苏姜猪A-FABP基因遗传多态性及其与肉质性状的关联分析

研究采用PCR-RFLP技术分析苏姜猪、姜曲海猪、长白猪群体A-FABP基因遗传多态性,并用统计学方法分析基因座多态与肉质性状的相关性。结果表明:3个猪种A-FABP基因内含子1均具有Bsm I酶切多态性,分别检测到3种基因型(AA、AB和BB),χ~2适合性检验显示,3个猪种A-FABP基因型分布处于Hardy-Weinberg平衡(P0.05)。多态信息含量(PIC)分析显示,3个品种猪均为中度多态;基因型与肉质性状关联分析显示,不同基因型肌内脂肪含量、大理石纹差异显著,大理石纹AA、AB型显著高于BB型(P0.05),肌内脂肪含量AA显著高于AB、BB型(P0.05)。结论显示,苏姜猪、姜曲海猪、长白猪A-FABP基因intron 1多态性对肉质性状影响显著。     

Prevalence and molecular analysis of Sarcocystis infections in cattle in Northwest Iran and the first global report of S. gigantea in cattle

Cattle are intermediate host for several species of Sarcocystis, including S. cruzi, S. hirsuta, and S. hominis with high prevalence worldwide. The present study aimed to determine the prevalence of Sarcocystis infection, species identification, and phylogenetic analysis of the parasite in cattle in Northwest Iran. The samples of diaphragm and esophagus from 290 cattle were collected from slaughterhouses in Northwest Iran and subjected to macroscopic, microscopic, and histopathology examinations, PCR-RFLP, sequencing and phylogenetic analyses. Tissue cysts of Sarcocystis spp. were detected in 92% of cattle by digestion and microscopic tests. Based on the PCR–RFLP and specific PCR, 87.9% and 1.03% of isolates were identified as S. cruzi, and S. hominis, respectively. Macrocyst was seen in a single sample that was identified as S. gigantea. The haplotype network exhibited the extension of the various haplotypes of S. cruzi between neighboring provinces in Northwest Iran. Heterogeneity analysis of S. cruzi 18S-rRNA sequences indicated genetic diversity among S. cruzi isolates (Haplotype diversity: 0.733-0.854) consisting 16 haplotypes; however, the nucleotide differences showed low diversity (0.01481 to 0.03351). Pair wise sequence distance matrix amongst S. cruzi sequences indicated an intra-species divergence of 0%-7.8% and identity of 92.6%-100%. Sarcocystis infection is highly prevalent in cattle in Northwest Iran, with the predominance of S. cruzi, and genetic variants of this species are unequivocally distributing in Northwest provinces. First global detection of S. gigantea in cattle reflects new insights of transmission dynamic and biology of this parasite in Iran.     

羊流产衣原体的分离鉴定及间接免疫荧光法检测

对收集的疑似衣原体感染的羊流产胎儿样本接种鸡胚卵黄囊进行分离培养,通过PCR-RFLP(限制性片段长度多态性分析)方法鉴定为流产衣原体。将收集到的流产衣原体阳性样本接种Hela229细胞,应用基于流产衣原体MOMP单克隆抗体的间接免疫荧光法,在Hela229细胞浆内可见呈绿色荧光的衣原体包涵体,进一步在病原学水平上确定发生在甘肃省肃南县羊流产的病原为流产衣原体。     

First molecular identification and subtype distribution of Blastocystis sp. isolated from hooded crows (Corvus cornix) and pigeons (Columba livia) in Tehran Province,Iran

