几种昆虫病毒交叉感染玉米螟的研究

银纹夜蛾NPV(P_aNPV)、柞蚕NPV(ApNPV)和赤松毛虫CPV(D_aCPV)可感染玉米螟。D_aCPV对玉米螟1龄幼虫的ID_(50)为5.9×10~5PIB/ml饲料,对幼虫的生长发育、蛹重、羽化率和成虫寿命有显著影响。D_aCPV主要侵染幼虫中肠柱状细胞,在细胞质中增殖;P_aNPV和A_pNPV主要侵染幼虫的体壁细胞和气管壁细胞。D_aCPV在玉米螟幼虫体内增殖后,多角体形态由正六角形变为锥形或四方形;成虫羽化时排出的蛹便可观察到多角体,病毒可传递给子代。利用其他昆虫病毒有防治玉米螟的可能性。     

荧光增白剂Tinopal LPW对蜀柏毒蛾核型多角体病毒增效作用研究

用1%TinopalLPW荧光增白剂作为蜀柏毒蛾核型多角体病毒增效剂对蜀柏毒蛾2龄幼虫进行室内毒力测定,结果表明1%TinonalLPW对ParocneriaorientaNPV有较强的增效作用。使用3.6×1011PIB/hm2 1%TinopalLPW、1.8×1011PIB/hm2 1%TinopalLPW、9.0×1010PIB/hm2 1%TinopalLPW和3.6×1011PIB/hm2、1.8×1011PIB/hm2、9.0×1010PIB/hm26种处理对林间越冬代2-3龄幼虫进行超低容量喷雾防治,结果表明除9.0×1010PIB/hm2 1%TinopalLPW表现出显著的增效作用外,其余剂量有增效作用但不显著。     

灰茶尺蠖核型多角体病毒对灰茶尺蠖的致病性研究

采取室内毒力测定和田间防效调查相结合的方法,对灰茶尺蠖核型多角体病毒(Ectropis grisescensnucleopolyhedrovir-us)的致病力进行了研究。毒力测定结果表明,该病毒对2龄灰茶尺蠖幼虫的剂量对数-死亡机率值回归方程为y=-0.8774 0.9498x,LC50=1.5417×106PIB.mL-1;田间用浓度为1×107、1.5×107和2×107PIB.mL-1的EgNPV防治灰茶尺蠖,防效分别达58.81%、89.51%和94.81%。     

文山松毛虫质型多角体病毒杀虫剂的研制

报道了文山松毛虫质型多角体病毒杀虫剂的研制,包括多角体的纯化、辅助剂的筛选、剂型研制、产品包装以及病毒杀虫剂的产品质量检测及生产等.该杀虫剂为乳剂,多角体浓度达2.5×109PIB/mL;所选辅助剂取材方便、容易配制,对环境没有污染.经安全检测证明,无致病菌,符合国家卫生标准,对试验动物小白鼠无毒性和致病性.生物测定用1×106PIB/mL感染2龄文山松毛虫幼虫,其死亡率平均为85.5%.     

三种主要环境因子对茶尺蠖核型多角体病毒毒力的影响

本文对温度、紫外线和茶园露水对茶尺蠖核型多角体病毒(EoNPV)毒力的影响进行了研究。结果表明,在18—26℃下,适于病毒感染各龄幼虫,幼虫病死率达100％;但在30、34℃下,除1龄初饲毒外,病死率、化蛹率和羽化率明显下降或上升,3龄初或其以后饲毒,74％以上幼虫能化蛹,89％以上蛹能羽化。离体病毒经45℃处理2天,毒力开始明显减退。紫外线照射2—8分钟,离体病毒毒力减退不明显;照射16分钟,毒力开始明显减退。紫外线对离体病毒的影响与温度及病毒浓度有关。茶园露水对离体病毒无明显影响。     

