甘南州蕨麻猪mtDNA D-环序列的起源及遗传多样性分析

为了研究甘南州蕨麻猪mtDNA D-环序列起源与遗传多样性,利用文献报道的1对引物对蕨麻猪mtDNA D-环序列进行PCR扩增及测序,对所得数据进行单倍型、遗传多样性、系统发育树和网络关系分析.分析结果显示,135头蕨麻猪mtDNA D-环序列表现出3种长度变异,其中4个序列长度为1 199 bp,3个序列长度是1 319 bp,127个序列长度为1 219 bp.对127个长度为1 219 bp的序列进行分析,发现83个单倍型.单倍型比例、单倍型多样度、核苷酸多样度和平均核苷酸差异数在卡加曼蕨麻猪最高,而在夏河蕨麻猪为最低.系统发育树和网络关系分析均将83个单倍型分为2个分支,说明蕨麻猪具有2个母系起源.与其他猪种mtDNA D-环序列比较,蕨麻猪与东南沿海野猪有较近的亲缘关系,没有发现东亚与东南亚野猪对蕨麻猪的起源有贡献的证据.     

Phylogenetic analysis of Myanmar indigenous chickens using mitochondrial D-loop sequence reveals their characteristics as a genetic resource

Myanmar indigenous chickens play important roles in food, entertainment, and farm business for the people of Myanmar. In this study, complete mitochondrial D-loop sequences (1232 bp) were analyzed using 176 chickens, including three indigenous breeds, two fighting cock populations, and three indigenous populations to elucidate genetic diversity and accomplish a phylogenetic analysis of Myanmar indigenous chickens. The average haplotype and nucleotide diversities were 0.948 ± 0.009 and 0.00814 ± 0.00024, respectively, exhibiting high genetic diversity of Myanmar indigenous chickens. Sixty-four haplotypes were classified as seven haplogroups, with the majority being haplogroup F. The breeds and populations except Inbinwa had multiple maternal haplogroups, suggesting that they experienced no recent purifying selection and bottleneck events. All breeds and populations examined shared haplogroup F. When 232 sequences belonging to haplogroup F (79 from Myanmar and 153 deposited sequences from other Asian countries/region) were analyzed together, the highest genetic diversity was observed in Myanmar indigenous chickens. Furthermore, Myanmar indigenous chickens and red junglefowls were observed in the center of the star-like median-joining network of 37 F-haplotypes, suggesting that Myanmar is one of the origins of haplogroup F. These findings revealed the unique genetic characteristic of Myanmar indigenous chickens as important genetic resources.     

Complete Mitochondrial DNA D-Loop Sequence and Genetic Diversity of Wuzhishan Pig

In order to research the genetic diversity and phylogenies of Wuzhishan pig, 56 complete mtDNA D-Loop were analyzed.55 sequences were amplified by polymerase chain reaction (PCR) and 1 sequence was downloaded from GenBank.The results showed that repetitive sequence "TTATAAAACAC" appeared in the conservative regions of 21 mtDNA D-Loop sequences.13 haplotypes and 14 variable sites were observed in 56 sequences.Haplotype diversity and nucleotide diversity (Hd =0.838 and Pi=0.00334) indicated that Wuzhishan pig had high genetic diversity.Two branches in the phylogenetic tree and Network analysis diagram indicated that there were two maternal origins in Wuzhishan pig.The phylogenetic tree of Wuzhishan pig and other 23 species showed that Wuzhishan pig had close genetic relationship with the pig breeds in the south region of the Yangtze river.     

