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神圣罗勒(Ocimum sanctum)的OscWRKY1调控苯丙烷类途径基因并促进拟南芥对病原体感染的抗性。

Ocimum sanctum, OscWRKY1, regulates phenylpropanoid pathway genes and promotes resistance to pathogen infection in Arabidopsis.

作者信息

Joshi Ashutosh, Jeena Gajendra Singh, Kumar Ravi Shankar, Pandey Alok, Shukla Rakesh Kumar

机构信息

Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, India.

Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, India.

出版信息

Plant Mol Biol. 2022 Oct;110(3):235-251. doi: 10.1007/s11103-022-01297-2. Epub 2022 Jul 2.

Abstract

OscWRKY1 from Ocimum sanctum positively regulates phenylpropanoid pathway genes and rosmarinic acid content. OscWRKY1 overexpression promotes resistance against bacterial pathogen in Arabidopsis. WRKY transcription factor (TF) family regulates various developmental and physiological functions in plants. PAL genes encode enzymes which are involved in plant defense responses, but the direct regulation of PAL genes and phenylpropanoid pathway through WRKY TF's is not well characterized. In the present study, we have characterized an OscWRKY1 gene from Ocimum sanctum which shows induced expression by methyl jasmonate (MeJA), salicylic acid (SA), and wounding. The recombinant OscWRKY1 protein binds to the DIG-labeled (Digoxigenin) W-box cis-element TTGAC[C/T] and activates the LacZ reporter gene in yeast. Overexpression of OscWRKY1 enhances Arabidopsis resistance towards Pseudomonas syringae pv. tomato Pst DC3000. Upstream activator sequences of PAL and C4H have been identified to contain the conserved W-box cis-element (TTGACC) in both O. sanctum and Arabidopsis. OscWRKY1 was found to interact with W-box cis-element present in the PAL and C4H promoters. Silencing of OscWRKY1 using VIGS resulted in reduced expression of PAL, C4H, COMT, F5H and 4CL transcripts. OscWRKY1 silenced plants exhibit reduced PAL activity, whereas, the overexpression lines of OscWRKY1 in Arabidopsis exhibit increased PAL activity. Furthermore, the metabolite analysis of OscWRKY1 silenced plants showed reduced rosmarinic acid content. These results revealed that OscWRKY1 positively regulates the phenylpropanoid pathway genes leading to the alteration of rosmarinic acid content and enhances the resistance against bacterial pathogen in Arabidopsis.

摘要

来自神圣罗勒的OscWRKY1正向调控苯丙烷类途径基因和迷迭香酸含量。OscWRKY1过表达增强了拟南芥对细菌病原体的抗性。WRKY转录因子(TF)家族调控植物的各种发育和生理功能。PAL基因编码参与植物防御反应的酶,但WRKY TF对PAL基因和苯丙烷类途径的直接调控尚未得到充分表征。在本研究中,我们鉴定了来自神圣罗勒的OscWRKY1基因,该基因受茉莉酸甲酯(MeJA)、水杨酸(SA)和创伤诱导表达。重组OscWRKY1蛋白与地高辛标记(洋地黄毒苷)的W盒顺式元件TTGAC[C/T]结合,并在酵母中激活LacZ报告基因。OscWRKY1的过表达增强了拟南芥对丁香假单胞菌番茄致病变种Pst DC3000的抗性。已确定PAL和C4H的上游激活序列在神圣罗勒和拟南芥中均含有保守的W盒顺式元件(TTGACC)。发现OscWRKY1与PAL和C4H启动子中存在的W盒顺式元件相互作用。使用病毒诱导的基因沉默(VIGS)沉默OscWRKY1导致PAL、C4H、COMT、F5H和4CL转录本的表达降低。OscWRKY1沉默的植株显示PAL活性降低,而OscWRKY1在拟南芥中的过表达株系显示PAL活性增加。此外,对OscWRKY1沉默植株的代谢物分析表明迷迭香酸含量降低。这些结果表明,OscWRKY1正向调控苯丙烷类途径基因,导致迷迭香酸含量改变,并增强拟南芥对细菌病原体的抗性。

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