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甘薯 IbMYB1 基因作为甘薯基因内载体系统的潜在可见标记。

The sweet potato IbMYB1 gene as a potential visible marker for sweet potato intragenic vector system.

机构信息

Bioindustrial Process Center, Korea Research Institute of Bioscience and Biotechnology, Jeonbuk 580-185, Korea.

出版信息

Physiol Plant. 2010 Jul 1;139(3):229-40. doi: 10.1111/j.1399-3054.2010.01365.x. Epub 2010 Feb 16.

Abstract

MYB transcription factors play important roles in transcriptional regulation of many secondary metabolites including anthocyanins. We cloned the R2R3-MYB type IbMYB1 complementary DNAs from the purple-fleshed sweet potato (Ipomoea batatas L. cv Sinzami) and investigated the expression patterns of IbMYB1 gene with IbMYB1a and IbMYB1b splice variants in leaf and root tissues of various sweet potato cultivars by reverse transcription-polymerase chain reaction. The transcripts of IbMYB1 were predominantly expressed in the purple-fleshed storage roots and they were also detectable in the leaf tissues accumulating anthocyanin pigments. In addition, transcript levels of IbMYB1 gene were up-regulated by treatment with methyl jasmonate or salicylic acid in leaf and root tissues of cv. White Star. To set up the intragenic vector system in sweet potato, we first evaluated the utilization of the IbMYB1 gene as a visible selectable marker. The IbMYB1a was transiently expressed in tobacco leaves under the control of a constitutive cauliflower mosaic virus 35S promoter, a root-specific and sucrose-inducible sporamin promoter, and an oxidative stress-inducible sweet potato anionic peroxidase2 promoter. We also showed that overexpression of IbMYB1a induced massive anthocyanin pigmentation in tobacco leaves and up-regulated the transcript levels of the structural genes in anthocyanin biosynthetic pathway. Furthermore, high-performance liquid chromatography analysis revealed that the expression of IbMYB1a led to production of cyanidin as a major core molecule of anthocyanidins in tobacco leaves. These results suggest that the IbMYB1 gene can be applicable to a visible marker for sweet potato transformation with intragenic vectors, as well as the production of anthocyanin as important nutritive value in other plant species.

摘要

MYB 转录因子在许多次生代谢物(包括花青素)的转录调控中发挥重要作用。我们从紫色果肉甘薯(Ipomoea batatas L. cv Sinzami)中克隆了 R2R3-MYB 型 IbMYB1 互补 DNA,并通过反转录-聚合酶链反应(RT-PCR)研究了 IbMYB1 基因及其 IbMYB1a 和 IbMYB1b 剪接变体在不同甘薯品种叶片和根组织中的表达模式。IbMYB1 的转录本主要在紫色肉质块根中表达,在积累花青素色素的叶片组织中也能检测到。此外,IbMYB1 基因的转录水平在 White Star 品种的叶片和根组织中用茉莉酸甲酯或水杨酸处理后上调。为了在甘薯中建立基因内载体系统,我们首先评估了 IbMYB1 基因作为可见选择标记的利用。IbMYB1a 在烟草叶片中瞬时表达,受组成型花椰菜花叶病毒 35S 启动子、根特异性和蔗糖诱导的蔗糖酶启动子以及氧化应激诱导的甘薯阴离子过氧化物酶 2 启动子的控制。我们还表明,IbMYB1a 的过表达诱导烟草叶片中大量花青素的形成,并上调花青素生物合成途径的结构基因的转录水平。此外,高效液相色谱分析表明,IbMYB1a 的表达导致烟草叶片中花氰定作为花青素的主要核心分子的产生。这些结果表明,IbMYB1 基因可用于甘薯基因内载体转化的可见标记,以及在其他植物物种中生产作为重要营养价值的花青素。

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