‘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.     

葡萄自交与杂交后代香气成分的遗传研究

 利用气相色谱- 质谱- 计算机联用系统(GC-MC-CS) 对两个自交组合、1个杂交组合亲本及其后代的香气成分进行研究。结果表明: 同一组合中不同香气成分的分离情况不同, 同种香气成分在不同组合中的分离情况也不完全相同。果实香气成分的遗传主要表现为分离( 1∶1、1∶3、3∶1、15∶1) 与不分离两种方式, 发生分离的成分有些是由单基因控制的, 有些是由多基因控制的。     

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。     

荧光PCR快速诊断一起新城疫

2005年1月底．广东省某规模养殖场的鸡出现了张口喘气、呼吸困难，有呼噜声、咳嗽，排黄绿色粪便等症状．部分鸡还有神经症状。发病约450只．发病率为1．2％。三天后鸡只陆续出现死亡．两天内死亡50只左右。解剖病死鸡，可见：喉头、气管粘膜明显出血，小肠轻度炎症。回肠、直肠、泄殖腔粘膜出血。临床无法确诊。经实验室快速诊断．为新城疫感染。     

犬胚胎移植手术路径研究性试验

犬胚胎移植手术路径的确定是移植能否成功的关键。我们选择了12例自然发情的母犬，分为供体组和受体组，每组6头。通过生物检测镜确定排卵时间，用腹腔镜观察卵巢黄体发育情况，同时于交配后第9、11、13天剖腹探查，确定冲卵时间。结果于交配后第11天冲洗供体犬左侧子宫角，保留右侧子宫角的两例犬受孕，移植给2头受体犬左侧子宫角。     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002     

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     

甘薯根腐病抗性在不同环境条件下的表现及遗传趋势

结果表明,甘薯根腐病发病轻重与环境条件有很大关系,表现为干旱少雨的年份发病较重,降雨量较多的年份发病较轻;土壤瘠薄的发病较重,肥力条件较好的则发病较轻;通常情况下,年份间品种的抗性表现较为一致,但遇特殊气候则年份间品种的抗性有一定的差异。对1150份甘薯品种资源及育种材料的根腐病抗性鉴定结果表明,高抗型材料占14.6％,抗病型占15.7％,感病型占26.0％,高感型占43.7％。对754份材料及亲本的抗性分析表明,不同的抗性组合后代中均可分离出高抗至高感类型的材料,杂交后代的抗性强弱随双亲抗性水平的提高而提高。中国自1970年以来采用品种间杂交和种间杂交育种技术,先后育成了一批高产、优质的高抗型优良品种。     

β- 氨基丁酸诱导甜( 辣) 椒抗疫病作用的研究

 报道了用β- 氨基丁酸( DL-β-aminon-butyric acid, BABA) 喷雾处理辣椒叶片和茎后的诱导抗疫病作用。研究证明: 高浓度BABA ( 1 000 g/mL) 对离体辣椒疫霉病菌无抗菌活性, 用其喷雾处理辣椒的茎叶所诱导的抗疫病作用可完全控制其危害; 用BABA 诱导处理后3 d 接种辣椒疫霉病菌, 辣椒植株开始表达出较高的诱导抗性, 这种抗病作用可持续20 d 以上, 并表现出与数量抗病性相似的特性。