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拟南芥AtPAP1转录因子诱导转基因短角蒲公英中花青素的产生。

Arabidopsis AtPAP1 transcription factor induces anthocyanin production in transgenic Taraxacum brevicorniculatum.

作者信息

Qiu Jian, Sun Shuquan, Luo Shiqiao, Zhang Jichuan, Xiao Xianzhou, Zhang Liqun, Wang Feng, Liu Shizhong

机构信息

The Key Laboratory of Biology and Genetic Resources of Rubber Tree, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, 571737, Hainan, China.

出版信息

Plant Cell Rep. 2014 Apr;33(4):669-80. doi: 10.1007/s00299-014-1585-8. Epub 2014 Feb 21.

Abstract

This study developed a new purple coloured Taraxacum brevicorniculatum plant through genetic transformation using the Arabidopsis AtPAP1 gene, which overproduced anthocyanins in its vegetative tissues. Rubber-producing Taraxacum plants synthesise high-quality natural rubber (NR) in their roots and so are a promising alternative global source of this raw material. A major factor in its commercialization is the need for multipurpose exploitation of the whole plant. To add value to the aerial tissues, red/purple plants of the rubber-producing Taraxacum brevicorniculatum species were developed through heterologous expression of the production of anthocyanin pigment 1 (AtPAP1) transcription factor from Arabidopsis thaliana. The vegetative tissue of the transgenic plants showed an average of a 48-fold increase in total anthocyanin content over control levels, but with the exception of pigmentation, the transgenic plants were phenotypically comparable to controls and displayed similar growth vigor. Southern blot analysis confirmed that the AtPAP1 gene had been integrated into the genome of the high anthocyanin Taraxacum plants. The AtPAP1 expression levels were estimated by quantitative real-time PCR and were highly correlated with the levels of total anthocyanins in five independent transgenic lines. High levels of three cyanidin glycosides found in the purple plants were characterized by high performance liquid chromatography-mass spectrum analysis. The presence of NR was verified by NMR and infrared spectroscopy, and confirmed that NR biosynthesis had not been affected in the transgenic Taraxacum lines. In addition, other major phenylpropanoid products such as chlorogenic acid and quercetin glycosides were also enhanced in the transgenic Taraxacum. The red/purple transgenic Taraxacum lines described in this study would increase the future application of the species as a rubber-producing crop due to its additional health benefits.

摘要

本研究通过基因转化,利用拟南芥AtPAP1基因培育出一种新的紫色短角蒲公英植株,该基因在其营养组织中过量产生花青素。产橡胶的蒲公英植株在其根部合成高质量的天然橡胶(NR),因此是这种原材料有前景的全球替代来源。其商业化的一个主要因素是需要对整株植物进行多用途开发。为了增加地上组织的价值,通过异源表达来自拟南芥的花青素色素1(AtPAP1)转录因子,培育出了产橡胶的短角蒲公英品种的红色/紫色植株。转基因植株的营养组织中总花青素含量平均比对照水平增加了48倍,但除色素沉着外,转基因植株在表型上与对照相当,生长活力相似。Southern杂交分析证实AtPAP1基因已整合到高花青素蒲公英植株的基因组中。通过定量实时PCR估计AtPAP1表达水平,并且在五个独立的转基因株系中与总花青素水平高度相关。通过高效液相色谱 - 质谱分析对紫色植株中发现的三种矢车菊素糖苷的高水平进行了表征。通过核磁共振和红外光谱验证了NR的存在,并证实转基因蒲公英株系中NR生物合成未受影响。此外,转基因蒲公英中其他主要的苯丙烷类产物如绿原酸和槲皮素糖苷也有所增加。本研究中描述的红色/紫色转基因蒲公英株系因其额外的健康益处,将增加该物种作为产橡胶作物的未来应用。

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