Temporal and Spatial Expression Analysis of Shoot-Regeneration Regulatory Genes during the Adventitious Shoot Formation in Hypocotyl and Cotyledon Explants of Tomato (CV. Micro-Tom)

Lee, Myoung Hui; Lee, Jiyoung; Jie, Eun Yee; Choi, Seung Hee; Jiang, Lingmin; Ahn, Woo Seok; Kim, Cha Young; Kim, Suk Weon

doi:10.3390/ijms21155309

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Temporal and Spatial Expression Analysis of Shoot-Regeneration Regulatory Genes during the Adventitious Shoot Formation in Hypocotyl and Cotyledon Explants of Tomato (CV. Micro-Tom)

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Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Korea

Department of Bioactive Materials, Chonbuk National University, Jeonju 54896, Korea

Department of Bioenergy Science and Technology, Chonnam National University, Gwangju 61186, Korea

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2020, 21(15), 5309; https://doi.org/10.3390/ijms21155309

Received: 20 June 2020 / Revised: 24 July 2020 / Accepted: 24 July 2020 / Published: 26 July 2020

(This article belongs to the Section Molecular Plant Sciences)

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Abstract

Enhancing the competence for plant regeneration in tissue culture studies is an important issue not only for efficient genetic transformation of commercial crops but also for the reproducibility of scientific reports. In this study, we investigated optimization of several tissue culture conditions including plant growth regulators, types and ages of explants, culture densities, and plant position in order to improve the competence of adventitious shoot formation of the tomato (Solanum lycopersicum cv. Micro-Tom). In addition, we examined the differential expression of D-type cyclin (CYCD3-1) and several shoot regeneration regulatory genes from hypocotyl and cotyledon explants of tomato during shoot organogenesis. A treatment of 1 mg L⁻¹ Zeatin and 0.1 mg L⁻¹ Indole-3-acetic acid (IAA) in Murashige and Skoog (MS) medium containing 3% sucrose was optimal for adventitious shoot formation from hypocotyl and cotyledon explants. The younger explants exhibited more shoot formation regardless of explant types. Additionally, those closest to the shoot apical meristem produced more shoots compared to the other regions in the hypocotyl and the cotyledon explants. Gene expression of CYCD3-1, SHOOT MERISTEMLESS (STM), and cytokinin dependent WUSCHEL (WUS) was significantly higher in younger explants than in older ones. Furthermore, an increase in CYCD3-1, STM, and WUS expression was evident at the distal part of hypocotyls and the proximal part of cotyledons compared to other regions. These differential gene expression profiles exhibited good agreement with the results of shoot formation obtained from diverse explants of tomato. These results suggest that temporal and spatial gene expression of shoot regeneration regulatory genes plays an important role in enhancing the competence and the reproducibility of adventitious shoot formation from tomato explants. View Full-Text

Keywords: tissue culture; plant regeneration; shoot formation; genes expression; reproducibility; tomato

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Lee, M.H.; Lee, J.; Jie, E.Y.; Choi, S.H.; Jiang, L.; Ahn, W.S.; Kim, C.Y.; Kim, S.W. Temporal and Spatial Expression Analysis of Shoot-Regeneration Regulatory Genes during the Adventitious Shoot Formation in Hypocotyl and Cotyledon Explants of Tomato (CV. Micro-Tom). Int. J. Mol. Sci. 2020, 21, 5309. https://doi.org/10.3390/ijms21155309

AMA Style

Lee MH, Lee J, Jie EY, Choi SH, Jiang L, Ahn WS, Kim CY, Kim SW. Temporal and Spatial Expression Analysis of Shoot-Regeneration Regulatory Genes during the Adventitious Shoot Formation in Hypocotyl and Cotyledon Explants of Tomato (CV. Micro-Tom). International Journal of Molecular Sciences. 2020; 21(15):5309. https://doi.org/10.3390/ijms21155309

Chicago/Turabian Style

Lee, Myoung H., Jiyoung Lee, Eun Y. Jie, Seung H. Choi, Lingmin Jiang, Woo S. Ahn, Cha Y. Kim, and Suk W. Kim. 2020. "Temporal and Spatial Expression Analysis of Shoot-Regeneration Regulatory Genes during the Adventitious Shoot Formation in Hypocotyl and Cotyledon Explants of Tomato (CV. Micro-Tom)" International Journal of Molecular Sciences 21, no. 15: 5309. https://doi.org/10.3390/ijms21155309

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Temporal and Spatial Expression Analysis of Shoot-Regeneration Regulatory Genes during the Adventitious Shoot Formation in Hypocotyl and Cotyledon Explants of Tomato (CV. Micro-Tom)

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