Specific down-regulation of PAL genes by artificial microRNAs in Populus trichocarpa

被引:0
|
作者
Rui Shi
Chenmin Yang
Shanfa Lu
Ronald Sederoff
Vincent L. Chiang
机构
[1] North Carolina State University,Forest Biotechnology Group, Department of Forestry and Environmental Resources
[2] Chinese Academy of Medical Sciences and Peking Union Medical College,Medicinal Plant Cultivation Research Center, Institute of Medicinal Plant Development
[3] North Carolina State University,Department of Forest Biomaterials
来源
Planta | 2010年 / 232卷
关键词
Artificial microRNA; Phenylalanine ammonia-lyase; transformation; Down-regulation; PolyA tailing-based real time RT-PCR; Quantitation;
D O I
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中图分类号
学科分类号
摘要
Artificial microRNAs (amiRNAs) are similar to microRNAs (miRNAs) in that they are able to reduce the abundance of specific transcripts in plants by RNA-Induced Silencing Complex (RISC)-mediated cleavage and degradation, but differ in that they are designed for specific targets. The long generation times of forest trees have limited the discovery of mutations by conventional genetics. AmiRNAs can create gene-specific transcript reduction in transgenic trees in a single generation and may have broad application for functional genomics of trees. In this paper, we describe the specific down-regulation of multiple genes in the phenylalanine ammonia-lyase (PAL) gene family of Populus trichocarpa using amiRNA sequences incorporated in a P. trichocarpa miRNA-producing precursor, ptc-MIR408. Two different amiRNA constructs were designed to specifically down-regulate two different subsets of PAL genes, revealing differential regulation within the gene family. Down-regulation of subset A (PAL2, PAL4 and PAL5) by amiRNA-palA led to an increase in transcript abundance of subset B (PAL1 and PAL3). The reciprocal effect was not observed.
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页码:1281 / 1288
页数:7
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