Differential gene expression of Eph‐ephrin A1 and LEPR‐LEP with high or low number of embryos in pigs during implantation

The objective of this study was to ascertain whether mRNA and protein expressions of implantation‐related genes (erythropoietin‐producing hepatocellular receptor–ligand A1, Eph‐ephrin A1 and leptin receptor–leptin, LEPR‐LEP) differed between pigs with high and low number of embryos, and whether these differences in gene expression might affect embryo implantation. Experimental pig groups (n = 24) for high and low number of embryos were prepared by altering the number of eggs ovulated in pre‐pubertal gilts treated with 1.5 × (High) or 1.0 × (Low) PG600 ([400 IU PMSG + 200 IU hCG]/dose, AKZO‐NOBEL). Gilts expressing oestrus were artificially inseminated twice and maintained in breeding and gestation until the reproductive tract was collected on day 22 of pregnancy. At slaughter, the reproductive tracts from each pregnant gilt from each treatment were immediately processed to collect samples for RNA and protein analysis. Within each gilt, three conceptus points were sampled, one from each horn and then a random conceptus within the tract. At each conceptus point, endometrial attachment site, chorion–allantois and embryo were collected and immediately frozen in liquid nitrogen. Number of corpus luteum (CL) (35.4 vs. 12.6) and total embryo number (18.8 vs. 10.2) were greater in the high‐embryo compared to the low‐embryo group, respectively (p < .05). Real‐time qPCR results showed that Eph‐ephrin A1 mRNA expression was less in the high‐embryo (p < .05) compared to the low‐embryo group. In addition, Western blotting analysis indicated that Eph‐ephrin A1 and LEP protein expression at endometrial attachment site in high‐embryo was less (p < .05) compared to low‐embryo group. It was also noted that mRNA expression of Eph‐ephrin A1 and LEPR‐LEP was greater in pregnant than non‐pregnant gilts (p < .05). Moreover, mRNA expression of Eph‐ephrin A1 (p < .05) and LEPR‐LEP was greatest at endometrial attachment site among all three tissues. There was a positive correlation between expressions of Eph‐ephrin A1, LEPR‐LEP and embryo length with the correlation coefficient 0.31–0.59. For Eph‐ephrin A1, the highest correlation coefficient appeared between Eph A1 expression and normal embryo number, between ephrin A1 expression and embryo length. For LEPR‐LEP, the highest correlation coefficient appeared between LEPR‐LEP expression and ovary weight (0.79 for both, p < .05), followed by embryo length and weight. The results of this study suggest that low expression of Eph‐ephrin A1 and LEPR‐LEP is somehow related to increased embryo number during implantation and that endometrial attachment site might be the main target tissue of these gene products. Yet, the increased expression of Eph‐ephrin A1 and LEPR‐LEP appeared associated with increased embryo growth (length and weight) and ovary weight, Eph‐ephrin A1 and LEPR‐LEP might play roles in the regulation of embryo implantation in pigs.     

紫花苜蓿9个盐响应相关基因表达模式

紫花苜蓿(Medicago sativa)是世界上种植面积最大、应用最广泛的豆科牧草。由于其耐盐性中等,其在我国北方地区的产量和种植受到土壤盐渍化的限制。因此提高紫花苜蓿的耐盐性具有重要的科学和生产意义。为此,以龙牧801紫花苜蓿(M.sativa‘Longmu 801’)为试验材料,采用实时荧光定量PCR技术分析了不同浓度NaCl胁迫下9个盐胁迫蛋白质组筛选出的盐响应相关基因的表达模式。结果显示,处理时间和处理浓度对9个基因的相对表达量均有显著性影响,表明这9个基因均在紫花苜蓿盐胁迫应答中发挥着一定作用。处理1h时,G6PI、ABP19a、Trx-h1、PR bet 1、FBPA、6PGDH和ALDH这7个基因的相对表达量在不同NaCl胁迫浓度处理的紫花苜蓿中均显著上调,RRM和GDPD在NaCl处理2h后开始上调。除G6PI基因,其他8个基因在0.4%NaCl处理下相对表达量显著高于0.2%和0.8%NaCl处理。这些基因参与糖代谢、信号转导和胁迫响应。以上结果表明,紫花苜蓿的耐盐性极其复杂,涉及到多基因的表达和代谢通路的调控。研究结果有助于全面研究并了解紫花苜蓿的盐响应相关基因的表达模式,促进紫花苜蓿耐盐分子育种。     

动物诊疗机构X光机使用监管问题分析

随着动物诊疗技术的发展,动物诊疗机构普遍安装使用了X光机。本文通过对《动物诊疗机构管理办法》《放射性同位素与射线装置安全和防护条例》等法规的梳理,探讨了当前动物卫生监督物诊疗机构对动物诊断机构使用X光机的监管内容,包括动物诊疗机构是否从事动物颅腔、胸腔和腹腔手术,是否使用X光机,以及对违法行为的处置。     

