稻曲病菌交配型基因MAT1-2-1和MAT1-2-8的特性分析

 有性生殖在真菌的生活史和进化过程中具有重要作用,而交配型基因是控制有性生殖的关键因子。前期研究发现稻曲病菌(Villosiclava virens)MAT1-2型菌株中包含MAT1-2-1和MAT1-2-8两个交配型基因,但是它们如何调控稻曲病菌有性生殖依然不清楚。本文研究了它们在不同侵染和生长发育时期的表达模式和编码的蛋白结构特性。研究表明MAT1-2-1在侵染不同阶段一直下调表达;而MAT1-2-8在侵染早期(5 dpi)上调表达,在侵染后期下调表达。与营养菌丝阶段比较,MAT1-2-1和MAT1-2-8在有性发育过程菌核形成、菌核萌发、子座原基形成和子座成熟4个阶段的表达量都是下降的,在菌核形成阶段表达量最低。生物信息学分析显示MAT1-2-1和MAT1-2-8具有磷酸化位点,为非分泌蛋白,无明显的跨膜结构域。蛋白同源比对分析表明MAT1-2-1与香柱菌(Epichloë typhina)的MAT1-2-1同源性最高,而MAT1-2-8与绿僵菌(Metarhizium)的MBR_08192蛋白同源性最高。进一步研究发现MAT1-2-1和MAT1-2-8能够互作,并分别主要定位在细胞核和细胞基质中。通过质谱技术鉴定到MAT1-2-1的一些候选互作蛋白,如假定Ran交换因子Prp20/Pim1(KDB12229.1)、假定rRNA处理蛋白Ebp2(KDB12923.1)及组蛋白H1(KDB12711.1)等。因此,以上结果为研究稻曲病菌交配型基因MAT1-2-1和MAT1-2-8调控有性生殖的生物学功能奠定了基础。     

Characterization of bovine ileal epithelial cell line for lectin binding,susceptibility to enteric pathogens,and TLR mediated immune responses

In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.     

Comparative effect of advanced soy products or corn protein concentrate with porcine meal on growth,body composition,and distal intestine histology of Florida pompano,Trachinotus carolinus

The present study was designed to investigate the effects of diets containing advanced soy products (enzyme‐treated soy and fermented soy) or corn protein concentrate (CPC) in combination with porcine meal (PM) to completely replace poultry byproduct meal (PBM) on growth performance, body composition, and distal intestine histology of Florida pompano, Trachinotus carolinus. Four experimental diets were formulated to be isonitrogenous and isolipidic, to contain 400 g/kg crude protein and 80 g/kg lipid. A reference diet (PBM diet [PBMD]) contained 150 g/kg PBM and 495 g/kg soybean meal (SBM), and three test diets were formulated replacing PBM with 15 g/kg of CPC (CPC diet [CPCD]) or replacing all SBM and PBM with 535 g/kg fermented soy (fermented soybean meal diet [FSBMD]) or 451.3 g/kg enzyme‐treated soy (enzyme‐treated soybean meal diet [ESBMD]). All three test diets were supplemented with 38 g/kg of PM. Diets were fed based on a percentage of bodyweight adjusted after sampling the fish every 2 weeks to triplicate groups of Florida pompano juveniles (mean weight 8.06 ± 0.22 g). After 8 weeks of feeding, fish fed CPCD and ESBMD performed equally well in terms of final body weight, thermal growth coefficient, and percentage weight gain in comparison to fish fed PBMD. In all cases, feeding FSBMD resulted in poor feed conversion and lower feed intake compared to other treatments. Protein retention efficiency, whole‐body proximate composition, phosphorus, sulfur, potassium, magnesium, calcium, sodium, and zinc contents were not significantly influenced by the dietary treatments. The results obtained in the present histological study showed no significant differences in the thickness of serous layer, muscular layer, and submucosal layer of the intestine among treatments. Fish fed CPCD showed a significant widening of the lamina propria with an increase of cellular infiltration and higher presence of goblet cells compared to other dietary treatment. Based on these results, 451 g/kg ESBM or combination of 150 g/kg of CPC and 495 g/kg SBM supplemented with 38 g/kg PM can be utilized to develop a practical diet for juvenile Florida pompano without impacting growth, nutritive parameters, and several distal intestine health parameters.     

