Proteomics provides insights into biological pathways altered by plant growth promoting bacteria and arbuscular mycorrhiza in sorghum grown in marginal soil.

Sorghum is an economically important crop, a model system for gene discovery and a biofuel source. Sorghum seedlings were subjected to three microbial treatments, plant growth promoting bacteria (B), arbuscular mycorrhizal (AM) fungi mix with two Glomus species (G. aggregatum and G. etunicatum), Funelliformis mosseae and Rhizophagus irregularis (My), and B and My combined (My+B). Proteomic analysis was conducted followed by integration with metabolite, plant biomass and nutrient data. Out of 366 differentially expressed proteins in sorghum roots, 44 upregulated proteins overlapping among three treatment groups showed positive correlation with sorghum biomass or element uptake or both. Proteins upregulated only in B group include asparagine synthetase which showed negative correlation with biomass and uptake of elements. Phosphoribosyl amino imidazole succinocarboxamide protein with more than 50-fold change in My and My+B groups correlated positively with Ca, Cu, S and sucrose levels in roots. The B group showed the highest number of upregulated proteins among the three groups with negative correlation with sorghum biomass and element uptake. KEGG pathway analysis identified carbon fixation as the unique pathway associated with common upregulated proteins while biosynthesis of amino acids and fatty acid degradation were associated with common downregulated proteins. Protein-protein interaction analysis using STRING identified a major network with thirteen downregulated proteins. These findings suggest that plant-growth-promoting-bacteria alone or in combination with mycorrhiza enhanced radical scavenging system and increased levels of specific proteins thereby shifting the metabolism towards synthesis of carbohydrates resulting in sorghum biomass increase and uptake of nutrients.

[1]  S. Ishida,et al.  14-3-3 Proteins Regulate Intracellular Localization of the bZIP Transcriptional Activator RSG Article, publication date, and citation information can be found at www.aspb.org/cgi/doi/10.1105/tpc.010188. , 2001, The Plant Cell Online.

[2]  A. G. Khan Role of soil microbes in the rhizospheres of plants growing on trace metal contaminated soils in phytoremediation. , 2005, Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements.

[3]  Rosane Minghim,et al.  InteractiVenn: a web-based tool for the analysis of sets through Venn diagrams , 2015, BMC Bioinformatics.

[4]  S. Robinson,et al.  The role of glutamate dehydrogenase in plant nitrogen metabolism. , 1991, Plant physiology.

[5]  U. Paszkowski,et al.  Contribution of the arbuscular mycorrhizal symbiosis to heavy metal phytoremediation , 2006, Planta.

[6]  R. David-Schwartz,et al.  Substantial roles of hexokinase and fructokinase in the effects of sugars on plant physiology and development. , 2014, Journal of experimental botany.

[7]  H. Hirt,et al.  Reactive oxygen species: metabolism, oxidative stress, and signal transduction. , 2004, Annual review of plant biology.

[8]  B. Hirel,et al.  Glutamate dehydrogenase in plants: is there a new story for an old enzyme? , 2003 .

[9]  J. Chory,et al.  Unraveling the paradoxes of plant hormone signaling integration , 2010, Nature Structural &Molecular Biology.

[10]  A. Copetta,et al.  Arbuscular mycorrhizal symbiosis affects the grain proteome of Zea mays: a field study , 2016, Scientific Reports.

[11]  R. Hell,et al.  Functional analysis of the cysteine synthase protein complex from plants: structural, biochemical and regulatory properties. , 2006, Journal of plant physiology.

[12]  D. De Vleesschauwer,et al.  Modulating plant primary amino acid metabolism as a necrotrophic virulence strategy , 2014, Plant signaling & behavior.

[13]  Hunseung Kang,et al.  An Expression Analysis of a Gene Family Encoding Plasma Membrane Aquaporins in Response to Abiotic Stresses in Arabidopsis Thaliana , 2004, Plant Molecular Biology.

[14]  Yongsheng Liu,et al.  The tomato DDI2, a PCNA ortholog, associating with DDB1-CUL4 complex is required for UV-damaged DNA repair and plant tolerance to UV stress. , 2015, Plant science : an international journal of experimental plant biology.

[15]  Joseph M. Dale,et al.  Empirical Analysis of Transcriptional Activity in the Arabidopsis Genome , 2003, Science.

