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两个与维管表达和块根形成相关的木薯启动子。

Two cassava promoters related to vascular expression and storage root formation.

作者信息

Zhang Peng, Bohl-Zenger Susanne, Puonti-Kaerlas Johanna, Potrykus Ingo, Gruissem Wilhelm

机构信息

Institute of Plant Sciences, ETH-Zentrum / LFW E 17, Universitätstrasse 2, 8092 Zürich, Switzerland.

出版信息

Planta. 2003 Dec;218(2):192-203. doi: 10.1007/s00425-003-1098-0. Epub 2003 Sep 10.

Abstract

Cassava ( Manihot esculenta Crantz) storage roots, organs accumulating large amounts of starch, develop from primary roots via secondary growth. The availability of promoters related to storage-root formation is a prerequisite for engineering root traits in cassava. Two cDNAs, c15 and c54, were identified from a storage-root cDNA library of cassava MCol1505 via differential screening. The transcripts of c15 and c54 were detected in storage roots but not in leaves by Northern analysis. Homology analysis of the deduced amino acid sequences showed that C15 is likely to be related to cytochrome P450 proteins, which are involved in the oxidative degradation of various compounds, while C54 may be related to Pt2L4, a cassava glutamic acid-rich protein. The promoter regions of c15 and c54 were isolated from the corresponding clones in a cassava genomic library. A 1,465-bp promoter fragment ( p15/1.5) of c15 and a 1,081-bp promoter region ( p54/1.0) of c54 were translationally fused to the uidA reporter gene, and introduced into cassava and Arabidopsis thaliana (L.) Heynh. The expression patterns of p15/1.5::uidA and p54/1.0::uidA in transgenic plants showed that both promoters are predominantly active in phloem, cambium and xylem vessels of vascular tissues from leaves, stems, and root systems. More importantly, strong beta-glucuronidase activity was also detected in the starch-rich parenchyma cells of transgenic storage roots. Our results demonstrate that the two promoters are related to vascular expression and secondary growth of storage roots in cassava.

摘要

木薯(Manihot esculenta Crantz)的贮藏根是积累大量淀粉的器官,通过次生生长从初生根发育而来。与贮藏根形成相关的启动子的可用性是改良木薯根性状的先决条件。通过差异筛选从木薯MCol1505的贮藏根cDNA文库中鉴定出两个cDNA,即c15和c54。通过Northern分析在贮藏根中检测到c15和c54的转录本,但在叶片中未检测到。对推导的氨基酸序列进行同源性分析表明,C15可能与细胞色素P450蛋白有关,该蛋白参与各种化合物的氧化降解,而C54可能与木薯富含谷氨酸的蛋白Pt2L4有关。从木薯基因组文库中的相应克隆中分离出c15和c54的启动子区域。将c15的一个1465 bp启动子片段(p15/1.5)和c54的一个1081 bp启动子区域(p54/1.0)与uidA报告基因进行翻译融合,并导入木薯和拟南芥(Arabidopsis thaliana (L.) Heynh.)。p15/1.5::uidA和p54/1.0::uidA在转基因植物中的表达模式表明,这两个启动子在叶、茎和根系维管组织的韧皮部、形成层和木质部导管中均主要具有活性。更重要的是,在转基因贮藏根富含淀粉的薄壁细胞中也检测到了强烈的β-葡萄糖醛酸酶活性。我们的结果表明,这两个启动子与木薯贮藏根的维管表达和次生生长有关。

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