Effects of EBV infection on growth and apoptosis of nasopharyngeal carcinoma cell line

AIM: To explore the effect of EBV infection on growth and apoptosis of nasopharyngeal carcinoma(NPC) cell line.METHODS: NPC cell line CNE1 was directly infected by Epstein Barr virus (EBV). The expression of EBV-latent membrane protein 1 (EBV-LMP1) and bcl-2 were detected by immunohistochemistry method (LSAB). The growth of NPC cells was identified by MTT method. Apoptotic carcinoma cells were detected by flow cytometry analysis and the terminal deoxynucletidyl transferase-medicated dUTP-biotin nick end labeling (TUNEL) methods. RESULTS: EBV-LMP1 was positive in CNE1 infected by EBV(E-CNE1). Compared with CEN1, the growth of E-CNE1 apparently increased (P＜0.01). No apoptotic carcinoma cells were detected and bcl-2 postive cells were 2%~3% respectively in 2 kinds of NPC cells.CONCLUSION: Growth of NPC cells is enhanced by EBV infection and EBV-LMP1 expression, but no influence on expression of bcl-2 and apoptosis of NPC cells.     

植物源杀菌剂大黄素甲醚对烟草病毒病的防治效果

为明确大黄素甲醚对烟草病毒病的防治效果,最佳施药时期及施用量。以0.1%大黄素甲醚水剂为试验药剂,8%宁南霉素水剂和20%盐酸吗啉胍可湿性粉剂为对照,开展田间小区试验。结果表明,0.1%大黄素甲醚水剂于移栽定植期施药防效最好,高浓度达62.81%,中、高浓度即制剂量80克/亩、100克/亩的防效与对照药剂均无显著性差异。田间观察发现大黄素甲醚对烟草有一定促生长作用。     

山西苹果茎痘病毒遗传多样性分析

苹果茎痘病毒是苹果安全生产的巨大威胁,严重影响苹果产量和质量。探讨苹果茎痘病毒(Apple stem pitting virus, ASPV)在山西省的分布、变异和多样性,可为制定可持续的病毒管理策略提供理论依据。从山西省11个地区采集苹果叶片样品进行RT-PCR检测。结合在线数据库,利用相关软件对文中研究获得的序列进行系统发育、遗传多样性、重组以及种群遗传分化等分析。RT-PCR检测结果显示从山西省11个地区采集的409份苹果叶片样品中,ASPV的检出率为36.33%(临漪)至55.37%(平遥)。选择15个ASPV检测为阳性的样品,分别克隆其CP基因并与22个不同的ASPV分离物进行核苷酸和氨基酸一致性分析,发现37个ASPV分离物的核酸一致率为70.27～96.31%,氨基酸一致率为37.04～96.64%,其中,Shanxi-2与JX673826.1的核酸一致率最高,为90.86%, Shanxi-2与MN617853.1的核酸一致率最低,为70.40%。基于CP的进化树分析结果显示37个ASPV分离物被划分为2个进化群体。遗传多样性分析结果表明ASPV种群发生了收缩,且在两个...     

烟草丛顶病毒RNA依赖RNA聚合酶(RdRp)介导的体外复制体系

通过重叠PCR扩增得到烟草丛顶病毒(Tobacco bushy top virus,TBTV)中国分离物RdRp的编码序列,构建以pMALC2X为基础载体的原核表达载体pMAL-TB-RdRp。0.5 mM IPTG诱导可特异性表达分子量约为120 kDa的MBP-RdRp融合蛋白。温度梯度实验显示,18℃下诱导表达的MBP-RdRp融合蛋白的可溶性比例较高,约17%;经亲和层析纯化的MBPRdRp可特异性识别TBTV正链和负链的3'末端序列,催化体外复制;对正负链的3'末端的体外复制效率存在差别,识别负链3'末端的体外复制效率明显高于正链3'末端。本研究创建的TBTV RdRp介导的体外复制体系为进一步研究TBTV基因组复制调控奠定了基础。     

