Rapid analysis of GBSS1 and Vinv genes expressed in potato tubers using microtubers produced in liquid culture medium
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ORIGINAL ARTICLE
Rapid analysis of GBSS1 and Vinv genes expressed in potato tubers using microtubers produced in liquid culture medium Yuhya Wakasa1 · Atsushi Kasai2 · Muneo Yamazaki1 · Yutaka Tabei1 · Mutsuo Tsuyama3 · Toshiya Igarashi3 · Tomokazu Okazaki4 · Kayo Yamamoto4 · Hideki Fujihara4 · Ayumi Kanno2 · Osamu Noro2 · Takeo Harada2 · Shinji Akada2 Received: 7 April 2020 / Accepted: 16 July 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Key message This study established a rapid method for the gene expression analysis in potato tubers. The use of microtubers would be useful for primary evaluation of tuber-expressed genes. Abstract In the development of transgenic potato or of potato with other genome modifications (e.g., genome editing or RNAdirected DNA methylation (RdDM) and so on) to improve tuber traits, analysis of the target gene is often difficult because of the long cultivation cycle (3–4 months), large areas required, numerous materials for plant cultivation, and considerable efforts needed to obtain transgenic tubers. We demonstrate here rapid and convenient analysis of gene expression in potato microtubers. Enough microtubers for expression analysis can be induced over about 4 weeks in a simple liquid medium in an Erlenmeyer flask. High-quality RNA and protein can be easily prepared from microtubers and used for northern blot, qRTPCR, and western blot analyses without further purification. We investigated the expression of two tuber-expressed genes (GBSS1 and Vinv) in microtubers derived from the wild-type and from lines derived from RdDM-mediated transcriptional gene silencing. As expected, the expression of both genes was similar between microtubers and normal tubers. Furthermore, we demonstrated that microtubers can be used in western blot and confocal immunofluorescent microscopy analyses. These results suggest that expression analysis using microtubers is a convenient tool for the analysis of tuber-expressed genes such as GBSS1 and Vinv in potato. Keywords Expression analysis · Microtuber · Solanum tuberosum L. · New plant breeding techniques · Transgenic potato
Introduction Communicated by Günther Hahne. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00299-020-02572-6) contains supplementary material, which is available to authorized users. * Yuhya Wakasa [email protected] 1
Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Kannondai 3‑1‑3, Tsukuba, Ibaraki 305‑8604, Japan
2
Faculty of Agriculture and Life Science, Hirosaki University, Bunkyo, Hirosaki, Aomori 036‑8561, Japan
3
Potato Research Center, Calbee Potato, Inc., Minami 3‑2, Higashimemuro, Memuro, Hokkaido 082‑0006, Japan
4
Research and Development Department II, Matsutani Chemical Industry Co., Ltd., Kita‑Itami 5‑3, Itami, Hyogo 664‑8508, Japan
The generation of genetically modified (GM) plants is a promising technology to improve crop traits. In 2018, many GM crops were commercially grown
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