鸡白细胞介素-6原核表达及双抗夹心ELISA方法的建立

应用分子生物学技术原核表达ChIL-6，以ChIL-6重组蛋白为免疫原，按免疫程序分别制备兔抗和鼠抗IL-6重组蛋白的多克隆抗体。应用此抗体建立双抗夹心EL-ISA方法后，为了优化此方法本试验对该方法的最佳试验条件、标准曲线、重复性和初步应用进行确定。结果显示，经SDS-PAGE和Western-blot分析，表明ChIL-6在大肠杆菌中正确表达，为了建立检测ChIL6的双抗夹心ELISA方法，本试验应用表达的重组蛋白制备兔抗和鼠抗多克隆抗体。建立的双抗夹心ELISA方法最佳反应条件为包被抗体的质量浓度为50mg／L，4℃过夜；酶标二抗稀释度为1：400，37。C1h；应用该方法检测感染金黄色葡萄球菌后的ChIL-6，其结果与本实验室之前应用人的ELISA试剂盒检测的结果相似。结果表明，本试验建立的检测ChlL-6的双抗夹心ELISA方法可用于临床。     

间接免疫荧光染色检测鸭源副黏病毒及在鸭体内的抗原定位

以抗新城疫病毒F蛋白的单克隆抗体为一抗,建立了间接免疫荧光染色法(IFA)检测石蜡切片中鸭副黏病毒(DPMV)的方法。以建立的IFA对DPMV人工感染鸭的各组织器官进行检测,结果显示:各组织器官均能检测到DPMV,但不同时间取样,阳性信号分布的器官不同。脾脏、胸腺、法氏囊、肠道、肝脏、肺脏DPMV的阳性检出率较高,表明这些器官为DPMV的主要靶器官。在所检测的阳性组织中,病毒抗原分布在细胞浆中。IFA检测石蜡切片中的DPMV具有直观、特异性强的优点,是对DPMV进行检测和抗原定位的良好方法。     

分泌抗猪瘟病毒单克隆抗体杂交瘤细胞株的建立

采用单克隆抗体制备技术,建立了分泌抗猪瘟病毒单克隆抗体的杂交瘤细胞株,液氮保存4年后复苏,细胞在HAT选择培养基中生长旺盛,分泌抗体稳定,分别命名为YNF1,YNF2,YNF3,YNF4,杂交瘤细胞染色体数介于96~115条。细胞培养上清和腹水的单抗间接ELISA效价分别为1∶80~1∶160和1∶5 000~1∶10 000。在间接ELISA和夹心ELISA检测中单抗与猪瘟病毒呈特异性反应,与正常的猪、兔肾组织提取液及其血清Ig无交叉。抗体蛋白属IgG类,亚类及轻链分别为IgG1/λ,IgG2a/κ,IgG2a/λ,IgG1/λ。结果表明,4株杂交瘤是分泌抗猪瘟病毒单克隆抗体的细胞株,可长期传代无限分泌单抗,是进一步研究猪瘟病毒和开发敏感、特异、快速诊断猪瘟试剂盒和试纸探针的免疫学特异性试剂来源。     

Resistance to the Potato Leafroll Virus (PLRV) in Diploid Potatoes

A high level of PLRV resistance has been found in four diploid genotypes originating from resistant ancestors widely utilized in European potato breeding. Plants of these genotypes were difficult to infect not only with aphids, but also with graft inoculation. Their resistance is associated with limited virus spread, but not with intolerance. The level of PLRV resistance in these genotypes appears to be comparable to a high level of resistance detected recently in some wild potato species. Evaluation of virus concentration after graft inoculation with PLRV was found useful in the selection of potato genotypes highly resistant to PLRV.     

棉花黄萎病菌毒素ELISA检测方法的建立及其应用

以纯化的棉花黄萎病菌毒素(PLPC)免疫新西兰白兔制备了PLPC特异性抗血清,建立了可特异性检测棉花黄萎病菌毒素的A蛋白双抗体夹心ELISA方法。应用建立的ELISA方法测定了病菌培养滤液、人工接种幼苗和大田病株不同组织中的毒素,结果表明:不同产毒能力菌株(VD 8和VD-5)在25℃下振荡培养3d后就能在培养滤液中检测到毒素,这比生物测定要早2～3d。将VD-8菌株的孢子以灌根方式接种泗棉3号幼苗5d后,就能从接种幼苗的茎杆和叶柄中检测到毒素,这比幼苗自然发病显症要早3～5d。在测定的30份大田病株茎杆、叶柄、叶脉样品中,阳性样品率为100%。这些结果表明建立的ELISA方法可用于菌株产毒能力的测定和大田棉花黄萎病的早期诊断。     

