转cry1F基因玉米花粉对腰带长体茧蜂存活和繁殖的影响

腰带长体茧蜂（Macrocentrus cingulum）是亚洲玉米螟（Ostrinia furnacalis）幼虫的主要寄生蜂,对亚洲玉米螟种群具有重要的控制作用。玉米花粉可以作为腰带长体茧蜂的补充营养。本研究利用转cry1F基因抗虫玉米花粉作为补充食物饲喂腰带长体茧蜂寄生亚洲玉米螟幼虫,研究其对腰带长体茧蜂存活和繁殖的影响,为转基因抗虫玉米的环境安全评价提供科学依据。结果表明,饲喂转cry1F基因抗虫玉米花粉后腰带长体茧蜂的寄生率、茧块重量、每茧块茧数、羽化率与饲喂对照玉米花粉相比无显著差异;在蜂蜜水+ Bt玉米花粉处理中虽然与对照玉米花粉处理相比雌蜂的寿命并没有显著差异,但雄蜂的寿命差异显著;在取食转cry1F基因抗虫玉米花粉的腰带长体茧蜂体内未检测到Bt蛋白成分。说明转cry1F基因抗虫玉米花粉对腰带长体茧蜂的存活和繁殖没有显著影响。     

腰带长体茧蜂对亚洲玉米螟幼虫的寄生功能反应

为评估腰带长体茧蜂Macrocentrus cingulum对亚洲玉米螟Ostrinia furnacalis的控害潜能,在实验室条件下（湿度60%~80%,光周期16L：8D）研究了20、24、28、32℃下腰带长体茧蜂对亚洲玉米螟幼虫的寄生功能反应和20、24、28℃下腰带长体茧蜂的自身密度干扰效应。结果表明：在20~32℃内,腰带长体茧蜂对亚洲玉米螟的寄生功能反应均符合Holling Ⅱ型;相同温度下,随着寄主亚洲玉米螟密度的增加,腰带长体茧蜂寄生量增加,但当寄主密度增加到一定数量后,寄生量趋于稳定。24℃的寄生效能最大,20℃最小。随着寄主亚洲玉米螟密度和腰带长体茧蜂密度的增加,搜寻效应逐渐降低。采用Hessell干扰模型,发现28℃时的搜寻常数Q和干扰效应m均为最大,腰带长体茧蜂个体间的干扰效应降低了其寄生效能。     

腰带长体茧蜂雌蜂触角的扫描电镜观察

采用扫描电镜技术对腰带长体茧蜂Microcentrus cingulum的触角感器进行了观察。结果表明,腰带长体茧蜂雌蜂触角上存在三种感器,分别为毛形感器、刚毛形感器、板形感器。对触角各种感器的形态和分布特点进行了描述,并就三类触角感器的功能进行了探讨。     

我国玉米主产区亚洲玉米螟越冬幼虫天敌调查

对我国玉米主产区 9省 1 1个点的亚洲玉米螟越冬幼虫寄生性天敌资源进行了调查。寄生性天敌昆虫主要有玉米螟厉寄蝇、腰带长体茧蜂、大螟钝唇姬蜂 ;病原微生物有白僵菌、苏云金杆菌、玉米螟微粒子虫和Nosemasp 等。总体上看 ,病原微生物的感染率高于寄生性天敌昆虫的寄生率 ,但不同种类在各地之间的分布有明显差异。白僵菌为东北春玉米区优势种 ,在吉林省的公主岭和通榆发病率达 74 8%～ 80 4 % ,可能与当地长期使用白僵菌进行玉米螟防治有关。苏云金杆菌在夏玉米区的寄生率明显较高 ,为 6 7%～ 1 3 7%。微孢子虫的感染率为 0～ 2 3 1 % ,玉米螟微粒子虫为优势种 ,主要流行在春玉米区 ,而在夏玉米区郑州寄生的微孢子虫为Nosema属一未知新种。腰带长体茧蜂分布范围较广 ,寄生率为 0～ 1 7 9% ,以夏玉米区寄生率较高。大螟钝唇姬蜂仅在赤峰、朔州和敦化有寄生 ,寄生率分别为 1 5%、0 5%和 0 8%。玉米螟厉寄蝇在除赤峰和衡水外的各点均有分布 ,寄生率为 0 7%～ 6 3% ,以山西朔州的寄生率为最高 ,并在该地所有天敌中为最高     

