Simple and Rapid Method for the Selection of Individual Rapeseed Plants Low in Glucosinolates

Abstract

A simple and rapid colorimetric method for the selection of individual rapeseed plants with low glucosinolate contents was developed using one of a pair of germinated cotyledons of a single seedling and a glucose-measuring method employed in clinical tests (CII-Test method). The CII-Test method can be used to estimate the amount of glucose released by the hydrolysis of glucosinolates extracted from a germinated cotyledon. In the present experiment, HPLC analysis showed that the total glucosinolate content of germinated cotyledons was not significantly different from that of seeds three days after seed sowing. When the glucosinolate content of the seeds and/or cotyledons was estimated by the glucosemeasuring CII-Test method and HPLC method, high positive correlations were recognized between both types of measurements. The CII-Test method exhibits some clear advantages in that 1) the glucosinolate content can be estimated in an individual plant, and 2) 150-500 samples can be estimated by one person in one day. These advantages indicate that the glucose measuring CII-Test method is suitable for efficient and accurate selection of rapeseed plants with low glucosinolate contents in the F 2 generation.

Development of an effective screening method for partial resistance to Alternaria brassicicola (dark leaf spot) in Brassica rapa

“…In Brassica species, leaves contain anti-pathogenic substances such as glucosinolates (Meah et al, 1988;Milford et al, 1989). The total glucosinolate content in cotyledons of seedlings varies between cultivars of rapeseeds (Ishida et al, 2003), and there is no correlation between the total glucosinolate content in true leaves and cotyledons (Kim et al, 2001). It may be necessary to investigate the inhibitory effect of anti-pathogenic substances in the two different leaf types in order to understand the lack of correlation of disease response between the detached leaf test and the cotyledon test.…”

Small variation of glucosinolate composition in Japanese cultivars of radish (<i>Raphanus sativus</i> L.) requires simple quantitative analysis for breeding of glucosinolate component

“…Contents and composition of the GSLs were analyzed using HPLC (LC-10A, Shimadzu Corp., Kyoto, Japan) with a column (150 × 4.6 mm, COSMOSIL 5C18-II, Nacalai Tesque Inc., Kyoto, Japan) following desulfation of GSLs with sulfatase (Sigma-Aldrich Corp.) using the methods described by Bjerg and Sørensen (1987) and Ishida et al (2003) . To quantify each GSL, relative response factors to the internal standard of sinigrin were used according to the International Organization for Standardization (1992) .…”

Variation in defence strategies in the metal hyperaccumulator plant Noccaea caerulescens is indicative of synergies and trade-offs between forms of defence

“…Samples were then incubated with 5 ml thioglucosidase (0.5 mg ml 21 ) from Sinapis alba seed (Sigma Aldrich) for 3 h at room temperature. This enzyme cleaves one molecule of glucose from each molecule of glucosinolate [54][55][56]. Glucose concentrations were measured using the Glucose (GO) Assay Kit (Sigma, UK) according to the manufacturer's specification.…”