五指山猪线粒体控制区序列分析及遗传多样性研究

为了研究五指山猪遗传多样性和系统地位,本研究采用PCR产物直接测序法得到55条五指山猪线粒体控制区(mtDNA D-Loop)全序列,并结合GenBank中发表的1条五指山猪mtDNA D-Loop全序列进行结构分析、遗传多样性研究及系统发育分析。结果发现,56条序列中出现21次保守区变异,即"TTATAAAACAC"序列重复,共定义13个单倍型,发现14个变异位点,单倍型多样度(Hd)和核苷酸多样度(Pi)分别为0.838和0.00334,表明五指山猪遗传多样性较丰富。五指山猪线粒体控制区序列构建的系统发育树和Network网络分析呈现两大分支,推测有两个母系起源,且与GenBank中其他23个猪种的聚类结果表明,五指山猪与长江流域以南地区的猪种有着较近的亲缘关系。     

温岭高峰牛线粒体DNA全基因组遗传多样性分析

[目的]通过测定温岭高峰牛线粒体DNA全基因组序列以分析温岭高峰牛的母系起源及遗传多样性。[方法]采用DNA提取、测序及生物信息学方法。[结果]通过对19头温岭高峰牛线粒体DNA全基因组序列分析,共发现263个变异位点,定义9种单倍型,单倍型多样度(Hd±SD)为0.778±0.096,核苷酸多样度(Pi±SD)为0.0017±0.0014,表明温岭高峰牛的遗传多样性较低。构建的NJ系统发育树和单倍型进化网络图表明温岭高峰牛有普通牛和瘤牛2种母系起源。[结论]温岭高峰牛线粒体DNA基因组的遗传多样性较低,有瘤牛和普通牛两个母系起源,但主要受瘤牛的影响。     

寿光鸡遗传多样性及其起源进化的研究

为了更好地对寿光鸡种质资源进行保护和开发利用,以线粒体基因组(mitochondrial genome,mt DNA) D-loop区为分子标记,检测了58只寿光鸡的遗传多样性,同时探讨了寿光鸡在5个红色原鸡亚种的进化地位。结果显示:寿光鸡mt DNA D-loop区全长1 231～1 232 bp,核苷酸多样性(Pi)为(0. 005 3±0. 001 35),单倍型多样性(Hd)为(0. 864±0. 018),平均核苷酸差异(K)为6. 525;群体共发现23处单核苷酸变异位点,并由此界定了8种单倍型;系统发育分析发现,寿光鸡主要分布在A、B、C和E 4个进化分支,构成比分别为43. 1%、36. 2%、12. 1%和8. 6%。结果表明:寿光鸡在线粒体水平上具有很高的遗传多样性,有4个母系来源,本研究为寿光鸡种质资源遗传评估、有效保种和选育利用提供了理论依据。     

泡状带绦虫线粒体部分基因克隆及序列比对分析

采集甘肃兰州市区宠物犬体内两株泡状带绦虫,运用分子生物学方法对其线粒体部分基因进行克隆,并使用基因分析软件Larsergene V7.1进行序列分析,结果发现这两株虫体同源性达到99.6%,与国内的泡状带绦虫广东虫株和甘肃虫株同源性分别达到98.7%和98.4%,鉴定为泡状带绦虫;与其他带科绦虫同源性均高于85%,低于88%。其中,与国内牛带绦虫同源性最高,为88%;与国内巨颈带绦虫同源性最低,只有85.1%。推断不同带科绦虫的cox1基因种间差异明显,在带科绦虫的分类上意义较大。     

The control of lymphoid leukosis in a flock White Plymouth Rock chickens

Summary

Lymphoid leukosis (LL) was successfully controlled in a commercial basic breeding line of White Plymouth Rock chickens. The control method has been developed for breeder flocks and consists of three elements: - In the flock under study, homogenates of embryos from all eggs collected during a number of I4‐day periods are tested for the presence of LL viruses.

- Only eggs from hens that have been shown not to shed virus in their eggs are used for the production of progeny. The offspring are reared in isolation during the first two months of life, at which time the age‐related resistance against tumour formation by LL viruses appears to be sufficiently developed.

- The chickens are subsequently inoculated intramuscularly with LL viruses of subgroups A and B transferred to a conventional chicken house.

The vaccination raises a solid immunity to horizontal LL virus exposure and, due to the age‐related resistance, tumour formation does not follow.