青枯雷尔氏菌胞外蛋白指纹多态性分析

 采用SDS-PAGE技术对70株不同来源及致病力青枯雷尔氏菌（Ralstonia solanacearum）进行胞外蛋白指纹多态性分析,研究结果表明,供试青枯雷尔氏菌菌株呈现丰富的胞外蛋白指纹多态性,多态性比率为100%。不同来源青枯雷尔氏菌分泌的胞外蛋白不同,EZ-Tn5TM插入诱变菌株电泳出20条不同大小蛋白条带,分子量集中在20~97 kD ,且菌株间蛋白条带相似或完全相同;60Co辐射诱变菌株共电泳出16条不同大小的蛋白条带,多数蛋白分子量44.3 kD;野生型菌株电泳的蛋白条带最多,共获得26条不同大小的蛋白条带。进一步对37株不同致病力的野生型菌株进行胞外蛋白含量测定,结果表明,不同致病力菌株胞外蛋白含量差异大,强致病力菌株分泌的胞外蛋白含量较高,为1.026～5.963μg/mL,无致病力菌株胞外蛋白含量较低,为0.083～0.761 μg/mL。     

亚洲玉米螟越冬幼虫主要寄生性天敌及病原菌致死率的考察

对我国主要玉米产区亚洲玉米螟Ostrinia furnacalis越冬代幼虫天敌和病原菌的寄生情况进行了调查。通过2年秋季采集14个省,20个点共计14984头玉米螟越冬幼虫,在室内(26±1)℃,RH(70%±10%),光周期16L:8D的环境中饲养。明确了主要病原菌苏云金芽孢杆菌Bacillus thuringiensis和白僵菌Beauveria bassiana及主要寄生性天敌昆虫腰带长体茧蜂Macrocentrus cingulum、玉米螟厉寄蝇Lydella grisescens和大螟钝唇姬蜂Eriborus terebranus在各地的寄生率。各地越冬幼虫平均感染苏云金芽孢杆菌死亡率为36.2%,感染白僵菌死亡率为14.4%。苏云金芽孢杆菌和白僵菌的感染率在东北春玉米区年度间有波动,而在夏玉米及其它玉米产区,苏云金芽孢杆菌的感染率显著高于白僵菌。腰带长体茧蜂和玉米螟厉寄蝇在各地普遍分布,为优势寄生性天敌昆虫,寄生致死率分别为7.0%和5.1%。大螟钝唇姬蜂仅在北方春玉米区的海伦、沈阳、应县和夏玉米区的保定等少数地区发生,且寄生率<1.0%。与天敌昆虫寄生死亡率相比,感染病原菌的死亡率显著较高,但年度间波动较大。由此可知,苏云金芽孢杆菌和白僵菌是玉米主产区亚洲玉米螟越冬幼虫的主要致死因子,且具有波动性;腰带长体茧蜂和玉米螟厉寄蝇是相对稳定的致死因子,大螟钝唇姬蜂具有区域特异性。     

北美车前种群内遗传多样性分析

目的:研究外来入侵种北美车前种群内遗传多样性水平,以期揭示其较强环境适应性在分子水平的根源。方法:用60条10碱基随机引物对生长于浙江师范大学校园内北美车前的9个不同样本进行了PCR扩增,筛选出7条有较高多态性检测能力的引物,用这7条随机引物对北美车前的RAPD指纹图谱进行了研究。结果:(1)共扩增到82条带,每条引物扩增带数在6~17条,平均带数为11.71条,其中60条带具有遗传多态性,约占总数的73.17%。(2)浙师大校园种群内的北美车前不同样本之间DNA指纹存在着较明显的差异,这种差异主要是受到了相对空间位置和极端土壤环境条件的影响。结论:(1)北美车前在种群内有较高的遗传多样性;(2)北美车前对极端土壤环境有较强适应性。     

