Abstract
The Orange (Or) gene is a gene mutation that can increase carotenoid content in plant tissues normally devoid of pigments. It affects plastid division and is involved in the differentiation of proplastids or non-colored plastids into chromoplasts. In this study, the de-etiolation process of the wild type (WT) cauliflower (Brassica oleracea L. var. botrytis) and Or mutant seedlings was investigated. We analyzed pigment content, plastid development, transcript abundance and protein levels of genes involved in the de-etiolation process. The results showed that Or can increase the carotenoid content in green tissues, although not as effectively as in non-green tissues, and this effect might be caused by the changes in biosynthetic pathway genes at both transcriptional and post-transcriptional levels. There was no significant difference in the plastid development process between the two lines. However, the increased content of antheraxanthin and anthocyanin, and higher expression levels of violaxanthin de-epoxidase gene (VDE) suggested a stress situation leading to photoinhibition and enhanced photoprotection in the Or mutant. The up-regulated expression levels of the reactive oxygen species (ROS)-induced genes, ZAT10 for salt tolerance zinc finger protein and ASCORBATE PEROXIDASE2 (APX2), suggested the existence of photo-oxidative stress in the Or mutant. In summary, abovementioned findings provide additional insight into the functions of the Or gene in different tissues and at different developmental stages.
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Acknowledgments
We sincerely thank Dr. Li for providing cauliflower homozygous Or mutant seeds. This work was supported by the China National Natural Sciences Foundation (Grant no. 30771167). Tianhu Sun was also supported by the Scientific Research Foundation of Graduate School of Nanjing University.
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10265_2013_579_MOESM2_ESM.tif
Fig. S1 Western blotting analysis of OR protein levels in curds and de-etiolated cotyledons of WT and Or mutant seedlings. For each sample, 80 μg of denatured total protein was separated on 12% SDS-PAGE gel and transferred onto nitrocellulose membrane. The primary antigen-specific antiserum was worked at 1:200 dilution and secondary HRP-conjugated goat-anti-mouse IgG antibody at 1:10000 dilution. After added the enhanced chemiluminescent substrate (Thermo), images were captured by the ChemiDoc MP Imaging System (Bio-RAD). The band is the 29 kD mature OR protein (TIFF 163 kb)
10265_2013_579_MOESM3_ESM.eps
Fig. S2 Anthocyanin content in WT and Or mutant seedlings grown under continuous white light. Seeds were sterilized and placed on 1/2 MS-agar plate. After 4 days’ synchronization, the seeds were germinated and grown under continuous white light. a WT (left) and Or mutant (right) seedlings grown under continuous white light. Anthocyanin accumulated mainly in the cotyledons in WT cauliflower seedlings while in Or mutant seedlings anthocyanin accumulated in both cotyledons and hypocotyls. b Anthocyanin content in the two lines grown under continuous white light for 5 days. Anthocyanin contents in Or mutant seedlings grown under continuous white light were similar or even lower than that in WT seedlings. Data are shown as mean ± SD and obtained from triplicates. *P < 0.05 and **P < 0.01 (EPS 4293 kb)
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Men, X., Sun, T., Dong, K. et al. Or mutation leads to photo-oxidative stress responses in cauliflower (Brassica oleracea) seedlings during de-etiolation. J Plant Res 126, 823–832 (2013). https://doi.org/10.1007/s10265-013-0579-x
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DOI: https://doi.org/10.1007/s10265-013-0579-x