Maternal Antibody Protected Chicks from Growth Retardation and Immunosuppression Induced by Early Reticuloendotheliosis Virus Infection

To determine if the maternal antibody from breeders vaccinated with cell culture-adapted reticuloendotheliosis virus （REV） could protect chicks from early REV infection, one-day-old chicks with or without anti-REV maternal antibodies were inoculated with REV, and then their growth rates and antibody titers to Newcastle disease virus （NDV） and avian influenza virus （AIV）, after vaccination with inactivated vaccines, were compared. This study indicated that REV infection could cause growth retardation and severely inhibit immune reactions to inactivated vaccines against NDV and Avian influenza virus （AIV, H9 and H5） in one-day-old broilers without maternal antibodies specific to REV. Maternal antibody from breeders vaccinated with an attenuated REV vaccine effectively protected REV-challenged birds from growth retardation and immunosuppression on antibody reactions to NDV and AIV vaccines. Four weeks after vaccination, the HI titers to NDV, AIV-H9, and AIV-H5 in maternal antibody positive and negative groups were 3.36 ＋- 2.04 versus 1.58± 1.69 （P〈0.01）, 6.27±3.87 versus 0.71 ± 1.60 （P〈0.01）, and 6.72±3.92 versus 0.54± 1.44 （P〈0.01）. Maternal antibodies from breeders vaccinated with REV vaccine could successfully protect chicks from REV infection and effectively prevent REV-induced growth retardation and immunosuppression in antibody responses to NDV and AIV.     

水稻不育突变株沈农05-9的形态和细胞学特征鉴定

以辽粳5号与丰锦杂交的高代系的突变株沈农05-9和其原始株系为材料,采用醋酸洋红染色压片技术及石蜡切片技术.进行了形态学特征、小孢子发生的细胞学特征观察.结果表明:该不育突变株花粉表现典败和染败,多数花粉母细胞减数分裂不形成二分体,而形成多核的花粉母细胞;花药表现干瘪且纤维化严重,绒毡层细胞在小孢子发育过程中提前解体.该不育突变株的减数分裂方式类似于萍乡型显性不育系及早稻昆植不育系.导致败育的直接原因为绒毡层细胞提前解体.     

栝楼的RAPD-PCR体系建立与优化

[目的]建立栝楼的RAPD-PCR体系并对该体系进行优化。[方法]以栝楼叶片为材料,采用CTAB法提取栝楼叶片基因组DNA,利用正交设计对RAPD-PCR体系进行优化。[结果]各因素水平变化对PCR反应影响大小依次为：Mg2＋、TaqDNA聚合酶、引物和dNTPs。通过试验分析,优化的栝楼RAPD反应体系为：在25μl反应体系中,含10×buffer2.5μl,Mg2＋2.0mmol/L,Taq酶1U,引物0.8μmol/L,dNTPs0.1mmol/L。反应程序为94℃预变性2mim;94℃变性30s,37℃退火温度40s,72℃延伸1.5mim,36次循环;72℃延伸10mim,4℃保存。[结论]该研究建立的栝楼RAPD反应体系稳定可靠,为栝楼性别鉴定、遗传多样性分析、亲缘关系分析等方面的研究提供了有效的方法。     

苏云金芽孢杆菌cry1Ac28基因的克隆及原核表达

研究旨在克隆苏云金芽胞杆菌(Bacillus thuringiensis,Bt)Q-12的cry1Ac28基因并在原核载体中表达。应用PCR-RFLP法鉴定出Bt Q-12中含有cry1Ac基因,根据已知的cry1Ac全长基因序列设计特异引物,以Bt Q-12基因组DNA为模板扩增cry1Ac全长基因,与大肠杆菌(Escherichia coli)表达载体pEB相连接获得含有cry1Ac全长基因的重组质粒pEB-cry1Ac,经过序列分析表明,该基因编码区为3 537 bp,编码1 178个氨基酸,分子质量为133.3176 ku,pI为4.885,为弱酸性蛋白质,亮氨酸(Leu)、丝氨酸(Ser)、谷氨酸(Glu)三种氨基酸含量最高,分别为8.06%、7.80%、7.72%,该基因在大肠杆菌BL21(DE3)菌株能够正常表达133.3176 ku蛋白。该基因核苷酸序列已在GenBank中登录,登录号为FJ610439,并由Btδ-内毒素基因国际命名委员会正式命名为cry1Ac28。为进一步研究cry1Ac28蛋白功能和活性打下了良好的基础。     

Construction of Tobacco Bax lnhibitor-1 ihpRNA Gene Silencing Vector and Transformation into Agrobacterium tumefaciens EHA105

The plasmid pGSA1285 was first modified by substituting its GUS sequence with the Chalcone synthase intron fragment from vector pFGC5941 to get the plant silencing expression vector that contained Kanamycin resistance site and was named as pGSA2285. Using PCR-based amplification, two different restriction sites at both ends of tobacco Bax inhibitor-1 (NtBI-1) gene were created, respectively, which made the construction of ihpRNA gene silencing vector more efficiently. Then, NtBI-1 genes were inserted into Multiple Cloning Site (MCS) of pGSA2285 respectively to form Bax inhibitor-1 ihpRNA gene silencing vector, named as pGSA4285, containing sense and anti-sense BI-1 sequence which was spliced by chalcone synthase intron. Combined PCR identification and enzyme restriction analyses, the results showed that Bax inhibitor-1 ihpRNA gene silencing vector had been constructed and transferred into Agrobacterium tumefaciens EHA105 successfully, which laid a foundation for the further study on the function of BI-1 in plant PCD regulation.     

