‘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.     

水稻三化螟蚁螟在稻株上的定向、着生习性及对水稻不同组织和人工合成“稻酮”的趋化性反应试验

本项工作的目的在于探求水稻三化螟Tryporyza incertulas(Walker)初孵幼虫在稻株上的定向特性,为鉴定水稻的不同生育期、品种植株间的抗螟性能做一些基本工作。其次,以人工合成的“稻酮”进行蚁螟趋化反应试验。工作系于1961—1964年进行。所得结果初步表明:1.在稻株上的定向部位:蚁螟侵入孔距稻株基部的相对高度为,分蘖期<10厘米;圆秆期10—20厘米,孕穗期后>30厘米。在生长期相同的植株上,籼稻>粳稻。入侵途径以叶鞘侧部的脉间蛀入为主,部分咬脉而入。2.蚁螟在分蘖期至圆秆期稻株上主动选择叶鞘部位入侵,并非由“向地性”所支配。3.蚁螟对各生育期水稻叶鞘组织的趋化反应与成虫的产卵选择性相吻合,即偏选分蘖期与孕穗期的稻株(或叶鞘)。4.在稻株叶鞘及孕穗穗苞内含有诱致蚁螟的物质,而在稻叶叶片组织内可能含有拒食作用的物质。5.蚁螟着生数量,在秧苗上为,17天秧>44天秧;在分蘖与圆秆期稻株上,籼稻>梗稻。6.一定浓度的人工合成“稻酮”能明显地引起蚁螟的嗅觉反应。     

新鲜蔬菜硝酸盐含量测定的改进试粉法

 为适应新鲜蔬菜硝酸盐快速检测的需要, 在现有试粉法的基础上对硝酸盐测定的试粉法进行了改良研究。结果表明, 本研究建立的直接以去离子水浸提蔬菜匀浆, 混合试粉配方为柠檬酸∶一水硫酸锰∶无水对氨基苯磺酸∶N - 1 - 萘乙二胺盐酸盐∶细锌粉= 30∶4∶1.6∶0.8∶1, 其加入量为0.1 g的改进试粉法, 对于溶液中硝酸盐含量在0～20 mg/L范围时, 显色吸光值与硝酸盐含量呈现良好的线性关系, 相关系数达0.9999, 方法回收率在97.7%～104.5%之间, 相对标准偏差2.71%。用本改进试粉法测定11种蔬菜的硝酸盐含量与国标法测定结果的t检验具有一致性。     

绵阳城市广场建设中的特色分析

在调查的基础上，介绍了四川绵阳市的广场，并以铁牛广场为例分析其特色和不足。     

施肥和灌溉对蔬菜品质影响的研究进展

综述了蔬菜品质的内涵及施肥、灌溉对蔬菜品质影响的研究进展。综合前人研究结果可以看出,适量增加施肥量通常可改善蔬菜商品品质,并可提高蔬菜产品中蛋白质、游离氨基酸、有机酸等营养物质指标的含量,但过量施肥则常导致总糖、可溶性糖、抗坏血酸等营养成分含量的降低,并可导致卫生品质及风味品质的降低。合理灌溉可以有效提高蔬菜各种品质指标,但对于许多果菜类而言,适当控制浇水量则有利于营养品质的提高。通常情况下,施肥与灌溉对蔬菜产品品质的影响结果受土壤基础肥力、肥料种类及其配比、蔬菜种类、产品成熟度等因素的影响较大。     

菇木年龄对香菇产量和营养成份含量的影响

用南酸枣、秃瓣杜英、马褂木、麻栎、锥栗、枫香、银荆等7个优良菇木树种的不同年龄树木木屑作为栽培基质，选用126香菇品种做供试菌株，通过香菇总产量、冬菇产量、香菇营养物质含量的对比试验，结果表明，这7种菇木林树种7年树龄木屑栽菇香菇的性能优于或不低于15年成年树龄，用南酸枣、秃瓣杜英、马褂木、麻栎、锥栗、枫香、银荆作为经营短轮伐期菇木林树种是可行的。     

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     

Indole-3-acetic acid biosynthesis in <Emphasis Type="Italic">Aciculosporium take</Emphasis>, a causal agent of witches' broom of bamboo

 Aciculosporium take (Ascomycota; Clavicipitaceae) is a causal agent of witches' broom of bamboo plants. The symptoms of this disease are believed to be induced by plant hormones, particularly auxins. Indole-3-acetic acid (IAA) was identified in cultures of this fungus in an l-tryptophan-supplemented liquid medium. IAA production was confirmed on 30 isolates of A. take from various hosts and locations at levels up to 1 mg/l. The biosynthetic pathway of IAA in A. take culture was examined by analyzing intermediate products and by feeding experiments. The results showed that the indole-3-pyruvic acid pathway (l-tryptophan → indole-3-pyruvic acid → indole acetaldehyde → IAA) was the dominant pathway in A. take. Received: June 3, 2002 / Accepted: July 25, 2002     

Inconspicuous restraint of rice seedling growth by root-infecting fungi in soil of a rice paddy field

 Rice seedling growth, estimated by plant height and root development and discoloration, was better in pasteurized soil than in unpasteurized soil obtained from a flooded rice field. Rice seedlings also grew better in sterilized soil modified by adding roots harvested from the pasteurized soil than in soil modified by adding roots harvested from the unpasteurized soil. The results demonstrate that seedling growth in the rice field soil was inhibited by soil microorganisms, even though no typical symptoms such as seedling blight or damping-off appeared. Pythium aristosporum is suggested to be involved in the inhibition. Thus, it appears that inconspicuous restraint of rice seedling growth could occur in soils of rice paddy fields. Received: May 20, 2002 / Accepted: October 16, 2002 Acknowledgments The authors thank Dr. T. Ichitani, former professor at Osaka Prefectural University, for providing an isolate of Pythium aristosporum for comparison, and Mr. Mitsuaki Sato of Akita Prefectural College of Agriculture for technical assistance.