Blastocystis is a common intestinal parasite among humans and animals such as non-human primates, pigs, cattle, birds, amphibians, and less frequently, rats, reptiles and insects. Since Blastocystis is a widely transmissible parasite between humans and mammals or birds, it is prominent to determine whether newly secluded non-human isolates are zoonotic. There are no comprehensive studies in Iran assessing the prevalence and molecular identification of Blastocystis infection in birds, especially in pigeons and crows. So, the aim of this study was to identify Blastocystis subtypes (STs) in crows and pigeons in Tehran province, Iran, using Nested PCR-RFLP and sequencing. Overall, 300 Blastocystis isolates from birds (156 pigeons and 144 crows) were subtyped by PCR, and the homology among isolates was then confirmed by RFLP analysis of the 18S rRNA gene. The prevalence of Blastocystis infection was detected 42.9% in pigeons and 44.4% in crows. All positive pigeons were owned by ST13 (100%). Among crows, 46 samples (71.8%) like pigeons were ST13, and 13 samples (20.3%) were ST14. Five samples (7.9%) remained unknown. This study was the first report of ST13 and ST14 of Blastocystis from birds. In the present study, our data revealed a high prevalence of Blastocystis sp. in pigeon’s and crow’s samples and the isolates from these birds were classified into two genetically distinct STs. Therefore, birds appear to be infected with various STs. It is important to determine the phylogenetic relationships between unknown STs from these birds and the multiple STs of Blastocystis.     

猪ECH1基因部分序列的遗传变异分析

为了进一步研究烯脂酰辅酶A水解酶1(enoyl CoA hydratase,ECH1)基因的生物学功能,研究采用克隆测序结合PCR-RFLP的方法分析了民猪ECH1基因的部分DNA序列,并对其中的1个点突变进行了3个猪种内的基因型频率和基因频率计算。结果表明:研究所检测的ECH1基因序列与网上已有序列相比存在8个单核苷酸多态性(SNPs)位点,其中有2个造成酶切位点的改变;民猪和大白猪在PCR-RFLP-BamHⅠ位点的A、B基因频率均接近0.5,而长白猪B为优势等位基因。     

淮南猪酸肉基因的多态性分析

为了研究淮南猪的种质特性,为淮南猪保种、可持续利用及种质创新提供依据,试验采用PCR-RFLP法,对96头淮南猪的RN基因进行了检测。结果表明,RN基因在淮南猪群体中存在隐性纯合子rn^+/rn^+和杂合子RN^-/rn^+,未发现显性纯合子RN^-/RN^-。其中,基因型rn^+/rn^+和RN^-/rn^+的频率分别为0.739 6和0.260 4;等位基因rn^+和RN^-的频率分别为0.869 8和0.130 2。卡方检验表明,rn^+和RN^-基因频率处于Hardy-Weinberg平衡状态,表明群体中RN基因没有受到选择的压力。     

Polymorphism of IGFBP2 Gene and Its Association with Meat Quality Traits in Pigs

This study was aimed to analyze the genetic variation of insulin-like growth factor binding protein 2 (IGFBP2) gene effect on meat quality traits in pigs. A pair of primer was designed,which based on DNA sequence of IGFBP2 gene (GenBank accession No.:BV727778). The genetic polymorphism of IGFBP2 gene was detected by PCR-RFLP in Sujiang pig, Jiangquhai pig and Landrace pig, and the association between IGFBP2 gene polymorphism and meat quality traits in three pig breeds was analyzed. The results showed that three genotypes (AA, AB and BB) were detected. The Chi-square test results showed that the genotype distribution of three pig breeds was in Hardy-Weinberg equilibrium (P > 0.05). The polymorphism information content (PIC) were moderately polymorphic in three pig breeds. According to the association analysis of different genotypes, significant difference was found in color value, intramuscular fat content and marbling, the color value (except for Jianquhai pig) and intramuscular fat content of AA genotype were significantly higher than AB and BB genotypes (P < 0.05), the marbling of AA and AB genotypes was significantly higher than BB genotype (P < 0.05). In conclusion,the polymorphism of IGFBP2 gene intron 3 had significant effects on meat quality traits, and was candidate gene for the potential impact on meat quality traits of pigs.     

