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Jointly silencing BoDWARF, BoGA20ox and BoSP (SELF-PRUNING) produces a novel miniature ornamental Brassica oleracea var. acephala f. tricolor variety

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Abstract

The ornamental Brassica oleracea var. acephala f. tricolor is a good winter and spring foliage plant. Plant architecture is an important agronomic trait of plants, especially for ornamental plants with high ornamental and economic value. In this study, three miniature-related genes, BoDWARF, BoGA20ox and BoSP (SELF-PRUNING), were cloned and their tissue-specific expression patterns were analyzed. The results showed that the three genes were all highly expressed in young leaves and flowers, followed by the lateral roots, seeds and stems. To further achieve the purpose of miniaturization of plants, an RNAi expression vector, jointly targeting BoDWARF, BoGA20ox and BoSP, was constructed and transformed into kale plants. Smaller plant size and slower growth and development speed of flowers and roots were observed in jointly silenced kales. Brassinosteroids and gibberellin contents in leaves and flower buds of transgenic plants were significantly decreased. Furthermore, the expressions of brassinolide-, gibberellin- and flowering-related genes were down-regulated by varying degrees in silenced plants. These results suggest that BoDWARF, BoGA20ox and BoSP play important roles in plant architecture, and that brassinolide and gibberellin are important hormones controlling plant growth and architecture. This miniaturization strategy of kale provides an efficient approach for cultivation of new varieties of ornamental plants and crops.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 31171968) and Natural Science Foundation of Chongqing of China (No. cstc, 2011BB1068), and the Fundamental Research Funds for the Central Universities (No. CDJXS11232244).

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  1. Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Campus A, 174 Shapingba Main Street, Chongqing, 400044, People’s Republic of China

    Qiaoli Xie, Guoping Chen, Lei Deng, Qin Liu, Yanjie Zhang & Zongli Hu

  2. Beijing Agro-Biotechnology Research Center, Beijing, 100089, People’s Republic of China

    Xuqing Chen

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  1. Qiaoli Xie
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Supplementary material 2 (DOC 42 kb)

11032_2014_20_MOESM3_ESM.tif

Supplemental Fig. 1 Multiple sequence alignment and phylogenetic analysis of BoDWARF and other cytochrome P450 family proteins. a. Multiple sequence alignment of BoDWARF and other cytochrome P450 family proteins. BoDWARF is marked with an asterisk. Identical amino acids are shaded in black, and similar amino acids are shaded in gray. b. Phylogenetic analysis of BoDWARF and other proteins were constructed by the neighbor-joining method, bootstrap analysis of 1000 replicates. BoDWARF is marked with an asterisk. Accession numbers and corresponding references for the proteins listed are as follows: SlDWARF (NM_001247334.1), AtDWARF (NP_566852.1), NtDWARF (XP_002330918.1), GmDWARF (XP_003554965.1), VvDWARF (ABB60086.1), ZmDWARF (AET06165.1), BoDWARF (Bol032288), CsDWARF (XP_004141472.1), BrDWARF (Bra020747). (TIFF 582 kb)

11032_2014_20_MOESM4_ESM.tif

Supplemental Fig. 2 Multiple sequence alignment and phylogenetic analysis of BoGA20ox and other GA20ox proteins. a. Multiple sequence alignment of BoGA20ox and other GA20ox proteins. BoGA20ox is marked with an asterisk. Identical amino acids are shaded in black, and similar amino acids are shaded in gray. b. Phylogenetic analysis of BoGA20ox and other GA20ox proteins were constructed by the neighbor-joining method, bootstrap analysis of 1000 replicates. BoGA20ox is marked with an asterisk. Accession numbers and corresponding references for the proteins listed are as follows: AtGA20ox (NM_118674.4), BoGA20ox (Bol042237), BrGA20ox (Bra013890), RsGA20ox (BAM73279.1), ZmGA20ox (NM_001254854.1), SlGA20ox (NM_001247141.1), PtGA20ox (JX305461.1). (TIFF 398 kb)

11032_2014_20_MOESM5_ESM.tif

Supplemental Fig. 3 Multiple sequence alignment and phylogenetic analysis of BoSP and other PEBP family proteins. a. Multiple sequence alignment of BoSP and other PEBP family proteins. BoSP is marked with an asterisk. Identical amino acids are shaded in black, and similar amino acids are shaded in gray. b. Phylogenetic analysis of the BoSP and other proteins were constructed by the neighbor-joining method, bootstrap analysis of 1000 replicates. BoSP is marked with an asterisk. Accession numbers and corresponding references for the proteins listed are as follows: AtTFL1 (U77674.1), BoSP (Bol026421), BrSP (Bra000506), SlSP (NM_001247045), AmCEN (S81193), CaSP (CAI38702), OsSP (NP_001173006), GhTF1 (ABW24963), ZmZCN2 (NP_001106241), GM-CEN like3 (ACU00129), VvTFL1a (XP_002276820), ScPEBP1 (1WPX_B), MmPEBP1 (AAG25635), HsPEBP1 (NP_002558). (TIFF 533 kb)

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Xie, Q., Chen, G., Chen, X. et al. Jointly silencing BoDWARF, BoGA20ox and BoSP (SELF-PRUNING) produces a novel miniature ornamental Brassica oleracea var. acephala f. tricolor variety. Mol Breeding 34, 99–113 (2014). https://doi.org/10.1007/s11032-014-0020-8

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