荧光增白剂Tinopal LPW对蜀柏毒蛾核型多角体病毒增效作用研究

用1%TinopalLPW荧光增白剂作为蜀柏毒蛾核型多角体病毒增效剂对蜀柏毒蛾2龄幼虫进行室内毒力测定,结果表明1%Tinonal LPW对Parocneria orienta NPV有较强的增效作用.使用3.6×1011PIB/hm2+1%TinopalLPW、1.8×1011PIB/hm2+ 1%TinopalLPW、9.0×1010PIB/hm2+1%TinopalLPW和3.6×1011PIB/hm2、1.8×1011PIB/hm2、9.0×1010PIB/hm26种处理对林间越冬代2-3龄幼虫进行超低容量喷雾防治,结果表明除9.0×1010PIB /hm2+1%TinopalLPW 表现出显著的增效作用外,其余剂量有增效作用但不显著.     

用单克隆抗体结合酶联免疫吸附试验检测云杉卷叶蛾幼虫体内的核多角体病毒

在ELISA间接法中,应用单克隆抗体检测感染云杉卷叶蛾核型多角体病毒(Choristoneura fumiferana nuclear polyhedrosis virus,CfNPV)的云杉卷叶蛾幼虫体内多角体蛋白和病毒粒子。三龄幼虫喂饲表层涂有CfNPV的人工饲料(2×10~5PIB/cm~2)后6小时,即可在幼虫抽提液中检出多角体蛋白抗原(每条幼虫含0.14μg)和病毒粒子抗原(每条幼虫含0.32μg),随后此两种抗原量逐渐增加,直至第5天。而用染色涂片镜检法,则在添食病毒后3天才可观察到有少量多角体,病虫的病症需5～6天后才出现。因此,本法是一种快速、特异和敏感的检测杆状病毒的方法,其敏感度为1ml含10ng提纯的病毒粒子或多角体蛋白都能被检测出来。     

不同温度对油桐尺蠖核型多角体病毒在卵巢细胞系中增殖的影响

本试验用油桐尺蠖核型乡角体病毒(Buzura Suppressaria Nuclear Polyhedro-sis Virus)感染油桐尺蠖卵巢细胞系,分别置于0℃、15℃、20℃、26℃、28℃、30℃、37℃下静止培养,观察细胞病理变化,测定细胞的感染百分率和多角体含量。试验结果表明,不同温度对病毒的感染率及其在细胞中复制有明显的影响。26℃是油桐尺蠖核型多角体病毒感染卵巢细胞并形成多角体的最适培养温度,感染率最高,多角体含量也最多。培养温度过低或过高,多角体都不能在细胞内复制。     

蓖麻蚕核型多角体病毒DNA转染四种昆虫细胞系的研究

用蓖麻蚕核型多角体病毒(PcrNPV)DNA转染草地贪夜蛾(SF)、家蚕(Bm)、斜纹夜蛾(SL)和菜粉蝶(Pr)四种昆虫细胞系,结果表明,PcrNPV DNA能在SF细胞内复制增殖并形成多角体,Pr细胞系对PcrNPV DNA转染不敏感;Bm和SL细胞出现病变症状,但在电镜下未观察到病毒粒子或多角体。     

温度和食物对稻虱缨小蜂发育、存活和繁殖的影响

在白背飞虱(Sogatella furcifera(Hovàrth))卵内,稻虱缨小蜂(Anagrus nilparvatae Pang et Wang)的发育起点温度为10.6℃,有效积温为162.3日度,随着温度上升,发育加快;寿命(y_1,D_(10.4)~0C)、产卵量(y_2,粒/♀)、产出卵率则在适宜温度范围(23—26℃)内最大,高低温区内均呈下降趋势,其方程分别为:y_1=exp(-0.04187+0.3612x-7.4654×10~(-3)x~2),y_2=exp(-1.9539+0.4563x-0.01001x~2),式中x为温度。温度对未产出卵量影响不显著;对子代雌性比的影响也不显著,其平均值为0.7631。温度主要通过对产卵量和产卵速率来影响繁殖力,理论上27.3℃时周限增长率λ最大,达1.2374倍/天。成虫期供蜜加水时的产卵量、产出卵率和寿命显著比仅供水时的高或长。此研究为预测害虫种群发展,充分利用天敌资源,达到综合管理害虫提供了科学依据。     

