德南牛全基因组变异解析和功能基因挖掘

[目的]研究德南牛的群体遗传结构和解析影响其重要性状的关键基因。[方法]通过主成分分析、群体进化树分析、群体遗传结构分析等方法,研究德南牛的遗传结构和与其他品种的亲缘关系;同时,利用全基因组SNP数据,计算德南牛与德国黄牛、南阳牛群体固定指数(Fst)和核苷酸多样性比值(π ratio)、复合似然比(CLR)、XP-EHH等多种选择信号指标,以筛选出德南牛基因组上受到选择的信号,鉴定出重要的候选基因。[结果] 德南牛具有欧洲普通牛(0.789)、东亚牛(0.176)、中国瘤牛(0.035)的混合血统。在θπ和CLR方法中鉴定出93个共有基因,使用FST和XP-EHH方法鉴定出17个共有基因。进行富集以及基因功能注释后,发现这些基因可能与生殖性能(TUBB3、NSD2、GRM1、ERBB4)、生长性状(RFX3、FGFR3、GABRB1、PDE4D)、环境适应性(PGR、GRIA4)和免疫系统反应(P2RY12、P2RY13、GPR87、P2RY14、GPR171)有关。[结论]本项研究发现德南牛血统受德国黄牛影响较多,且挖掘了德南牛基因组内可能具有经济意义的性状相关的选择特征,为德南牛种群的选育提高提供理论支撑。     

美洲鲥鱼嗜水气单胞菌病原的分离与鉴定

2021年,山东省临沂市一养殖场养殖的美洲鲥鱼(Alosa sapidissima)突发疾病并出现严重死亡,日死亡率高峰期达到2.5%,累积死亡率约为90%。患病鱼主要症状为体表出血、溃疡,解剖可见腹腔腹水、肝脏暗红,并伴有肠炎。组织病理学检测发现,病鱼肝脏出现弥散性坏死,嗜碱性粒细胞增多和细胞肿胀空泡变性;脾脏出现出血性贫血性坏死灶、核破裂和核固缩;肾脏淋巴细胞坏死脱落,肾小体毛细血管球萎缩,近端小管和远端小管内的细胞出现不同程度的细胞结构消失。发病鱼肉眼和显微镜观察未见明显寄生虫,利用PCR方法检测鲤疱疹病毒2型(Cyprinid herpesvirus 2)、鲈鱼蛙病毒(largemouth bass ranavirus)等淡水鱼类常见病毒均为阴性。细菌分离培养结果显示,从发病鱼的肝脏、肾脏和脾脏中分离得到形态一致的优势菌,命名为AS-AH2101。经16S rRNA测序比对和生理生化鉴定,确定AS-AH2101为嗜水气单胞菌(Aeromonas hydrophila)。毒力基因检测结果显示,AS-AH2101携带气溶素(aerA)、溶血素(hlyA)、丝氨酸蛋白酶(ahpA)、...     

基于转录组测序筛选乌苏里白鲑肌肉生长关键候选基因研究

为了挖掘调控乌苏里白鲑(Coregonus ussurinsis Berg)肌肉生长的关键候选基因,本研究对不同生长速度乌苏里白鲑的肌肉组织进行转录组测序,以期为乌苏里白鲑群体选育提供基础数据。首先,在同等条件下(从混合池中)随机选择乌苏里白鲑F2个体进行实验分组(快长组和慢长组)。接着分别从快长组[体重为(219.20±38.66) g]和慢长组[体重为(74.30±17.86) g]中随机选取10尾样本,取其背部肌肉进行转录组测序。以FDR (false discovery rate)<0.05且|log2(FC)|>1 (FC, fold change)为条件筛选差异表达基因并对其进行GO (gene ontology)和KEGG (Kyoto encyclopedia of genes and genomes)富集分析,并通过qPCR验证转录组数据的准确性。转录组测序结果显示,共筛选出2 211个差异表达基因,与慢长组相比,快长组中583个差异基因表达上调及1 628个基因表达下调。GO功能注释结果显示,差异基因主要参与细胞过程和结合过程,差异基因显著富集到251条KEGG通路(P<0.05),其中,MAPK信号通路(MAPK signaling pathway)、PI3K-Akt信号通路(PI3K-Akt signaling pathway)、紧密连接(tight junction)、胰岛素信号通路(insulin signaling pathway)、糖酵解/糖异生(glycolysis/gluconeogenesis)和PPAR信号通路(PPAR signaling pathway)参与细胞生长。之后结合功能注释结果和KEGG,鉴定出肌浆/内质网钙ATP酶基因atp2a1和atp2a2、葡萄糖-6-磷酸脱氢酶基因g6pc、生长因子结合蛋白1基因igfbp1以及肌球蛋白重链基因myh1、myh4、myh6、myh7、myh9和myh13等可能与肌肉生长密切相关的基因。本研究共筛选出10个可能与乌苏里白鲑肌肉生长相关的关键候选基因,为今后乌苏里白鲑分子标记辅助育种提供了基础数据。     

