• Journal of Plant Biotechnology
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• v.4 no.2
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• pp.59-65
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• 2002

ADP-glucose pyrophosphorylase (AGPase) catalyzes the key regulatory step in starch biosynthesis. Two cDNA clones encoding AGPase subunits were isolated from the leaf cDNA library of Chinese cabbage (Brassica campestris L. spp. pekinensis). One was designated as BCAGPS for the small subunit and the other as BCAGPL for the large subunit. Both cDNAs have uninterrupted open reading frames deriving 57 kDa and 63 kDa polypeptides for BCAGPS and BCAGPL, respectively, which showed significant similarity to those of other dicot plants. Also, However, the deduced amino acid sequence of BCAGPL has a unique feature. That is, it contains two regions (Rl and R2) lacking in all other plant enzymes. This is the first report of BCAGPL containing Rl and R2 among plant large subunits as well as small subunits. From the genomic Southern analysis and BAC library screening, we inferred the genomic status of BCAGPS and BCAGPL gene.

• Plant Biotechnology Reports
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• v.4 no.3
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• pp.201-211
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• 2010

We present a highly effective T-DNA inserted gene screening method as part of a reverse genetics model system using the Chinese cabbage (Brassica rapa L. spp. pekinensis). Three-step two-dimensional (2D) matrix strategies are potentially accurate and useful for the identification of specific T-DNA inserted mutants from a large population. To construct our Chinese cabbage model, we utilized a forward genetics screening approach for the abnormal phenotypes that were obtained from transgenic plants of Brassica rapa generated with Agrobacteria tumefaciens containing the pRCV2 vector. From one transgenic plant with an abnormal phenotype, we observed that the st1 gene (which is related to senescence-associated process proteins) contained a T-DNA fragment, and that its expression level was decreased. This T-DNA insert was then used as a control to construct an effective screening pool. As a result, the optimum template concentration was found to be 0.1-1 ng in our PCR strategy. For other conditions, positive changes to the Gibbs free energy prevented the formation of oligo dimers and hairpin loop structures, and autosegment extension gave better results for long fragment amplification. Using this effective reverse genetics screening method, only 23 PCR reactions were necessary to select a target gene from a pool of 100 individual DNAs. Finally, we also confirmed that the sequence we obtained from the above method was identical to the flanking sequence isolated by rescue cloning.

• Korean Journal of Agricultural Science
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• v.41 no.2
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• pp.125-133
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• 2014

The variation of glucosinolates (GSLs) in Chinese cabbage ('Sinhongssam') (Brassica rapa L. spp. pekinensis) cultivated under lights to control plant growth conditions was evaluated at different development stages. Under experimental conditions in plant factory system, plant growth conditions including light, temperature, and nutrients were designed to enhance GSLs. The variation of glucosinolates (GSLs) in Chinese cabbage ('Sinhongssam') (Brassica rapa L. spp. pekinensis) cultivated under lights to control plant growth conditions was evaluated at different development stages. Under experimental conditions in plant factory system, plant growth conditions including light, temperature, and nutrients were designed to enhance GSLs. The contents of GSLs were quantified in Chinese cabbage according to different light sources (Red+White, RW; Red+Blue+White, RBW, Fluorescence lamp, FL) at development stages (28, 42, and 56 days after sowing, DAS) using HPLC. Nine GSLs including five aliphatic (progoitrin, sinigrin, glucoalyssin, gluconapin, and glucobrassicanapin) three indolyl (glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrassicin), and one aromatic (gluconasturtiin) GSLs were identified based on peak retention time in previous results of our laboratory. GSL contents were higher in RBW (36.55) and lower in FL ($15.24{\mu}mol/g/\;DW$). Results revealed that GSL contents were higher under controlled photoperiods (20/4 h) ($58.35{\mu}mol/g\;DW$) and controlled light intensity ($160{\mu}mol/m^2/s$) ($34.02{\mu}mol/g\;DW$), respectively. Lower amount of progoitrin and comparatively higher amount of glucobrassicin and gluconasturtiin was noted in Chinese cabbage cultivated under FL light (2.38, 9.82, and 2.10) at 42 DAS, photoperiod 20/4 h (3.16, 2.52, and 1.30) at 28 DAS, and light intensity at $130{\mu}mol/m^2/s$ (2.28, 2.24, and $1.51{\mu}mol/g\;DW$) at 42 DAS. Therefore FL light, photoperiod (20/4 h), and light intensity ($130{\mu}mol/m^2/s$) were considered as most suitable for the enhancement of GSLs in Chinese cabbage.