Establishment of a library of membrane and effector proteins of ´Candidatus P. mali´ to identify host-(apple)-pathogen interactions and to disrupt them

建立“Candidatus P. mali”的膜和效应蛋白库，以识别宿主-（苹果）-病原体相互作用并破坏它们

• 批准号: 261316402
• 负责人: Dr. Kajohn Boonrod
• 金额: --
• 依托单位: AlPlanta Institute for Plant Research
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/261316402?language=en
• 关键词: Establishment; library; membrane; effector; proteins

Candidatus Phytoplasma mali (´Ca. P.mali´) is a plant pathogenic phloem colonizing prokaryote causing apple proliferation, an economically important disease in apple in all European production areas with moderate climates. The disease is transmitted by insect vectors belonging to the genus Cacopsylla. Up to now the apple proliferation disease is controlled only by the application of insecticides. Until recently phytoplasmas have resisted all attempts of cultivation in cell free media, therefore these pathogens are poorly characterized on a physiological and biochemical basis. Recently the genome of ´Ca. P. mali´ and three other phytoplasmas genomes were completely sequenced. With a size of approximately 600.000 bp and 497 open reading frames the genome of `Ca. P. mali´ differs in size and organisation from the three other known phytoplasma genomes.First results indicate that both immunodominant membrane proteins and putative effector proteins of phytoplasmas are relevant in the pathogenesis of phytoplasma diseases. Therefore we will establish a yeast two hybrid (Y2H) library of membrane and effector proteins of ´Ca. P. mali´ ("Prey") to identify interactions of host-(apple-) proteins with phytoplasma proteins based on a screening with a Y2H library deriving from a cDNA library of Malus x domestica cv Golden ("Bait") .The identification of these interacting proteins will lead to a better understanding of the pathogenesis of the apple proliferation disease and moreover, enable the development of innovative management strategies. In consequence we will select aptamers via phage display which are able to disturb and/or disrupt these interactions. Moreover we will evaluate strategies for the application of these aptamers.

Phytoplasma mali（'Ca.苹果疫霉（P.mali '）是一种能侵染苹果韧皮部的原核生物，在欧洲所有气候温和的苹果产区都是一种重要的经济病害。该疾病由属于Cacopsylla属的昆虫媒介传播。目前，苹果增殖病的防治主要依靠农药。直到最近植原体一直抵制在无细胞培养基中培养的所有尝试，因此这些病原体在生理和生化基础上的特征很差。最近的基因组'钙。苹果疫霉和其他三种植原体的基因组被完全测序。Ca基因组大小约为600.000 bp，有497个开放阅读框。苹果疫霉在大小和组织上与其他三种已知植原体基因组不同。初步结果表明，植原体的免疫显性膜蛋白和推定的效应蛋白与植原体疾病的发病机制有关。因此，我们将建立'Ca的膜和效应蛋白的酵母双杂交（Y2 H）库。利用来自苹果（Malus x apartica cv Golden）cDNA文库的Y2 H文库（“Bait”）筛选寄主（苹果）蛋白与植原体蛋白的相互作用，鉴定这些相互作用蛋白将有助于更好地了解苹果增殖病的发病机理，并有助于开发创新的管理策略。因此，我们将通过噬菌体展示选择能够干扰和/或破坏这些相互作用的适体。此外，我们将评估这些适体的应用策略。