寨卡病毒SYBR GreenⅠreal-time PCR方法的建立及应用

为建立一种针对寨卡病毒的快速诊断方法,本研究根据寨卡病毒的3’端保守基因序列,设计合成1对引物,建立了检测寨卡病毒的荧光定量PCR方法。结果显示:所建立的检测方法的Ct值与标准品在1.41×10^1~1.41×10^10^ copies/μL具有良好的线性关系,相关性为1,斜率为-3.502;灵敏性结果显示,该方法的检测限度为1.41×10^1 copies/μL,是普通PCR的10000倍;特异性结果显示,对CHIKV、DENV和JEV无特异性扩增,特异性强;重复性试验结果显示,组内和组间变异系数均小于1%,重复性好。本研究建立的SYBR Green I real-time PCR检测方法,可用于寨卡病毒感染的快速诊断。     

品种间大豆脂肪氧化酶活性差异及β-胡萝卜素漂白性研究

为了解东北地区大豆品种间大豆脂肪氧化酶(LOX)活性差异规律,整理适于东北地区种植的大豆种质资源307份(6份半野生品种,17份地方品种及4份国外品种,其余为栽培品种),统一种植,秋季收获后,测定各品种成熟籽粒脂肪氧化酶总活性。分析品种间脂肪氧化酶总活性分布规律、大豆脂肪氧化酶总活性与β-胡萝卜素漂白性之间关系。结果表明,307份大豆品种间脂肪氧化酶总活性差异极显著,平均值为21.68 U·mL~(-1),最小值为2.51 U·mL~(-1),最大值为42.38 U·mL~(-1)。根据脂肪氧化酶总活性差异,通过聚类分析将307份大豆品种分为3类。低活性品种占总量43.65%,脂肪氧化酶总活性平均值为15.25 U·mL~(-1);中等活性品种占总量43.97%,脂肪氧化酶活性平均值为24.64 U·mL~(-1);高活性品种占总量12.38%,脂肪氧化酶总活性平均值为33.87 U·mL~(-1)。脂肪氧化酶总活性与油分含量呈正相关,与蛋白含量呈负相关,与漂白性呈正相关。     

1株H1N1亚型猪流感病毒的全基因组特征及其对小鼠的致病性

旨在了解河南省猪流感病毒的流行情况及其遗传进化和基因组特征。2018年4月，从河南省某一出现疑似流感症状猪群中采集鼻拭子样品150份用于分离病毒，对分离病毒的全基因组进行序列测定和分析。同时感染6周龄BALB/c小鼠，研究其对小鼠的致病性。结果显示，获得1株H1N1亚型病毒[命名为A/swine/Henan/NY20/2018（H1N1）]。遗传进化表明，其HA和NA基因属于欧亚类禽H1N1分支，PB2、PB1、PA、NP和M基因属于2009甲型H1N1分支，NS基因属于经典H1N1分支。HA蛋白的裂解位点序列为PSIQSR↓GL，具有低致病性流感病毒的分子特征，在小鼠肺和鼻甲有效复制并能引起肺组织病理学变化。本研究分离到1株3源重排H1N1亚型病毒，对小鼠呈现一定致病力，提示应进一步加强对SIV的监测。     

宁南霉素等药剂对番茄黄化曲叶病毒病的田间防效

[目的]评价宁南霉素、艾德拉和碧护3种药剂混合使用防治番茄黄花曲叶病毒病的效果。[方法]通过田间试验研究8%宁南霉素、艾德拉绿(花、果)与碧护组成的药剂对番茄黄化曲叶病毒病的防治效果。[结果]8%宁南霉素、艾德拉绿(花、果)与碧护组成的试验药剂对番茄黄化曲叶病毒病具有很好的田间防治效果,第3次施药后10、20、30 d防效均在82%以上,与对照药剂番茄黄化曲叶病毒灵差异达极显著水平,且其产量比清水对照增加25 747.5 kg/hm~2,保产率为103.90%。[结论]宁南霉素、艾德拉和碧护混合使用对番茄黄花曲叶病毒病具有较好的防治效果,为番茄黄花曲叶病毒病的有效防治提供了理论依据。     

