then clustered with Yili geese. Genetic differences can be distinguish between different varieties in FactorⅠ, corresponding to differences of origin between China goose and foreign breeds; FactorⅡcorresponds to the difference size of geese. The results analysed by Structure program by the of 13 groups consistent with the results of genetic distance reflects the local breeds genetic structure and relationship, also consistent with 13 local breeds’ geographical distribution, formation history and gene flow. we determined the real kind of contemplating the class based on theΔK method,13 varieties can obviously divided into two groups; and 11 China local varieties can be divide three groups.5.681bp fragment of mitochondrial DNA COI gene were analysed in 79 individual of the 11 varieties. The results are as follows:11 groups Hd (Haplotype diversity)was 0.6219±0.036; Pi(Nucleotide diversity) was 0.00120±0.00011 and have 6 haplotypes. H2 and H3 haplotypes were main haplotype,and frequency were 51.9% and 31.65%, respectively, and H4, H5 and H6 haplotype existed in only someone individual. Rhin haplotype was most(4), Yili geese was least, only one.677 conserved sites,4 variable sites,4 parsimony informative sites, and 2 single-informative sites were found by Mega4 software for multiple sequence alignment. which can be seen base content was no serious bias. Haplotype diversity in Rhin group was highest level (0.8100±0.0169), followed by Grey Goose Geese (0.7140±0.0151), and Yili haplotype diversity was 0. Nucleotide diversity in Grey Goose and Rhine geese were significantly higher than that in Landes and Yili geese; In China local domestic goose breeds, Huoyan geese and Wanxi White geese was significantly higher than other domestic goose breeds and swan goose. The net nucleotide divergence (Da) between grey goose and other two geese (Yili goose and Zhedong White goose) groups were highest, Da between swan goose and other two breeds (Yili goose and Zhedong). The degree of genetic differentiation between Yili goose and the swan goose larger than Yili goose and gray goose. The degree of genetic differentiation were small in China local geese breeds except Yili goose, Indicated that most of variation existed within local varieties.6.32 COI gene sequences of subfamily of geese were analysed showed that average content of bases (T, C, A, G) were 23.8%,34.6%,24.9%,16.7%, the average content of purine close to pyrimidine content, indicated that there was no significant base bias.24 haplotypes were detected in 31 mtDNA CoI sequences, the magpie geese as outgroup subfamilies (Anseranatinae) form a separate haplotype, all breeds’ Hd were 0.9819; domestic goose, swan geese and swan geese sequence was downloaded share the same haplotype, Yili geese, Landes geese and gray geese share the same haplotype.7. The results showed that COI gene sequences were analysed by 10 pairs of goose microsatellite primers as follows:the degree of genetic differentiation between swan geese and Rhine geese was highest (0.386). the genetic distance vaule between Rhin geese and Yili geese was 0.1484, between Landes geese and Yili geese was 0.2099, between Rhin geese and gray geese was 0.2850, between Landes geese and gray geese was 0.3031. The genetic distance between China local goose breeds and the collections of all the breeds are far, but closer to swan geese (0.1768). the genetic distance between Landes geese and swan geese was largest (0.855), the genetic distance between Landes geese and China local goose breeds was 0.854. Accordingly, the earliest time of species differentiation between them were also reached 777.2 years and 776.1 years. differentiation The divergence time between Yili goose and Europe geese later than between Yili goose goose and China local geese. The results of cluster analysis showed that:swan goose cluster with Chinese geese, Rhin and Landes firstly cluster together, and then clustered together with Yili geese and gray geese. On each branch was obtained a higher support rate.8. According to the results of geese varieties of jiangsu province were analyzed based on 10 pair of goose microsatellite seats, The geese varieties of jiangsu province can be divided into four major categories, one kind is taihu goose:including taihu goose in goose field of suzhou, taihu lake goose of Nanjing agricultural university, and currently Yancheng taihu goose; The second is four seasons goose in Lishui and Jurong, and Hongze geese which contain blood of eastern zhejiang white geese and Wanxi white geese; Three is sichuan white geese were introduced to breed and promotion, actually Changshu breeding is not taihu lake goose, but sichuan white geese, they Outward promotion sichuan white geese; Four is Yangzhou geese, which was breed by Hybrid breeding Sichuan white goose through the introduction and taihu goose, so the goose genetic was rich, can self-contained to continue breeding and promotion. And variable eyes goose and annals of goose still free in jiangsu province raising goose outside. |
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