[16]  Hisashi Ito,et al.  The sugar-metabolic enzymes aldolase and triose-phosphate isomerase are targets of glutathionylation in Arabidopsis thaliana: detection using biotinylated glutathione. , 2003, Plant & cell physiology.

[17]  H. Yoon,et al.  Co-expression of monodehydroascorbate reductase and dehydroascorbate reductase from Brassica rapa effectively confers tolerance to freezing-induced oxidative stress , 2013, Molecules and cells.

[18]  A. Ziemienowicz,et al.  Proliferating cell nuclear antigen (PCNA): a key factor in DNA replication and cell cycle regulation. , 2011, Annals of botany.

[19]  R. Meagher,et al.  De Novo Purine Synthesis in Arabidopsis thaliana (II. The PUR7 Gene Encoding 5[prime]-Phosphoribosyl-4-(N-Succinocarboxamide)-5-Aminoimidazole Synthetase Is Expressed in Rapidly Dividing Tissues) , 1996, Plant physiology.

[20]  G. Adam,et al.  INTRODUCTION OF A MODIFIED RIBOSOMAL PROTEIN L3 GENE AS A STRATEGY TO INCREASE TRICHOTHECENE TOXIN RESISTANCE IN PLANTS , 2000 .

[21]  Martina B. Zell,et al.  Loss of cytosolic NADP-malic enzyme 2 in Arabidopsis thaliana is associated with enhanced susceptibility to Colletotrichum higginsianum. , 2012, The New phytologist.

[22]  R. Ferl,et al.  Interaction of a plant 14-3-3 protein with the signal peptide of a thylakoid-targeted chloroplast precursor protein and the presence of 14-3-3 isoforms in the chloroplast stroma. , 2000, Plant physiology.

[23]  Bernard R. Glick,et al.  Plant Growth-Promoting Bacteria: Mechanisms and Applications , 2012, Scientifica.

[24]  Role of polygalacturonic acid and the cooperative effect of caffeic and malic acids on the toxicity of Cu(II) towards triticale plants (× Triticosecale Wittm) , 2015, Biology and Fertility of Soils.

[25]  Qing Yang,et al.  Expression of a wild eggplant ribosomal protein L13a in potato enhances resistance to Verticillium dahliae , 2013, Plant Cell, Tissue and Organ Culture (PCTOC).

[26]  S. Tyerman,et al.  New potent inhibitors of aquaporins: silver and gold compounds inhibit aquaporins of plant and human origin , 2002, FEBS letters.

[27]  F. Medina,et al.  The nucleolar structure and nucleolar proteins as indicators of cell proliferation events in plants , 2005 .

[28]  Lenwood S Heath,et al.  Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. , 2002, Journal of experimental botany.

[29]  H. El-Maarouf-Bouteau,et al.  From intracellular signaling networks to cell death: the dual role of reactive oxygen species in seed physiology. , 2008, Comptes rendus biologies.

[30]  Y. Yamazaki,et al.  Functional Isolation of Novel Nuclear Proteins Showing a Variety of Subnuclear Localizationsw⃞ , 2005, The Plant Cell Online.

[31]  Davide Heller,et al.  STRING v10: protein–protein interaction networks, integrated over the tree of life , 2014, Nucleic Acids Res..

[32]  N. Tuteja,et al.  Mitogen-activated protein kinase signaling in plants under abiotic stress , 2011, Plant signaling & behavior.

[33]  B. Hwang,et al.  Pepper asparagine synthetase 1 (CaAS1) is required for plant nitrogen assimilation and defense responses to microbial pathogens. , 2011, The Plant journal : for cell and molecular biology.

[34]  A. Prescha,et al.  Repression of the 14-3-3 gene affects the amino acid and mineral composition of potato tubers. , 2002, Journal of agricultural and food chemistry.

[35]  K. Shah,et al.  Effect of cadmium on lipid peroxidation, superoxide anion generation and activities of antioxidant enzymes in growing rice seedlings , 2001 .

[36]  Faten Dhawi,et al.  Mycorrhiza and heavy metal resistant bacteria enhance growth, nutrient uptake and alter metabolic profile of sorghum grown in marginal soil. , 2016, Chemosphere.

[37]  S. Duplessis,et al.  Mitogen-Activated Protein Kinase Signaling in Plant-Interacting Fungi: Distinct Messages from Conserved Messengers[W] , 2012, Plant Cell.

[38]  Patrik Rydén,et al.  OnPLS integration of transcriptomic, proteomic and metabolomic data shows multi-level oxidative stress responses in the cambium of transgenic hipI- superoxide dismutase Populus plants , 2013, BMC Genomics.

