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A. Schots J. De Boer A. Schouten J. Roosien J. F. Zil Verentant H. Pomp L. Bouwman-Smits H. Overmars F. J. Gommers B. Visser W. J. Stiekema J. Bakker 《European journal of plant pathology / European Foundation for Plant Pathology》1992,98(2):183-191
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 (VL) and heavy chain (VH) 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. 相似文献
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枣胚培技术体系的建立 总被引:8,自引:1,他引:8
以‘六月鲜’枣为试材, 对花后12~82 d的幼胚进行培养。结果表明, 适宜于胚挽救的最小胚龄为花后50 d左右; 在胚形成前进行离体培养, 蔗糖浓度以7%为宜, 而胚形成后离体培养以5%蔗糖为宜; 暗培养可促进离体胚的生长发育, 且有利于生根; 植物生长调节剂及有机添加物极显著地影响着胚培效果。花后70~72 d胚龄的胚适宜培养基为1 /2MS + IBA 0.4 (mg/L, 单位下同) +BA 0.8 +LH 500 + GA35 + 5%蔗糖。胚培苗经扩繁后转入1 /2 MS + IBA 1.0 + IAA 0.06 + 2%蔗糖培养基上, 获得了健壮生根苗,生根苗移栽成活率达85%以上。 相似文献
6.
Comparison of morphology, viability, and function between blood and milk neutrophils from peak lactating goats 下载免费PDF全文
Sui Zhi Tian Chai Ju Chang Chih Chi Chiang Huo Cheng Peh Mu Chiou Huang Jai-Wei Lee Xin Zhao 《Canadian journal of veterinary research》2005,69(1):39-45
The morphological features of blood and milk neutrophils from peak lactating goats were compared using light microscopy, scanning electron microscopy and flow cytometry in order to investigate the cytological changes of neutrophils after migration into the mammary gland. The kinetics of reactive oxygen intermediates (ROI) generation and gelatinase release of blood and milk neutrophils, with or without stimulation of phorbol 12-myristate, 13-acetate ester (PMA), were used to characterize their responses to inflammatory stimuli. Neutrophils isolated from goat milk were highly segmented and contained multi-lobed nuclei. Ultrastructurally, milk neutrophils were more ruffled on the surface compared to blood neutrophils. Approximately 30% of milk neutrophils were undergoing cell death, either necrosis or apoptosis, in contrast to 8% of blood neutrophils. The ROI production of activated milk neutrophils peaked earlier than blood neutrophils, but the duration and the intensity were much less. Neutrophils from both sources augmented the release of gelatinase in response to PMA (1 ng/mL). However, the amount of gelatinase released from milk neutrophils was lower (P < 0.05) than that of blood neutrophils. In summary, more neutrophils become apoptotic and necrotic in the mammary gland, presumably due to spontaneous aging, the process of diapedesis, and the interaction with milk components. Milk neutrophils have impaired functionalities in comparison with blood neutrophils. The information is relevant when studying mammary gland immunity and related diseases, such as mastitis. 相似文献
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Tsutomu MATSUMOTO Yuichiro NARA Hiromitsu FURUYA Harumi TAKAHASHI Kiichi TAIRAKO Hideki YAMAMOTO 《Journal of General Plant Pathology》2002,68(4):382-384
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 相似文献
8.
Shohei MATSUURA Shigeru HOSHINO Hideaki HAYASHI Tetsuyuki KOHGUCHI Kyoji HAGIWARA Toshihiro OMURA 《Journal of General Plant Pathology》2002,68(1):99-102
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 相似文献
9.
甜椒始花期氮素分配动态的研究 总被引:2,自引:1,他引:2
利用15N 示踪技术研究了营养液培养甜椒始花期吸收的氮素在体内的动态分配规律。结果表明: 根与果实中氮的含量在始采期以后保持稳定; 而叶片中氮的含量则随生育进程迅速下降, 盛采期时与果实和根相近, 且两倍高于茎和侧枝。始花期由根吸收的标记氮主要贮存在叶片中, 2 周后向新生器官的运转率高达50. 6%, 4 周后达到57. 1%, 6 周后为58. 0%, 说明越是新近吸收的氮素越容易被再度运转到其它器官, 随着在体内时间的延长, 氮的再运转能力逐渐降低。开花后2 周收获的果实中标记氮占始花期吸收总量的3. 24%, 第3、4、6 周分别为11. 12%, 9. 49%和4. 75%。果实是甜椒体内氮的强力库, 氮素竞争力最强。 相似文献
10.
梨果实若干性状的遗传倾向 总被引:5,自引:0,他引:5
以梨12 个杂交组合和138 个栽培品种为试材, 研究几个果实性状的遗传倾向, 结果如下: 栽培品种果实含糖量集中在6 %~9 % , 平均为7. 39 %; 果实含糖量表现为数量性状, 呈正态分布, 杂种后代果实含糖量主要分布在6 %~10 % , 平均为7. 8 % , 遗传传递力平均为90. 8 %。栽培品种含酸量介于0. 08 %~0. 74 % , 平均为0. 238 %; 杂种后代果实含酸量为数量性状, 组合遗传传递力平均为69. 2 %。梨果实挥发性物质共检测出5 种酯类、8 种醇类和乙烯、乙醛、丙酮类共16 种化合物。杂种后代果实中发现了其亲本果实中所没有的戊醇成分。果实肉质性状表现为质量性状, 脆肉对软肉为显性。杂种后代表现为果点变大、密, 果柄变长, 果汁变少, 果皮厚度相当于亲中值。 相似文献