菜蛾盘绒茧蜂多分DNA病毒EP-1-like基因克隆、原核表达与多克隆抗体制备方法

本实验提取被菜蛾盘绒茧蜂寄生后24h的小菜蛾幼虫体内总RNA,反转录合成cDNA,以此为模板PCR扩增出菜蛾盘绒茧蜂多分DNA病毒(Cotesia vestalis polydnavirus,CvBV)EP-1-like基因,将其分别克隆到表达载体pET30a、pET28a中,转化宿主菌BL21(DE3),获得单克隆重组质粒pET-30a-EP1L和pET-28a-EP1L。经IPTG诱导,pET-28a-EP1L成功表达了约34.8kDa的包涵体蛋白。将诱导条件优化以后,采用割胶回收的方法纯化包涵体,将纯化后的表达蛋白免疫新西兰大耳兔制备多克隆抗体。ELISA分析表明制备的抗体效价达1:128000,Western blot检测证明抗体具有良好的免疫反应特异性。     

菜蛾盘绒茧蜂多分DNA病毒对寄主小菜蛾幼虫体内部分组织的影响

利用显微注射方法，观察了菜蛾盘绒茧蜂多分DNA病毒（CpBV）对其寄主小菜蛾幼虫部分组织的影响。结果表明，CpBV进入寄主体内后能够影响其精巢、前胸腺和中肠等的变化。当被注射CpBV后，寄主精巢体积的增长受到抑制，并且有被分解的现象；颜色也随之逐渐变淡．直至呈现透明。前胸腺大小在感染CpBV 12h时就已经变小。中肠发育受阻，被分解，分布的气管出现萎缩现象，颜色由正常发育时的绿色变为黄色，个别会呈现红色，约20％中肠出现红色斑点。凝胶电泳分析表明，小菜蛾幼虫注射过CpBV后，其精巢、中肠和前胸腺中蛋白种类均减少，特别是精巢，仅有一条蛋白条带（78．8kD）较为清晰，蛋白条带明显变淡，说明蛋白含量明显下降；但前胸腺81．2kD的蛋白条带在感染CpBV后要比对照更为清晰。中肠酯酶活性测定表明，感染CpBV后中肠酯酶的活力受到抑制，酯酶活性减弱。     

黄瓜绿斑驳花叶病毒北京和山东分离物的生物学测定及其基因组比较

为揭示黄瓜绿斑驳花叶病毒的分子流行学特征,通过RT-PCR和cDNA克隆技术获得并分析了从北京甜瓜和山东西瓜上分离的2个黄瓜绿斑驳花叶病毒(CGMMV)分离物CGMMV-Beijing和CGMMV-Shandong的全序列.结果表明CGMMV-Beijing和CGMMV-Shandong的基因组分别由6423和6427个核苷酸组成,包括5'及3'端非编码区和4个开放性的阅读框,分别编码129kDa和186kDa复制相关蛋白、29kDa的移动蛋白及17.4kDa的外壳蛋白.寄主范围测定显示在供试的6科11种植物中CGMMV-Beijing和CGMMV-Shandong均可侵染葫芦科的葫芦、南瓜和西瓜以及茄科的本氏烟和藜科的苋色藜,不侵染供试的其它植物.CGMMV-Beijing和CGMMV-Shandong基因组的核苷酸同源性为99.5%,186kDa蛋白的核酸和氨基酸同源性分别为99.7%和99.8%,移动蛋白的核酸和氨基酸同源性分别为99.3%和98.1%,外壳蛋白的核苷酸同源性为100%.多序列比对分析结果显示,CGMMV的各分离物之间的分子差异并不明显,核苷酸同源性均在98%以上.将CGMMV-Beijing和CGMMV-Shandong与侵染葫芦科植物的烟草花叶病毒属其它成员比较,结果显示其亲缘关系较远,基因组的核苷酸同源性介于56.7%～59.6%,外壳蛋白的氨基酸同源性在44.7%～54.0%之间.综合分析显示,CGMMV分离物之间的差异并不明显,可能具有共同的侵染来源.     