细叶远志愈伤组织的诱导

[目的]探讨影响细叶远志（Polygala tenuifolia Willd.）愈伤组织诱导的因素,建立细叶远志愈伤组织培养方法。[方法]采用组织培养方法,比较不同外植体、pH、温度、光照、激素种类及配比对细叶远志愈伤组织诱导的影响。[结果]在培养基MS上,子叶的诱导率最高,为88%;叶片和茎诱导生成的愈伤组织较少;不同pH对愈伤组织生长质量的影响差异不明显;愈伤组织诱导最适温度为28℃;66%的光照培养诱导率高于33%的光照培养;1.0 mg/L6-BA＋0.1 mg/L NAA的激素配比的诱导率最高。[结论]细叶远志愈伤组织的适宜培养基为MS＋1.0 mg/L6-BA＋0.1 mg/LNAA＋30 g/L蔗糖＋5 g/L琼脂（pH6.8）,培养温度为28℃,66%的光照培养18 h和黑暗培养6 h交替进行。     

凌源地区百合新品种引种栽培试验研究

为了在凌源地区引进推广新的百合品种,辽宁省农业科学院引进14个百合新品种在凌源试验栽培。通过主要性状的调查分析,确定适宜凌源地区栽培的百合品种有LA杂种系的纳帮;OT杂种系的罗宾娜、黄色风暴、木门、曼尼萨;OO杂种系的赞布拉星、能量。     

章鱼胺对营养性肥胖小鼠减肥作用的研究

采用高脂饲料建立小鼠肥胖模型,以小鼠体重、Lee's指数、脂肪湿重、脂肪系数、血清总胆固醇(TC)、甘油三脂(TC)及高密度脂蛋白(HDL-C)等指标的变化,研究章鱼胺对小鼠的减肥作用.结果表明,肥胖对照组小鼠较普通对照组小鼠的体重、脂肪湿重、脂肪系数、血清总胆固醇(TC)、甘油三脂(TG)及高密度脂蛋白(HDL-C)含量均显著增加(P<0.05),表明营养肥胖模型构建成功;章鱼胺各剂量组及阳性对照组中小鼠的体重、脂肪湿重、脂肪系数、血清总胆固醇(TC)与模型组存在显著性差异(P<0.05),其中Lee's指数和甘油三脂(TG)的中、高剂量组与模型组存在显著性差异(P<0.05);章鱼胺对营养性肥胖小鼠有一定的减肥降脂作用.     

高良姜多糖对酪氨酸酶抑制作用的研究

[目的]研究高良姜多糖对酪氨酸酶活性的抑制作用。[方法]以新鲜的高良姜和马铃薯为原料,通过比色法、色差仪法、Photoshop图像灰度值分析等测定高良姜多糖对马铃薯酪氨酸酶活性的抑制率,以及对马铃薯匀浆褐变度和反应体系颜色值变化的影响。[结果]试验得出,高良姜多糖对酪氨酸酶的活力有显著的抑制作用,其IC50为0.315 mg/m L,能显著降低反应体系的褐变度。[结论]高良姜多糖具有较强的酪氨酸酶抑制活性,可望用于美白护肤、果汁护色、鲜切果蔬保鲜等领域,值得进一步研究。     

基于氧化石墨烯生物免疫传感器检测副溶血弧菌的研究

副溶血弧菌(Vibrio parahaemolyticus)是最为严重的水产动物致病菌之一,准确即时的检测手段是预防控制该菌传播的关键所在.通过量子点(QDs)标记副溶血弧菌鞭毛蛋白抗体(Ab),利用生物免疫传感器技术实现副溶血弧菌的快速、特异性检测.结果显示,QDs-Ab的荧光通过加入氧化石墨烯(G0)产生淬灭,构建了捕获目标细菌的探针,氧化石墨烯最适淬灭浓度为60 ng/ml.细菌捕获探针中加入副溶血弧菌后,能够检测到重新恢复强度的绿色荧光.副溶血弧菌的检出限达到104 CFU/mL.针对副溶血弧菌设计的生物免疫传感器的特异性,选取灿烂弧菌、溶藻胶弧菌、迟缓爱德华氏菌和嗜水气单胞菌作为对照,结果显示,对照组不能明显引起荧光强度的改变,而试验组却能显著提高荧光强度.该研究建立的基于荧光能量共振转移(FRET)的具有高灵敏度和特异性的生物传感器检测方法在细菌的早期诊断中有良好的应用潜质.