Reduced quenching and extraction time for mammalian cells using filtration and syringe extraction

Bort, Juan A. Hernández; Shanmukam, Vinoth; Pabst, Martin; Windwarder, Markus; Neumann, Laura; Alchalabi, Ali; Krebiehl, Guido; Koellensperger, Gunda; Sonntag, Denise; Altmann, Friedrich; Heel, Christine; Borth, Nicole

doi:10.1016/j.jbiotec.2014.04.014

Cited by 16 publications

(1 citation statement)

References 42 publications

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“…Leaves were taken from the same part (leaf number 17) to be used as samples. The sample was prepared using a quenching or freezing technique using liquid nitrogen in the eld to stop the metabolic process (Bort et al 2014). The sample was then ground using a mortar and liquid nitrogen.…”

Section: Sample Preparation and 1 H Nmr Analysismentioning

confidence: 99%

1H NMR metabolomics analysis of leaf tissue from oil palms against Ganoderma boninense

Rahmadi,

Syukur,

Widodo

et al. 2024

Preprint

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Introduction Breeding for oil palm resistance against basal stem rot caused by Ganoderma boninense is challenging and lengthy. Advanced oil palm gene pools are very limited hence it is assumed that parental palms have experienced genetic drift and lost their resistance genes against Ganoderma. High-throughput selection criteria should be developed. Metabolomic analysis using 1H Nuclear Magnetic Resonance (NMR) is known to be easy to prepare and the produced metabolite could be used as a diagnostic tool for disease incidents in various host-pathogen combinations. Objectives The study objective is to identify metabolite variations of Dura (D) and Pisifera (P) parental palms with different resistance against Ganoderma and moderately resistant Tenera (DxP) using 1H NMR analysis Methods Leaf tissues of seven different oil palm categories consisting of: resistant, moderate, and susceptible Dura (D), moderate and susceptible Pisifera (P), resistant Tenera/Pisifera (T/P) progenies, and moderately resistant DxP variety progenies were sampled and determined their metabolites using NMR spectroscopy. Results Twenty-nine types of metabolites were identified and most of the metabolites fall in the monosaccharides, amino acid, and fatty acids compound classes. The PCA, PLS-DA, and heatmap multivariate analysis indicated that there were two identified groups of resistances based on their metabolites. The first group consisted of resistant T/P, moderate P, resistant D, and moderately resistant DxP. In contrast, the second group consisted of susceptible P, moderate D, and susceptible D. Glycerol and ascorbic acid were detected as a biomarker candidate by OPLS-DA to differentiate moderately resistant DxP against susceptible D and P. The pathway analysis suggested that the glycine, serine, and threonine metabolism and taurine and hypotaurine metabolism were involved in the oil palm defense mechanism against Ganoderma. Conclusion Metabolomic study with 1H NMR was able to describe the metabolites composition that could differentiate the characteristics of oil palm resistance against basal stem rot caused by Ganoderma boninense. These metabolites revealed in this study had enormous potential to become support tools to breed new oil palm varieties with higher resistance against basal stem rot.

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Section: Sample Preparation and 1 H Nmr Analysismentioning

confidence: 99%