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在转基因水稻植株的整个生长阶段进行六个干旱诱导启动子的功能分析。

Functional analysis of six drought-inducible promoters in transgenic rice plants throughout all stages of plant growth.

机构信息

School of Biotechnology and Environmental Engineering, Myongji University, Yongin 449-728, Korea.

出版信息

Planta. 2010 Aug;232(3):743-54. doi: 10.1007/s00425-010-1212-z. Epub 2010 Jun 22.

Abstract

There are few efficient promoters for use with stress-inducible gene expression in plants, and in particular for monocotyledonous crops. Here, we report the identification of six genes, Rab21, Wsi18, Lea3, Uge1, Dip1, and R1G1B that were induced by drought stress in rice microarray experiments. Gene promoters were linked to the gfp reporter and their activities were analyzed in transgenic rice plants throughout all stages of plant growth, from dry seeds to vegetative tissues to flowers, both before and after drought treatments. In fold induction levels, Rab21 and Wsi18 promoters ranged from 65- and 36-fold in leaves to 1,355- and 492-fold in flowers, respectively, whereas Lea3 and Uge1 were higher in leaves, but lower in roots and flowers, as compared with Rab21 and Wsi18. Dip1 and R1G1B promoters had higher basal levels of activity under normal growth conditions in all tissues, resulting in smaller fold-induction levels than those of the others. In drought treatment time course, activities of Dip1 and R1G1B promoters rapidly increased, peaked at 2 h, and remained constant until 8 h, while that of Lea3 slowly yet steadily increased until 8 h. Interestingly, Rab21 activity increased rapidly and steadily in response to drought stress until expression peaked at 8 h. Thus, we have isolated and characterized six rice promoters that are all distinct in fold induction, tissue specificity, and induction kinetics under drought conditions, providing a variety of drought-inducible promoters for crop biotechnology.

摘要

在植物中,特别是在单子叶作物中,用于诱导应激基因表达的有效启动子很少。在这里,我们报告了在水稻微阵列实验中鉴定的六个基因 Rab21、Wsi18、 Lea3、Uge1、Dip1 和 R1G1B,它们受到干旱胁迫的诱导。将基因启动子与 gfp 报告基因连接,并在转基因水稻植株的整个生长阶段(从干种子到营养组织到花)分析其活性,包括干旱处理前后。在折叠诱导水平上,Rab21 和 Wsi18 启动子在叶片中的诱导倍数分别为 65-和 36-倍,在花中的诱导倍数分别为 1355-和 492-倍,而 Lea3 和 Uge1 在叶片中的诱导倍数较高,但在根和花中的诱导倍数较低,与 Rab21 和 Wsi18 相比。Dip1 和 R1G1B 启动子在正常生长条件下在所有组织中具有较高的基础活性,导致折叠诱导水平小于其他启动子。在干旱处理时间过程中,Dip1 和 R1G1B 启动子的活性迅速增加,在 2 小时时达到峰值,并保持恒定直到 8 小时,而 Lea3 启动子缓慢但稳定地增加直到 8 小时。有趣的是,Rab21 活性迅速而稳定地响应干旱胁迫,直到 8 小时达到表达峰值。因此,我们已经分离和表征了六个水稻启动子,它们在折叠诱导、组织特异性和干旱条件下的诱导动力学方面都有所不同,为作物生物技术提供了多种干旱诱导启动子。

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