No excretion of LL viruses could be detected in three generations of White Plymouth Rock chickens to which the three elements of the control procedure were applied. Clinical disease was not observed in any of the chickens under notice.     

The control of lymphoid leukosis in a flock White Plymouth Rock chickens

Summary Lymphoid leukosis (LL) was successfully controlled in a commercial basic breeding line of White Plymouth Rock chickens. The control method has been developed for breeder flocks and consists of three elements: - In the flock under study, homogenates of embryos from all eggs collected during a number of I4-day periods are tested for the presence of LL viruses. - Only eggs from hens that have been shown not to shed virus in their eggs are used for the production of progeny. The offspring are reared in isolation during the first two months of life, at which time the age-related resistance against tumour formation by LL viruses appears to be sufficiently developed. - The chickens are subsequently inoculated intramuscularly with LL viruses of subgroups A and B transferred to a conventional chicken house. The vaccination raises a solid immunity to horizontal LL virus exposure and, due to the age-related resistance, tumour formation does not follow. No excretion of LL viruses could be detected in three generations of White Plymouth Rock chickens to which the three elements of the control procedure were applied. Clinical disease was not observed in any of the chickens under notice.     

新批准的西南地区5个山羊遗传资源mtDNA D-loop遗传多样性与亲缘关系研究

为了研究新批准的西南地区5个山羊遗传资源的遗传多样性及其与其他羊种的亲缘关系,试验利用设计的引物对山羊的mtDNA D-loop序列进行了PCR扩增和测序,对序列数据进行了核苷酸多样性、单倍型和亲缘关系分析。结果表明:在西南地区新批准的5个山羊遗传资源中,大足黑山羊、黔北麻羊和酉州乌羊平均多态位点数比较接近,均高于渝东黑山羊和贵州黑山羊;大足黑山羊、黔北麻羊和酉州乌羊遗传多样性高于贵州黑山羊和渝东黑山羊;从亲缘关系来看,渝东黑山羊与板角山羊和南江黄羊亲缘关系较近,酉州乌羊与板角山羊亲缘关系最近,贵州3个羊种(贵州白山羊、贵州黑山羊和黔北麻羊)遗传距离近,大足黑山羊与乐至黑山羊遗传距离较近。说明渝东黑山羊和贵州黑山羊遗传多样性较低,需要加强遗传资源的保护和提纯;新批准的5个山羊遗传资源的遗传距离和亲缘关系与其所处的地理位置存在密切关系。     

A Genetic Diversity Analysis of Mitochondrial DNA D-loop Region in Four High Quality Chicken Breeds

This study was conducted to elucidate the genetic diversity of mitochondrial DNA (mtDNA) D-loop region in Qingyuan partridge chicken group 1,Qingyuan partridge chicken group 2,Yangshan chicken and Qingyuan Yellow feather black-bone chicken.The specific primers were designed according to mtDNA D-loop region of Gullus gullus spadiceus (accession No.:NC_007235.1) in GenBank.The sequence was analyzed after PCR amplification and sequencing,and the haplotype number,polymorphism number,haplotype diversity,nucleotide diversity and nucleotide mean difference were counted.The evolution divergence among breeds was calculated by Mega 5.10 software,and the phylogenetic tree was constructed.The results showed that the length of mtDNA D-loop region in four high quality chicken breeds was 591 bp,and 549 bp were used for subsequent analysis.The content of A,T,C and G were 27.2% to 27.3%,30.1% to 30.4%,29.5% to 29.8% and 12.8% to 12.9%,respectively,and the average content of G+C was 42.5%.There were 92 polymorphic sites which contained 14 singleton variable sites and 78 parsimony informative sites,and the percentage of transitions and transversions were 89.13% (82/92) and 10.87% (10/92),respectively.The haplotype diversity ranged from 0.682 to 0.835,and the nucleotide diversity ranged from 0.00849 to 0.01167.There were 32 haplotypes in all sequences,which could be divided into clades A,B,C and E,however,most of the individuals belonged to clades B (51.2%) and E (37.6%).The phylogenetic tree results showed that four high quality chicken breeds could be classified as 4 branches which were consistent with the haplotypes classification results.The results indicated that the four high quality chicken populations from Qingyuan had relatively high haplotype and nucleotide diversity and likely shared two or more common maternal lineages.     