BBTV两个株系DNA组分1的克隆及序列分析

 对在生物学特性特别是寄主范围上存在明显差异的NSP株系和NS株系的代表分离物(广州天河分离物和高州分离物)的DNA组分1进行了克隆和序列分析,结果表明:2个代表分离物的DNA组分1全序列、ORF及其编码的氨基酸序列的变异率分别为3.2%、3.1%和2.8%,从而进一步确证了可以用寄主范围来作上述2个株系的鉴定。此外,这2个株系的DNA组分1、ORF及其编码的氨基酸序列分别与肖火根等报道的中国(广东)分离物、以及亚洲组和南太平洋组各分离物进行比较,结果表明:NS株系与肖火根等报道的中国(广东)分离物的亲缘关系很密切;这2个株系与亚洲组各分离物的差异均较小,而与南太平洋组各分离物的差异均较大,它们应属亚洲组。     

番茄褪绿病毒病在山东设施西葫芦上的发生及潜在危害

番茄褪绿病毒tomato chlorosis virus (ToCV)是我国蔬菜生产的重要新发病毒,其寄主范围逐年扩大。2019年10月项目组在山东寿光西葫芦温室调查时发现部分叶片呈现黄化、脉间褪绿,类似于ToCV侵染症状,并伴有烟粉虱发生。利用特异性引物检测发现,扩增的目的片段与GenBank中登录的侵染番茄的ToCV基因序列(登录号：KC887998.1)同源性高达99.58%,充分说明西葫芦植株已被ToCV侵染。通过对病毒病发生规律和烟粉虱虫口数量调查发现,西葫芦定植后1个月表现侵染症状的植株为2.0%,定植后2个月达到4.2%,定植后4个月后高达68.2%,病毒发生呈指数增长,而烟粉虱虫口数量却维持较低密度。从济南和德州采集的西葫芦疑似病叶和烟粉虱中也检出ToCV病毒,说明该病毒可能已经在山东省设施西葫芦主要种植区普遍发生,并经烟粉虱广泛传播,需引起高度重视。     

Identification of divergent variants of Grapevine virus A

Eight isolates of Grapevine virus A (GVA), which induced different symptoms in leaves of Nicotiana benthamiana, were recovered from various grapevines. The dsRNA patterns of two isolates, which consistently induced mild vein clearing (referred here as mild isolates of GVA) were similar, but different from those of other isolates of GVA. Analysis based on overall nucleotide (nt) sequence identity in the 3 terminal part of the GVA genome, comprising part of ORF3 (putative movement protein, MP), entire ORF4 (capsid protein, CP), entire ORF5 and part of 3 UTR, revealed that GVA isolates separate into three groups (I, II, III), sharing 91.0–99.8% nt sequence identity within groups and 78.0–89.3% nt sequence identity between groups. Mild isolates of the virus were group III and shared only 78.0–79.6% nt sequence identity with the other isolates. The comparison of predicted amino acid sequences for MP and CP revealed many amino acid alterations, revealing distinct local net charges of these proteins for mild isolates of the virus. Based on both conserved and divergent nt regions in the CP and ORF5, oligonucleotide primers were designed for the simultaneous RT-PCR detection of all GVA isolates and for the specific detection of the most divergent virus variants represented here by mild isolates of the virus.     

Intra-specific variation and inheritance of BYDV-PAV transmission in the aphid <Emphasis Type="Italic">Sitobion avenae</Emphasis>

Vector efficiency of 44 clonal lines (clones) of Sitobion avenae belonging to 31 different genotypes (distinct patterns for five microsatellite loci) originating from Western France was evaluated by transmitting the isolate PAV4 of BYDV-PAV to barley seedlings. Variation in transmission rates from 3.7% to 92.5% was observed, with significant effects of the aphid clone, of the plant species on which clones were collected, and of the reproductive mode of the clones. When genotypes are considered instead of clones, a continuum in transmission rates was observed. A subset of S. avenae clones was tested for transmission of one (10 clones) and 13 (4 clones) other BYDV-PAV isolates, and a clear clone effect modulated by an isolate effect was observed. Crosses were made between clones with different vectoring phenotypes and their F1 progeny were tested for PAV4 transmission. The narrow sense heritability of the PAV transmission character was rather high in the F1 families (h²=0.5) and the segregation analyses suggested an oligo/polygenic determinism of this character. The possibility of generating new transmission variants by sexual reproduction and its consequences on transmission mechanism studies and on BYD epidemics are discussed.