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监测遭受渐进性干旱胁迫的拟南芥土培根和地上部的转录组变化

Monitoring Transcriptomic Changes in Soil-Grown Roots and Shoots of Arabidopsis thaliana Subjected to a Progressive Drought Stress.

作者信息

Bashir Khurram, Rasheed Sultana, Matsui Akihiro, Iida Kei, Tanaka Maho, Seki Motoaki

机构信息

Plant Genomic Network Research Team, RIKEN Center for Sustainable Resource Sciences, Yokohama, Kanagawa, Japan.

CREST, JST, Saitama, Japan.

出版信息

Methods Mol Biol. 2018;1761:223-230. doi: 10.1007/978-1-4939-7747-5_17.

Abstract

Numerous experiments have been performed in Arabidopsis to monitor changes in gene expression that occur in response to a variety of abiotic and biotic stresses, different growth conditions, and at various developmental stages. In addition, gene expression patterns have also been characterized among wild-type and mutant genotypes. Despite these numerous reports, transcriptional changes occurring in roots of soil-grown plants subjected to a progressive drought stress have remained undocumented. To fill this gap, we established a system that allows one to establish water-deficit conditions and to collect root and shoot samples with minimal damage to the root system. Arabidopsis plants are grown in a ceramic-based granular soil and subjected to progressive drought stress by withholding water. Root and shoot samples were collected separately, RNA was purified, and a microarray analysis of drought-stressed roots and shoots was performed at 0, 1, 3, 5, 7, and 9 days after the onset of drought stress treatment. Here, we describe the detailed protocol used to analyze the transcriptomic changes occurring in roots and shoots of soil-grown Arabidopsis subjected to a progressive drought stress.

摘要

在拟南芥中已经进行了大量实验,以监测因各种非生物和生物胁迫、不同生长条件以及不同发育阶段而发生的基因表达变化。此外,野生型和突变基因型之间的基因表达模式也已得到表征。尽管有这些大量报道,但遭受渐进性干旱胁迫的土壤种植植物根系中发生的转录变化仍未得到记录。为了填补这一空白,我们建立了一个系统,该系统能够营造缺水条件,并在对根系造成最小损害的情况下收集根和地上部分样本。拟南芥植株种植在基于陶瓷的颗粒状土壤中,通过停水使其遭受渐进性干旱胁迫。分别收集根和地上部分样本,纯化RNA,并在干旱胁迫处理开始后的第0、1、3、5、7和9天对受干旱胁迫的根和地上部分进行微阵列分析。在此,我们描述了用于分析遭受渐进性干旱胁迫的土壤种植拟南芥根和地上部分发生的转录组变化的详细方案。

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