贵州威宁黄牛线粒体DNA D-loop区全序列分析

为了解贵州威宁黄牛的遗传多样性及遗传背景，测定了19个个体的线粒体DNAD-loop区全序列。威宁黄牛D-loop区全序列中，A＋T平均含量为61．4％，G＋C含量为38．6％。经比对，共检测到威宁黄牛D-loop区8种单倍型，核苷酸多态位点45个，其中7种为普通牛血统的单倍型，1种为瘤牛血统的单倍型，表明威宁黄牛同时受到普通牛和瘤牛的影响。在威宁黄牛19个个体中，其单倍型多样度为0．715，核苷酸歧异度（π值）为2．415％，表明威宁黄牛品种的遗传多样性丰富。     

贵州黎平黄牛线粒体DNA D-loop区全序列初步分析

为了解贵州黎平黄牛的遗传多样性及遗传背景,我们测定了20头黄牛的线粒体DNA D-loop区序列。结果检测到8种单倍型,其中4种为普通牛血统的单倍型,4种为瘤牛血统的单倍型,表明黎平黄牛同时受普通牛和瘤牛的影响。该研究对于黎平黄牛的保种及持续利用有着重要的理论指导意义。     

Genetic diversity of Bangladeshi native chickens based on complete sequence of mitochondrial DNA D-loop region

ABSTRACT

1. The aim of this study was to explore genetic diversity and possible origin of Bangladeshi (BD) native chickens. The complete mtDNA D-loop region was sequenced in 60 chickens representing five populations; naked neck, full feathered, Aseel, Hilly and autosomal dwarf. The 61 reference sequences representing different domestic chicken clades in China, India, Laos, Indonesia, Myanmar, and other Eurasian regions were included. The mtDNA D-loop sequence polymorphism and maternal origin of five BD populations were analysed.

2. A total of 35 polymorphic sites, and 21 haplotypes were detected in 60 mtDNA D-loop sequences. The haplotype and nucleotide diversity of the five populations were 0.921 ± 0.018 and 0.0061 ± 0.0019, respectively. Both mtDNA network and phylogenetic analysis indicated four clades (four haplogroups) in BD populations (21 haplotypes) along with 61 reference haplotypes. Clade E contained the most individuals (20) and haplotypes (11) of BD chickens, followed by clade D (17, 6), clade C (12, 2) and clade F (11, 2), respectively.

3. The higher number of unique haplotypes found in Yunnan, China, suggested that the origin of BD chickens was in this region. The haplotypes from different haplogroups were introduced in Bangladeshi chickens from India, China and Myanmar. The phylogenetic tree showed a close relationship of BD chickens with the clusters from India, China, Myanmar and Laos, and indicated the dispersion of BD chickens from these sources. The phylogenetic information revealed high genetic diversity of BD chickens because of their origin from different lineages with high genetic variation and distance, which was determined from four cluster and neighbour-joining trees.

4. In conclusion, BD populations had high genetic diversity. The mtDNA network profiles and phylogenetic trees showed multiple maternal origins of BD chickens from India, China, Myanmar and Laos.     