不同品种猪PBD-124基因多态性及差异表达研究

【目的】 克隆获得猪β-防御素-124（porcine beta-defensin-124,PBD-124）基因CDS区并探究其多态性,分析PBD-124基因在不同品种猪及同种猪不同组织内的表达情况。【方法】 采用RT-PCR方法扩增并克隆猪PBD-124基因CDS区,利用PCR-RFLP酶切法对大白猪、民猪和野杂猪PBD-124基因的Bln Ⅰ酶切位点进行多态性检测,利用实时荧光定量PCR方法检测该基因在不同品种猪肝脏、脾脏和血液内的表达差异。【结果】 试验成功克隆出猪PBD-124基因CDS区,长423 bp,共编码140个氨基酸,测序结果发现其存在c.257 G>A和c.263 T>G 2个突变位点,因2个突变位点间隔过近,可能存在连锁,仅对第1个突变位点进行酶切,大白猪中检测到GG、GA、AA 3种基因型,而民猪和野杂猪中仅检测到GA和AA 2种基因型,3个群体中AA基因型均为优势基因型,大白猪、民猪和野杂猪的AA基因型频率分别为0.5261、0.9412和0.6452,且3个群体均处于Hardy-Weinberg平衡（P>0.05）。3个群体的多态性均不高,大白猪处于中度多态（0.25<PIC<0.5）,民猪和野杂猪则处于低度多态（PIC<0.25）。实时荧光定量PCR结果显示,PBD-124基因在大白猪、民猪、野杂猪的脾脏、肝脏及血液中均有表达,且存在表达差异。【结论】 猪PBD-124基因CDS区长423 bp,共编码140个氨基酸,与参考序列比对发现2个突变位点,均未引起氨基酸突变;3个群体多态性均不高;PBD-124基因在不同品种猪及同种猪不同组织间表达均存在差异。     

Overcoming hybridization barriers between pea and some of its wild relatives

Pea would benefit from the plasticity and adaptability of its cross-incompatible relatives Pisum fulvum and Lathyrus sativus L., and we have tested reciprocal sexual crossings by manually cross-pollinating plants of genotypes of these three species. Studies of in situ germination of pollen grains on stigmata showed that pollen tubes were generally unable to germinate or could not reach the ovary. A few putative hybrid pods were nevertheless harvested, with one grain per pod germinated in vitro, then micropropagated for flow cytometry, isoenzyme, molecular (ribosomal ITS PCR-RFLP) and genomic in situ hybridization (GISH) studies. One such grain was recovered from an inter-generic cross of P. sativum x L. sativus and four from an inter-specific P. sativum x P. fulvumcross. A strong cross-incompatibility was shown between pea and grass pea, where the putative hybrid turned out to be pea. Conversely, with the interspecific, P. sativum x P. fulvum cross, flow cytometry and isoenzymes with leaf tissues strongly suggested hybridity, while molecular approaches and GISH confirmed the production of inter-specific hybrids, and without the need for a wild type P. sativum accession as a bridging cross.     

A molecular technique for selection of self-compatible varieties of Japanese pear (Pyrus pyrifolia Nakai)

The pear cultivar ‘Osa-Nijisseiki’ (S₂S^sm ₄; sm = stylar-part mutant) has been used as a parent to breed self-compatible cultivars that produce excellent fruits. However, determination of the self-compatibility of ‘Osa-Nijisseiki’ offspring requires a lot of time, 6 years or more, by conventional cross breeding. We have designed a rapid reliable method for the identification of self-compatible varieties of ‘Osa-Nijisseiki’ offspring based on the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) with S-allele specific restriction endonucleases. By using this method, 8 self-compatible varieties were selected among 16 selections resulting from a cross between the self-compatible cultivar ‘Osa-Nijisseiki’ (S₂S^sm ₄) and the self-incompatible cultivars ‘Niitaka’ (S₃S₉), ‘Whasan’ (S₃S₅), ‘Chuwhangbae’ (S₄S₆). The S-genotypes of 16 ‘Osa-Nijisseiki’ offsprings were also determined. This revised version was published online in August 2006 with corrections to the Cover Date.