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| Cloning and Characterization of R2R3-MYB Transcription Factor Gene GhTT2 in Brown Cotton (Gossypium hirsutum) |
| MEI Jun, MU Rong-Rong, NIU Qing-Qing, WANG Xiao-Li, ZHAO Yan-Yan, CAI Fang-Fang, SUN Yu-Qiang* |
| School of Life Sciences and Medicine/Plant Genomics & Molecular Improvement of Colored Fiber Laboratory, Zhejiang Sci-Tech University, Hangzhou 310018, China |
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Abstract Natural colored cottons, including brown fiber cotton and green fiber cotton which can be spun into garments directly without chemical printing and dyeing, are special upland cottons (Gossypium hirsutum) with green environmental protection and high ecological benefits. Due to the accumulation of different anthocyanins, procyanidins and their derivatives in the fibers, colored cottons show different colors. It have been confirmed that proanthocyanidins and their derivatives are thought to be responsible for the color formation of brown fibers. Based on previous transcriptome data, this study cloned a R2R3-MYB (v-myb avian myeloblastosis viral oncogene homolog, MYB) transcription factor gene GhTT2 (transparent testa 2), which was predominantly expressed during the development stage of brown cotton fibers. The structure, expression pattern and subcellular localization of GhTT2 gene and coding protein were predicted and analyzed by phylogenetic tree construction, amino acid sequence multiple alignment, prokaryotic expression, qRT-PCR, tobacco (Nicotiana benthamiana) transient transformation, and yeast (Saccharomyces cerevisiae) hybridization assay. The results showed that the open reading frame of GhTT2 gene was 765 bp, encoding 254 amino acids, with predicted relative molecular mass 28.863 kD, isoelectric point 7.423, and a typical R2R3-MYB domain. The prokaryotic expression analysis showed that expression of GhTT2 could be efficiently induced under the condition of 37 ℃ and 1 mmol/L isopropyl β-D-thiogalactoside (IPTG). The results of qRT-PCR showed that, compared with white cotton, GhTT2 was predominantly expressed during the development stage of brown cotton fibers, and its expression in fibers was significantly higher than that in other tissues. The recombinant protein GhTT2-GFP was located in the nucleus, which conformed to the characteristics of typical transcription factors. Yeast transformation test showed that GhTT2 protein had obvious transcriptional activation activity. The study provides basic material for further verifying the biological function of GhTT2 in the color formation of brown fibers and the improvement of the color diversity of colored cotton fibers.
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Received: 07 December 2021
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Corresponding Authors:
* Corresponding author,sunyuqiang@zstu.edu.cn
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