Application of high-performance liquid chromatography with photodiode array detection to the metabolic profiling of plant isoprenoids

Fraser, Paul D.; Pinto, M. E. S.; Holloway, Daniel E.; Bramley, Peter M.

doi:10.1046/j.1365-313x.2000.00896.x

Cited by 368 publications

(220 citation statements)

References 30 publications

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“…Sample preparation, analysis of carotenoids and chlorophylls by high-performance liquid chromatography (HPLC) were performed essentially according to a previous report [44]. The samples were subjected to HPLC analysis using a reverse-phase C30, 5 mm column (250 × 4.6 mm 2 ) coupled to a 20 × 4.6 mm C30 guard column (YMC Inc., Wilmington, NC, USA).…”

Section: Analysis Of Carotenoids and Chlorophyllsmentioning

confidence: 99%

The Arabidopsis Spontaneous Cell Death1 gene, encoding a ζ-carotene desaturase essential for carotenoid biosynthesis, is involved in chloroplast development, photoprotection and retrograde signalling

et al. 2007

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Carotenoids, a class of natural pigments found in all photosynthetic organisms, are involved in a variety of physiological processes, including coloration, photoprotection, biosynthesis of abscisic acid (ABA) and chloroplast biogenesis. Although carotenoid biosynthesis has been well studied biochemically, the genetic basis of the pathway is not well understood. Here, we report the characterization of two allelic Arabidopsis mutants, spontaneous cell death1-1 (spc1-1) and spc1-2. The weak allele spc1-1 mutant showed characteristics of bleached leaves, accumulation of superoxide and mosaic cell death. The strong mutant allele spc1-2 caused a complete arrest of plant growth and development shortly after germination, leading to a seedling-lethal phenotype. Genetic and molecular analyses indicated that SPC1 encodes a putative z-carotene desaturase (ZDS) in the carotenoid biosynthesis pathway. Analysis of carotenoids revealed that several major carotenoid compounds downstream of SPC1/ZDS were substantially reduced in spc1-1, suggesting that SPC1 is a functional ZDS. Consistent with the downregulated expression of CAO and PORB, the chlorophyll content was decreased in spc1-1 plants. In addition, expression of Lhcb1.1, Lhcb1.4 and RbcS was absent in spc1-2, suggesting the possible involvement of carotenoids in the plastid-to-nucleus retrograde signaling. The spc1-1 mutant also displays an ABA-deficient phenotype that can be partially rescued by the externally supplied phytohormone. These results suggest that SPC1/ZDS is essential for biosynthesis of carotenoids and plays a crucial role in plant growth and development.

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Section: Analysis Of Carotenoids and Chlorophyllsmentioning

confidence: 99%

The Arabidopsis Spontaneous Cell Death1 gene, encoding a ζ-carotene desaturase essential for carotenoid biosynthesis, is involved in chloroplast development, photoprotection and retrograde signalling

et al. 2007

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“…HPLC was performed in a Varian ProStar model 410 apparatus, a ProStar model 240 pump, and a ProStar 330 photodiode array detector, essentially as described in Ref. 33 (see accompanying paper (31) for details). In some experiments, a combination of octadecyl silica and poly(ethylene) powder media was used, likewise allowing separation of both porphyrins and chlorins in the same HPLC run (34).…”

Section: Methodsmentioning

confidence: 99%

In Situ Conversion of Protochlorophyllideb to Protochlorophyllide a in Barley

Reinbothe¹,

Pollmann²,

Reinbothe³

2003

Journal of Biological Chemistry

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“…For compounds that include double bonds or aromatic substructures, the absorption of UV light can be used. LC/UV profiles of isoprenoids (carotenes, xanthophylls, ubiquinones, tocopherols and plastoquinones) have been used to characterize transgenic and mutant tomato genotypes and for screening Arabidopsis mutants (Fraser et al, 2000). By using arrays of photodiodes, complete UV spectra are acquired at sensitivities capable of producing read-outs with samples as small as 1 ng injected onto the column.…”

Section: Metabolite Profilingmentioning

confidence: 99%

Metabolomics — the link between genotypes and phenotypes

Fiehn

2002

Functional Genomics

1,624

1,337

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Metabolites are the end products of cellular regulatory processes, and their levels can be regarded as the ultimate response of biological systems to genetic or environmental changes. In parallel to the terms 'transcriptome' and 'proteome', the set of metabolites synthesized by a biological system constitute its 'metabolome'. Yet, unlike other functional genomics approaches, the unbiased simultaneous identification and quantification of plant metabolomes has been largely neglected. Until recently, most analyses were restricted to profiling selected classes of compounds, or to fingerprinting metabolic changes without sufficient analytical resolution to determine metabolite levels and identities individually. As a prerequisite for metabolomic analysis, careful consideration of the methods employed for tissue extraction, sample preparation, data acquisition, and data mining must be taken. In this review, the differences among metabolite target analysis, metabolite profiling, and metabolic fingerprinting are clarified, and terms are defined. Current approaches are examined, and potential applications are summarized with a special emphasis on data mining and mathematical modelling of metabolism.

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Application of high-performance liquid chromatography with photodiode array detection to the metabolic profiling of plant isoprenoids

Cited by 368 publications

References 30 publications

The Arabidopsis Spontaneous Cell Death1 gene, encoding a ζ-carotene desaturase essential for carotenoid biosynthesis, is involved in chloroplast development, photoprotection and retrograde signalling

The Arabidopsis Spontaneous Cell Death1 gene, encoding a ζ-carotene desaturase essential for carotenoid biosynthesis, is involved in chloroplast development, photoprotection and retrograde signalling

In Situ Conversion of Protochlorophyllideb to Protochlorophyllide a in Barley

Metabolomics — the link between genotypes and phenotypes

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