Separation and identification of soybean leaf proteins by two-dimensional gel electrophoresis and mass spectrometry

Xu, Chenping; Garrett, Wesley M.; Sullivan, Joe H.; Caperna, Thomas J.; Natarajan, Savithiry

doi:10.1016/j.phytochem.2006.09.002

Cited by 52 publications

(47 citation statements)

References 42 publications

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“…Proteomics databases and reference maps for Medicago truncatula (Lei et al, 2005) and soybean (Xu et al, 2006) have been constructed using two-dimensional (2D) gel electrophoresis. The 2D gels have also been employed in the identification of proteins expressed during seed filling (Mooney and Thelen, 2004;Hajduch et al, 2005), symbiotic nitrogen fixation (El Yahyaoui et al, 2004), for the detection of glycoproteins using iodine-labeled lectins (Basha and Roberts, 1981), and to explore the consequences of biotic stresses (Lei et al, 2005).…”

mentioning

confidence: 99%

The Nematode Resistance Allele at the rhg1 Locus Alters the Proteome and Primary Metabolism of Soybean Roots

et al. 2009

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Heterodera glycines, the soybean cyst nematode (SCN), causes the most damaging chronic disease of soybean (Glycine max). Host resistance requires the resistance allele at rhg1. Resistance destroys the giant cells created in the plant's roots by the nematodes about 24 to 48 h after commencement of feeding. In addition, 4 to 8 d later, a systemic acquired resistance develops that discourages later infestations. The molecular mechanisms that control the rhg1-mediated resistance response appear to be multigenic and complex, as judged by transcript abundance changes, even in near isogenic lines (NILs). This study aimed to focus on key posttranscriptional changes by identifying proteins and metabolites that were increased in abundance in both resistant and susceptible NILs. Comparisons were made among NILs 10 d after SCN infestation and without SCN infestation. Two-dimensional gel electrophoresis resolved more than 1,000 protein spots on each gel. Only 30 protein spots with a significant (P , 0.05) difference in abundance of 1.5-fold or more were found among the four treatments. The proteins in these spots were picked, trypsin digested, and analyzed using quadrupole time-of-flight tandem mass spectrometry. Protein identifications could be made for 24 of the 30 spots. Four spots contained two proteins, so that 28 distinct proteins were identified. The proteins were grouped into six functional categories. Metabolite analysis by gas chromatography-mass spectrometry identified 131 metabolites, among which 58 were altered by one or more treatment; 28 were involved in primary metabolism. Taken together, the data showed that 17 pathways were altered by the rhg1 alleles. Pathways altered were associated with systemic acquired resistance-like responses, including xenobiotic, phytoalexin, ascorbate, and inositol metabolism, as well as primary metabolisms like amino acid synthesis and glycolysis. The pathways impacted by the rhg1 allelic state and SCN infestation agreed with transcript abundance analyses but identified a smaller set of key proteins. Six of the proteins lay within the same small region of the interactome identifying a key set of 159 interacting proteins involved in transcriptional control, nuclear localization, and protein degradation. Finally, two proteins (glucose-6-phosphate isomerase [EC 5.3.1.9] and isoflavone reductase [EC 1.3.1.45]) and two metabolites (maltose and an unknown) differed in resistant and susceptible NILs without SCN infestation and may form the basis of a new assay for the selection of resistance to SCN in soybean.

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confidence: 99%

The Nematode Resistance Allele at the rhg1 Locus Alters the Proteome and Primary Metabolism of Soybean Roots

et al. 2009

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“…As result, a total of 192 proteins from transgenic soybean seeds were identified, 179 of them identified within the pH range from 4 to 7, and 13 of them identified within the 3-10 pH range. Regarding the pH range from 4 to 7, 49% of the spots present in the gel were identified in the database showing good efficiency with a similar study involving soybean published in the literature [80]. Regarding their biological functions, 50% were related to storage function, 18% related in growth/cell division process, 9% involved in metabolic/ energy process, 6% related to protein transport, 4% corresponding to proteins involved in the disease/defense category and 21% in the category of non-classified proteins.…”

Section: Two-dimensional Gel Electrophoresis: 2-d Pagementioning

confidence: 56%

“…The determination of an organism metallome involves separation techniques associated to microanalytic processes, such as mass spectrometry. These are the two key steps for general proteomics: separation and posterior identification of the proteins [80].…”

Section: Metallomics Studies Involving Hplc Coupled To Icp-msmentioning

confidence: 99%

Comparative Studies Involving Transgenic and Non-Transgenic Soybean: What is Going On?

Arruda¹,

Azevedo²,

Barbosa³

et al. 2013

A Comprehensive Survey of International Soybean Research - Genetics, Physiology, Agronomy and Nitrogen Relationships

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“…Of the 79 spots, 56 had been identified previously (Xu et al, 2006). The remaining 23 spots were digested with trypsin and subjected to matrix-assisted laser desorption/ ionization-time of flight (MALDI-TOF) MS. Only one spot (spot 3, Rubisco activase) could be identified in this way.…”

Section: Identification Of Differentially Accumulated Proteinsmentioning

confidence: 99%

Impact of solar Ultraviolet-B on the proteome in soybean lines differing in flavonoid contents

Sullivan

Garrett

et al. 2008

Phytochemistry

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Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to systematically investigate the impact of solar ultraviolet-B (UV-B) radiation on the soybean leaf proteome. In order to investigate the protective role of flavonoids against UV-B, two isolines of the Clark cultivar (the standard line with moderate levels of flavonoids and the magenta line with reduced flavonoids) were grown in the field with or without natural levels of UV-B. The 12-day-old first trifoliates were harvested for proteomic analysis. More than 300 protein spots were reproducibly resolved and detected on each gel. Statistical analysis showed that 67 protein spots were significantly (P < 0.05) affected by solar UV-B. Many more spots were altered by UV-B in the magenta line than in the standard line. Another 12 protein spots were not altered by UV-B but showed significantly (P < 0.05) different accumulations between the two lines, and for most spots the line-specific differences were also observed under UV-B exclusion. Most of the differentially accumulated spots were identified by mass spectrometry. The proteins were quite diverse, and were involved in metabolism, energy, protein destination/storage, protein synthesis, disease/defense, transcription, and secondary metabolism. The results suggest that high levels of flavonoids lead to a reduction in UV-B sensitivity at the proteomic level. Published by Elsevier Ltd.

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Separation and identification of soybean leaf proteins by two-dimensional gel electrophoresis and mass spectrometry

Cited by 52 publications

References 42 publications

The Nematode Resistance Allele at the rhg1 Locus Alters the Proteome and Primary Metabolism of Soybean Roots

The Nematode Resistance Allele at the rhg1 Locus Alters the Proteome and Primary Metabolism of Soybean Roots

Comparative Studies Involving Transgenic and Non-Transgenic Soybean: What is Going On?

Impact of solar Ultraviolet-B on the proteome in soybean lines differing in flavonoid contents

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