东方蜜蜂线粒体DNA应用进展

【研究目的】为掌握线粒体DNA（mtDNA）在东方蜜蜂研究中的取得的进展。【方法】本文总结了蜜蜂mtDNA结构特征、mtDNA标记研究技术和东方蜜蜂非编码区和编码区标记的应用情况。【结果】RNA^leu-COⅡ区和部分COⅡ区的酶切和测序均发现了丰富的多态性。RNA^leu-COⅡ区和部分编码区标记能较好地解决泰国、菲律宾东方蜜蜂的种群遗传分化问题。根据RNA^leu-COⅡ区的系统发育关系将东方蜜蜂分成5个类型,并推测了他们的地理发育关系。【结论]mtDNA标记是解决东方蜜蜂遗传学和进化学问题较好的标记,增加样本量和研究的区段能解决mtDNA存在的不足。     

Genetic Diversity Analysis of Three Newly Discovered Native Chicken Breeds in Yunnan Using mtDNA D-loop Region Sequences

This study was aimed to evaluate the information of their genetic background of Frizzle chicken (FM),Naked-neck chicken (CB) and YN chicken (YN) that were three newly discovered native chicken genetic resources with excellent characteristics,which had been found in Nujiang prefecture and mountainous area of Yunnan province.The variation in a total of 168 individuals sampled from the three chicken populations was assessed using the mitochondrial DNA (mtDNA) D-loop region sequences as genetic marker.The results showed that there were a total of 27 haplotypes were defined in the three native chicken as well as the haplotype diversity of these three chicken were 0.947,0.938 and 0.596,respectively.The nucleotide diversity of these three chicken were 0.01268,0.01434 and 0.00239,respectively.Phylogenetic tree displayed that all 168 individuals distributed in maternal lineage A,B,C,E,F and G.Frizzle chicken contained lineages E,F and G,and Naked-neck chicken contained all 6 lineages,of which the main lineages were E,F and G.YN chicken included lineages E,F and G,and lineage E was the highest percentage in this population.Also,it found that YN chicken was closely related to Gallus gallus murghi,White Plymouth Rock chicken,White Leghorn chicken and New Hampshires chicken,while Naked-neck chicken was closely related to Gallus gallus spadiceus,Gallus gallus jabouillei,Gallus gallus gallus Linnaeus,Indonesian cockfight and Laos chicken.The Frizzle chickens shared more haplotypes with Naked-neck chicken,and the evolutionary relationship between the two species was closer.This study provided a basis for the origin and genetic assessment of three newly discovered Yunnan local chicken breeds.     

Study on Genetic Diversity and System Evolution of mtDNA D-Loop Region in Xianglushan Chicken

This study was aimed to analyze the genetic diversity and phylogenetic evolution in Xianglushan chicken.The full-length sequences of mitochondrial DNA D-Loop (mtDNA D-Loop) region of 121 Xianglushan chickens were analyzed by PCR amplification combined with bidirectional sequencing,and the composition,variation and maternal origin of mtDNA D-Loop region were discussed.The results showed that the total length of mtDNA D-Loop region in Xianglushan chicken was 1 231-1 233 bp.The contents of A,T,C and G were 26.62%,33.55%,26.49% and 13.34%,respectively.The content of T was the highest,the content of G was the lowest,and the content of A+T was significantly higher than that of G+C,it indicated that region might have certain base hobbies.Analysis of 121 full-length sequences were found to coexist in 11 haplotypes and 26 mutation sites,of which 2 were single polymorphic information sites,24 were simple information sites,4 bases were inserted and 2 bases were missing.The genetic diversity analysis results showed that the haplotype diversity was 0.814,the nucleotide diversity was 0.00447,the average nucleic acid difference was 5.494,which indicated that the genetic diversity of Xianglushan chicken was relatively rich,the effect of preservation was better,which had a certain breeding space.Tajima's D was 0.39378 and the test results were not significant (P>0.10),in line with neutral mutations.The cluster analysis results showed that Xianglushan chicken and Gallus gallus gathered as one,indicating that Xianglushan chicken originated from Gallus gallus, and there were 4 branches inside the branch,indicating that Xianglushan chicken had many matrilineal origins.The results could provide some reference data for the protection and exploitation of pheasant germplasm resources in Xianglushan chicken.     

