抗氰烯菌酯假禾谷镰孢突变体的诱导及其生物学特性

为评估引起小麦茎基腐病的病原菌假禾谷镰孢Fusarium pseudograminearum对氰烯菌酯的抗性风险,对5株敏感菌株进行了室内药剂驯化,获得33株抗性突变体,突变频率为16.5%,其对氰烯菌酯的抗性水平范围为7.39～1 665.76倍,3株表现低抗,4株表现中抗,26株表现高抗;发现在myosin-5基因上存在11种抗性突变类型,其中217位的丝氨酸突变为亮氨酸(S217L)、420位的谷氨酸突变为赖氨酸(E420K)和135位的丙氨酸突变为苏氨酸(A135T)为主要突变类型,其比例分别为45.5%、15.2%和9.1%。S217L型抗性突变体的产孢量显著下降,菌丝生长速率和致病力与亲本菌株无显著差异。E420K型抗性突变体的菌丝生长速率和致病力显著下降,产孢量与亲本菌株无显著差异。A135T型抗性突变体的菌丝生长速率和产孢量与亲本菌株无显著差异。研究结果表明假禾谷镰孢在药剂选择压力下易形成氰烯菌酯的抗性群体,对氰烯菌酯存在中到高等的潜在抗性风险,其myosin-5的点突变与其对氰烯菌酯的抗性相关。     

不同播期对威芋5号产量及生长发育的影响

以马铃薯威芋5号为试验材料,研究在威宁县不同播期对威芋5号生理指标和产量的影响。试验结果表明,3月23日播种,威芋5号产量最高,可达2 116.46 kg/667 m2,纯收入为1 134.81元/667 m2,经济效益显著,而不同播期对威芋5号生育期影响不大。     

Study on Synergistic Effect of VA5 Immunopotentiator on Pigeon NDV Inactivated Vaccine

This study was designed to evaluate the synergistic effect of VA5 immunopotentiator on pigeon ND4416 inactivated vaccine.160 healthy pigeons were randomly divided into four groups,including ND4416 strain inactivated vaccine group (NDV group),ND4416 strain inactivated vaccine and immunopotentiator (VA5) mixed group (NDV+VA5 group),La Sota inactivated vaccine group (La Sota group) and normal saline as the control group (C group),to assess their vaccine efficacy against virulent pigeon NDV by serological analysis and animal testing.Pigeons sera were collected at different time points after immunization and measured the HI antibody titer of each group.The results showed that VA5 immunopotentiator significantly improved the serum antibody level of pigeons immunized with pigeon Newcastle disease inactivated vaccine (P<0.05).In addition,comparative test of spleen lymphocyte transformation was conducted at various time points after immunization.The results indicated that VA5 effectively stimulated the lymphocyte transformation of immune pigeon.Pigeons in each groups were challenged with ND4416 strain at the 30th,90th and 180th d after immunization.The results presented that the NDV+VA5 group had 100% protection rate and higher than La Sota group.The duration of immunization test showed that the antibody titer of NDV+VA5 group reached peak 11.20log₂ at the 21st d,remained 7.50log₂ at 180th d,and the protection rate remained 100% at 180th d.It indicated that VA5 immunopotentiator sustained the immune duration of pigeon NDV vaccine up to 180 d.Moreover,the in vitro detoxification test results suggested that VA5 immunopotentiator reduced the in vitro detoxification cycle of pigeons after challenge.Overall,this study suggested VA5 immunopotentiator could significantly improve the immune efficacy of pigeon Newcastle disease inactivated vaccine,which provided a basis for further enhancing the efficacy of Newcastle disease vaccine,as well increased experimental data for the application of immunopotentiators.     

