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Transcriptional changes of gibberellin oxidase genes in grapevines with or without gibberellin application during inflorescence development

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Abstract

The concept that gibberellin (GA) application on seeded grapevines induces seedlessness has been known for decades in viticulture. GA was applied to inflorescence clusters of seeded diploid grapevine cultivar ‘Tamnara’ (Vitis spp.) at 14 days before full bloom (DBF). Morphological and molecular effects of GA application were examined on the induction of parthenocarpic fruit development. With GA application, ovaries were enlarged and pollen tube growth was completely inhibited. Vitis GA oxidase enzymes, key determinants for GA level, were characterized through phylogenetic analysis with Arabidopsis GA oxidase enzymes. Five VvGA 20-oxidase (VvGA20ox), three VvGA 3-oxidase (VvGA3ox), and nine VvGA 2-oxidase (VvGA2ox) family proteins, and one VvGA methyltransferase (VvGAMT) and one Vitis cytochrome P450 714A1 proteins were identified, and their expression patterns were analyzed during inflorescence development from 14 DBF to 5 days after full bloom (DAF). VvGA2ox1, VvGA20ox3, and VvGA3ox2 were the most abundantly expressed genes in each gene family at 7, 5, and 2 DBF, respectively. Following GA application at 14 DBF inducing seedlessness, GA catabolic genes such as VvGAMT2, VvGA2ox3, and VvGA2ox4 were up-regulated at 12 DBF, full bloom, and 5 DAF, respectively. Conversely, most GA biosynthetic genes, VvGA20oxs and VvGA3oxs, were down-regulated at near full bloom, and the timing of their peak expression was changed. These results suggest that GA application at pre-bloom changes the GA biosynthesis into GA catabolic pathway at near full bloom by altering the transcription level and timing of GA oxidase genes during grapevine inflorescence development.

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Acknowledgments

This work was supported by a grant from the Next-Generation BioGreen 21 Program (PJ008213), Rural Development Administration, Suwon, Republic of Korea.

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Authors and Affiliations

  1. Fruit Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Suwon, 440-706, Republic of Korea

    Chan Jin Jung, Youn Young Hur, Sung-Min Jung, Jung-Ho Noh, Gyung-Ran Do, Seo-June Park, Jong-Chul Nam, Kyo-Sun Park & Hae-Sung Hwang

  2. Department of Plant Science, Seoul National University, Seoul, 151-921, Republic of Korea

    Youn Young Hur, Doil Choi & Hee Jae Lee

  3. Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, Republic of Korea

    Doil Choi & Hee Jae Lee

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  1. Chan Jin Jung
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Correspondence to Hee Jae Lee.

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C. J. Jung and Y. Y. Hur contributed equally to this work.

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Table S1. Primers used for qRT-PCR

10265_2013_623_MOESM2_ESM.tif

Fig. S1. qRT-PCR efficiency plots for GA metabolic genes. Mean quantification cycle (C T) values obtained from 10-fold serial dilution series of each gene plotted against the logarithm of cDNA template concentration. The amplification efficiency (E) was calculated by E = [10(−1/S) − 1] x 100, where S was the slope of the linear regression line (TIFF 74333 kb)

10265_2013_623_MOESM3_ESM.tif

Fig. S2. Sequence analysis of VvGA20ox and VvGA3ox family proteins. a Comparison of VvGA20oxs and AtGA20oxs. Both consensus sequences LPWKET for GA substrates binding and NYYPXCXXP for the common co-substrate, 2-oxoglutarate, binding of Vitis GA20oxs are denoted with dotted lines. b Comparison of VvGA3oxs and AtGA3oxs. ▼, Fe2+ binding site; ▽, 2-oxoglutarate binding site. The Fe2+ 2-ODD domain is underlined with a solid line (TIFF 125051 kb)

10265_2013_623_MOESM4_ESM.tif

Fig. S3. Sequence analysis of VvGA2ox, VvGAMT, and VvCYP714A family proteins. a Comparison of VvGA2oxs and AtGA2oxs. ▼, Fe2+ binding site; ▽, 2-oxoglutarate binding site. The Fe2+ 2-ODD domain is underlined with a solid line. b Comparison of VvGAMT2 with AtGAMT1 and AtGAMT2. The S-adenosyl-L-methionine-dependent methyltransferase domain is underlined. c Comparison of VvCYP714A1, AtCYP714A1, and AtCYP714A2. ▼, heme binding site. The conserved domain is underlined. Highly conserved amino acids are in black and similar amino acids are gray colored (TIFF 218303 kb)

10265_2013_623_MOESM5_ESM.pdf

Fig. S4. Comparative nucleotide sequence analysis of Vitis and Arabidopsis GA biosynthesis genes. a Comparison of VvGA20oxs and AtGA20oxs. b Comparison of VvGA3oxs and AtGA3oxs (PDF 6968 kb)

10265_2013_623_MOESM6_ESM.pdf

Comparative nucleotide sequence analysis of Vitis and Arabidopsis GA catabolic genes. a Comparison of VvGA2oxs and AtGA2oxs. b Comparison of VvGAMT2 with AtGAMT1 and AtGAMT2. c Comparison of VvCYP714A1, ELA1, and ELA2 (PDF 9793 kb)

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Jung, C.J., Hur, Y.Y., Jung, SM. et al. Transcriptional changes of gibberellin oxidase genes in grapevines with or without gibberellin application during inflorescence development. J Plant Res 127, 359–371 (2014). https://doi.org/10.1007/s10265-013-0623-x

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