异硫氰酸丙烯酯毒杀松材线虫作用机制初步研究

对异硫氰酸丙烯酯毒杀松材线虫作用机制进行了初步研究。测定了其对线虫虫体形态、运动行为、体内总糖和蛋白质含量的影响,以及处理前后线虫体内过氧化氢酶(CAT)、乙酰胆碱酯酶(AChE)和谷胱甘肽-S-转移酶(GST)活性变化。结果表明:经异硫氰酸丙烯酯处理后的松材线虫虫体形态严重扭曲,运动形态出现由正常的S形变化为多节弯曲和螺旋状卷曲等异常状况,线虫行动随时间的推移变得迟缓,最终呈C形、新月形或直接死亡。随着异硫氰酸丙烯酯处理质量浓度的增大,线虫体内总糖和蛋白质含量显著上升,糖和蛋白质的代谢发生紊乱;线虫体内CAT、AChE和GST活性显著降低,且均低于对照组,表明线虫体内氧化和抗氧化作用失衡;细胞受到极大的影响,神经系统遭到破坏,解毒能力明显降低。研究结果可为研究利用异硫氰酸丙烯酯作为植物源杀线虫剂提供依据。     

6株牛传染性鼻气管炎病毒BICP4基因序列的同源性分析

牛传染性鼻气管炎病毒的立即早期启动基因IER4.2编码感染细胞蛋白BICP4基因。各种疱疹病毒的BICP4基因的Ⅰ，Ⅲ，Ⅴ区变异性较大，并为其特征区。为了比较分离自不同牛种，不同部位的5种病毒株BICP4蛋白I区基因的差异性，本文根据已发表的IBBV K22毒株重复序列IRs中BICP4的基因序列设计了一特异性引物，对各毒株进行PCR扩增，均得到约540bp的片段，将其克隆，测序并连同K22株进行同源性比较。结果表明，6株病毒的核苷酸序列及氨基酸序列同源性均在98%以上，说明这些毒株重复序列中的早期基因高度保守，在一定程度上也显示出IBRV的高度保守性。     

Effect of Heat Stress on Grain Growth, Starch Synthesis and Protein Synthesis in Grains of Wheat (Triticum aestivum L.) Varieties Differing in Grain Weight Stability

The effect of heat stress on the components of grain weight was analysed in wheat ( Triticum aestivum L.) varieties differing in grain weight stability. The wheat varieties PBW154, Sonalika and Hindi62 were raised in the field (New Delhi, India; 77°12' E, 28°40' N, 228.6 m a.s.l.) at three dates of sowing: 19 November 1992 (DOS I), 14 December 1992 (DOS II) and 18 January 1993 (DOS III). The late-sown crop (DOS III) experienced 6–8 °C warmer temperatures during grain development than the crop sown at the normal time (DOS I). The heat susceptibility index (S) revealed that grain weight was less susceptible to heat in Sonalika and PBW154 than in Hindi62. Heat stress reduced both the grain growth duration (GGD) and the grain growth rate (GGR). The grain weight reduction in PBW154 and Sonalika was mainly due to a reduction in GGR, while that of Hindi62 was due to a reduction in GGD. In vivo studies on starch and protein synthesis in excised endosperm at 15, 25 and 35 °C revealed that both processes were more thermotolerant in Hindi62 than in PBW154. The grain starch content was stable in Hindi62 while that in PBW154 was significantly reduced under heat stress. The grain nitrogen content at maturity increased in both varieties under heat stress. It was concluded that the heat susceptibility of grain weight in Hindi62 was mainly due to a reduction in GGD, although GGR, starch and protein synthesis were more thermotolerant in developing grains of Hindi62 than in those of PBW154.