DNA去甲基化参与提高白菜型冬油菜抗寒性的生理机制

利用来自不同育种环境的10份白菜型冬油菜材料,通过半致死温度的测定分析其抗寒性与环境的关系,并利用不同浓度DNA甲基化抑制剂5-azaC处理,分析DNA去甲基化对白菜型冬油菜DNA整体甲基化水平及低温胁迫下苗期生理特性的影响。结果表明,不同育种环境选育的材料,其半致死温度具有显著差异,其中材料2018-FJT、DT-7、DT-9与MXW-1的半致死温度分别为-16.04℃、-15.98℃、-15.63℃、-15.04℃;材料CT-2360、CT-2380、CT-2400、CT-2420、CT-2440、CT-2460的半致死温度分别为-11.32℃、-11.6℃、-11.42℃、-11.44℃、-12.97℃、-13.28℃。依据半致死温度,10个白菜型冬油菜抗寒性由强到弱依次为:2018-FJTDT-7DT-9MXW-1CT-2460CT-2400CT-2380CT-2420CT-2440CT-2360。抗寒性的形成与育成环境的纬度和年平均气温呈极显著正相关。选择抗寒性及产地不同的4个材料CT-2360、MXW-1、2018-FJT、DT-7进行生理测定,结果表明,1 000μmol·L~(-1) 5-azaC处理能显著抑制幼苗的根生长,4个材料的根长较对照减少93.14%～95.06%;低温下5-azaC处理对抗寒性较弱材料CT-2360幼苗的相对电导率、丙二醛含量以及强抗寒性材料DT-7幼苗的脯氨酸、可溶性蛋白含量影响最显著(P0.05);低温处理过程中,4个材料的SOD、POD、CAT活性均上升,以低温处理5d最显著(P0.05),其中2018-FJT的SOD活性增幅最高,为45.85%,DT-7的POD活性增幅最大,为460%,CT-2360的CAT活性增幅最显著,为321.02%。HPLC分析发现常温下抗寒性较弱材料CT-2360的甲基化水平为77.48%,高于其他3个抗寒性强的材料;经过5-azaC处理后发生明显的去甲基化作用,证明抗寒能力受DNA去甲基化的调控。     

南方水稻籼改粳与“青森5号事件”——20世纪50年代发生的一起重大引种事故

建国初期,各级政府习惯用战争年代开展"群众运动"的做法指导农业生产,"青森5号事件"是一起典型的违反科学规律和自然规律、不尊重科学家意见,盲目引种造成水稻大面积减产事例。《人民日报》发表社论,针对北种南移、远程调运造成减产的问题,批评在种子工作中存在的盲目调种、急于求成、强迫命令现象。最后由政府承担了"盲目引种"造成损失的责任。"青森5号事件"催生了农作物品种区域试验制度,以及"自繁、自选、自留、自用,辅之以调剂"(通称为"四自一辅")种子工作方针。     

AIV H5N1亚型高免卵黄抗体的制备及理化特性研究

应用禽流感灭活疫苗(H5N1,Re-1株)免疫20周龄的高产来航蛋鸡,制备抗AIV H5N1高免卵黄抗体(IgY).在比较了3种缓冲液对IgY活性的影响后,确定采用水稀释法提取IgY.同时在模拟的胃肠内环境下对IgY的各种理化性质进行了试验.结果表明,纯化的IgY在pH 2.0和胃蛋白酶终浓度为1.5 mg/mL的环境下,以及在pH 8.0和胰蛋白酶终浓度为2.5 mg/mL的环境下,37℃作用1 h后其ELISA抗体效价均无明显下降;同时,热稳定性试验表明,IgY在60℃以下作用30 min,其ELISA抗体效价也无明显下降;中和试验测得IgY的中和效价为1:640,具有较好的中和H5N1亚型AIV的能力.以上结果表明,本试验所制备的抗禽流感H5N1亚型高免IgY具有较高的抗体效价和中和效价,同时还具有一定的耐蛋白酶消化的作用,可用于H5N1亚型禽流感的预防和治疗.     

2016-2020年华北地区猪繁殖与呼吸综合征病毒分子流行病学调查

利用RT-PCR方法对2016-2020年华北地区692份疑似猪繁殖与呼吸综合征(PRRS)临床样本进行检测,获得368份阳性样品,阳性率为53.18%,并对其进行了ORF5基因扩增和测序,开展遗传变异和分子流行病学分析。结果显示,共获得33条完整的ORF5基因序列,除了HB-XT株为600 bp,其余全长均为603 bp。ORF5基因遗传进化分析表明33株均为Ⅱ型,其中有9株分布在Lineage 8,24株分布在Lineage 1。33株之间的氨基酸序列同源性为80.1%～99.5%,与JX-A1株和NADC30株的同源性分别为81.1%～99.5%和85.6%～94.5%。氨基酸分析结果表明,33株均在诱骗表位和中和表位发现了突变;在33株中共发现7个N-糖基化位点,其中包括相对保守的N44和N51位点,以及主要位于抗原表位的N30,N32,N33,N34和N35位点。结果表明,2016-2020年华北地区HP-PRRSV毒株逐渐被类NADC30毒株所取代,成为优势毒株,且ORF5基因变异差异较大,使PRRSV流行变得更加复杂。因此,本研究对PRRS分子流行病学以及疫苗的选择和防控...     

