Abiotic stress and tissue-specific reference genes for quantitative reverse transcription PCR analysis in Korean native watermelons, Citrullus lanatus ‘Black-King’ and ‘Speed-Plus-Honey’

• Published in: Applied biological chemistry Vol. 61; no. 5; pp. 593 - 598
• Main Authors: Kim, Soo Jin, Jung, Il Lae, Lee, Hye-Eun, Lee, Ji-Hoon
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.10.2018; Springer Nature B.V; 한국응용생명화학회
• Subjects: Abscisic acid; Actin; Applied Microbiology; Biological Techniques; Bioorganic Chemistry; Chemistry; Chemistry and Materials Science; Citrullus lanatus; Cultivars; Dehydrogenase; Dehydrogenases; Drought; Elongation; Environmental stress; Gene expression; Genes; Genomes; Glyceraldehyde; Honey; Isocitrate dehydrogenase; NADP; NADP-isocitrate dehydrogenase; Nucleotide sequence; Reverse transcription; rRNA 18S; Seedlings; Sodium chloride; Stresses; Tissues; Ubiquitin; Ubiquitin-conjugating enzyme; Water melons; 농학; Reference gene; Watermelon; RT-qPCR; Loading control gene
• ISSN: 2468-0834; 2468-0842
• DOI: 10.1007/s13765-018-0391-3

A wide variety of research on watermelon has been conducted, and such studies have been motivated by the published genome sequence database of watermelon. Screening of proper reference genes is the primary step for normalization in gene expression analysis. Based on previous studies conducted on Arabidopsis and cucumber, we selected eight candidate reference genes of ClACT , ClEF1α , ClGAPDH , ClIDH , ClLUG , ClPTB , ClUBC2 , and Cl18SrRNA , respectively, encoding β-Actin, elongation factor 1-α, glyceraldehyde-3-phosphate-dehydrogenase, NADP-isocitrate dehydrogenase, leunig, polypyrimidine tract-binding protein1, ubiquitin-conjugating enzyme E2, and 18S ribosomal RNA from watermelon ( Citrullus lanatus ). The expression levels of these eight genes were evaluated by RT-qPCR under plant hormone-treatment (100 μM ABA) and abiotic stresses such as drought, cold (4 °C), and high salt concentration (250 mM NaCl). The expression patterns of these eight genes were further compared across different types of watermelon tissues such as flower, leaf, tendril, stem, root, and whole seedling. Our results showed that expressions of ClACT and ClEF1α , respectively in the Korean native watermelon cultivars Citrullus lanatus ‘Black-King’ and ‘Speed-Plus-Honey’ were least affected by the environmental stresses regardless of tissue types. Here, we suggest two ideal reference genes for watermelon RT-qPCR-based gene expression study.