Dynamics of the IgG3 responses following immunisation of BALB/c mice with somatic and excretory/secretory antigens from various Trichinella species

Comparison of the dynamics and antigen recognition profiles of IgG3 following immunisation with larval crude extracts (LCE) and excretory-secretory (ES) products from muscle larvae of different species of Trichinella (T. spiralis, T. nativa, T. britovi, T. nelsoni and genotype T6) was made in BALB/c mice. High levels of gG3 response were obtained in ELISA following immunisation with LCE from all species with maximum levels achieved between days 59 and 64 post-immunisation (p.i.) and maintained up to the limit of the observation (day 164). Antigen recognition profiles as measured by western-blot showed dense and numerous bands in the range 45-64 kDa that were consistent from week 5th with variation in epitope recognition among the different species. Following immunisation with ES antigens a significant decrease in IgG3 response was observed for all species especially for T. nativa in comparison to LCE. Antigen recognition on ES antigens showed three main bands in the range of 45-60 kDa for all species excepting T. nelsoni and T. britovi where an additional band was also present. These results clearly show that IgG3 epitopes are more abundant in somatic (LCE) than in ES products of Trichinella muscle larvae and that quantitative as well as qualitative variations exist among different species.     

逆境胁迫下植物LEA蛋白的研究进展

在逆境胁迫条件下,植物体内会产生诱导蛋白,LEA蛋白（Late embryogenesis abundant protein）是其中最常见的一类.本文就逆境胁迫下植物体内LEA蛋白的产生、特性、生理作用和基因的表达调控等作一简要阐述,并对LEA蛋白今后的研究方向及运用前景进行了展望.     

Effects of momilactone on the protein expression in Arabidopsis germination

Although momilactone is known to play an important role in the allelopathy of rice (Oryza sativa L.), there is no information available about the mode of action of momilactone on the allelopathy of Arabidopsis. The present research describes the allelopathic activity of momilactone A and B on the germination of Arabidopsis thaliana and the effects of momilactone A and B on protein expression during germination. Momilactone A and B inhibited the germination of Arabidopsis at concentrations >30 and 10 μmol L^?1, respectively. The protein bands of 18, 19 and 21 kDa on sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) were abundant in the momilactone A‐ and B‐treated Arabidopsis compared with the control. The protein bands of 18, 19 and 21 kDa were identified by the N‐terminal amino acid sequencing as cruciferin 2, cruciferina and cruciferin 3, respectively. Cruciferins and cruciferina are important storage proteins and play an important role as the initial source of nitrogen for seed germination. These results suggest that momilactone A and B may inhibit the germination of Arabidopsis seeds by inhibiting the degradation process of cruciferin and cruciferina.     

葡萄座腔菌(Botryosphaeria dothidea)全基因组分泌蛋白的预测及功能分析

葡萄座腔菌(Botryosphaeria dothidea)是引起多种果树和林木枝干溃疡病和果实腐烂病的重要病原菌.为明确葡萄座腔菌的致病机理,采用SignalP、WoLF PSORT、TMHMM、GPI-SOM等软件,对葡萄座腔菌全基因组14 998条蛋白序列进行了分泌蛋白预测和功能分析.结果表明B.dothidea...     

Comparison of the protein profiles of salivary gland extracts derived from three species of unfed and partially fed ixodid ticks analysed by SDS-PAGE

Salivary gland extracts (SGE) from unfed and 5 days fed adult female Ixodes ricinus (Linnaeus, 1758); Haemaphysalis inermis (Birula, 1895) and Dermacentor reticulatus (Fabricius, 1794) ticks were prepared. The protein content after feeding increased by 10.6, 8.7 and 6.8 times, respectively. Extracts were equilibrated to the same protein content and submitted to SDS-polyacrylamide gel electrophoresis followed by computer analysis of the scanned gels. Relative differences in protein profiles of extracts obtained from unfed and partially fed ticks were found in all species and some of them were similar in all three species used in the study. Results demonstrate that the increase of the protein content in salivary glands during the feeding does not occur proportionally. Some proteins are synthesised preferentially (67.1 kDa, 13.5 kDa) but other bands (in range of 15-16 kDa) present in the SGE derived from unfed ticks are less discernible in that of fed ticks.     

