Li Lin-lin, Li Tian-lai, Jiang Guo-bin, Jin Hua, Zou Ji-xiang
Ying Yong Sheng Tai Xue Bao. 2015 Nov;26(11):3497-502.
In this study, we investigated the effect of exogenous calcium and salicylic acid (SA) on Botrytis cinerea resistance in tomato seedlings. We treated a tomato strain susceptible to Botrytis cinerea with foliar spraying of water, SA, SA+CaCl2 and SA+EGTA (Ca2+ chelating agent) for one to five days. During the treatment, leaves were collected to analyze the reactive oxygen species (ROS) content, phenylalanine ammonia lyase (PAL) activity, chintase and β-1,3-glucanase levels, and the expression of pathogenesis related protein 1, 2, 3 (PR1, PR2, PR3). Three days after infection, the disease index was 74.8 in control plants, and 46.9, 38.5 and 70.3 in SA, SA+Ca and SA+ EGTA treated plants, respectively. SA treatment significantly increased ROS leaf accumulation, and activities of PAL, chintase and β-1,3-glucanase. These values were further enhanced in SA+Ca treated plants, but decreased in SA+EGTA treated plants. Application of SA significantly increased the expression levels of PR1, PR2a and PR3b, which were further elevated by the combination treatment with Ca2+. These effects were counteracted by the combination treatment of SA and EGTA. The transcription levels of PR2b and PR3a were up-regulated by 1-2 folds, and PR1, 2a and 3b by 2-5 folds in SA- and SA+Ca-treated plants relative to control. These data suggested that application of Ca2+ could synergistically increase SA-induced resistance to B. cinerea. The resistance was associated with ROS accumulation, therefore the increase in resistance might be through ROS ability to increase the activity of defense-related enzymes and expression levels of PR1, PR2a and PR3b.
在本研究中,我们调查了外源钙和水杨酸(SA)对番茄幼苗抗灰霉病能力的影响。我们用清水、SA、SA + CaCl₂ 和 SA + EGTA(Ca²⁺ 螯合剂)对一株对灰霉病敏感的番茄品系进行叶面喷施处理,持续 1 至 5 天。在处理过程中,采集叶片以分析活性氧(ROS)含量、苯丙氨酸解氨酶(PAL)活性、几丁质酶和 β-1,3-葡聚糖酶水平,以及病程相关蛋白 1、2、3(PR1、PR2、PR3)的表达。接种三天后,对照植株的病情指数为 74.8,而 SA、SA + Ca 和 SA + EGTA 处理的植株病情指数分别为 46.9、38.5 和 70.3。SA 处理显著增加了叶片中 ROS 的积累以及 PAL、几丁质酶和 β-1,3-葡聚糖酶的活性。这些值在 SA + Ca 处理的植株中进一步提高,但在 SA + EGTA 处理的植株中降低。SA 的施用显著增加了 PR1、PR2a 和 PR3b 的表达水平,Ca²⁺ 联合处理使其进一步升高。SA 和 EGTA 的联合处理抵消了这些效应。相对于对照,SA 和 SA + Ca 处理的植株中 PR2b 和 PR3a 的转录水平上调了 1 - 2 倍,PR1、2a 和 3b 上调了 2 - 5 倍。这些数据表明,Ca²⁺ 的施用可协同增强 SA 诱导的对灰霉病的抗性。这种抗性与 ROS 的积累有关,因此抗性的增加可能是通过 ROS 增强防御相关酶活性以及 PR1、PR2a 和 PR3b 表达水平的能力实现的。