贵州下司犬线粒体DNA控制区核苷酸序列分析

采用聚合酶链式反应和直接测序的方法,分析了贵州下司犬线粒体D-loop区序列。结果显示,该犬的线粒体D-loop全长1260bp,碱基序列中A、T含量明显高于G、C含量。其中有一段以串连方式排列、重复29次的序列,重复单元为TACACGT(A/G)CG。对该串连重复序列上游673bp的碱基序列进行分析,比较已知犬种相应碱基序列之间的差异,聚类分析结果显示下司犬与亚洲犬种的亲缘关系较近。     

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.     

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.     

黄河下游同域山羊种群mtDNA D-环多态性及系统发育

对黄河下游7个山羊种群共59个个体的mtDNA D-环533 bp序列进行分析,发现59个多态位点,其中单一多态位点21个,简约信息位点38个,分别占全序列的11.7%、3.94%和7.13%。确定了37种单倍型,崂山奶山羊1个个体还发现了1处5碱基的缺失。各群体单倍型多样度为0.785 7~0.969 7,核苷酸多样度为1.27%~1.73%,遗传多样性较丰富。构建系统发育树将37种单倍型分为2个分支,由此推断黄河下游山羊资源至少具有2个母系起源,角猾羊与A类型聚为1支,7个群体具有角猾羊起源,未发现捻角山羊对7个群体有贡献的证据。错配分析表明,A类型经历过群体扩张,B类型未经历过群体扩张。并通过遗传距离分析了各群体的遗传分化。     

云南龙陵黄山羊线粒体DNA限制性酶切分析

 采用碱性变性法提取来自于龙陵县不同地区的18只黄山羊个体的线粒体DNA(mtDNA),并用ApaⅠ,AvaⅠ,BamHⅠ,BclⅠ,BglⅠ,BglⅡ,ClaⅠ,DraⅠ,EcoRⅠ,EcoRⅤ,HaeⅠ,HindⅢ,KpnⅠ,PstⅠ,PvuⅡ,SacⅠ,SalⅠ,SmaⅠ,StuⅠ和XhoⅠ等20种限制性内切酶进行酶切分析.结果发现龙陵黄山羊线粒体DNA的分子量大小约为15.8Kb;不同酶的酶切位点分别为:DraⅠ有7个酶切位点,AvaⅡ有6个酶切位点,EcoRⅤ和StuⅠ共有5个酶切位点,HindⅢ和HeaⅡ有4个酶切位点,BamHⅠ,BglⅡ,PstⅠ和PvuⅡ有3个酶切位点,ApaⅠ,ClaⅠ有2个酶切位点,其余有1个酶切位点.     

东昌湖野生鲤鱼线粒体DNA RFLP分析

利用差速离心法及RNase消化法制备并纯化了采自山东东昌湖野生鲤(Cyprinus carpio haematopterus)的肝脏及性腺线粒体DNA,用7种限制性内切酶对其mtDNA进行酶解,经琼脂糖凝胶电泳分离酶解片段,用Gel-5000凝胶图像分析系统进行采集和分析。结果显示,东昌湖野生鲤的mtDNA分子大小为(16.62±0.15)kb,BglⅠ、BglⅡ、EcoRⅠ、HindⅢ、PstⅠ、KpnⅠ和BamHⅠ的酶切点分别为3或2、1、3或4、4、1、0、3;其中,BglⅠ、EcoRⅠ和BamHⅠ内切酶具有限制性多态性,多态酶比例为42.86%,并检测出12个限制性态型,可归并为5种单倍型,各单倍型间的遗传距离0.0075～0.0365,揭示了东昌湖野生鲤存在较高的种内多态性。试验结果与以前报道的其他地区鲤属线粒体DNA酶切结果相比较,表明鲤属mtDNA在限制性酶切位点数量及其长度上存在地域差异。     

