Determination of the activity signature of key carbohydrate metabolism enzymes in phenolic-rich grapevine tissues

TY - JOUR

T1 - Determination of the activity signature of key carbohydrate metabolism enzymes in phenolic-rich grapevine tissues

AU - Covington, Elizabeth Dunn

AU - Roitsch, Thomas Georg

AU - Dermastia, Marina

PY - 2016

Y1 - 2016

N2 - Physiological studies in plants often require enzyme extraction from tissues containing high concentrations of phenols and polyphenols. Unless removed or neutralized, such compounds may hinder extraction, inactivate enzymes, and interfere with enzyme detection. The following protocol for activity assays for enzymes of primary carbohydrate metabolism, while based on our recently published one for quantitative measurement of activities using coupled spectrophotometric assays in a 96-well format, is tailored to the complexities of phenolic- and anthocyanin-rich extracts from grapevine leaf. As a case study we applied the protocol to grapevine leaf samples infected with plant pathogenic bacteria 'Candidatus Phytoplasma solani', known to alter carbohydrate metabolism in grapevine. The described adaptations may be useful for determination of metabolic fingerprints for physiological phenotyping of other plant species with inherently high levels of phenolic compounds.

AB - Physiological studies in plants often require enzyme extraction from tissues containing high concentrations of phenols and polyphenols. Unless removed or neutralized, such compounds may hinder extraction, inactivate enzymes, and interfere with enzyme detection. The following protocol for activity assays for enzymes of primary carbohydrate metabolism, while based on our recently published one for quantitative measurement of activities using coupled spectrophotometric assays in a 96-well format, is tailored to the complexities of phenolic- and anthocyanin-rich extracts from grapevine leaf. As a case study we applied the protocol to grapevine leaf samples infected with plant pathogenic bacteria 'Candidatus Phytoplasma solani', known to alter carbohydrate metabolism in grapevine. The described adaptations may be useful for determination of metabolic fingerprints for physiological phenotyping of other plant species with inherently high levels of phenolic compounds.

KW - AGPase

KW - Carbohydrates

KW - Invertases

KW - Panel of enzyme activity assays

KW - Phytoplasma

KW - Sucrose synthase

U2 - 10.17344/acsi.2016.2484

DO - 10.17344/acsi.2016.2484

M3 - Journal article

AN - SCOPUS:85003875024

SN - 1318-0207

VL - 63

SP - 757

EP - 762

JO - Acta Chimica Slovenica

JF - Acta Chimica Slovenica

IS - 4

ER -