Monitoring of methyl jasmonate-responsive genes in Arabidopsis by cDNA macroarray: self-activation of jasmonic acid biosynthesis and crosstalk with other phytohormone signaling pathways.

Yuko Sasaki, E. Asamizu, D. Shibata, Yasukazu Nakamura, T. Kaneko, K. Awai, Masayuki Amagai, C. Kuwata, T. Tsugane, T. Masuda, H. Shimada, K. Takamiya, H. Ohta, S. Tabata
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引用次数: 302

Abstract

Jasmonates mediate various physiological events in plant cells such as defense responses, flowering, and senescence through intracellular and intercellular signaling pathways, and the expression of a large number of genes appears to be regulated by jasmonates. In order to obtain information on the regulatory network of jasmonate-responsive genes (JRGs) in Arabidopsis thaliana (Arabidopsis), we screened 2880 cDNA clones for jasmonate responsiveness by a cDNA macroarray procedure. Since many of the JRGs reported so far have been identified in leaf tissues, the cDNA clones used were chosen from a non-redundant EST library that was prepared from above-ground organs. Hybridization to the filters was achieved using alpha-33P-labeled single-strand DNAs synthesized from mRNAs obtained from methyl jasmonate (MeJA)-treated and untreated Arabidopsis seedlings. Data analysis identified 41 JRGs whose mRNA levels were changed by more than three fold in response to MeJA. This was confirmed by Northern blot analysis by using eight representatives. Among the 41 JRGs identified, 5 genes were JA biosynthesis genes and 3 genes were involved in other signaling pathways (ethylene, auxin, and salicylic acid). These results suggest the existence of a positive feedback regulatory system for JA biosynthesis and the possibility of crosstalk between JA signaling and other signaling pathways.
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利用cDNA大阵列监测拟南芥茉莉酸甲基应答基因:茉莉酸生物合成的自激活及其与其他植物激素信号通路的串扰。
茉莉酸盐通过细胞内和细胞间信号通路介导植物细胞的防御反应、开花和衰老等多种生理事件,大量基因的表达似乎受到茉莉酸盐的调控。为了了解拟南芥(Arabidopsis thaliana)茉莉酸响应基因(JRGs)的调控网络,我们利用cDNA macroarray程序筛选了2880个茉莉酸响应基因的cDNA克隆。由于迄今为止报道的许多JRGs都是在叶片组织中鉴定出来的,因此所使用的cDNA克隆是从地上器官制备的非冗余EST文库中选择的。利用茉莉酸甲酯(MeJA)处理和未处理的拟南芥幼苗的mrna合成α - 33p标记的单链dna进行杂交。数据分析确定了41个JRGs,其mRNA水平对MeJA的反应改变了三倍以上。通过8个代表的Northern blot分析证实了这一点。在所鉴定的41个JRGs中,5个基因为JA生物合成基因,3个基因参与其他信号通路(乙烯、生长素和水杨酸)。这些结果表明JA生物合成存在正反馈调控系统,JA信号通路与其他信号通路之间可能存在串扰。
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