槟榔黄化病叶片差异表达蛋白筛选与鉴定

Screening and Identification of Differentially Expressed Proteins of leaf of Areca infeced by Yellow-leaf disease

以相同品种,树龄、长势一致的黄化病槟榔和健康槟榔叶片为试材,采用TCA(三氯乙酸)/丙酮法制备蛋白质样品,结合双向电泳-质谱结合技术,分析病原菌-槟榔互作后差异表达蛋白。结果表明,双向电泳SDS-PAGE胶中黄化病槟榔叶片与健康叶片平均蛋白质点数分别为1081个和960个,其中差异明显的点34个。选择5个差异蛋白点进行质谱(MALDI-TOF-MS)鉴定,并进行数据库查询,结果鉴定了2个蛋白质,分别为核酮糖-1,5-二磷酸羧化/加氧酶和蕈状支原体高同源蛋白,这些蛋白质可能参与了槟榔黄化病发生和发展过程。另外3个蛋白点在数据库中未检索到同源性和匹配率较高的蛋白质,认为是未知蛋白。

Preparation of protein samples of the leaves by the method of TCA （trichloroacetic acid）/acetone,taking leaves of yellowing-leaf disease and healthy areca as material which was consistent with species,age and growth.Differentially expressed proteins after interaction of pathogen and host was analyzed by two-dimensional electrophoresis combined to mass spectrometry technology.The results showed that average of protein point was of 1 081 and 960 on two-dimensional electrophoresis SDS-PAGE for yellow leaf disease and health of betel leaf,respectively,of which differentially expressed proteins are of 34.Five differentially expressed proteins spots identified through spots mass spectrometry （MALDI-TOF-MS） and database queries.Two proteins was identified,namely,ribulose 1,5-bisphosphate carboxylase of/oxygenase and the mushroom-shaped mycoplasma high homologous proteins,the proteins may be involved in taking place and development of yellow disease of areca.Another three samples were not retrieved in the database and matching higher homology protein,considered to be unknown proteins.