DAFNAE, University of Padova, Agripolis, 35020 Legnaro PD, Italy.
J Hazard Mater. 2012 Nov 15;239-240:192-205. doi: 10.1016/j.jhazmat.2012.08.060. Epub 2012 Sep 1.
A differential display cDNA-AFLP derived technique was used to identify gene transcripts regulated by chromium (Cr) in relation to sulfur (S) nutrition in Brassica juncea. Twelve-day old plants were grown with 200 μM sulfate (+S), without sulfate (-S), with 200 μM sulfate plus 200 μM chromate (+S+Cr), or without sulfate plus 200 μM chromate (-S+Cr). Forty-four combinations of degenerate primers were assayed, which allowed the detection of 346 Transcript-Derived Fragments (TDFs) differentially regulated by Cr and S at times 0, 10 min, 1 h, 24 h. Eight sulfate transporters were identified, whose transcript abundance was dependent on the levels of plant S-compounds. For some of these transporters, a tight coordinated regulation of gene expression was observed in response to Cr. The MapMan analysis revealed a differential pattern of gene expression between +S+Cr and -S+Cr plants for several other transcripts and highlighted an overlap among responses to metals, defence against pathogens and senescence, hence suggesting the existence of common mechanisms of gene regulation. Among the identified gene transcripts, those involved in S metabolism and proteolitic processes may represent potential targets of genetic engineering in efforts to increase Cr accumulation and tolerance in plant species employed in phytoremediation techniques.
采用差异显示 cDNA-AFLP 技术,鉴定与油菜(Brassica juncea)硫(S)营养相关的铬(Cr)调节的基因转录本。12 天大的植物在 200μM 硫酸盐(+S)、无硫酸盐(-S)、200μM 硫酸盐加 200μM 铬酸盐(+S+Cr)或无硫酸盐加 200μM 铬酸盐(-S+Cr)中生长。对 44 种简并引物进行了检测,可检测到 346 个受 Cr 和 S 时间 0、10 分钟、1 小时、24 小时调节的转录衍生片段(TDFs)。鉴定了 8 种硫酸盐转运蛋白,其转录丰度依赖于植物 S 化合物的水平。对于其中一些转运蛋白,观察到它们的基因表达受到 Cr 的紧密协调调节。MapMan 分析显示,+S+Cr 和 -S+Cr 植物之间的基因表达存在差异,这表明几种其他转录本之间存在差异,并且对金属、病原体防御和衰老的反应存在重叠,这表明存在共同的基因调控机制。在鉴定的基因转录本中,那些参与 S 代谢和蛋白水解过程的转录本可能代表在植物修复技术中用于增加 Cr 积累和耐受性的植物遗传工程的潜在靶点。