Proteome of germinating and dormant axillary buds of sugarcane and analysis of sprouting-related proteins

M. M. Benez, R. C. Maranho, G. B. Maranho, Adeline Neiverth, M. F. Santos, Ana Lúcia O. Carvalho, A. Gonela, C. A. Mangolin, M. F. P. S. Machado
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引用次数: 1

Abstract

The germination potential of the axillary buds is crucial for the installation of new sugarcane crop and is highly relevant for the maintenance of the crop during the regrowth cycles. In this way, 1-DE-UPLC-ESI-Q-TOF system of mass spectrometry in a shotgun approach evaluated total and differential proteome of germinating and dormant axillary buds of cultivar RB867515 in second ratoon. The current paper investigates whether differently expressed proteins can be detected in germinating axillary buds or not. The proteome of germinative and dormant axillary buds were obtained using TCA/acetone in triplicates with subsequent proteins and polypeptides pre-separation by SDS-PAGE and UPLC. Mass spectra were obtained via MS/MS and were confronted with Viridiplante mass spectra from NCBIprot, SwissProt and UniProtKB databases, using Mascot tool from Matrix Science. A total of 159 proteins were identified. In total, 46 proteins were exclusively detected in the germinating buds, while 41 proteins were exclusively reported in the dormant buds and 72 proteins were common in germinating and dormant buds. Different proteins, involved with biotic and abiotic stress, were detected at equivalent proportions in germinating (29.5%) and dormant (31.7%) buds, while contrasting proportions of proteins involved with energy metabolism were detected in germinating (22.7%) and dormant (12%) buds. Budding potential in the third cut seems to be determined by specific proteins related to energy metabolism and steps of the biosynthesis of primary and secondary metabolites for shoot growth and development. Seven enzymes (Aconitase, Succinate dehydrogenase, Pyruvate dehydrogenase, Fructose-6-phosphate 1-phosphotransgerase, Chacone synthase, Chalcone isomerase, and Monodehydroascorbate), one protein subunits (RuBisCo large subunit-binding protein) and one protein of Argonaute protein family (Tudor-Sn) supposedly involved with germinating potential of axillary buds and two enzymes (Hexokinase and Aldehyde dehydrogenase) supposedly involved with non-budding were highlighted.
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甘蔗萌发和休眠腋芽蛋白质组学及发芽相关蛋白分析
腋芽的萌发势对甘蔗新作物的种植至关重要,对作物在再生周期中的维持也有很大关系。采用霰弹枪法的1-DE-UPLC-ESI-Q-TOF质谱系统对RB867515胚芽萌发和休眠腋芽的总蛋白质组和差异蛋白质组进行了分析。本文研究了在萌发的腋芽中是否可以检测到不同表达的蛋白。利用TCA/丙酮法获得萌发和休眠腋芽的蛋白质组,随后用SDS-PAGE和UPLC对蛋白质和多肽进行预分离。质谱通过MS/MS获得,并利用Matrix Science的Mascot工具与NCBIprot、SwissProt和UniProtKB数据库中的Viridiplante质谱进行比对。共鉴定出159个蛋白。在萌发芽中只检测到46种蛋白,在休眠芽中只检测到41种蛋白,在萌发芽和休眠芽中共有72种蛋白。在萌发芽(29.5%)和休眠芽(31.7%)中检测到的与生物和非生物胁迫有关的蛋白质比例相当,而在萌发芽(22.7%)和休眠芽(12%)中检测到的与能量代谢有关的蛋白质比例则不同。第三个切口的出芽势似乎是由与能量代谢有关的特定蛋白质和新梢生长发育的初级和次级代谢物的生物合成步骤决定的。强调了7种酶(乌头酶、琥珀酸脱氢酶、丙酮酸脱氢酶、果糖-6-磷酸1-磷酸转氨酶、查尔酮合成酶、查尔酮异构酶和单脱氢抗坏血酸)、1种蛋白亚基(RuBisCo大亚基结合蛋白)和1种Argonaute蛋白家族蛋白(tuddor - sn)和2种酶(己糖激酶和醛脱氢酶)可能与腋芽萌发电位有关。
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