Gene and enhancer trap transposable elements reveal oxygen deprivation-regulated genes and their complex patterns of expression in Arabidopsis
Gene and enhancer trap transposable elements reveal oxygen deprivation-regulated genes and their complex patterns of expression in Arabidopsis
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DOI：
10.1093/aob/mcf119
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发表时间：
2003-01-01
期刊：
影响因子：
4.2
通讯作者：
Bailey-Serres, J
中科院分区：
文献类型：
--
作者：
Baxter-Burrell, A;Chang, R;Bailey-Serres, J
利用转座子标记技术，利用玉米Ds-GUS构建体分离拟南芥中缺氧诱导的基因。将800个基因捕获（DsG）和600个增强子捕获（DsE）系的幼苗在垂直放置的平板上生长1周，缺氧长达24小时，并对GUS活性进行染色。缺氧诱导65个品系幼苗基因表达的复杂模式。对15个株系的插入位点和表型进行了检测。令人惊讶的是，没有一个插入到编码已知厌氧多肽的基因中。在编码调节、酶、线粒体蛋白输入和未知功能的蛋白质的基因内或附近，以及编码推定的受体样激酶和推定的传感器组氨酸激酶的基因附近鉴定插入。四行有显着降低ADH活性后，24小时的缺氧，其中三个表现出降低的胁迫耐受性。野生型ADH水平的两个线是低氧不耐。奇怪的是，几个线有显着较高的ADH活性后12小时的缺氧，但降低了胁迫耐受性。咖啡因治疗，增加ADH比活性在野生型幼苗有氧条件下，足以增加GUS染色的15行中的7个，提供证据表明，这些基因可能是由胞质钙水平调节。这些结果证明了Ds-GUS标记系统在鉴定响应氧剥夺和钙第二信使而被调节的基因中的有效性。(C)2003年《植物学年鉴》（Annals of Botany Company）
Transposon tagging with modified maize Ds-GUS constructs was used to isolate genes induced by oxygen deprivation in Arabidopsis thaliana. Seedlings of 800 gene-trap (DsG) and 600 enhancer-trap (DsE) lines were grown on vertically positioned plates for 1 week, oxygen deprived for up to 24 h and stained for GUS activity. Oxygen deprivation induced intricate patterns of gene expression in seedlings of 65 lines. The insertion site and phenotypes of 15 lines were examined. Surprisingly, none of the insertions were into genes that encode known anaerobic polypeptides. Insertions were identified within or adjacent to genes encoding proteins of regulatory, enzymatic, mitochondrial protein import and unknown function, as well as adjacent to genes encoding a putative receptor-like kinase and putative sensor-histidine kinase. Four lines had significantly lower ADH activity after 24 h of oxygen deprivation and three of these showed reduced stress tolerance. Two lines with wild-type levels of ADH were low-oxygen intolerant. Paradoxically, several lines had significantly higher ADH activity after 12 h of oxygen deprivation but reduced stress tolerance. Caffeine treatment, which increased ADH specific activity in wild-type seedlings under aerobic conditions, was sufficient to increase GUS staining in seven of the 15 lines, providing evidence that these genes may be regulated by cytosolic calcium levels. These results demonstrate the effectiveness of the Ds-GUS tagging system in the identification of genes that are regulated in response to oxygen deprivation and a calcium second messenger. (C) 2003 Annals of Botany Company.