梨属叶绿体非编码区trnL-trnF和accD-psaI特征及其在系统发育研究中的应用价值

 对梨属19个种共26个供试样本及两个苹果属外类群样本的叶绿体非编码区trnL-trnF和accD-psaI进行了序列分析。排列后的序列长度分别为927 bp和944 bp。两个区域组合后共有36个核苷酸替换（substitution）和11个插入/缺失（indel）。将所有indel处理为一次变异事件并编码为替换,用于简约法系统树和NeighborNet系统发育网络（phylogenetic networks）的构建。研究结果表明：indel为系统发育分析提供了可靠的信息;相比trnL-trnF,accD-psaI因进化速率较快,更适合应用于种及种以下分类水平的梨属植物系统关系研究;NeighborNet系统发育网络表明系统树上部分较低的分支支持率由冲突的信息引起,其余则由信息位点不足引起。此外结果还揭示了可疑种间杂交种的母系祖先。     

三江黄牛全基因组数据分析

【目的】研究三江黄牛群体遗传多样性,从基因组层面讨论其群体遗传变异情况。【方法】提取50个体基因组总DNA,等浓度等体积混合,构建混合样本DNA池,利用CovarisS2进行随机打断基因组DNA,电泳回收长度500 bp的DNA片段,构建DNA文库。应用Illumina HiSeq 2000测序,最终得到测序数据。利用BWA软件将短序列比对到牛参考基因组(UMD 3.1),来检测三江黄牛基因组突变情况。SAMtools、Picard-tools、GATK、Reseqtools对重测序数据进行分析,Ensemb1、DAVID、dbSNP数据库对SNPs和indels进行注释。【结果】全基因组重测序分析共计得到77.8 Gb序列数据,测序深度为25.32×,覆盖率为99.31%。测序得到778 403 444个reads和77 840 344 400个碱基,比对到参考基因组(UMD 3.1)的reads为673 670 505,碱基为67 341 451 555,匹配率分别为86.55%和86.51%,成对比对上的reads数为635 242 898(81.61%),成对比对上的碱基数为63 512636 924(81.59%);共确定了20 477 130个SNPs位点和1 355 308个indels,其中2 147 988个SNPs(2.4%)和90 180个indels(6.7%)是新发现的。总SNPs中,鉴定出纯合SNPs989 686(4.83%),杂合SNPs19 487 444(95.17%),纯合/杂合SNP比为1:19.7。转换数为14 800 438个,颠换为6 680 058个,转换/颠换(TS/TV)为2.215。剪切位点突变SNP727个,开始密码子变非开始密码子SNP117个,提前终止密码子的SNP 530个,终止密码子变非终止密码子SNP88个。检测到非同义突变数为57 621,同义突变为83 797,非同义/同义比率为0.69。检测到非同义SNPs分布在9 017个基因上,其中发现567个基因与已报道的重要经济性状相符,肉质、抗病、产奶、生长性状、生殖等相关基因的数量分别为471、77、21、10、8个,其中包括功能相重叠的基因;indels数据中,缺失数量为693 180(51.15%),插入数量为662 148(48.85%),纯合indels数量为161 198(11.89%),杂合indels数量1 194 110(88.11%),大部分的变异都位于基因间隔区和内含子区;三江黄牛全基因组杂合度(H)、核苷酸多样性(Pi)及theta W分别为7.6×10~(-3)、0.0 039、0.0 040,说明其遗传多样性较为丰富。三江黄牛群体Tajima'D为-0.06 832,推测可能由于群体内存在不平衡选择所致。【结论】本研究为进一步分析与经济性状相关的遗传学机制和保护三江黄牛品种遗传多样性提供了基因组数据支持。     

BSE‐associated polymorphisms in the prion protein gene: an investigation

The aim of this study was to determine the frequency of the 12‐bp and 23‐bp indel polymorphisms in the prion protein gene (PRNP) in cattle and to investigate the association between these frequencies and the occurrence of bovine spongiform encephalopathy (BSE). There was no significant difference in the 12‐bp indel frequency between the BSE animals and control group. For the 23‐bp indel, the BSE animals had a significantly lower + + (insins) genotype frequency and + allele frequency compared with the control animals. The ? ? / ? ? genotype frequency in the BSE animals was not significantly higher when compared with the control animals. One ? allele increased the risk of BSE by a factor of 1.55 (i.e. by 55%) for the 12‐bp indel and by a factor of 2.10 for the 23‐bp indel. When both indels are considered, one ? allele increased the risk of BSE by a factor of 1.54.     

Detection of prion gene promoter and intron1 indel polymorphisms in Anatolian water buffalo (Bubalus bubalis)

Bovine spongiform encephalopathy (BSE) is a fatal disease caused by miss folded prion protein. Studies in the cattle, comparing genetic data from BSE diseased and healthy animals have shown that indel polymorphisms in the promoter and intron 1 of PRNP gene were associated with disease susceptibility. Several studies were conducted to find out allele and genotypic frequencies of indel polymorphisms in promoter and intron 1 of the cattle PRNP gene. Unlike domestic cattle and bison, no indel polymorphisms of the PRNP promoter and intron 1 were examined in any population of the water buffalo (Bubalus bubalis). Aim of this study was to analyse frequencies of allele, genotype, and haplotype of the indel polymorphisms (23 bp indel in promoter and 12 bp indel in intron 1) in prion protein coding gene (PRNP) of water buffalo. Therefore a PCR based procedure, previously used in cattle to detect indel polymorphisms of PRNP promoter and intron 1 locus, was applied to 106 Anatolian water buffalo DNAs. Our results have revealed high frequency of in variants and in23/in12 haplotype for PRNP promoter and intron 1 indel polymorphisms in water buffalo. The results of the study have demonstrated that frequencies of allele, genotype, and haplotype of the indel polymorphisms in PRNP gene of the Anatolian water buffalo are significantly different those from cattle and bison PRNP indel polymorphisms.     

