Abstract
In addition to Circadian clock associated 1 (CCA1) and Late elongated hypocotyl (LHY), Reveille/LHY-CCA1-LIKE (RVE/LCL) genes are known as clock regulators in plants. So far, the association of RVE expression with anthocyanin accumulation has been reported only in Arabidopsis and pear. In this study, Brassica oleracea RVE5 (BoRVE5) was selected as a candidate gene to develop a PCR marker to predict anthocyanin accumulation under low temperature (LT) condition, based on the observation that the lengths of AT simple sequence repeats in 3′ untranslated region of BoRVE5 tend to inversely correlates with anthocyanin amount. Among BoRVE5 genomic DNA sequences of purple cabbages and green cabbages—both high anthocyanin accumulators (HAAs) and low anthocyanin accumulators (LAAs) of green cabbages under LT condition, sequence polymorphisms changing its amino acid sequence were identified between LAAs and HAAs/purple cabbages. We named the LAA-type gene as BoRVE5a and HAA-type as BoRVE5b, and found their expression levels are associated with anthocyanin accumulation phenotypes. Using these polymorphisms, a molecular marker discriminating LAAs from HAAs and purple cabbages was developed and validated using cabbage lines with diverse phenotypes, including 57 green and 59 purple cabbages. This marker is expected to be even more valuable if used in combination with other markers for anthocyanin phenotypes, such as those developed from BoMYBL2-1 or BoDFR1.
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Data availability
Accession numbers of BoRVE5 genomic DNA sequences are as follows: MW394167 (from line 2409), MW394166 (from line 842), MW394168 (from line 2437), MW394171 (from line 09WH45), MW394164 (from line 154), MW394165 (from line 337), MW394169 (from line 7S4-51), MW394172 (from line B90), MW394173 (from line B98), MW394170 (from line 7S4-63).
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Acknowledgements
We thank HanKook Seed Co. and Asia Seed Co. for providing plant samples for this research. This work was supported by a grant from the Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry, and Fisheries (Grant 213007-05-4-SB620), Republic of Korea, and Ministry of Science and ICT (No. 2020-0-01441, Artificial Intelligence Convergence Research Center at Chungnam National University), Republic of Korea.
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Conceptualization: Y.H.; Methodology: J.Y.A., C.S.J., H.S.; Formal analysis and investigation: J.Y.A., C.S.J., H.S., H.Y.; Writing—original draft preparation: Y.H.; Writing—review and editing: H.Y., Y.H.; Funding acquisition:, H.Y., Y.H.; Resources: C.S.J., Y.H.
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Ahn, J.Y., Song, H., Jang, C.S. et al. Polymorphisms found in Brassica oleracea REVEILLE/LHY-CCA1-LIKE 5 (BoRVE5/BoLCL4) are associated with anthocyanin accumulation in cabbage. Euphytica 217, 180 (2021). https://doi.org/10.1007/s10681-021-02913-w
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DOI: https://doi.org/10.1007/s10681-021-02913-w