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Transformation of Solanum tuberosum plastids allows high expression levels of β-glucuronidase both in leaves and microtubers developed in vitro

机译:马铃薯茄质体的转化允许在体外发育的叶片和微块茎中高水平表达β-葡萄糖醛酸苷酶

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摘要

Plastid genome transformation offers an attractive methodology for transgene expression in plants, but for potato, only expression of gfp transgene (besides the selective gene aadA) has been published. We report here successful expression of β-glucuronidase in transplastomic Solanum tuberosum (var. Desiree) plants, with accumulation levels for the recombinant protein of up to 41% of total soluble protein in mature leaves. To our knowledge, this is the highest expression level reported for a heterologous protein in S. tuberosum. Accumulation of the recombinant protein in soil-grown minitubers was very low, as described in previous reports. Interestingly, microtubers developed in vitro showed higher accumulation of β-glucuronidase. As light exposure during their development could be the trigger for this high accumulation, we analyzed the effect of light on β-glucuronidase accumulation in transplastomic tubers. Exposure to light for 8 days increased β-glucuronidase accumulation in soil-grown tubers, acting as a light-inducible expression system for recombinant protein accumulation in tuber plastids. In this paper we show that plastid transformation in potato allows the highest recombinant protein accumulation in foliar tissue described so far for this food crop. We also demonstrate that in tubers high accumulation is possible and depends on light exposure. Because tubers have many advantages as protein storage organs, these results could lead to new recombinant protein production schemes based on potato.
机译:质体基因组转化为植物中的转基因表达提供了一种有吸引力的方法,但是对于马铃薯,仅公开了gfp转基因的表达(除了选择性基因aadA)。我们在这里报告了β-葡萄糖醛酸苷酶在转质体茄(var。Desiree)植物中的成功表达,其中重组蛋白的积累水平高达成熟叶中总可溶性蛋白的41%。据我们所知,这是报道的马铃薯中异源蛋白的最高表达水平。如以前的报道所述,重组蛋白在土壤种植的微型块茎中的积累非常低。有趣的是,体外发育的微块茎显示出更高的β-葡萄糖醛酸苷酶积累。由于在它们发育期间暴露于光可能是这种高积累的触发因素,我们分析了光对转质体块茎中β-葡萄糖醛酸苷酶积累的影响。光照8天可增加土壤生长的块茎中β-葡萄糖醛酸苷酶的积累,可作为光诱导表达系统来重组块茎质体中的蛋白质。在本文中,我们表明,迄今为止,对于这种粮食作物,马铃薯中的质体转化使重组蛋白在叶组织中的积累最高。我们还证明在块茎中高积累是可能的,并且取决于光照。由于块茎具有作为蛋白质储存器官的许多优点,这些结果可能会导致基于马铃薯的新的重组蛋白质生产方案。

著录项

  • 来源
    《Planta》 |2012年第4期|p.807-818|共12页
  • 作者单位

    Laboratorio de Biotecnología Vegetal, Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET), Vuelta de Obligado 2490, 2do. Piso, C1428ADN, Ciudad Autónoma de Buenos Aires, Argentina;

    Laboratorio de Agrobiotecnología, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Intendente Güiraldes 2160, Ciudad Universitaria, C1428EGA, Ciudad Autónoma de Buenos Aires, Argentina;

    Facultad de Ciencias Exactas y Naturales, Departamento de Fisiología, Biología Molecular y Celular, Universidad de Buenos Aires, Intendente Güiraldes 2160 Pab. II, Ciudad Universitaria, C1428EGA, Ciudad Autónoma de Buenos Aires, Argentina;

    Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Bernal, Buenos Aires, Argentina;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Potato; Tuber; Plastid transformation; Tobacco; Chloroplast; β-Glucuronidase;

    机译:马铃薯;Tuber;质体转化;烟草;叶绿体;β-葡萄糖醛酸苷酶;

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