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CEF1/OsMYB103L is involved in GA-mediated regulation of secondary wall biosynthesis in rice

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Abstract

Although the main genes in rice involved in the biosynthesis of secondary wall components have been characterized, the molecular mechanism underlying coordinated regulation of genes expression is not clear. In this study, we reported a new rice variety, cef1, showed the culm easily fragile (CEF) without other concomitant phenotypes. The CEF1 gene encodes a MYB family transcription factor OsMYB103L, was cloned based on map-based approach. Bioinformatics analyses indicated that CEF1 belongs to the R2R3-MYB subfamily and highly similar to Arabidopsis AtMYB103. Expression pattern analysis indicated that CEF1 is mainly expressed in internodes and panicles. Biochemical assays demonstrated that OsMYB103L is a nuclear protein and shows high transcriptional activation activity at C-terminus. OsMYB103L mediates cellulose biosynthesis and secondary walls formation mainly through directly binding the CESA4, CESA7, CESA9 and BC1 promoters and regulating their expression. OsMYB103L may also function as a master switch to regulate the expression of several downstream TFs, which involved in secondary cell wall biosynthesis. Furthermore, OsMYB103L physically interacts with SLENDER RICE1 (SLR1), a DELLA repressor of GA signaling, and involved in GA-mediated regulation of cellulose synthesis pathway. Our findings revealed that OsMYB103L plays an important role in GA-regulating secondary cell wall synthesis, and the manipulation of this gene provide a new strategy to help the straw decay in soil.

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Acknowledgments

This work was supported by grants from National Natural Science Foundation of China (Grant 31301297), The Science and Technology Service program of Chinese Academy of Sciences (Grant KFJ-EW-STS-083), the State Key Laboratory of Plant Cell and Chromosome Engineering(Grant PCCE-KF-2014-03) and the Ministry of Agriculture of China for Transgenic Research (Grant 2014ZX08009-35B).

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    Authors and Affiliations

    1. Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences and Anhui Province, Hefei, 230031, China

      Yafeng Ye, Binmei Liu, Meng Zhao, Kun Wu, Weimin Cheng, Zan Liu & Yuejin Wu

    2. Institute of Technical Biology and Agriculture Engineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, 230031, China

      Yafeng Ye, Binmei Liu & Yuejin Wu

    3. State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China

      Kun Wu, Xiangbin Chen, Qian Liu & Xiangdong Fu

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    1. Yafeng Ye
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    Correspondence to Xiangdong Fu or Yuejin Wu.

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    Yafeng Ye and Binmei Liu have contributed equally to this work.

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    Supplemental Figure 1

    CEF1 is a R2R3-MYB family transcription factor. A, Prediction of the domain structure of CEF1. B, Phylogenetic tree of the CEF1 homologs in Arabidopsis and rice. CEF1 and AtMYB103 have a very high homology. C, CEF1/MYB103L is a Nuclear-localized protein. A rice protoplast cell expressing MYB103L-GFP, indicating that CEF1/MYB103L is a Nuclear-localized protein. DAPI is a Nuclear dye (TIFF 542 kb)

    Supplemental Figure 2

    Transcriptional activation of CEF1/MYB103L. A, Transactivation activity of MYB103L in a yeast assay. Transformants harbouring pBD-MYB103L, the positive control pGAL4 and the negative control pBD were streaked onto SD-Trp or SD-Trp, His, Ade medium to determine growth. B, Effector and reporter constructs used in the Arabidopsis protoplast transient assay (TIFF 645 kb)

    Supplemental Figure 3

    Expression of cellulose synthesis related genes. A, the expression of OsCESA genes in NIL-CEF1 and NIL-cef1. B, the expression of CESA4, 7, 9 in NIL-CEF1 and NIL-cef1 p35S::myc-MYB103L transgenic plants. The Actin1 was used as internal control. All data given as mean ± SE (n = 3) (TIFF 436 kb)

    Supplemental Figure 4

    CEF1/OsMYB103L expression in overexpression (OX) transgenic plants (NIL-cef1 p35S::myc-MYB103L) as determined by qRT-PCR. The Actin was used as internal control. Error bars, SE of three biological replicates (TIFF 157 kb)

    Supplemental Figure 5

    MYB103L can’t binding SND2, MYB42/85, MYB52/54 promoters. Yeast one-hybrid assay showing the activity of LacZ reporters driven by BC1, SND2, MYB42/85, MYB52/54 promoters and activated by activation domain (AD) fusion effectors. The empty pB42AD and pLacZi were used as negative control (TIFF 469 kb)

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    Ye, Y., Liu, B., Zhao, M. et al. CEF1/OsMYB103L is involved in GA-mediated regulation of secondary wall biosynthesis in rice. Plant Mol Biol 89, 385–401 (2015). https://doi.org/10.1007/s11103-015-0376-0

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