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改变蒲公英(俄罗斯蒲公英)小橡胶粒子蛋白 TkSRPP3 的水平会导致橡胶代谢的定性和定量变化。

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

机构信息

Department of Biochemistry and Molecular Biology, University of Nevada, Reno, NV 89557, USA.

出版信息

Phytochemistry. 2012 Jul;79:46-56. doi: 10.1016/j.phytochem.2012.04.015. Epub 2012 May 19.

Abstract

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.

摘要

已经鉴定出几种参与天然橡胶生物合成的蛋白质,其中一种是小橡胶粒子蛋白(SRPP),最初在巴西橡胶树中被鉴定为与细胞质小泡(即橡胶粒子)相关的丰富蛋白。虽然之前的体外研究表明 SRPP 在橡胶生物合成中发挥作用,但缺乏体内证据支持这一假说。为了解决这个问题,采用转基因方法在 Taraxacum kok-saghyz(俄罗斯蒲公英或 Tk)中确定改变 SRPP 水平是否会影响橡胶生物合成。发现三种蒲公英 SRPP 在蒲公英橡胶粒子中高度丰富。最丰富的与粒子相关的 SRPP,TkSRPP3,其表达的时间和空间模式与蒲公英中天然橡胶积累的模式一致。为了证实其在橡胶生物合成中的作用,采用过表达和 RNAi 方法改变了俄罗斯蒲公英中的 TkSRPP3 表达。虽然 TkSRPP3 过表达株系的根中橡胶含量略高于对照,但 TkSRPP3 RNAi 株系的根中橡胶含量显著下降,并且产生的橡胶分子量明显低于对照株系。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,还表明其在调节顺式-1,4-聚异戊二烯聚合物的分子量方面具有功能。

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