Carbonate extraction process for the metabolic, isozymic and proteomic profiling of rose-scented geranium (Pelargonium sp.), a hyper-acidic plant.
Rose-scented geranium (Pelargonium sp.) is a valuable monoterpene-yielding plant. It has been well characterised phytochemically through the isolation of >270 secondary metabolites, however, there is hardly any biochemical or metabolic information concerning this plant. Initial attempts to investigate its metabolism failed to produce any enzyme activity in the tissue extracts prepared in routine extraction buffers owing to the intrinsic properties of the tissue matrix. It was recognised that cellular hyper-acidity (cell sap pH approximately 3.0) gave rise to very low protein levels in the extracts, thus prohibiting detection of activities of even primary metabolic enzymes that are usually abundantly present in plants. Tissue extraction in Tris solution without pH adjustment (as used for studies involving citrus and banana) led to little or no improvement. Therefore, a novel approach using sodium carbonate solution as an efficient extraction system for enzymes and proteins from the plant was studied. Functionality of the carbonate extraction has been demonstrated through its effectiveness, a several-fold superior performance, in yielding protein, monitoring primary metabolism and secondary metabolic enzymes, and isozymic and polypeptide profiling. The process may also be helpful in the reliable analysis of other acidic plant tissues.
Sangwan
Rajender Singh
RS
Central Institute of Medicinal and Aromatic Plants, PO CIMAP, Kukrail Picnic Spot Road, Lucknow-226015, India. sangwan.lab@gmail.com
Sangwan
Neelam Singh
NS
Sharma
Pankaj Kumar
PK
Chaurasiya
Narayan Das
ND
Mishra
Siddhartha Kumar
SK
Tyagi
Bali Ram
BR
Srivastava
Avdhesh Kumar
AK
D000975:Antioxidants / Q000378:metabolism; D050260:Carbohydrate Metabolism; D002254:Carbonates / Q000737:chemistry*; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D007527:Isoenzymes; D031316:Pelargonium / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D013729:Terpenes / Q000378:metabolism
D000975:Antioxidants; D002254:Carbonates; D007527:Isoenzymes; D010940:Plant Proteins; D020543:Proteome; D013729:Terpenes