Rapid analysis of GBSS1 and Vinv genes expressed in potato tubers using microtubers produced in liquid culture medium
| Autor: | Hideki Fujihara, Muneo Yamazaki, Yutaka Tabei, Tomokazu Okazaki, Osamu Noro, Ayumi Kanno, Shinji Akada, Kayo Yamamoto, Atsushi Kasai, Takeo Harada, Yuhya Wakasa, Toshiya Igarashi, Mutsuo Tsuyama |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine Transgene Blotting Western Plant Science Biology 01 natural sciences Genome Tissue Culture Techniques 03 medical and health sciences Western blot Gene Expression Regulation Plant Gene expression medicine Northern blot Gene Silencing Gene Plant Proteins Solanum tuberosum medicine.diagnostic_test Gene Expression Profiling fungi food and beverages RNA General Medicine Plants Genetically Modified Culture Media Plant Tubers 030104 developmental biology Biochemistry Microscopy Fluorescence RNA Plant DNA methylation Agronomy and Crop Science Plant Shoots 010606 plant biology & botany |
| Zdroj: | Plant cell reports. 39(11) |
| ISSN: | 1432-203X |
| Popis: | 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. In the development of transgenic potato or of potato with other genome modifications (e.g., genome editing or RNA-directed 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, qRT-PCR, 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. |
| Databáze: | OpenAIRE |
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