葡萄抗、感白粉病植株叶片蛋白质组差异比较

摘要

【Objective】This research aimed to find disease-susceptible and resistant protein by proteomics comparison of susceptible and resistant grape leaves.【Method】With the material of disease resistant plant 6-12-4 and susceptible plant 6-12-2,their leaves were artificially inoculated by grape powdery mildew and then picked 0 ,2 and 4 days after inoculation.The total proteim were extracted from the sample leaves and their proteomic differences were analysed by two dimension electrophoresis.Differntially expressed protein dots were recycled,purified and identified by MALDITOF Mass method.【Result】Totally 26 differentially expressed protein spots were obtained,10 of the most differentially expressed proteins were identified by MS.7 out of 10 were positive,and the rest negative.Peptide fragment sequencing result showed that among these 7 positive proteins,two positive spots were hypothetical proteins,and the other two unnamed proteins,spot 5411 was oxygen evolving complex 33 ku photosystem II protein,spot 5625 was ribulose-1,5-bis-phosphate carboxylase/oxygenase activase 1,Rubisco;spot 7121 was pathogensis related protein,PR10.【Conclusion】These proteins were somehow associated with the disease-resistant and disease-susceptible characteristics of grape.%【目的】运用蛋白质组学比较研究白粉病不同抗病性葡萄蛋白质组构成的差异，找出白粉病感病蛋白和抗病蛋白。【方法】以抗病葡萄植株“6-12-4”和感病植株“6-12-2”为材料，在人工接种白粉菌后0（接种前），2，4 d取叶片样品，提取其总蛋白，通过双向电泳技术，找到不同抗病性葡萄接种白粉菌后不同时间表达有差异的蛋白质点，对其中表达差异明显的蛋白质点进行回收和纯化，并进行MALDI-TOF质谱鉴定。【结果】从供试样品中共获得26个差异表达的蛋白质点，其中10个表达有明显差异的蛋白质经质谱鉴定后有7个为阳性斑点，3个为阴性斑点。蛋白质肽质量指纹鉴定结果表明，7个阳性斑点中有2个是假定蛋白，2个是未命名蛋白，差异蛋白质点5411为33 ku光系统Ⅱ放氧复合体外周蛋白，差异蛋白质点5625为核酮糖1,5-二磷酸羧化酶/加氧酶，差异蛋白质点7121为病程相关蛋白PR10。【结论】这些差异表达的蛋白可能与葡萄植株抗、感白粉病的特性有一定关联。