Effects of gamma irradiation on in vitro pollen germination of different Cucumis species

Summary Pollen grains of 14 Cucumis accessions were irradiated with 0, 1, 2, 3 or 4 kGy acute gamma rays and germinated in vitro directly afterwards. Pollen germination was significantly reduced by increasing irradiation dose for all species, except C. melo var. agrestis. Pollen tube growth was generally reduced likewise. Pollen of two C. anguria subspecies was most sensitive to irradiation. Sensitivity of the pollen with respect to pollen tube growth in relation to irradiation dose was inversely related to total DNA amount per nucleus. In vitro germination was not related to DNA amount per nucleus. Results show that the examined Cucumis species, especially C. melo var. agrestis, are sufficiently resistant to irradiation to be used as donor species for in vivo egg cell transformation of the cucumber.     

Genetics of resistance to five strains of turnip mosaic virus in Chinese cabbage

Summary Inheritance of resistance to turnip mosaic virus (TuMV) strains C1, C2, C3, C4, and C5 in Chinese cabbage (Brassica campestris subsp. pekinensis) was evaluated using monoclonal antibodies. Crosses were made between a resistant line, 0–2, and four susceptible line. Seoul (SE),SSD31 (SS), Cheongbang (CH), and Yaki 1 ho (YA), to determine the inheritance of resistance of 0–2 in different genetic backgrounds. Resistance to TuMV was controlled by a single dominant gene or double dominant genes depending on the strain and cross. The resistance genes of 0–2 were modified by susceptible parents such that a single dominant gene was involved in the SS×0–2 combination, but double dominant genes in the SE×0–2 against TuMV-C3 or TuMV-C5. ELISA tests using inoculated and noninoculated leaves in the same plant suggested that the dominant resistance genes inhibit virus movement rather than virus multiplication.     

关于甘薯属甘薯组中 A 群和 B 群之间可交配性的研究

用对甘薯组 A 群的种 I.tiliacea 和 I.gracilis 与 B 群的种 I.trifida 和 I.littoralis 杂交观察授粉后的胚胎发生的情况,发现授粉后7小时,花粉粒均能在柱头上发芽,花粉管也能通过花柱到达胚珠。在授粉后两天的子房内,可看到已受精、正受精、未受精等受精过程的各个阶段。此时以 B 群 K_(233-1)为母本的卵细胞受精率可达80%     

单克隆抗体在植物病毒学中的应用

单克隆抗体是由杂交瘤细胞分泌的针对单一抗原决定簇的抗体。单克隆抗体在植物病毒学中的应用非常广泛，包括对植物病毒的检测、种下分类级别划分、抗原决定簇鉴定、非结构蛋白功能分析、体内病毒含量测定及病毒纯化等。     

Fusion Expression and Purification of Marek’s disease virus Tegument Protein VP16 and Its Antibody Preparation

马立克氏病病毒(Marek’s disease virus ,MDV)UL48基因编码的蛋白与单纯疱疹病毒1型(Herpes simplex virus ,HSV)衣壳蛋白VP16为同源物，将其克隆入pET-32a载体中，获得pET-VP16, 转化Escherichia coli BL21感受态细胞，经IPTG诱导表达，SDS-PAGE电泳显示:融合蛋白获得可溶性表达，表达蛋白的相对分子量约为67 kD；将表达的融合蛋白过His·Bind柱，获得纯化的蛋白，将该蛋白免疫家兔，制备多克隆抗体。通过间接ELISA和Western blot鉴定制备的多克隆抗体的效价和特异性，结果表明，该抗体具有较高的特异性，间接ELISA效价大于2×10－5。     

浅谈中国蛋禽产业的发展方向

简要阐述了中国蛋禽产业的发展现状及其发展过程中存在的问题。通过对发展现状和存在问题的分析，初步论述了中国蛋禽业应沿着产业化和可持续化的方向发展。同时，还要充分利用先进的科研成果，处理好蛋禽产业的发展与环境保护的关系。     

Sandwich ELISA system for cartilage oligomeric matrix protein in equine synovial fluid and serum