牡蛎中GⅡ型诺如病毒巢式RT-PCR检测方法的优化与评价

为了解决目前巢式PCR法检测牡蛎样品中GII型诺如病毒时存在的非特异性扩增、假阴性等问题,本研究对NCBI数据库中在线获得的所有GII型诺如病毒序列进行系统分析,重新设计出针对GII型诺如病毒检测的通用型巢式PCR引物(NG2OF/NG2OR),并从基因型覆盖度、特异性、灵敏度和实际样品检测的稳定性几个方面对新设计的引物进行评估。结果显示：(1)新引物的特异性好,只对GII型诺如病毒进行有效扩增,且不受牡蛎样品本底的影响。(2)新引物灵敏度高,最低可检测至26.4个拷贝。(3)在对92个牡蛎样品进行检测时,新引物的阳性检出率为28.3%,比经典引物高出7.5%。因此,新建立的GII型诺如病毒巢式PCR检测方法在检测牡蛎样品时特异性更强,灵敏度更高,为牡蛎中诺如病毒的检测提供了一种新的、可靠的技术手段。     

玉米苗期耐碱性鉴定方法研究

在三叶一心期,用一定浓度梯度的Na2CO3溶液（5～25mmol&#183;L^-1）对玉米幼苗进行胁迫处理,观察其生长情况,7d后测定地上部分株高、鲜重、干重等生理指标。研究表明,各项生理指标均随Na2CO3溶液浓度的增加而降低,并在各碱浓度间、自交系间达极显著差异。综合苗情及各生理指标的变化,初步确定可以将25mmol&#183;L^-1 Na2CO3溶液作为玉米苗期耐碱性筛选的理想浓度,将该浓度下的苗情及株高、鲜重、干重及含水量变化率作为玉米苗期耐碱性筛选的鉴定指标。     

2019年湖南省部分地区鸡、鸭、猪源大肠杆菌耐药性调查

旨在调查2019年湖南省部分地区不同来源的大肠杆菌对抗生素的耐药水平,为养殖合理用药提供参考。从4个市畜禽养殖场采集猪、鸡和鸭粪便285份,使用麦康凯琼脂培养基和伊红美蓝培养基对大肠杆菌筛选,利用基质辅助激光解析电离飞行时间质谱进行鉴定,采用微量肉汤稀释法检测16种抗生素对大肠杆菌的最小抑菌浓度(MIC)。共分离出203株大肠杆菌(鸡源146株、鸭源20株和猪源37株),对四环素和氨苄西林耐药率最高,分别达到89.2%和84.7%;对头孢他啶、奥格门丁、黏菌素耐药率较低,分别为2.5%、2.0%和0.5%;所有菌株均对美罗培南敏感。抗3种及以上抗生素的菌株数目占比为87.7%,而仅5.42%的菌株对所有检测的抗生素敏感。另外,来源于减抗示范养殖场的大肠杆菌对大部分抗生素的耐药率,与来源于非减抗养殖场大肠杆菌相比未见显著差异。上述结果表明,湖南省动物源大肠杆菌的耐药水平仍处在较高水平,重视兽药使用管理并持续开展对大肠杆菌的耐药性监测十分必要。     

卷丹转基因体系构建及岷江百合LrCCoAOMT的导入

以卷丹(Lilium lancifolium Thunb.)无菌小鳞片为试材,通过切片处理、优化农杆菌侵染浓度与时间以及重悬液和共培养基成分,构建农杆菌介导的高效遗传转化体系。结果表明,MS+1.5 mg·L^-16-BA+0.5 mg·L^-1 NAA+30 g·L^-1蔗糖是切片芽分化的最佳培养基,添加100 mg·L^-1抗坏血酸能有效抑制褐化并促进不定芽增殖。MS+2.0 mg·L^-1 NAA+30 g·L^-1蔗糖是生根诱导的最佳培养基。抗生素敏感性测试发现,培养基中添加Kan 100 mg·L^-1或Hyg 75 mg·L^-1结合Cef 400 mg·L^-1适宜抗性筛选。GUS染色分析表明,以去除大量元素的改良MS+100μmol·L^-1AS和去除大量元素的改良MS+1.0mg·L^-16-BA+1.0mg·L^-1NAA+100...     

‘品丽珠’葡萄不同营养系酚类物质差异分析

筛选山东蓬莱产区酿酒葡萄‘品丽珠’黄烷醇、花色素苷含量较高的优良营养系,以期为发挥其酿酒性能提供理论依据。以蓬莱产区品丽珠母本及3个芽变营养系(214、327、623)为试材,采用分光光度计法分析葡萄果皮和种子中总酚、总单宁、总花色素含量,利用高效液相色谱法分析酚酸类、黄酮醇类、黄烷醇类、花色苷单体及白藜芦醇含量。研究结果表明,黄烷醇是种子和果皮中主要的单体酚,其中儿茶素含量最高;二甲花翠素葡萄糖苷是果皮中主要的花色苷。在相同的栽培管理条件和自然条件下,品丽珠不同营养系间的酚类物质差异较明显,相同营养系不同年份间也有显著差异。其中营养系623果皮和种子的总酚、种子的单宁和黄烷醇以及果皮和种子中的酚酸类、单体花色苷含量显著高于其他营养系和母本。     

茭白新品种"金茭2号"的选育及其栽培技术

介绍了金茭2号的选育背景、选育过程、产量表现及其特征特性和主要栽培技术。