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体外分泌蛋白质组学分析鉴定甘蔗黑粉病菌的假定致病相关蛋白

In vitro secretomic analysis identifies putative pathogenicity-related proteins of Sporisorium scitamineum - The sugarcane smut fungus.

作者信息

Barnabas Leonard, Ashwin N M R, Kaverinathan Kalimuthu, Trentin Anna Rita, Pivato Micaela, Sundar Amalraj Ramesh, Malathi Palaniyandi, Viswanathan Rasappa, Carletti Paolo, Arrigoni Giorgio, Masi Antonio, Agrawal Ganesh Kumar, Rakwal Randeep

机构信息

Division of Crop Protection, ICAR-Sugarcane Breeding Institute, 641 007 Coimbatore, India.

Department of Agronomy, Food, Natural Resources, Animals and Environment, University of Padova, Via dell'Università 16, 35020 Legnaro, Padova, Italy.

出版信息

Fungal Biol. 2017 Mar;121(3):199-211. doi: 10.1016/j.funbio.2016.11.004. Epub 2016 Dec 8.

Abstract

Sporisorium scitamineum, the sugarcane smut pathogen, relies predominantly on its secretome to successfully colonise its host, in accordance with other related smut fungi. Considering the significance of deciphering its secretome, we have examined alterations in the in vitro secretome of S. scitamineum in response to synthetic and sugarcane meristem tissue-amended growth media, so as to identify host signal responsive secretory proteins. Secretory proteins that were differentially abundant and exclusively secreted in response to host extract media were identified by two-dimensional gel electrophoresis coupled with MALDI-TOF/TOF MS. Of the 16 differentially abundant and exclusively secreted proteins, nine proteins were identified. Among which, six were related to cell wall modification, morphogenesis, polysaccharide degradation, and carbohydrate metabolism. In planta gene expression profiling indicated that five in vitro secreted proteins were expressed in distinct patterns by S. scitamineum during different stages of infection with relatively higher expression at 1 day after inoculation, suggesting that these proteins could be aiding S. scitamineum at early time points in penetration and colonisation of sugarcane cells. The present study has provided insights into the alterations occurring in the secretome of S. scitamineum at in vitro conditions and has resulted in the identification of secretory proteins that are possibly associated with pathogenicity of the sugarcane smut fungus.

摘要

甘蔗黑粉病菌(Sporisorium scitamineum)与其他相关黑粉菌一样,主要依靠其分泌组成功侵染寄主。鉴于解析其分泌组的重要性,我们研究了甘蔗黑粉病菌在体外对合成培养基和添加甘蔗分生组织的生长培养基的响应过程中分泌组的变化,以鉴定宿主信号响应分泌蛋白。通过二维凝胶电泳结合基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF/TOF MS)鉴定了在宿主提取物培养基中差异丰富且特异性分泌的分泌蛋白。在16种差异丰富且特异性分泌的蛋白中,鉴定出了9种蛋白。其中,6种与细胞壁修饰、形态发生、多糖降解和碳水化合物代谢有关。植物体内基因表达谱分析表明,5种体外分泌蛋白在甘蔗黑粉病菌侵染的不同阶段呈现不同的表达模式,在接种后1天表达相对较高,这表明这些蛋白可能在甘蔗黑粉病菌侵染甘蔗细胞的早期阶段发挥作用。本研究深入了解了甘蔗黑粉病菌在体外条件下分泌组的变化,并鉴定出了可能与甘蔗黑粉病菌致病性相关的分泌蛋白。

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