中国地方猪种线粒体基因组遗传多样性及其系统发育分析

为从分子水平上探究中国地方猪种遗传多样性和分类地位,本试验采用生物信息学方法比较了6个类型共22个中国地方猪种的线粒体基因组全序列,分析了其多态性,并构建了6个类型猪种线粒体D-loop区单倍型的网络中介图以及基于线粒体D-loop序列、Cytb基因、完整编码区序列的系统进化树。结果表明,6个类型22个猪种中共检测到了144个多态位点,22种单倍型,说明地方猪种具有丰富的遗传多样性;地方猪线粒体基因组序列中核苷酸变异以转换为主,且Ti/Tv大于转换/颠换比临界值（2.0）,变异位点均符合中性突变。6个类型猪种间遗传距离均较小,且有共享单倍型。系统进化树结果表明,6种类型地方猪种主要聚为两个支系。表明线粒体D-loop序列及Cytb基因均可作为研究种内系统发育、起源进化的分子标记。     

Genetic Diversity and Phylogenetic Relationships of the Complete Mitochondrial Genome in Chinese Indigenous Pig Breeds

In order to explore the genetic diversity and the classification status of the Chinese domestic pigs at the molecular level, the complete mitochondrial DNA (mtDNA) genomic sequences of 22 Chinese indigenous pig breeds from 6 types were downloaded from GenBank for the analysis using the bioinformatics method. The polymorphism of nucleic acid sequences was analyzed,the molecular phylogenetic tree based on D-loop sequences, Cytb gene, complete coding region mtDNA sequences and the Median-Joining network of the mtDNA D-loop haplotypes were constructed for 22 pig breeds from 6 types. The results showed that a total of 144 mutation sites among 22 pig breeds were detected, 22 haplotypes were discovered, which indicated that Chinese indigenous pig breeds had abundant genetic diversity. According to the complete mtDNA genomic sequences analysis, which suggested that the main mutations was transition, the transition/transversion ratio was greater than 2.0 and all mutations were in line with the neutral mutation. Our findings clearly demonstrated that there was a near genetic distance among 6 types, meanwhile, the shared haplotypes were found. The phylogenetic tree showed that Chinese indigenous pigs from 6 types were diverged from two ancestors.The results indicated that mtDNA D-loop and Cytb gene might serve as molecular markers for phylogenesis, origin and evolution.     

Differences in genetic structure assessed using Y‐chromosome and mitochondrial DNA markers do not shape the contributions to diversity in African sires

Up to 173 African sires belonging to 11 different subpopulations representative of four cattle groups were analysed for six Y‐specific microsatellite loci and a mitochondrial DNA fragment. Differences in Y‐chromosome and mtDNA haplotype structuring were assessed. In addition, the effect of such structuring on contributions to total genetic diversity was assessed. Thirty‐five Y‐chromosome and 71 mtDNA haplotypes were identified. Most Y‐chromosomes analysed (73.4%) were of zebu origin (11 haplotypes). Twenty‐two Y‐haplotypes (44 samples) belonged to the African taurine subfamily Y2a. All mtDNA haplotypes belonged to the “African” taurine T1 haplogroup with 16 samples and nine haplotypes belonging to a recently identified subhaplogroup (T1e). Median‐joining networks showed that Y‐chromosome phylogenies were highly reticulated with clear separation between zebu and taurine clusters. Mitochondrial haplotypes showed a clear star‐like shape with small number of mutations separating haplotypes. Mitochondrial‐based F_ST‐statistics computed between cattle groups tended to be statistically non‐significant (p > .05). Most F_ST values computed among groups and subpopulations using Y‐chromosome markers were statistically significant. AMOVA confirmed that divergence between cattle groups was only significant for Y‐chromosome markers (Φ_CT = 0.209). At the mitochondrial level, African sires resembled an undifferentiated population with individuals explaining 94.3% of the total variance. Whatever the markers considered, the highest contributions to total Nei's gene diversity and allelic richness were found in West African cattle. Genetic structuring had no effect on patterns of contributions to diversity.