Association of growth traits with a structural variation downstream of the KCNJ11 gene: a large population-based study in chickens

ABSTRACT

1. The potassium voltage-gated channel subfamily J member 11 gene (KCNJ11) is involved in the insulin secretion pathway. Studies have shown that mutation in this gene is associated with muscle weakness. The objective of the present study was to establish the association between KCNJ11 gene polymorphism and chicken growth performance and to analyse its expression pattern.

2. A novel 163-bp insertion/deletion (indel) polymorphism was identified in the region downstream of the KCNJ11 gene in 2330 individuals from ten populations by polymerase chain reaction (PCR). An F₂ resource population was used to investigate the genetic effects of the chicken KCNJ11 gene. Association analysis showed that the indel was significantly associated with chicken growth traits and that the phenotypic value of the ins-ins (II) genotype is higher than that of the ins-del (ID) and del-del (DD) genotypes.

3. Gene expression for different genotypes showed that birds carrying the II allele had a higher expression level than the DD genotypes. Analysis of tissue and spatiotemporal expression patterns indicated that the KCNJ11 gene was highly expressed in muscle tissues, with the highest levels in muscle tissue at one week of age, and that a 10% crude protein diet reduced the expression of this gene, average daily gain and muscle fibre diameter.

4. The results suggested that this novel 163-bp indel has the potential to become a new target for marker-assisted selection.     

Indel variants within the PRL and GHR genes associated with sheep litter size

Growth hormone and prolactin belong to the class of peptide hormones that have a wide range of regulatory functions. In this study, polymorphisms of growth hormone receptor (GHR) and prolactin (PRL) genes were analysed as candidate genes, which are responsible for the litter size in Australian White (AUW) sheep. According to the statistical analyses results, the polymorphism information content (PIC) values of the PRL-P1-ins-23 bp, GHR-P2-del-23 bp and GHR-P8-del-23 bp were 0.371, 0.366 and 0.375, respectively, which indicates the high genetic polymorphism in AUW sheep. Moreover, all indel loci are not conformed to the HWE (p < .05). Further, our findings revealed that the PRL-P1-ins-23 bp polymorphism in the ovine PRL gene was significantly related to the first parity litter size (p = .001) and the DD genotype displaying the highest genotypic mean. Meanwhile, the GHR-P2-del-23 bp and GHR-P8-23 bp indels in the ovine GHR gene were significantly correlated with first parity litter size (p < .05), and the individuals with the genotype II showed significantly higher litter size than others. Collectively, these results demonstrated that our findings could be useful for future sheep breeding strategies based on the molecular-assisted selection (MAS).     

催乳素基因5''-调控区24-bp插入/缺失变异与非就巢鸡血浆催乳素水平及产蛋性能的关系

以笼养绿壳蛋鸡和农大3号矮小型蛋鸡为试验材料，探索蛋鸡在非就巢状态下催乳素基因5’-调控区24-bp插入／缺失（-377--354bp）变异与血浆PRL水平和产蛋性能的关系。结果表明，该位点不同基因型（PRL＋／＋、＋／-和-／-型）对非就巢蛋鸡300日龄前的血浆PRL水平和300日龄（40周龄）产蛋性能均无显著影响（P〉0．05），表明鸡在非就巢状态下，PRL调控区的24-bp插入／缺失变异与早期血浆PRL水平和产蛋性能无直接关系。     

催乳素基因5′-调控区24-bp插入/缺失变异与非就巢鸡血浆催乳素水平及产蛋性能的关系

以笼养绿壳蛋鸡和农大3号矮小型蛋鸡为试验材料,探索蛋鸡在非就巢状态下催乳素基因5′-调控区24-bp插入/缺失(-377~-354 bp)变异与血浆PRL水平和产蛋性能的关系。结果表明,该位点不同基因型(PRL / 、 /-和-/-型)对非就巢蛋鸡300日龄前的血浆PRL水平和300日龄(40周龄)产蛋性能均无显著影响(P>0.05),表明鸡在非就巢状态下,PRL调控区的24-bp插入/缺失变异与早期血浆PRL水平和产蛋性能无直接关系。     

Polymorphisms of the growth hormone gene in domesticated red sea bream populations (Pagrus major) based on minisatellite genotypes and nucleotide sequences

The genetic diversity of the growth hormone gene in domesticated red sea bream (pmaGH) was evaluated using a minisatellite DNA marker located in intron 3 (pmaGH22) and nucleotide sequences. The number of alleles of pmaGH22 was largely decreased in domesticated strains of red sea bream, and the possibility of selection pressures was also detected based on the analysis of the Hardy–Weinberg equilibrium in some strains. However, each strain inherited a small number of alleles of pmaGH22, and the entire domesticated population (combining all strains) showed a large number of alleles (n = 17), similar to the allelic richness of the wild population (n = 18.5). Based on nucleotide sequencing analysis, three synonym mutations were found in the coding regions, and also several SNPs and indels were found in the noncoding regions. In addition, four genealogies of growth hormone haplotypes were identified based on principal coordinate analysis, and these genealogies of pmaGH partly reflected allele size ranges of pmaGH22. Several haplotypes shared alleles of pmaGH22, and also fragment size homoplasy in pmaGH22 was suspected. These alleles of pmaGH22 and the haplotypes will be a useful indicator for divergence of pmaGH and for broodstock individual selection with minimum inbreeding effect.