福州市周边蛋鸡场禽白血病亚群的血清流行病学检测

应用ELISA方法对分别采集自福州市周边30个蛋鸡场的597羽海兰蛋鸡、512羽罗曼蛋鸡、486羽伊莎蛋鸡和553羽特佳蛋鸡共2148羽的血样进行禽白血病病毒(ALV)亚群ALV-A/B和ALV-J抗体检测.结果表明:样品ALV-A/B抗体阳性率由高至低依次为:特佳蛋鸡群11.39%(63/553)、罗曼蛋鸡群11.32%(58/512)、海兰蛋鸡群8.54%(51/597)和伊莎蛋鸡群7.61%(37/486);样品ALV-J抗体阳性率由高至低依次为:特佳蛋鸡群5.24%(29/553)、罗曼蛋鸡群4.10%(21/512)、伊莎蛋鸡群3.51%(13/486)和海兰蛋鸡群3.18%(19/597);而样品ALV-A/B抗体总阳性率为9.73%(209/2148),高于ALV-J抗体总阳性率3.82%(82/2148),其中有0.70%(15/2148)的ALV-A/B和ALV-J抗体双阳性样品;蛋鸡场ALV-A/B抗体阳性率为70.00%(21/30),高于ALV-J抗体阳性率36.7%(11/30),其中有16.7%(5/30)的ALVA/B和ALV-J抗体双阳性样品.结论:福州市周边蛋鸡场不同品系和不同鸡群间存在不同程度的ALV流行,相比ALV-J,ALV-A/B流行更普遍,且存在ALV-A/B和ALV-J抗体双阳性样品,应予重视.     

有机肥、碱性物质对龙舌兰麻H.11648幼苗营养生长影响

以玄武岩发育而成酸性粘土、龙舌兰麻H.11648幼苗为试材,以施用充足化肥为对照,施用有机肥、碱性物质(碳酸钙、白云石粉)作为其他处理,测量各处理对龙舌兰麻H.11648幼苗株高、叶宽、叶片数影响,为龙舌兰麻H.11648合理施用有机肥和碱性物质提供依据。结果如下： 1、用碱性物质碳酸钙或白云石粉调节的土壤pH值6.0、6.5处理的龙舌兰麻H.11648幼苗株高、叶宽、叶片数增长率与对照(pH5.5)没有显著差异,龙舌兰麻H.11648幼苗在玄武岩发育土壤上生长酸害阈值为pH值5.5；2、有机肥处理对龙舌兰麻H.11648幼苗株高、叶片数增长率没有显著影响,但4%、8%有机肥显著增加了叶宽(增长率分别比对照提高62.7%、76.7%),有机肥显著促进了龙舌兰麻H.11648幼苗生长。     

利用CRISPR/Cas9系统编辑日本结缕草ZjSGR基因技术研究

日本结缕草(Zoysia japonica)是一种常见的具有众多优良性状的暖季型草种,秋冬季节在我国北方地区会较早出现叶片脱绿现象,在一定程度上限制了日本结缕草在北方的大面积使用。ZjSGR是从结缕草中分离出来的一种衰老诱导基因,负调控叶片滞绿性状。本研究利用CRISPR/Cas9系统,通过基因枪介导的遗传转化方法实现对日本结缕草ZjSGR基因的编辑,经测序验证获得1株单碱基缺失突变植株。突变植株叶片深绿,在正常生长温度(28~30℃)条件下,和对照相比,叶绿素含量更高(P<0.05)。本研究为进一步阐述ZjSGR基因在日本结缕草滞绿性状调控中的作用机理提供了基础研究材料,也为培育日本结缕草滞绿品种育种工作奠定了基础。     