Isolation and characterization of lectins from the AG-D group of binucleate <Emphasis Type="Italic">Rhizoctonia</Emphasis> species

Isolates from 18 anastomosis groups (AGs) of binucleate Rhizoctonia were screened for lectin activity. Eight AGs (AG-B, AG-D, AG-F, AG-G, AG-H, AG-I, AG-R and AG-U) had low to moderate lectin activities. Among these, members of AG-D and AG-I had the highest activity. Partially purified lectins from AG-D preferentially agglutinated human blood type A to type B and O. Mucin and galactose were the most potent inhibitors among the tested carbohydrates. The molecular masses of these lectins ranged from 12.7 kDa for the monomer to 62 kDa for the pentamer type. Proline, alanine, glutamic acid, aspartic acid, leucine, threonine, serine and tyrosine were the major amino acid components of these lectins. Lectins from AG-D were stable at 4–50°C and from pH 6.0 to 10.0. When assayed with isoelectric focusing, these lectins gave bands at pI 9.30. Specificity of lectins from AG-D to galactose and its derivatives suggest a possible recognition role in this fungal species.     

Spiroplasma citri Spiralin Acts In Vitro as a Lectin Binding to Glycoproteins from Its Insect Vector Circulifer haematoceps

ABSTRACT In order to understand the molecular mechanisms underlying transmission of Spiroplasma citri by the leafhopper Circulifer haematoceps, we screened leafhopper proteins as putative S. citri-binding molecules using a spiroplasma overlay assay of protein blots (Far-western assay). Insect proteins were separated by one- or two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis, blotted, and probed with S. citri proteins. In this in vitro assay, we found that spiroplasma proteins exhibited affinity for seven leafhopper proteins. The interactions between S. citri proteins and insect proteins with molecular masses of 50 and 60 kDa were found to be sugar sensitive. These insect proteins were identified as high mannose N-glycoproteins, which support an interaction of glycoprotein-lectin type with S. citri proteins. Lectin detection in S. citri has revealed only one protein of 24 kDa. Using a leafhopper protein overlay assay on an S. citri protein blot, one spiroplasma protein with a similar molecular mass of 24 kDa was shown to display an insect protein-binding capacity. This protein was identified as the spiralin, which is the most abundant membrane protein of S. citri. Far-western experiments performed with purified spiralin and insect glycoproteins confirmed the binding of spiralin to the insect glycoproteins of 50 and 60 kDa. Thus, the spiralin could play a key role in the transmission of S. citri by mediating spiroplasma adherence to epithelial cells of insect vector gut or salivary gland.     

Antigenic characterization of rat louse Polyplax spinulosa

Number and relative molecular weights of proteins in whole body homogenates of the louse, Polyplax spinulosa were determined using SDS-PAGE. 13 protein components from the total of 28 bands were defined as Con A binding glycoproteins. In immunoblotting sera of lice infested laboratory rats of BN strain specifically recognized 3 major protein components with r.m.w. of 77,000, 105,000 and 230,000 Da and from 6 to 8 minor protein components with r.m.w. of 31,000-180,000 Da.     

寄生蜂毒液蛋白的研究进展

寄生蜂毒液蛋白具有抑制寄主血细胞免疫、调控寄主生长发育、抑制寄主生殖系统发育、调节寄主内分泌等功能,但一直以来对其组成成分知之甚少.近年来,不仅研究了寄生蜂毒液蛋白的生化特性,而且通过分离纯化、酶活测定、基因克隆等方法明确了寄生蜂毒液的部分蛋白组分.根据这些毒液蛋白性质和功能,可将其分为麻痹毒素、抗菌活性物质、酶类、酶的抑制剂等,它们在害虫生防、药理学和生理学研究中有着广阔的应用前景.该文作者就国内外对寄生蜂毒液蛋白组成成分的研究进展进行了综述,并对其应用前景进行了简要探讨.