亲环素基因在烟草不同器官及叶片不同发育时期的表达规律研究

通过对烟草中3个亲环素家族基因（CyPl、CyP2、CyP40）表达模式的研究,结果显示：在不同发育时期中,CyPI在烟草移栽后40、50、60d时表达量较高,CyP2则是在移栽后30、40、60d表达量较高,而CyP40只在移栽后60d表达量较高。在不同组织部位及器官的表达中,3个亲环素家族基因在中部叶和下部叶中表达均较强。在烟草根中主要表达CyPl和CyP40基因,CyP40还在花中表达较高,而CyP2基因的主要表达器官为叶片。     

紫花苜蓿细胞质雄性不育系相关microRNA的鉴定

紫花苜蓿（Medicago sativa L.）是一种多年生豆科苜蓿属牧草,利用紫花苜蓿细胞质雄性不育系育种有着重要意义,相关microRNA的鉴定工作是研究其分子机制的重要内容。本试验以培育的紫花苜蓿细胞质雄性不育系MSGN1A及其同型保持系MSGN1B的花蕾为试验材料,采用small RNA测序技术对MSGN1A及MSGN1B在花药发育阶段的鉴定差异表达microRNA以及差异表达microRNA靶基因功能进行注释。结果表明：共计检测到1 478个miRNA,其中known miRNA 1 351个,novel miRNA 127个;筛选出差异表达miRNA 130个,包括90个上调表达的miRNA和40个下调表达的miRNA;差异表达micro RNA靶基因主要涉及的通路有：“代谢过程”、“细胞过程”、“信号生物过程”、“植物激素信号传导”、“氧化磷酸化”和“淀粉蔗糖代谢”等。差异表达microRNA靶基因的功能主要是调控花药的发育,其次是调控花期以及信号传导,而差异表达microRNA靶向的转录因子中多为控制花期、花药发育和信号传导等过程的因子,对不育系的败育起到一定的影响。     

Postulation of leaf-rust resistance genes in Czech and Slovak barley cultivars and breeding lines

Leaf‐rust resistance (Rph) genes in 61 Czech and Slovak barley cultivars and 32 breeding lines from registration trials of the Czech Republic were postulated based on their reaction to 12 isolates of Puccinia hordei with different combinations of virulence genes. Five known Rph genes (Rph2, Rph3, Rph4, Rph7, and Rph12) and one unknown Rph gene were postulated to be present in this germplasm. To corroborate this result, the pedigree of the barley accessions was analysed. Gene Rph2, as well as Rph4, originated from old European cultivars. The donor of Rph3, which has been mainly used by Czech and Slovak breeders, is ‘Ribari’ (‘Baladi 16’). Rph12 originates from barley cultivars developed in the former East Germany. Rph7 in the registered cultivar ‘Heris’ originates from ‘Forrajera’. A combination of two genes was found in 10 cultivars. Nine heterogeneous cultivars were identified; they were composed of one component with an identified Rph gene and a second component without any resistance gene. No gene for leaf rust resistance was found in 17 of the accessions tested. This study demonstrates the utility of using selected pathotypes of P. hordei for postulating Rph genes in barley.     

