小麦穗发芽抗性相关Vp1基因启动子的分离及功能验证

doi:10.3724/SP.J.1006.2011.01743

Abstract

Abstract: Viviparous1 (Vp1) plays an important role in regulating embryo development, seed dormancy, and germination in higher plants. Although we previously demonstrated a close correlation between Vp1B allelic variation and the different levels of tolerance to pre-harvest sprouting (PHS) in common wheat, little is known about the potential cis-elements in Vp1B promoter region, which may be involved in the control of PHS by Vp1B. In this study, a 2 232 bp Vp1B upstream sequence was isolated. In silico analysis revealed the presence of nine ABRE, two DREB, six MYB, three GARE, one TCA-E, two TGACG-motif, four SKn-1, and one RYREPE cis-elements in the isolated sequence.Based on this, we constructed various plasmid vectors with 5′-truncated Vp1B promoters fused with the GUS (encoding β-glucuronidase) reporter gene. Transient expression assay in wheat callus indicated that Vp1B promoter activity was inducible by low-temperature, ABA, or GA treatment. However, the level of induced GUS activity declined as promoter length decreased. Subsequently, six truncated promoter reporter fusions were introduced into the genome of durum wheat by Agrobacterium-mediated transformation and stable transgenic lines were obtained. Analysis of the transgenic lines indicated that Vp1B promoter could drive GUS expression in the anther, axis,aleurone and root, but not in the leaves, stems and nodes. However, in the plants with the transgenic Vp1B promoter fragment longer than 660 bp, GUS expression could be induced in the nodes by ABA treatment.

Key words: Pre-harvest Sprouting, Vp1 promoter, Functional analysis, Transgenic wheat

SUN Yong-Wei, NIE Li-Na, MA Wei-Zhi, XU Zhao-Shi, JIA Lan-Qin. Cloning and Functional Analysis of Viviparous-1 Promoter in Wheat[J].Acta Agron Sin, 2011, 37(10): 1743-1751.

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Cloning and Functional Analysis of Viviparous-1 Promoter in Wheat

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