螺旋对称病毒衣壳参数的研究

本文从数学角度分析和研究了螺旋对称病毒衣壳参数、整理出衣壳参数２０多个,对每一个参数的概念、含义等给予了明确的认定。提示了参数间的关系,推导出了一些计算参数值的公式。     

Polypeptide composition of flacherie viruses of the silkworm

The purified flacherie viruses of the silkworm, Bombyx mori, (FVS I, FVS II, FVS III, and FVS IV) were iodinated by using chloramine-T. The iodinated FVSes were purified by sucrose density gradient centrifugation or 2.4% polyacrylamide gel electrophoresis. FVS IV was found in the sedimentation analysis of FVS I, FVS II, and FVS IV. Electrophoretic patterns of FVS IV showed that it was a mixture of components having identical mobilities with FVS I, EVS IIa, and FVS IIb. FVS IV was a decomposed particle of FVS I, FVS II, and/or FVS III. All of these particles contained three polypeptides with molecular weights of about 51,000, 31,000, and 12,000 daltons. FVS I composed of six polypeptides with molecular weights of 67,000, 51,000, 39,000, 31,000, 14,000, and 12,000 daltons. The maturation process of FVS I was discussed and was suggested as the following process, FVS IIb→FVS IIa→FVS I. It is not clear whether FVS III is an intermediate for FVS IIa to convert into FVS I, or FVS III is a decomposed particle of FVS I.     

山东省玉米矮花叶病毒的生物学特性及基因组全序列测定

对山东省玉米矮花叶病毒原分离物(SD)进行了寄主范围、血清学等普通生物学鉴定,测定了该病毒的基因组核苷酸全序列。该病毒基因组RNA由9596个核苷酸组成(不包括3’—polyA的长度),整个基因组按一个ORF编码一个3063个氨基酸的多聚蛋白。序列比较表明,该病毒分离物(SD)与玉米矮花叶病河南分离物(HN,EMBL登录号：AF94510)核苷酸全序列同源性最高,为98．2％,与已报道的甘蔗花叶病毒(SCMV)7个分离物同源性也高达79．5％-98．2％,但与玉米矮花叶病毒保加利亚分离物(MDMV—B8,AJ001691)和高梁花叶病毒萧山甘蔗分离物(SrMV—XoS,AJ310197)的同源性仅为67．8％和69．3％,与约翰逊草花叶病毒(226920,JGMV)差异最大。系统进化树分析也表明,该病毒与SCMV分离物位于同一进化簇,而与MDMV进化关系很远。     

Comparison of sequence homology of poly(A) and non-poly(A) containing 34S RNA of AKR murine leukemia virus.

AKR MuLV 70S RNA was separated on Poly(U)-Sepharose into poly(A) and non-poly(A) containing 34S subunits. The ratio of the two fractions was 2:1, respectively. Both fractions were hybridized to AKR MuLV [³H]cDNA, and the hybrids were assayed by nuclease S₁ and cesium sulfate centrifugation. The poly(A) and non-poly(A) subunits hybridized to [³H]cDNA to the same extent (80%), with identical $\text{CO}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$ values; and the hybrids of both fractions had identical T_m values (81°C in 0.15 M NaCl). These results demonstrate that the poly(A) and non-poly(A) containing subunits of the AKR genome have identical or very similar base sequences in the heteropolymeric regions.     

