GC-MS analysis of headspace and liquid extracts for metabolomic differentiation of citrus Huanglongbing and zinc deficiency in leaves of 'Valencia' sweet orange from commercial groves.
BACKGROUND
Citrus Huanglongbing (HLB) is considered the most destructive citrus disease worldwide. Symptoms-based detection of HLB is difficult due to similarities with zinc deficiency.


OBJECTIVE
To find metabolic differences between leaves from HLB-infected, zinc-deficient, and healthy 'Valencia' orange trees by using GC-MS based metabolomics.


METHODS
Analysis based on GC-MS methods for untargeted metabolite analysis of citrus leaves was developed and optimized. Sample extracts from healthy, zinc deficient, or HLB-infected sweet orange leaves were submitted to headspace solid phase micro-extraction (SPME) and derivatization treatments prior to GC-MS analysis.


RESULTS
Principal components analysis achieved correct classification of all the derivatized liquid extracts. Analysis of variance revealed 6 possible biomarkers for HLB, of which 5 were identified as proline, β-elemene, (-)trans- caryophyllene, and α-humulene. Significant (P < 0.05) differences in oxo-butanedioic acid, arabitol, and neo-inositol were exclusively detected in samples from plants with zinc deficiency. Levels of isocaryophyllen, α-selinene, β-selinene, and fructose were significantly (P < 0.05) different in healthy leaves only.


CONCLUSIONS
Results suggest the potential of using identified HLB biomarkers for rapid differentiation of HLB from zinc deficiency.
IFAS, Citrus Research and Education Center, University of Florida, Lake Alfred, FL, USA.
Juan Manuel
JM
Cevallos-Cevallos
Rosalía
R
García-Torres
Edgardo
E
Etxeberria
José Ignacio
JI
Reyes-De-Corcuera
D015415:Biomarkers; D002957:Citrus / Q000737:chemistry / Q000378:metabolism* / Q000382:microbiology; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D055432:Metabolomics / Q000379:methods*; D010935:Plant Diseases / Q000145:classification* / Q000382:microbiology; D010936:Plant Extracts / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry / Q000378:metabolism / Q000382:microbiology; D025341:Principal Component Analysis; D052617:Solid Phase Microextraction / Q000379:methods; D015032:Zinc / Q000378:metabolism*
D015415:Biomarkers; D010936:Plant Extracts; D015032:Zinc