鸡FKBP5基因的克隆、生物信息学及组织表达谱分析

本研究旨在对鸡FK506结合蛋白5(FK506 binding protein 5,FKBP5)基因进行克隆和生物信息学分析,并检测其在鸡不同组织中的表达量。以鸡脾脏cDNA为模板,通过PCR方法扩增和克隆鸡FKBP5基因完整CDS区序列,并进行同源性比对及系统进化树构建;使用在线软件分析FKBP5蛋白的理化性质、疏水性、跨膜结构、信号肽、二级结构和三级结构;利用实时荧光定量PCR检测其在肢体内外翻畸形(valgus-varus deformity,VVD)组肉鸡和正常组肉鸡组织中的表达量,并绘制组织表达谱。结果显示,鸡FKBP5基因CDS区序列全长1350 bp,编码449个氨基酸;同源性比对和进化树分析表明,鸡FKBP5基因氨基酸序列与鹌鹑、鸭、壁虎、中华鳖、小鼠、人、猪、斑马鱼的同源性分别为98.4%、96.2%、85.8%、87.4%、81.1%、85.2%、86.1%和61.2%,与鹌鹑、鸭的亲缘关系最近,爬行类和哺乳类动物次之,与斑马鱼(鱼类)亲缘关系最远。鸡FKBP5蛋白分子质量为50.43 ku,理论等电点(pI)为5.94,半衰期为30 h,肽链N端为蛋氨酸(Met),不稳定系数为23.80,属于稳定蛋白;跨膜区和信号肽预测结果显示,FKBP5蛋白不属于跨膜蛋白和分泌型蛋白。结构域分析结果表明,该蛋白包括2个FKBP型肽基脯氨酰异构酶(FKBP-type peptidylprolyl isomerases)、3个四肽重复(TPR)序列,主要包含α-螺旋(46.77%)、延伸链(12.47%)和无规则卷曲(40.76%),且该蛋白与HSPA2、PPID、STIP1、HSP90AA1和HSP90AB1等互作蛋白具有很强的相关性。实时荧光定量PCR结果显示,FKBP5基因在鸡各组织中广泛表达,其中在软骨和法氏囊中表达量较高,在发病组肉鸡组织中的表达量均高于正常组肉鸡,且在心脏、腿肌、法氏囊、胸腺、软骨中表达量差异显著(P<0.05),在脾脏中表达量差异极显著(P<0.01)。本试验结果可为肉鸡骨骼疾病及腿部健康选育提供参考。     

Sublytic C5b-9 induces protective autophagy in cultured podocytes

AIM: In podocytes, autophagy occurs at a high basal level and dysregulated autophagy is associa-ted with a variety of podocytopathies. This paper is to investigate the role of autophagy in sublytic C5b-9-induced podocyte injury. METHODS: Sublytic complement C5b-9 stimulation was used as an in vitro model. Autophagosomes were confirmed using monodansylcadaverine (MDC) staining. Immunoblotting was used to measure the change of autophagy-related markers. Cellular morphological changes were observed by Wright-Giemsa staining. Immunofluorescence staining and confocal microscopy were used to detect the expression and distribution of nephrin. The cell viability was assessed by methylthiazol tetrazolium (MTT) assay. The cell apoptosis was assessed by Annexin V-fluorescein isothiocyanate/PI staining. RESULTS: For ensuring sublytic complement injury, the maximal amounts of anti-podocyte antiserum and 160×-diluted normal human serum were used without inducing cell lysis (defined as >5% LDH release). Sublytic C5b-9 promoted autophagy of podocytes in vitro. The proautophagic effect of sublytic C5b-9 manifested in the form of accumulated MDC-labeled vesicles and enhanced the expression of LC3-Ⅱ. Autophagy inhibitor 3-methyladenosine (3-MA) promoted sublytic C5b-9-induced podocyte morphological abnormalities. Compared with the sublytic C5b-9-injured podocytes, 3-MA exposure further decreased the expression of nephrin. 3-MA enhanced sublytic C5b-9-induced podocyte apoptosis. CONCLUSION: Sublytic C5b-9 attack induces autophagy, which may play a protective role against complement-mediated podocyte injury.     

NO/ADMA在布鲁菌侵染小鼠机体过程中的响应特征

本试验旨在研究布鲁菌侵染小鼠过程中一氧化氮(NO)的作用,以及NO和非对称性二甲基精氨酸(ADMA)在此过程的相互作用关系。以布鲁菌标准疫苗株M5侵染小鼠,首先用小鼠的血清进行虎红平板凝集(RBPT)和试管凝集试验(SAT),再用Griess试剂法和ELISA法测定对照组和侵染组不同组织和血清中的NO和ADMA含量,对小鼠肝脏和脾脏的各个侵染时间段进行CFU计数观察。结果显示,用布鲁菌M5侵染小鼠后,RBPT和SAT在14d时检测到有阳性反应。侵染组和空白对照组NO总含量上变化不大,但侵染组血清中的NO含量随时间出现下降,而肝脾中NO含量随时间出现上升,其他各组织NO含量波动不明显。而ADMA的总趋势与NO相反。CFU计数结果显示,小鼠肝脾内布鲁菌的数量在14d时一直处于增长状态,28d时布鲁菌的数量下降,表明布鲁菌的分裂繁殖和机体NO的含量存在一定的关系。