Measurement of the distribution of non‐structural carbohydrate composition in onion populations by a high‐throughput microplate enzymatic assay

Revanna, Roopashree; Turnbull, Matthew H.; Shaw, Martin; Wright, Kathryn; Butler, R.C.; Jameson, Paula E.; McCallum, John

doi:10.1002/jsfa.6062

Cited by 12 publications

(14 citation statements)

References 53 publications

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“…Those containing two to nine fructose units have been termed fructo‐oligosaccharides; however, the degree of polymerisation can extend to between 10 and 15 units . No distinction was made in the present study between the types of polymers present because fructan was determined by analysis as fructose following enzymatic hydrolysis . Spectroscopically, fructans contain the same types of bonds as their constituent monomers, apart from the β‐(2 → 1) linkages.…”

Section: Resultsmentioning

confidence: 83%

“…Non‐structural water‐soluble carbohydrates (glucose, fructose, sucrose and fructans) were determined in enzymatic microplate assays . Two industry sweetness indices were subsequently determined from these concentration data: a PFR sweetness index (PFR‐SI) expressed in glucose equivalents, calculated as Glucose + 2 × Fructose + 1.45 × Sucrose + 0.5 × Fructan, and the National Onion Laboratory sweetness index (NOL‐SI) expressed as sucrose equivalents, calculated as 0.75 × Glucose + 1.75 × Fructose + Sucrose …”

Section: Methodsmentioning

confidence: 99%

“…This has enabled recurrent selection for very high DM levels in dehydrator breeding programmes but is more difficult to apply in varieties with coloured scales. We previously reported that microplate‐based enzymatic analyses can provide useful quantitative estimates of the relative content of mono‐ and disaccharides and fructan in onion extracts …”

Section: Introductionmentioning

confidence: 99%

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Quantification of free sugars, fructan, pungency and sweetness indices in onion populations by FT‐MIR spectroscopy

et al. 2018

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Section: Resultsmentioning

confidence: 83%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Quantification of free sugars, fructan, pungency and sweetness indices in onion populations by FT‐MIR spectroscopy

et al. 2018

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“…Samples of these families were grown at Lat 42 deg S near Christchurch, New Zealand. Cured bulbs were phenotyped for red bulb colour and freeze-dried samples were analyzed for fructan and hexose content as described elsewhere [56]. DNA was isolated from fresh leaf material or freeze-dried bulb tissue as described previously [57].…”

Section: Methodsmentioning

confidence: 99%

A Toolkit for bulk PCR-based marker design from next-generation sequence data: application for development of a framework linkage map in bulb onion (Allium cepa L.)

et al. 2012

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BackgroundAlthough modern sequencing technologies permit the ready detection of numerous DNA sequence variants in any organisms, converting such information to PCR-based genetic markers is hampered by a lack of simple, scalable tools. Onion is an example of an under-researched crop with a complex, heterozygous genome where genome-based research has previously been hindered by limited sequence resources and genetic markers.ResultsWe report the development of generic tools for large-scale web-based PCR-based marker design in the Galaxy bioinformatics framework, and their application for development of next-generation genetics resources in a wide cross of bulb onion (Allium cepa L.). Transcriptome sequence resources were developed for the homozygous doubled-haploid bulb onion line ‘CUDH2150’ and the genetically distant Indian landrace ‘Nasik Red’, using 454™ sequencing of normalised cDNA libraries of leaf and shoot. Read mapping of ‘Nasik Red’ reads onto ‘CUDH2150’ assemblies revealed 16836 indel and SNP polymorphisms that were mined for portable PCR-based marker development. Tools for detection of restriction polymorphisms and primer set design were developed in BioPython and adapted for use in the Galaxy workflow environment, enabling large-scale and targeted assay design. Using PCR-based markers designed with these tools, a framework genetic linkage map of over 800cM spanning all chromosomes was developed in a subset of 93 F2 progeny from a very large F2 family developed from the ‘Nasik Red’ x ‘CUDH2150’ inter-cross. The utility of tools and genetic resources developed was tested by designing markers to transcription factor-like polymorphic sequences. Bin mapping these markers using a subset of 10 progeny confirmed the ability to place markers within 10 cM bins, enabling increased efficiency in marker assignment and targeted map refinement. The major genetic loci conditioning red bulb colour (R) and fructan content (Frc) were located on this map by QTL analysis.ConclusionsThe generic tools developed for the Galaxy environment enable rapid development of sets of PCR assays targeting sequence variants identified from Illumina and 454 sequence data. They enable non-specialist users to validate and exploit large volumes of next-generation sequence data using basic equipment.

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“…Albeit reliable, these enzymatic activity methods for food materials are time consuming and require intensive labor, limiting the number of samples that can be analyzed daily. Lately, methods have been developed utilizing spectrophotometry by microplates, because it may have the same sensitivity, requires smaller amounts of sample and reactants and is less time consuming, since several tests can be run simultaneously in a spectrophotometer with a plate reader (Murshed et al, 2008;Revanna et al, 2013;Drtiz et al, 2014;Bobo-García et al, 2015). Consequently, the objective of this work was to propose and to validate a reliable microplate assay for determining PPD and PDD activities in fruits and vegetables.…”

Section: Introductionmentioning

confidence: 99%

Validation of spectrophotometric microplate methods for polyphenol oxidase and peroxidase activities analysis in fruits and vegetables

Siguemoto

Gut

2017

Food Sci. Technol

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Enzymes polyphenol oxidase (PPD) and peroxidase (PDD) play important roles in the processing of fruits and vegetables, since they can produce undesirable changes in color, texture and flavor. Classical methods of activity assessment are based on cuvette spectrophotometric readings. This work aims to propose, to validate and to test microplate spectrophotometric methods. Samples of apple juice and lyophilized enzymes from mushroom and horseradish were analyzed by the cuvette and microplate methods and it was possible to validate the microplate assays with satisfactory results regarding linearity, repeatability, accuracy along with quantitation and detection limits. The proposed microplate methods proved to be reliable and reproducible as the classical methods besides having the advantages of allowing simultaneous analysis and requiring a reduced amount of samples and reactants, which can beneficial to the study of enzyme inactivation in the processing of fruits and vegetables.Keywords: microplate; polyphenol oxidase; peroxidase; enzymatic assay; apple juice.Practical Application: Rapid assessment of PDD/PPD activities in fruits and vegetables with reduced amounts of reactants.

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Measurement of the distribution of non‐structural carbohydrate composition in onion populations by a high‐throughput microplate enzymatic assay

Cited by 12 publications

References 53 publications

Quantification of free sugars, fructan, pungency and sweetness indices in onion populations by FT‐MIR spectroscopy

Quantification of free sugars, fructan, pungency and sweetness indices in onion populations by FT‐MIR spectroscopy

A Toolkit for bulk PCR-based marker design from next-generation sequence data: application for development of a framework linkage map in bulb onion (Allium cepa L.)

Validation of spectrophotometric microplate methods for polyphenol oxidase and peroxidase activities analysis in fruits and vegetables

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