Determination of the absolute hand of the helix in the nucleocapsid of haemagglutinating virus (Japan)

The basic hand of the helix of the nucleocapsid of haemagglutinating virus (Japan) was determined to be left-handed from observation of serrated-smooth asymmetry in tilted specimens examined in the electron microscope according to Finch's (1972) technique. Absolute determination of the helical hand was made by comparison with a tilted model of the virus nucleocapsid and by comparing this with the helical sense of the tobacco mosaic virus particle determined by the same method.The left-handed sense of the helix in the nucleocapsid was recognized by examination of cases showing opened-out turns of the helices. Under these conditions the far side of the particle was found to be contrasted predominantly. These conditions are discussed.     

正廿面体病毒的圆球度及其与等效球间关系的研究

本文根据形态计量学原理,对正廿面体病毒的圆球度进行了推导和计算．同时揭示了病毒内部各种等效球间的关系是按等比级数排列的,其比值近似等于廿面体圆球度（Φ）,即0.9552．     

北柴胡茎叶总黄酮抗流感病毒的作用

北柴胡 (BupleurumchinenseDC .)茎叶总黄酮 (TFB) 0 .0 9g kg剂量组对乙型流感病毒 (B 京防 98 76 )感染小鼠具有明显的保护作用 (P <0 .0 5 ) ,0 .0 3g kg和 0 .0 9g kg剂量组能明显降低乙型流感病毒 (B 京防 98 76 )感染小鼠肺指数值 (P <0 .0 5 ,P <0 .0 1) ,肺指数抑制率分别为 2 0 .5 %和 2 5 .4 %。组织病理学检查结果表明 ,TFB各剂量组与模型组比较 ,肺部病变均明显减轻 ,其中以TFB高剂量组肺部总体病变程度最轻 ,疗效最好。     

从PRRSV BJ-4株基因组全长cDNA获得感染性病毒

对已构建的覆盖猪繁殖与呼吸综合征病毒(PRRSV)BJ-4全长cDNA的6个重组质粒进行测序,并对部分点突变进行定点回复突变,将突变片段顺次连接,获得了全长cDNA克隆pWSK-DCBA。通过体外转录获得病毒基因组RNA,将RNA与脂质体混合后直接转染MARC-145细胞,获得拯救病毒(rV68)。rV68能在MARC-145细胞上稳定传代,并可引起PRRSV特征性的细胞病变(CPE)。增殖动态分析表明,rV68在MARC-145细胞上的生长有所迟滞,达到最高滴度的培养时间比亲本病毒延迟12h,但滴毒无显著差异(P>0.05)。结果表明,构建的BJ-4全长cDNApWSK-DCBA具有感染性,为研究中国PRRSV的分子致病与免疫机制、新型疫苗等奠定了基础。     

Construction and immunogenicity of recombinant pseudorabies virus expressing the modified GP5m protein of porcine reproduction and respiratory syndrome virus

Pseudorabies virus (PRV),an alpha-herpesvirus,has been developed as a live viral vector for animal vaccines.However,the PRV recombinant virus TK-/gE-/GP5+expressing GP5 of porcine reproductive and respiratory syndrome virus (PRRSV),based on the PRV genetically depleted vaccine strain TK-/gE-/LacZ+,scarcely stimulated the vaccinated animals to produce neutralizing antibodies against PRRSV.To develop a booster-specific immune response of such PRV recombinants,the ORF5m gene (the modified ORF5 gene having better immune responses)was substituted for the ORF5 gene and introduced into PRV TK-/gE-/LacZ+,resulting in a PRV recombinant named TK-/gE-/GPSm+,which expressed the modified GPSm protein.The recombinant virus was confirmed using PCR,Southern blotting and Western blotting.TK-/gE-/GPSm+and TK-/gE-/GP5+expressing the authentic GP5 protein were inoculated into Balb/c mice to evaluate their immune responses.The results indicated that the protecting neutralization antibodies (the 3/6 vaccinated mice obtained 1:16)and cell immune responses induced by TK-/gE-/GPSm+against PRRSV were higher than that induced by TK-/gE-/GP5+.Thus,the development of the new PRV recombinant expressing the modified GP5m protein as a candidate vaccine established the basis for the study of bivalent genetic engineering vaccines against PRRSV and PRV.