[39]  G. Cramer,et al.  Proteomic analysis indicates massive changes in metabolism prior to the inhibition of growth and photosynthesis of grapevine (Vitis vinifera L.) in response to water deficit , 2013, BMC Plant Biology.

[40]  L. Harris,et al.  Transgenic corn plants with modified ribosomal protein L3 show decreased ear rot disease after inoculation with Fusarium graminearum , 2012 .

[41]  A. Copetta,et al.  Maize development and grain quality are differentially affected by mycorrhizal fungi and a growth-promoting pseudomonad in the field , 2013, Mycorrhiza.

[42]  Gavin Lingiah,et al.  Function and specificity of 14-3-3 proteins in the regulation of carbohydrate and nitrogen metabolism. , 2003, Journal of experimental botany.

[43]  C. Stange,et al.  Superoxide dismutase is a critical enzyme to alleviate oxidative stress in Aloe vera (L.) Burm. plants subjected to water deficit , 2012 .

[44]  F. Dupont,et al.  Metabolic pathways of the wheat (Triticum aestivum) endosperm amyloplast revealed by proteomics , 2008, BMC Plant Biology.

[45]  A large decrease of cytosolic triosephosphate isomerase in transgenic potato roots affects the distribution of carbon in primary metabolism , 2012, Planta.

[46]  Kefeng Li,et al.  Effect of multiple metal resistant bacteria from contaminated lake sediments on metal accumulation and plant growth. , 2011, Journal of hazardous materials.

[47]  Faten Dhawi,et al.  Mycorrhiza and PGPB modulate maize biomass, nutrient uptake and metabolic pathways in maize grown in mining-impacted soil. , 2015, Plant physiology and biochemistry : PPB.

[48]  K. Shinozaki,et al.  Arabidopsis RPT2a encoding the 26S proteasome subunit is required for various aspects of root meristem maintenance, and regulates gametogenesis redundantly with its homolog, RPT2b. , 2011, Plant & cell physiology.

[49]  Double mutation in tomato ribosomal protein L3 cDNA confers tolerance to deoxynivalenol (DON) in transgenic tobacco. , 2007, Pakistan journal of biological sciences : PJBS.

[50]  J. Kurepa,et al.  Proteasome regulation, plant growth and stress tolerance , 2009, Plant signaling & behavior.

[51]  I. Fridovich,et al.  DNA strand scission by enzymically generated oxygen radicals. , 1981, Archives of biochemistry and biophysics.

[52]  Mohammad Pessarakli,et al.  Reactive Oxygen Species, Oxidative Damage, and Antioxidative Defense Mechanism in Plants under Stressful Conditions , 2012 .

[53]  Kanako Sasaki,et al.  Two solanesyl diphosphate synthases with different subcellular localizations and their respective physiological roles in Oryza sativa , 2010, Journal of experimental botany.

[54]  M. J. Coon,et al.  Inactivation of key metabolic enzymes by mixed-function oxidation reactions: possible implication in protein turnover and ageing. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[55]  M. Maeshima,et al.  iTRAQ Analysis Reveals Mechanisms of Growth Defects Due to Excess Zinc in Arabidopsis1[W][OA] , 2011, Plant Physiology.

[56]  Shweta Sharma,et al.  Methylglyoxal, Triose Phosphate Isomerase, and Glyoxalase Pathway: Implications in Abiotic Stress and Signaling in Plants , 2015 .

[57]  J. Thioulouse,et al.  Fluorescent pseudomonads occurring in Macrotermes subhyalinus mound structures decrease Cd toxicity and improve its accumulation in sorghum plants. , 2006, The Science of the total environment.

[58]  M. Zwieniecki,et al.  The physiological response of Populus tremula x alba leaves to the down-regulation of PIP1 aquaporin gene expression under no water stress , 2013, Front. Plant Sci..

[59]  C. Petri,et al.  Ectopic expression of cytosolic superoxide dismutase and ascorbate peroxidase leads to salt stress tolerance in transgenic plums. , 2013, Plant biotechnology journal.

[60]  R. McCulley,et al.  Tall fescue cultivar and fungal endophyte combinations influence plant growth and root exudate composition , 2015, Front. Plant Sci..

[61]  R. David-Schwartz,et al.  Hexose Kinases and Their Role in Sugar-Sensing and Plant Development , 2013, Front. Plant Sci..