Briggs Amy G, Adams-Phillips Lori C, Keppler Brian D, Zebell Sophia G, Arend Kyle C, Apfelbaum April A, Smith Joshua A, Bent Andrew F
Department of Biology, Beloit College, Beloit, WI, United States of America.
Department of Plant Pathology, University of Wisconsin-Madison, Madison, WI, United States of America.
PLoS One. 2017 Dec 28;12(12):e0190268. doi: 10.1371/journal.pone.0190268. eCollection 2017.
Pharmacological inhibition of poly(ADP-ribose) polymerase (PARP) or loss of Arabidopsis thaliana PARG1 (poly(ADP-ribose) glycohydrolase) disrupt a subset of plant defenses. In the present study we examined the impact of altered poly(ADP-ribosyl)ation on early gene expression induced by the microbe-associate molecular patterns (MAMPs) flagellin (flg22) and EF-Tu (elf18). Stringent statistical analyses and filtering identified 178 genes having MAMP-induced mRNA abundance patterns that were altered by either PARP inhibitor 3-aminobenzamide (3AB) or PARG1 knockout. From the identified set of 178 genes, over fifty Arabidopsis T-DNA insertion lines were chosen and screened for altered basal defense responses. Subtle alterations in callose deposition and/or seedling growth in response to those MAMPs were observed in knockouts of At3g55630 (FPGS3, a cytosolic folylpolyglutamate synthetase), At5g15660 (containing an F-box domain), At1g47370 (a TIR-X (Toll-Interleukin Receptor domain)), and At5g64060 (a predicted pectin methylesterase inhibitor). Over-represented GO terms for the gene expression study included "innate immune response" for elf18/parg1, highlighting a subset of elf18-activated defense-associated genes whose expression is altered in parg1 plants. The study also allowed a tightly controlled comparison of early mRNA abundance responses to flg22 and elf18 in wild-type Arabidopsis, which revealed many differences. The PARP inhibitor 3-methoxybenzamide (3MB) was also used in the gene expression profiling, but pleiotropic impacts of this inhibitor were observed. This transcriptomics study revealed targets for further dissection of MAMP-induced plant immune responses, impacts of PARP inhibitors, and the molecular mechanisms by which poly(ADP-ribosyl)ation regulates plant responses to MAMPs.
对聚(ADP - 核糖)聚合酶(PARP)的药理学抑制或拟南芥PARG1(聚(ADP - 核糖)糖水解酶)的缺失会破坏植物防御的一个子集。在本研究中,我们研究了聚(ADP - 核糖基)化改变对由微生物相关分子模式(MAMPs)鞭毛蛋白(flg22)和延伸因子Tu(elf18)诱导的早期基因表达的影响。严格的统计分析和筛选确定了178个基因,其MAMP诱导的mRNA丰度模式因PARP抑制剂3 - 氨基苯甲酰胺(3AB)或PARG1基因敲除而改变。从这178个已鉴定的基因中,选择了五十多个拟南芥T - DNA插入系,并筛选其基础防御反应的改变。在At3g55630(FPGS3,一种胞质叶酰聚谷氨酸合成酶)、At5g15660(含有一个F - 盒结构域)、At1g47370(一个TIR - X(Toll样白细胞介素受体结构域))和At5g64060(一种预测的果胶甲酯酶抑制剂)的基因敲除植株中,观察到对这些MAMPs的胼胝质沉积和/或幼苗生长的细微改变。基因表达研究中过度富集的基因本体(GO)术语包括elf18/parg1的“先天免疫反应”,突出了elf18激活的防御相关基因的一个子集,其在parg1植株中的表达发生了改变。该研究还对野生型拟南芥中对flg22和elf18的早期mRNA丰度反应进行了严格对照的比较,揭示了许多差异。PARP抑制剂3 - 甲氧基苯甲酰胺(3MB)也用于基因表达谱分析,但观察到该抑制剂具有多效性影响。这项转录组学研究揭示了用于进一步剖析MAMP诱导的植物免疫反应、PARP抑制剂的影响以及聚(ADP - 核糖基)化调节植物对MAMPs反应的分子机制的靶点。