‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

黄瓜低温弱光耐受性机理及其应用研究的主要进展

选择低温弱光抗性水平不同的欧洲温室型、欧亚杂交型、华北温室型、华北露地型黄瓜(Cucumis sati- vus L.)的16个品种为试验材料,以光合代谢为切入点,开展苗期光合速率、光补偿点、1,5-二磷酸核酮糖羧化酶 (rubisco)活性和叶绿素a荧光动力学曲线的系统研究,结果表明:单一偏低温和偏低温弱光协效应未明显伤害光合系统,而临界低温可能伤及PSⅡ;偏低温和弱光组合时,弱光对黄瓜幼苗的影响起主导作用,弱光与临界低温组合时,临界低温起主导作用;偏低温弱光(单一弱光)下的叶面积增长量和临界低温下的冷害指数可分别作为评价黄瓜对偏低温弱光(单一弱光)耐受性和临界低温耐受性的比较稳定可靠的指标,据此设计了相关的评价指标体系。用117个重组自交系为材料,以弱光下叶面积增长量为指标进行弱光耐受性评价并以RAPD、AFLP、SSR分子图谱为基础,通过区间作图,检测到5个与黄瓜弱光耐受性有关的QTLs。采用mRNA差异显示银染技术克隆得到黄瓜冷敏型品种津研4号低温锻炼中特异表达基因的cDNA克隆(ccr18),其大小为639 bp。在基因组中以单拷贝或低拷贝形式存在。序列同源性比较表明它与拟南芥[Arabidopsis thaliana(L．)Heynh．]染色体ⅣBAC库中的 F14P3基因组序列具有88％的同源性。     

用外翻肠囊法研究高钙条件下有机锰在肉仔鸡小肠中的吸收特点

本试验用外翻肠囊法研究高钙对肉仔鸡不同肠段(十二指肠、空肠和回肠)吸收不同形态锰的影响,以比较有机锰和无机锰在高钙条件下吸收特点的差异。采用单因子完全随机设计,培养液中添加的4种锰源分别为:硫酸锰、弱络合强度的蛋氨酸锰(Mn-MetE)、中等络合强度的氨基酸锰(Mn-AAA)和强络合强度的氨基酸锰(Mn-AAB)。为了扣除内源的影响,设置1个零添加锰水平处理。将40只31日龄AA肉公鸡随机分到以上5个组,每组8个重复,每个重复1只鸡,每只鸡的十二指肠、空肠和回肠分别作为相应肠段外翻肠囊的一个重复。结果表明:1)体外培养回肠肠囊对锰的吸收率显著高于十二指肠(P<0.01);2)回肠肠囊对强络合强度的Mn-AAB中锰的吸收率显著高于硫酸锰或中等络合强度的Mn-AAA(P<0.05)。本试验结果表明:在高钙条件下,络合形态的有机锰在体外培养的肉仔鸡肠囊中的吸收率显著高于无机形态的锰;强络合强度有机锰源中的锰能更强地抵抗小肠内的解离,比弱或中等络合强度的有机锰源更有利于锰的吸收。     

HPLC法检测发酵玉米粉中的黄曲霉毒素B1、B2、G1、G2

发酵玉米粉用甲醇∶水(80∶20,V∶V)提取黄曲霉毒素(AFT),经免疫亲合柱(IAC)分离纯化、在线光化学衍生器柱后衍生化后,反相高效液相色谱(HPLC)荧光检测器测定AFT(B1、B2、G1、G2)含量。4种毒素在0.5ng/g和2ng/g2个水平下,样品加标回收率平均值分别为B188.0%和93.6%、B285.2%和85.3%、G192.4%和91.5%、G289.6%和85.6%;5次测定的标准差分别为:B12.5和6.3、B23.5和6.0、G12.8和9.0、G22.3和6.9;4种毒素的方法检出限均达到0.05ng/g。     

Analysis of gene expression profile of multidrug resistant MCF/DOX cell line after benflumetol derivative LY980503 treatment

AIM:To investigate the effect of LY980503(a benflumetol derivative)on multidrug resistance of tumor cell line using DNA microarray.METHODS:Total RNA was extracted from multidrug resistant MCF/DOX cell line. cDNA microarray containing 320 cDNAs was used to detect the gene expression profile.RESULTS:9 down-regulated genes and 1 up-regulated gene were identified after multidrug resistant MCF/DOX cells were treated with LY980503.CONCLUSION:LY980503 can effectively reverse the resistance of MCF/DOX to DOX in vitro by adjusting the expression of multi-genes.     

灭线磷分析方法浅探

本文介绍了在5％OVl01／Gas ChromQ，长度为2m的玻璃填充柱上，以莠去津为内标物，用FID检测器对灭线磷进行定量分析，分析方法的标准偏差为0．1384％，变异系数为0．15％，回收率范围99.35%-100.28%，线性相关系数为0．9998。     

Inhibitory effects of LDL-ACM complex on subcutaneous implanted tumors in nude mice()

AIM: In order to evaluate the applicable value of LDL as a targeted vehicle for chemotherapeutic agents, we investigated and compared the inhibitory effects of LDL-ACM complex and free ACM on nude mice's subcutaneous implanted tumors derived from gastric cancer cell lines, SGC-7901 and NKM-45. METHODS: LDL-ACM complex was prepared and the tumor model of nude mice was established by subcutaneous implantation of SGC-7901 and NKM-45. Then, the groups of nude mice developed subcutaneous implanted tumors were received either LDL-ACM complex or free ACM. Subsequently, the tumor size, weight and leukemia cell counts were measured and the rates of tumor-inhibition and the survival were compared among the groups. RESULTS: The inhibitory effects of LDL-ACM complex on the tumors, especially on SGC-7901 implanted tumors were much more obvious than that of free ACM. It was also indicated that the action of LDL-ACM complex was mediated by LDL receptor. CONCLUSION: These results showed that LDL-ACM complex had significant inhibitory effects on the implanted tumors and the effect might be mediated by LDL receptor.     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001     

十种常用农药与球孢白僵菌的生物学相容性

球孢白僵菌孢子粉与10种常用农药相容性的测定结果显示,随着孢子浓度上升,所试农药对孢子的抑制作用均有不同程度的增强。在1/10田间常规使用浓度下,百菌清和代森锰锌均能抑制或杀死孢子(萌发率<1％)。除阿维菌素外,所有杀虫剂均与白僵菌孢子相容,在常规使用浓度的10倍稀释液中孢子萌发率达90％以上。吡虫啉、蚜虱灵、灭多威和氟虫腈与孢子的相容性最好,其中吡虫啉和蚜虱灵对孢子萌发率的影响不明显随药剂浓度的变化而变化,即使在田间常规使用浓度下孢子萌发率也在95％以上,而阿维菌素与白僵菌的相容性极差。因此,应用白僵菌制剂防治害虫,选择生物学相容性好的农药以低剂量与白僵菌制剂混用,既可使菌剂增效,又可大幅度降低化学药剂用量。