Reasons for performing study: Measurement of cartilage oligomeric matrix protein (COMP) in serum has potential for diagnosis of equine osteoarthritis (OA), but clinical use is currently limited by the lack of specificity of an inhibition ELISA as well as by baseline increases due to exercise. Improved methods for ELISA with increased antigen specificity and sensitivity are therefore required for reliable measurement. Hypothesis: Measurement of the serum level of COMP by sandwich ELISA allows identification of horses with OA. Methods: New monoclonal antibodies (mAbs) were elicited against equine cartilage COMP, their epitopes were determined and a sandwich ELISA was developed. The concentrations of COMP in synovial fluid (SF; n = 100) and sera (n = 100) from OA cases were measured by sandwich ELISA as well as by inhibition ELISA and compared with concentrations in normal joints (n = 95) and horses (n = 50). Results: Immunoblots of enzymatically cleaved COMP showed that the new mAbs recognised different epitopes located on a 20 kDa fragment between K⁶³ and K²³⁸ of the EGF‐like repeats. Inhibition ELISA with any mAb detected significantly increased levels of COMP in OA SF compared with normal SF, whereas no significant difference was detected between serum levels of COMP in OA and normal horses. Conversely, sandwich ELISA with the combination of unlabelled 2A11 × biotinylated 11F10 mAbs detected a significant increase in COMP levels in both serum and SF from OA cases compared with levels in normal animals. Conclusions and potential relevance: Measurement of serum COMP with sandwich ELISA may be useful in identifying horses with OA.     

牛病毒性腹泻病毒(BVDV)抗原捕获ELISA检测方法的建立

用牛病毒性腹泻病毒(BVDV)单克隆抗体包被酶标板,以兔多克隆抗体作为夹心抗体,建立BVDV抗原捕获ELISA(AC-ELISA)检测方法,优化反应条件并对该方法的稳定性等指标进行了测试和评价。结果表明,单抗最佳包被质量浓度为5μg/mL,兔多抗血清最佳质量浓度为10μg/mL;单抗在4℃包被12~24 h,多抗在37℃作用1 h为双抗体的最佳反应条件;酶标抗体最适稀释度1∶10000,最适作用时间为1 h;采用1%BSA和1%明胶分别在抗体包被后和加入待检抗原反应后进行两次封闭效果好。用AC-ELISA方法检测临床采集的11份牛腹泻病料和12份健康牛组织样品,同时以病毒分离和RT-PCR检测方法做对比,3种方法符合率很高。研究表明AC-ELISA方法稳定性好,可用于BVDV的临床快速检测。     

栗蚕卵发育起点温度和有效积温的测定

栗蚕(Dictypoea japonica)是珍贵的野生泌丝昆虫资源。对解除滞育的栗蚕卵分别通过15、18、21、24℃恒温处理暖种,测定栗蚕卵的发育起点温度为(10.2±0.6)℃、孵化有效积温为(181.7±11.6)℃.d,表明一旦给予越冬后解除滞育的栗蚕卵9.6℃以上的温度条件,就可能使其开始发育,当有效积温达到170℃.d时便可孵化。该结果可作为栗蚕卵的人工越冬保护及暖种技术处理的理论依据之一。     

猪生长分化因子11的原核表达及抗体制备

本研究旨在获得猪GDF-11的特异性抗体,以便于进一步研究猪GDF-11的功能.对猪GDF-11的编码区进行了克隆、表达载体的构建和转化、原核诱导表达和纯化以及将GDF-11注射到小鼠中并分离抗体.从猪胚肾中分离总RNA,经RT-PCR扩增得到猪GDF-11基因的编码区,插入pMD18-T载体中.再经限制性内切酶BarnH Ⅰ和EcoRⅠ双酶切,将回收的GDF-11酶切片段插入原核表达载体pGEX-4T-2中,获得重组原核表达质粒pGEX-4T-2-GDF-11.重组质粒经测序鉴定后转化大肠杆菌BL21(DE3),并经IPTG诱导产生了65 ku的GST-GDF-11融合蛋白.融合蛋白经凝血酶酶切,分离纯化出42 ku左右的GDF-11蛋白;然后注射小鼠,制备了多克隆抗体,其滴度最高可达1:25 600,而且特异性好,表明得到了猪GDF-11的多克隆抗体.