二倍体和四倍体泥鳅线粒体ＮＤ ５／６基因多态性的ＰＣＲ ＲＦＬＰ分析

采用PCR-RFLP分析方法,对洞庭湖、武汉两地的二倍体和四倍体泥鳅线粒体DNAND-5/6基因多态性及4个群体遗传变异和遗传关系进行了研究。结果表明,120尾泥鳅mtDNAND-5/6扩增片段长度均为2.2kb;从13种限制性内切核酸酶中筛选出6种多态性内切酶(HaeⅢ、DraⅠ、RsaⅠ、TaqⅠ、HinfⅠ、MspⅠ),对扩增产物进行酶切,共检测到66种单倍型。泥鳅群体内单倍型多样性和核苷酸多样性分别为0.7679～0.9385和0.01041～0.03212;其中,洞庭湖二倍体核苷酸多样性最高(0.03212),武汉二倍体其次(0.02452),二倍体群体内核苷酸多样性高于四倍体。群体间的核苷酸多样性(π)大小为0.02588～0.04144,平均值为(0.031737±0.000005)。群体间的核苷酸歧化距离(δ)大小为0.00462～0.01617,平均值为(0.010659±0.000003);其中,武汉二倍体和四倍体之间的核苷酸歧化距离最大(0.01617),武汉四倍体和洞庭湖二倍体之间的核苷酸歧化距离最小(0.00462)。MonteCarlo模拟x2检验表明,4个群体间的单倍型频率存在极显著差异(P<0.0001)。UPGMA聚类分析表明,武汉四倍体、洞庭湖二倍体和四倍体聚为一支,亲缘关系较近;武汉二倍体为单独一支。     

基于3种线粒体基因的珠母贝属系统进化关系

为了研究珠母贝属的分类地位和系统演化情况,通过聚合酶链式反应技术(PCR)扩增并直接测序出雷州半岛珠母贝属中的马氏珠母贝、大珠母贝、珠母贝、斑珠母贝和黑珠母贝的3种线粒体基因(12S r RNA、16S r RNA和COⅠ)部分序列,分析其碱基组成和种间遗传距离。同时结合Gen Bank上发表的白珠母贝序列,构建珠母贝属的分子系统树。结果显示,5种珍珠贝的3种基因部分序列碱基AT含量大于GC含量,与其他无脊椎动物线粒体DNA序列基本一致,序列都处于高度饱和的状态。系统分析显示,构建的分子系统树与传统的形态分类基本一致。在黑珠母贝和白珠母贝亲缘关系上,3种线粒体DNA片段在碱基组成、种间遗传距离和系统进化树上都显示黑珠母贝和白珠母贝非常相近,可以认为是同一种中的2个亚种。在斑珠母贝的亲缘关系上,系统分析显示斑珠母贝与黑珠母贝和白珠母贝更为接近。研究表明,大珠母贝和珠母贝在系统进化中较早的分离出来,是一个比较原始的种类。3种分子标记对马氏珠母贝亲缘关系分析上存在一定差异,根据现有的数据尚不足以得出结论,需进一步做分子系统研究并结合形态特征和解剖结构进行分析。     

橘小实蝇不育雄虫与野生雄虫间遗传分化

对橘小实蝇不育雄虫和福建7个不同地区的野生雄虫共37个个体的线粒体细胞色素氧化酶I（mtDNA COI）基因进行了部分序列测定。在获得的650bp序列中,共有72个变异位点、32个单倍型,其中共享单倍型3种;对所有单倍型聚类分析发现,单倍型在系统树中的分布散乱,没有显示出明显的地理分布族群;结合遗传变异指数（Fst）和净遗传距离（Da）分析认为橘小实蝇不育雄虫与野生虫源间、不同来源的野生虫源间已存有一定程度的遗传分化,但分化程度较低。研究认为应用昆虫不育技术防治橘小实蝇,在饲养过程中需针对不同防治区引进野生虫源复壮,以确保不育雄虫和野生虫源相匹配,提高防治效果。     

小麦质核互作型雄性不育系线粒体DNA变异性的RAPD分析(英文)

在杂交小麦选育中,为了更好地利用细胞质雄性不育性,研究与不断挖掘新的不育细胞质类型及其对应育性恢复基因以改良现有不育资源至关重要.基于这一目的,本文对具有山羊草属细胞质的四类不育系线粒体DNA(mtDNA)进行了RAPD分析,分别比较了具有同一细胞质背景的山羊草、雄性不育系,以及该类不育系与恢复系组配的可育杂种F1的mtDNA的变异性.结果显示,供试山羊草与其对应细胞质雄性不育系在mtDNA上存在明显多态性,表明不育系在质核互作的影响下很可能已导致mtDNA发生变异:而不育系与对应的可育杂种F1在mtDNA上也存在多态性,同样表明育性恢复核基因对不育系进行育性恢复的过程中亦可能引起mtDNA发生相应变异;mtDNA变异很可能涉及到不育系育性本质的改变.