Selection for Seed Yield in Lolium perenne L.*

Components of seed yield were estimated for each set of 100 plants derived from six varieties of Lolium perenne. Significant differences between the varieties were found for all components of seed yield, but the range was broader within each variety. An indirect selection of seed yield using the character seeds per spike produced a selection response similar to that obtained by direct selection on high seed yield per plant. A direct and indirect selection of characters estimated in small, drilled plots of topcross off-springs (F₁) was less successful than the selection based on the single plants. Spearman rank correlation coefficients between the single plants and their offsprings did not show significant agreement for seed yield. But altogether the results indicated that the increase of seed yield/area was about 6 % when 10 % of the best plants were selected.     

Interactions between different genotypic tissues in citrus graft chimeras

The heritability of tolerance of wheat F₃ lines to competition from annual ryegrass (Lolium rigidum), and its relationship to morphological traits, were determined using crosses between wheat cultivars with good and poor competitive ability. Forty F₂-derived F₃ lines from a cross between two late flowering varieties (Machete × Spear) and 40 from a cross between early flowering varieties (Wilgoyne × Gutha) were grown in the field with and without annual ryegrass. There was significant genetic variation between lines within each of the two populations in a number of aspects of plant growth and development, including yield in monoculture. The estimates of heritability for % yield loss due to competition were 0.25 and 0.57 in the two crosses respectively, indicating that selection for high tolerance to competition in the F₃ generation or later should be effective. Fairly strong relationships between height and % yield loss and between leaf length and % yield loss suggest that these may be useful auxiliary traits when selecting for low % yield loss. However, differences between crosses in the magnitude and sign of genetic and phenotypic correlation between traits indicate that competitive ability is a complex character influenced by many factors. This revised version was published online in August 2006 with corrections to the Cover Date.     

Indirect Regulation Pricing Methods of Public Utility

The author analyses the advantages and disadvantages of common indirect regulation pricing methods of public utility goods. It is pointed out that it results in high trade-cost and A-J effect binging in an excess of investment when capital yield method be used. It is not reasonable enough in setting PRI-X of PRI-X pricing method. An integrated indirect regulation pricing method is put forward which is to form a two-tier incentive mechanics both in expanding producing ability and improving efficiency. In initial stages of new public project, capital yield method should be used in order to activate investment motive. Upper-limit pricing method should be used in anaphase in order to improve the producing efficiency.     

GA1,3,4,7酶联免疫检测法（ELISA）的建立

用碳二亚胺法将赤霉素Ａ４与卵清蛋白连接，形成ＧＡ４－ＯＶＡ复合物用于包板，利用ＧＡ４－Ｍｅ与ＧＡ４－０ＶＡ对兔抗ＧＡ４－ＰＡｂｓ竞争结合反应，建立了ＧＡ１，３，４，７间接酶闻免疫检测法。其灵敏度，检测范围和最小检测样品量分别为４５ｐｇ，０．２－２００ｎｇ和５０－５００ｍｇ，样品的平均回收率为９７％，批内和批间的变异系数分别为４．２％和４．８％。     

Selection for Increased Seed Nitrogen Accumulation in Common Bean: Implications for Improving Dinitrogen Fixation and Seed Yield

Estimates of N₂ fixation in segregating populations of bean plants based on ¹⁵N-isotope methods are technically demanding and expensive; therefore, indirect measures based on related traits including total seed N were used to select for improved N₂ fixation and yield. In 1985, six populations of F₂-derived F₃ families resulting from six parental lines crossed to a common tester were grown in field trials on a low-N soil. In 1986, 25 selected half-sib families and two populations of full-sib F₄ families were grown under similar conditions. Parents and a non-nodulating soybean line were included both years. Narrow sense (H_NS) heritability estimates based on parent-offspring regressions ranged from 0.57 for total seed N to 0.39 for shoot biomass in one population, but were near zero for all measured traits in a second population. Among the criteria used to identify parental lines with superior potential for producing progenies with high total seed N, testcross population means combined with estimates of realized heritability were the most reliable. Selection of the best F₃ families for total seed N resulted in F₄ families with increased total seed N and higher seed yields, while seed protein percentage was unchanged. When plants are grown on low-N soils, selection for total seed N offers a useful alternative to selection for increased N₂ fixation based on ¹⁵N-isotopic or total-N-difference method.