Lab Plant-Microbe Interactions, Department of Crop Protection, Institute for Sustainable Agriculture, Agencia Estatal Consejo Superior de Investigaciones Científicas (CSIC) Córdoba, Spain.
Front Microbiol. 2014 Sep 5;5:427. doi: 10.3389/fmicb.2014.00427. eCollection 2014.
Pseudomonas fluorescens PICF7, a native olive root endophyte and effective biocontrol agent (BCA) against Verticillium wilt of olive, is able to trigger a broad range of defense responses in root tissues of this woody plant. In order to elucidate whether strain PICF7 also induces systemic defense responses in above-ground organs, aerial tissues of olive plants grown under non-gnotobiotic conditions were collected at different time points after root bacterization with this endophytic BCA. A suppression subtractive hybridization (SSH) cDNA library, enriched in up-regulated genes, was generated. This strategy enabled the identification of 376 ESTs (99 contigs and 277 singlets), many of them related to response to different stresses. Five ESTs, involved in defense responses, were selected to carry out time-course quantitative real-time PCR (qRT-PCR) experiments aiming to: (1) validate the induction of these genes, and (2) shed light on their expression pattern along time (from 1 to 15 days). Induction of olive genes potentially coding for lipoxygenase 2, catalase, 1-aminocyclopropane-1-carboxylate oxidase, and phenylananine ammonia-lyase was thus confirmed at some time points. Computational analysis also revealed that different transcription factors were up-regulated in olive aerial tissues (i.e., JERF, bHLH, WRKY), as previously reported for roots. Results confirmed that root colonization by this endophytic bacterium does not only trigger defense responses in this organ but also mounts a wide array of systemic defense responses in distant tissues (stems, leaves). This sheds light on how olive plants respond to the "non-hostile" colonization by a bacterial endophyte and how induced defense response can contribute to the biocontrol activity of strain PICF7.
荧光假单胞菌 PICF7 是一种土生土长的橄榄根内生菌,也是橄榄黄萎病的有效生物防治剂(BCA),能够在木本植物的根组织中引发广泛的防御反应。为了阐明菌株 PICF7 是否也能诱导地上器官的系统防御反应,在非无菌条件下生长的橄榄植株的气生组织在根部用这种内生 BCA 进行细菌接种后不同时间点进行了收集。生成了一个富含上调基因的抑制性消减杂交(SSH)cDNA 文库。该策略使鉴定出 376 个 EST(99 个连续序列和 277 个单序列),其中许多与对不同胁迫的反应有关。选择了 5 个参与防御反应的 EST 进行时间过程定量实时 PCR(qRT-PCR)实验,旨在:(1)验证这些基因的诱导,(2)阐明它们的表达模式随时间的变化(从 1 到 15 天)。因此,在某些时间点证实了橄榄基因潜在编码脂氧合酶 2、过氧化氢酶、1-氨基环丙烷-1-羧酸氧化酶和苯丙氨酸氨解酶的诱导。计算分析还表明,不同的转录因子在橄榄气生组织中被上调(即 JERF、bHLH、WRKY),这与以前在根部的报道一致。结果证实,这种内生细菌对根的定殖不仅会在该器官中引发防御反应,而且还会在远处的组织(茎、叶)中引发广泛的系统防御反应。这揭示了橄榄植物如何对细菌内生菌的“非宿主”定殖做出反应,以及诱导的防御反应如何有助于 PICF7 菌株的生物防治活性。