草地贪夜蛾Sf9细胞对饥饿诱导自噬的响应

为研究草地贪夜蛾对饥饿诱导自噬的响应。从草地贪夜蛾EST数据库中筛选到一段核苷酸序列,设计特异性引物,RT-PCR技术从草地贪夜蛾Sf9细胞中扩增该核苷酸序列,经过生物信息学分析该序列与昆虫自噬相关基因ATG8同源性最高,命名为SfATG8基因,并构建了pIEX-4-mCherry-EGFP-SfATG8载体转染Sf9细胞。Sf9细胞经饥饿(PBS)诱导4 h后,Lyso-Tracker染色、Western Blotting、共聚焦显微镜检测细胞对自噬的响应程度。结果表明,经饥饿(PBS)诱导4 h的Sf9细胞中出现Sf Atg8-PE的表达,共聚焦显微镜检测到Sf9细胞膜上的自噬小体。以上证据证实饥饿(PBS)能诱导Sf9细胞发生自噬,为深入研究Sf9细胞对自噬的响应奠定了基础。     

H3N2亚型猪流感病毒HA和HA1蛋白的原核表达

为原核表达H3N2亚型猪流感病毒(SIV)的HA和HA1蛋白,通过RT-PCR方法获得SIV的HA和HA1基因,克隆至原核表达载体pMAL-c5X,并转化至原核表达菌Rosetta(DE3)中,表达菌经IPTG诱导表达后进行SDS-PAGE和Western blot分析。结果显示,成功表达了H3亚型SIV的HA和HA1蛋白,目的蛋白均以可溶性蛋白和包涵体两种形式存在,并与H3N2亚型SIV的HA单克隆抗体反应,说明HA和HA1蛋白具有良好的反应原性。     

Infections Investigation of ALV and REV in Guangxi Local Poultry Breeds and Sequence Analysis of gp85 Gene of ALV Isolates

In order to investigate the infection status of avian leukosis virus (ALV) and avian reticuloendotheliosis virus (REV) in the major local breeds of Qinzhou,Guangxi,totally 953 samples of egg white,cloaca swab and serum of Ma duck,Shitou goose,Tiejiao-Ma chicken,turkey and pigeon were collected from the representing flocks and detected by the commercial ELISA kits.ALV was isolated for the ALV p27 positive samples by culturing on DF-1 cells,and gp85 gene was sequenced.The results showed that the detections of ALV were negative in the samples except those of Tiejiao-Ma chicken,while REV antibody was found positive in Ma duck,Tiejiao-Ma chicken and turkey.The nucleotide sequences of gp85 gene of two isolates shared 94.5% identity with each other,and shared 86.9% to 94.9% with reference strains.The amino acid sequences of gp85 gene of two isolates shared 91.5% identity with each other,and shared 84.0% to 91.6% with reference strains.There were many variable sites in the hyper variable region hr1 and hr2,and the vr2 and vr3 variable regions were relatively conservative.Phylogenetic tree analysis showed that the two isolates shared the highest homology with SCAU11-XG strain.     

Effects of different NS genes of avian influenza viruses and amino acid changes on pathogenicity of recombinant A/Puerto Rico/8/34 viruses

To examine the effects of the NS1 and NEP genes of avian influenza viruses (AIVs) on pathogenicity in mice, we generated recombinant PR8 viruses containing 3 different NS genes of AIVs. In contrast to the reverse genetics-generated PR8 (rPR8) strain and other recombinant viruses, the recombinant virus rPR8-NS(0028), which contained the NS gene of A/chicken/KBNP-0028/2000 (H9N2) (0028), was non-pathogenic to mice. The novel single mutations of 0028 NS1 to corresponding amino acid of PR8 NS1, G139D and S151T increased the pathogenicity of rPR8-NS(0028). The replacement of the PL motifs (EPEV or RSEV) of pathogenic recombinant viruses with that of 0028 (GSEV) did not reduce the pathogenicity of the viruses. However, a recombinant virus with an EPEV-grafted 0028 NS gene was more pathogenic than rPR8-NS(0028) but less than rPR8. The lower pathogenicity of rPR8-NS(0028) might be associated with the lower virus titer and IFN-β level in the lungs of infected mice, and be attributed to G139, S151 and GSEV-PL motif of NS1 gene of 0028. In conclusion we defined new amino acid residues of NS1 related to mice pathogenicity and the presence of pathogenic NS genes among low pathogenic AIVs may encourage continuous monitoring of their mammalian pathogenicity.