Proper reprogramming of imprinted and non‐imprinted genes in cloned cattle gametogenesis

Epigenetic abnormalities in cloned animals are caused by incomplete reprogramming of the donor nucleus during the nuclear transfer step (first reprogramming). However, during the second reprogramming step that occurs only in the germline cells, epigenetic errors not corrected during the first step are repaired. Consequently, epigenetic abnormalities in the somatic cells of cloned animals should be erased in their spermatozoa or oocytes. This is supported by the fact that offspring from cloned animals do not exhibit defects at birth or during postnatal development. To test this hypothesis in cloned cattle, we compared the DNA methylation level of two imprinted genes (H19 and PEG3) and three non‐imprinted genes (XIST, OCT4 and NANOG) and two repetitive elements (Satellite I and Satellite II) in blood and sperm DNAs from cloned and non‐cloned bulls. We found no differences between cloned and non‐cloned bulls. We also analyzed the DNA methylation levels of four repetitive elements (Satellite I, Satellite II, Alpha‐satellite and Art2) in oocytes recovered from cloned and non‐cloned cows. Again, no significant differences were observed between clones and non‐clones. These results suggested that imprinted and non‐imprinted genes and repetitive elements were properly reprogramed during gametogenesis in cloned cattle; therefore, they contributed to the soundness of cloned cattle offspring.     

Nuclear receptor and target gene mRNA abundance in duodenum and colon of dogs with chronic enteropathies

Nuclear receptors (NR), such as constitutive androstane receptor (CAR), pregnane X receptor (PXR) and peroxisome proliferator-associated receptors alpha and gamma (PPAR, PPARγ) are mediators of inflammation and may be involved in inflammatory bowel disease (IBD) and food responsive diarrhea (FRD) of dogs. The present study compared mRNA abundance of NR and NR target genes [multi drug-resistance gene-1 (MDR1), multiple drug-resistance-associated proteins (MRD2, MRD3), cytochrome P450 (CYP3A12), phenol-sulfating phenol sulfotransferase (SULT1A1) and glutathione-S-transferase (GST A3-3)] in biopsies obtained from duodenum and colon of dogs with IBD and FRD and healthy control dogs (CON; n = 7 per group). Upon first presentation of dogs, mRNA levels of PPAR, PPARγ, CAR, PXR and RXR in duodenum as well as PPARγ, CAR, PXR and RXR in colon were not different among groups (P > 0.10). Although mRNA abundance of PPAR in colon of dogs with FRD was similar in both IBD and CON (P > 0.10), PPAR mRNA abundance was higher in IBD than CON (P < 0.05). Levels of mRNA of MDR1 in duodenum were higher in FRD than IBD (P < 0.05) or CON (P < 0.001). Compared with CON, abundances of mRNA for MRP2, CYP3A12 and SULT1A1 were higher in both FRD and IBD than CON (P < 0.05). Differences in mRNA levels of PPAR and MRP2 in colon and MDR1, MRP2, CYP3A12 and SULT1A1 in duodenum may be indicative for enteropathy in FRD and (or) IBD dogs relative to healthy dogs. More importantly, increased expression of MDR1 in FRD relative to IBD in duodenum may be a useful diagnostic marker to distinguish dogs with FRD from dogs with IBD.     

小尾寒羊多胎性能候选基因的研究进展

小尾寒羊具有常年发情、性成熟早和多胎性能的高繁殖力特性,平均每胎产羔2.6只,是我国优良的地方绵羊品种,也是我国独特的遗传资源。目前在BooroolaMerino羊、Inverdale羊和Hanna羊中已找到影响多胎性能的主基因,但国内对小尾寒羊多胎性能遗传机制的研究主要集中在常规的选育和分子标记方面,也有一些以候选基因法从分子水平上对小尾寒羊多胎机制进行研究,仍没有确立控制小尾寒羊高繁特性的主效基因,本文就其研究现状进行综述,为小尾寒羊主效基因的研究提供有益的参考。