‘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.     

早生桑品种育151、育237的育成

采用杂交方法育成早生桑品种育151、育237,具有发芽早、产叶量高,叶质较优、抗黄化型萎缩病较强等特性,发芽期比早青桑捉早2-3天.比湖桑32号早8天左右.产叶量提高6-41%.     

黄瓜全雌性基因连锁的AFLP和SCAR分子标记

 本研究以全雌品种‘戴多星’自交系和弱雌品种‘北京截头’自交系为双亲杂交获得F₁ ,然后得到F₂ 性型分离群体, 利用分离群体分组分析法(Bulked Segregant Analysis, BSA) 构建全雌和弱雌两个基因池, 筛选了64对AFLP选择性引物EcoR I-NN +Mse I-NNN组合, 发现EcoR I-TG +Mse I-CAC引物组合在全雌基因池中扩增出一条分子量为234 bp的特异带。经F₂ 代单株验证, 该特异条带能在全雌单株中稳定出现。以MAP MAKER (Version 310) 软件分析, 该标记与全雌性位点的连锁距离在617 cM。命名该连锁标记为TG/CAC₂₃₄。将该特异条带回收、克隆、测序, 设计特异SCAR引物, 再对F₂ 代单株基因组DNA进行扩增, 仅在全雌单株中扩增出1条分子量为166 bp 的特异带, 表明已成功地将与黄瓜全雌性连锁的AFLP标记转化为操作简便、表现稳定的SCAR标记, 该标记命名为SA₁₆₆。     

蝴蝶兰组织培养中的褐化控制研究

 系统研究了吸附剂(活性炭、PVP) 与柠檬酸、半胱氨酸、谷胱甘肽、维生素C和硫代硫酸钠5种抗氧化剂对蝴蝶兰组培中褐化的影响。结果表明: 活性炭能有效控制褐化和促进生长, 但不利于分化; 谷胱甘肽控制褐化的效果虽不及活性炭, 但综合效果最佳。     

不同化学物质打破葡萄、桃休眠的研究

以保护地栽培的葡萄、桃为试材,对氰氨类、细胞分裂素、赤霉酸和KNO3打破休眠的效果进行了研究。结果表明,1.5%单氰氨、11%石灰氮可明显促使葡萄提前萌芽,提早着色9-10d;50mg/LGA3和100mg/L6-BA对打破休眠基本无效。TDZ与单氰氨混用提高萌芽率的效果最好。在2个葡萄品种上,均没有观察到单氰氨或石灰氮处理对果穗或果实品质性状的不良影响。在华光油桃上,没有观察到化学物质对提早开花或成熟有明显的作用。对葡萄和桃上应用化学物质打破休眠的有关问题进行了讨论。     

日光温室内外金太阳杏光合特性的比较研究

为了探明杏对设施栽培的生理反应,对日光温室内外金太阳杏光合特性进行了比较研究。结果表明,日光温室栽培条件下,金太阳杏单位重量叶片的叶绿素含量增加;光(CO2)饱和点和光(CO2)补偿点降低鸦露地晴天净光合速率Pn日变化呈明显的双峰曲线,温室内也呈双峰线,但双峰出现原因不同。并且研究了影响光合作用的主要生态因子及变化规律,发现净光合速率与胞间CO2浓度、蒸腾作用、气温、叶温、光合有效辐射等因子相关。     

安徽省鸡免疫抑制性疾病的流行病学调查

采集了安徽省6个主要养鸡地区65群鸡的185只病鸡共986份组织样品，对可引起免疫抑制性疾病的5种最常见病毒进行了PCR检测。结果，传染性腔上囊病病毒（IBDV）、鸡传染性贫血病毒（CIAV）、马立克氏病病毒（MDV）、禽白血病病毒（ALV）和网状内皮增生症病毒（REV）的群阳性率和个体阳性率分别为73．08％和40．91％、46．15％和25．95％、41．54％和17．84％、18．46％和7．57％、13．85％和4．86％，其中被检鸡之二重或多重混合感染的总阳性率为24．33％。调查结果证实，免疫抑制性疾病在安徽省的商业鸡群中普遍存在，并与鸡群疾病多而复杂、损失大相关。     

Malignant catarrhal fever-like disease in sheep after intranasal inoculation with ovine herpesvirus-2.

A malignant catarrhal fever (MCF)-like disease was induced experimentally in 3 sheep after aerosol inoculation with ovine herpesvirus-2 (OvHV-2). Each of 3 OvHV-2-negative sheep was nebulized with 2 ml of nasal secretions containing approximately 3.07 X 10(9) OvHV-2 DNA copies from a sheep experiencing an intensive viral-shedding episode. Ovine herpesvirus-2 DNA became detectable by polymerase chain reaction in the peripheral blood leukocytes of all 3 sheep within 3 days, and all 3 seroconverted between 6 and 8 days postinfection (PI). The sheep developed clinical signs, with copious mucopurulent nasal discharge and fever around 14 days PI. One of the 3 clinically affected sheep was euthanized at 18 days PI. Major lesions at necropsy were multifocal linear erosions and ulcers in mucosa of the cheeks, tongue, pharynx, and proximal esophagus and mild disseminated pneumonia. Microscopically, there was extensive moderate superficial histiocytic-lymphocytic rhinitis with epithelial dissociation and degeneration. Moderate multifocal histiocytic bronchointerstitial pneumonia was associated with loss of terminal bronchiolar epithelium. Lymphocytic vasculitis was present only in the lung. The remaining 2 sheep recovered clinically, approximately 25 days PI. The study revealed that clinical signs and lesions resembling MCF can develop when uninfected sheep are exposed to a high dose of aerosolized OvHV-2.     

樟子松在西北干旱沙区的光合日变化特征

在甘肃省民勤治沙站 ,用 Li- 62 0 0便携式光合仪分别测定人工栽植的 4龄、9龄和 2 0龄樟子松的净光合速率日进程 ,其结果表明 :在干旱沙区 ,9龄以上樟子松的光合速率在午前随光强和气温而上升 ,午后下降。其 4龄、9龄和 2 0龄樟子松的最大净光合速率分别为 8.7μmol· m- 2 · s- 1、62 .3μmol· m- 2 · s- 1、1 9.7μmol· m- 2 · s- 1。     

Association of polymorphism of Fc receptor γ chain gene at position-29 in promoter with the susceptibility to systemic lupus erythematosus in southern Chinese

AIM:To detect the association between the polymorphism of Fc receptor γ chain gene at position-29 in promoter and systemic lupus erythematosus(SLE).METHODS:The genotypes at position -29 in promoter of Fc receptor γ chain gene were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 180 patients with SLE and 140 ethnically matched controls in southern China.RESULTS:The frequencies of TT genotype(33.3%) and T allele (54.4%) at position -29 in patients with SLE were significantly higher than those in controls (17.2% and 42.9%, respectively), whereas, the frequencies of GG genotype (24.4%) and G allele (45.6%) in patients with SLE were remarkably lower than those in controls (31.4% and 57.1%, respectively) (P＜0.05). The TT genotype and T allele at position -29 were not associated with lupus nephritis in SLE patients (P＞0.05).CONCLUSION:Our results indicate that the T allele at position -29 in promoter of Fc receptor gene probably contributes to the susceptibility to SLE, but does not play a role in the occurrence of lupus nephritis.