Extraction of Total Protein from Axillary Buds of Sugarcane (Saccharum spp.) for Proteomic Analysis

Maranho, Rone Charles; Benez, Mariana Mancini; Maranho, Gustavo Barizon; Fernandes, Vanessa Neves de Azevedo; Gonela, Adriana; Mangolin, Claudete Aparecida; Machado, Maria de Fátima Pires da Silva

doi:10.1007/s12355-017-0520-z

Cited by 2 publications

(3 citation statements)

References 9 publications

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“…Total proteins were extracted from the axillary buds (200 mg; eight axillary buds) of each plant using the modified TCA-acetone method for sugarcane (Maranho et al, 2018). Following extraction, the proteins were quantified by fluorimetry (Qubit Fluorometer 1.0; Invitrogen, Carlsbad, CA, USA) using the Qubit Protein Assay Kit (Invitrogen).…”

Section: Protein Extraction Quantification and Digestionmentioning

confidence: 99%

Proteome in regrowth cycles of sugarcane: Absence of proteins to tolerate adverse growth conditions may be related to reduced agricultural productivity

Maranho

Benez

et al. 2023

Acta Sci. Agron.

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One of the main objectives of sugarcane plantations is to increase their longevity without decreasing agricultural productivity. In the present study, we analyzed the proteome of the axillary buds of ‘RB966928’ to investigate possible changes in the number of proteins at different cutting stages. Using tryptic digestion followed by ultra-performance liquid chromatography coupled with high-resolution time-of-flight mass spectrometry, 122 proteins were identified from the proteome of the axillary buds of ‘RB966928’. Of the 122, respectively 97 and 95 proteins were detected at the first and fifth cutting stages, of which 27 and 25 proteins were unique to the respective stage. Proteins that prevent the misfolding of polypeptides generated under stress were exclusively detected at the first cutting stage. Meanwhile, proteins associated with stress responses and disease resistance were exclusively detected at the fifth cutting stage. The present proteomic analysis in the regrowth cycles and axillary bud development of ‘RB966928’ significantly advanced our understanding of the biological processes linked to the reduction of agricultural productivity of sugarcane with the advancement of cutting age. Absence of proteins to tolerate adverse growth conditions at the fifth cutting stage may be related to reduced agricultural productivity, in addition to environmental stress, soil compaction, nutrient availability, cultural practices, and pests or pathogen attacks at different phenological stages of crops.

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Section: Protein Extraction Quantification and Digestionmentioning

confidence: 99%

Proteome in regrowth cycles of sugarcane: Absence of proteins to tolerate adverse growth conditions may be related to reduced agricultural productivity

Maranho

Benez

et al. 2023

Acta Sci. Agron.

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“…The extraction of total proteins following method described by Maranho et al (2017) provided an average amount of 1.526 μg•mL -1 and 1.237 μg•mL -1 proteins in the germinating and dormant axillary buds, respectively. Protein pattern obtained by 1DE SDS-PAGE revealed proteins with molecular weights varying between 120 and 10 kDa.…”

Section: Protein Extraction Of Axillary Buds and Protein Profile By Smentioning

confidence: 99%

“…Axillary buds (200 mg) were used in triplicates from each germinating and dormant buds. Total proteins were extracted by modified TCA/acetone method for sugarcane axillary buds (Maranho et al, 2017). After extraction, proteins were quantified by a fluorometer (Qubit® Fluorometer 1.0) with Qubit® Protein Assay quantification kit from Life Technologies™.…”

Section: Extraction Quantification and Pre-fractioning Of Total Protmentioning

confidence: 99%

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

Benez

Maranho

et al. 2019

Plant Omics

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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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Extraction of Total Protein from Axillary Buds of Sugarcane (Saccharum spp.) for Proteomic Analysis

Cited by 2 publications

References 9 publications

Proteome in regrowth cycles of sugarcane: Absence of proteins to tolerate adverse growth conditions may be related to reduced agricultural productivity

Proteome in regrowth cycles of sugarcane: Absence of proteins to tolerate adverse growth conditions may be related to reduced agricultural productivity

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

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