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Altered levels of the Taraxacum kok-saghyz (Russian dandelion) small rubber particle protein, TkSRPP3, result in qualitative and quantitative changes in rubber metabolism

机译:蒲公英蒲公英小橡胶颗粒蛋白TkSRPP3水平的改变导致橡胶代谢的质和量变化

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摘要

Several proteins have been identified and implicated in natural rubber biosynthesis, one of which, the small rubber particle protein (SRPP), was originally identified in Hevea brasiliensis as an abundant protein associated with cytosolic vesicles known as rubber particles. While previous in vitro studies suggest that SRPP plays a role in rubber biosynthesis, in vivo evidence is lacking to support this hypothesis. To address this issue, a transgene approach was taken in Taraxacum kok-saghyz (Russian dandelion or Tk) to determine if altered SRPP levels would influence rubber biosynthesis. Three dandelion SRPPs were found to be highly abundant on dandelion rubber particles. The most abundant particle associated SRPP, TkSRPP3, showed temporal and spatial patterns of expression consistent with patterns of natural rubber accumulation in dandelion. To confirm its role in rubber biosynthesis, TkSRPP3 expression was altered in Russian dandelion using over-expression and RNAi methods. While TkSRPP3 over-expressing lines had slightly higher levels of rubber in their roots, relative to the control, TkSRPP3 RNAi lines showed significant decreases in root rubber content and produced dramatically lower molecular weight rubber than the control line. Not only do results here provide in vivo evidence of TkSRPP proteins affecting the amount of rubber in dandelion root, but they also suggest a function in regulating the molecular weight of the cis-1, 4-polyisoprene polymer.
机译:已经鉴定出几种蛋白质并参与了天然橡胶的生物合成,其中一种小橡胶粒子蛋白质(SRPP)最初在巴西橡胶树中被鉴定为与胞质囊泡相关的丰富蛋白质,被称为橡胶粒子。尽管先前的体外研究表明SRPP在橡胶生物合成中起作用,但缺乏体内证据支持这一假设。为了解决这个问题,在蒲公英(Taraxacum kok-sa​​ghyz)(俄罗斯蒲公英或Tk)中采用了转基因方法,以确定改变的SRPP水平是否会影响橡胶的生物合成。发现三种蒲公英SRPP在蒲公英橡胶颗粒上含量很高。与颗粒相关的最丰富的SRPP TkSRPP3表现出与蒲公英中天然橡胶积累模式一致的时空表达模式。为了证实其在橡胶生物合成中的作用,使用过度表达和RNAi方法在俄罗斯蒲公英中改变了TkSRPP3的表达。虽然TkSRPP3过表达品系的根部橡胶含量略高,但相对于对照而言,TkSRPP3 RNAi品系的根部橡胶含量显着降低,并且分子量明显低于对照品系。这里的结果不仅提供了TkSRPP蛋白影响蒲公英根中橡胶含量的体内证据,而且还暗示了调节cis-1,4-聚异戊二烯聚合物分子量的功能。

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