乙烯对匍匐翦股颖ISR抗病反应中AsA-GSH循环及胼胝质沉积的影响

doi:10.11686/cyxb2017429

摘要/Abstract

摘要： 以匍匐翦股颖品种“Penn-A4”为试验材料,侵染立枯丝核菌后,经丁二醇(BDO)诱导产生系统抗性(ISR),喷施不同浓度的乙烯合成抑制剂氯化钴(CoCl₂)和促进剂1-氨基环丙烷羧酸(ACC)后,检测抗坏血酸-谷胱甘肽(AsA-GSH)循环中抗坏血酸(AsA)和还原型谷胱甘肽(GSH)含量及相关酶活性的变化,并观察匍匐翦股颖ISR反应中胼胝质沉积的变化。结果表明,乙烯抑制剂处理下,匍匐翦股颖幼苗中AsA含量较低,抗坏血酸过氧化物酶(APX)活性下降,大量GSH被催化还原为氧化型谷胱甘肽(GSSG),GSSG大量积累,同时谷胱甘肽还原酶(GR)活性较低,GSSG经GR少量还原为GSH。乙烯促进剂处理下,AsA含量较高,APX活性升高,GSSG在高活性GR作用下催化还原为GSH,使得GSH大量积累。因此在匍匐翦股颖ISR抗病反应中,高浓度乙烯促进AsA和GSH的大量积累,它们不仅参与了活性氧的代谢平衡,同时也作为信号分子在ISR抗病反应中起着重要作用。匍匐翦股颖感染褐斑病后,胼胝质主要沉积在厚壁细胞、韧皮部、木质部和表皮组织,其中厚壁细胞胼胝质沉积最多,表皮组织沉积最少。此外,胼胝质沉积面积在不同乙烯信号分子处理间存在差异,但随着处理时间的延长差异不显著。一定浓度的乙烯对ISR反应中胼胝质的沉积具有一定的影响,随着处理时间的延长显著增加,在100 μmol·L^-1ACC处理5 d后,胼胝质沉积总面积仅为9.916 mm²,处理10 d后升至最高,为38.396 mm²,但在病害侵染后期,胼胝质数量减少,在100 μmol·L^-1ACC处理15 d后,胼胝质沉积总面积降至20.052 mm²,反映了乙烯信号分子对胼胝质沉积的影响是一种短期效应,短期内可提高匍匐翦股颖植株抗病性,具有信号分子的短时效特点。研究结果为探清匍匐翦股颖ISR抗病响应中ET信号分子如何调控抗病生理特性提供了理论基础。

关键词: 匍匐翦股颖, AsA-GSH循环, 胼胝质, 乙烯, 丁二醇, ISR抗病反应, 石蜡切片

Abstract: Creeping bentgrass seedlings cv. ‘Penn-A4’ infected with Rhizoctonia solani, which was produced by the induced-systemic-resistance (ISR) disease-resistance action induced by butanediol (BDO), was sprayed with different concentrations of the ethylene synthesis inhibitor, CoCl₂ and ethylene synthesis promoter, 1-amino cyclopropane carboxylic acid (ACC). Subsequently, changes inascorbic acid (AsA) and glutathione (GSH) contents and related enzyme activity in theascorbate-glutathione (AsA-GSH) cycle in seedlings were measured. Paraffin sectioning and fluorescence microscopy techniques were used to assess the effects of ethylene on callose deposition during ISR disease-resistance reactions. Under low ethylene concentrations AsA content was low and ascorbate peroxidase (APX) activity was reduced. A large amount of GSH was catalytically reduced to oxidize glutathione (GSSG) which accumulated. Glutathione reductase (GR) activity was low; GSSG is reduced to GSH by GR, consequently GSH remained at low levels. However, under high ethylene concentrations, AsA content was high and APX activity was higher. GSSG was catalytically reduced to GSH which accumulated. Therefore, in ISR disease-resistance reaction, high concentrations of ethylene induced the accumulation of AsA and GSH, which involved not only in the metabolic balance of reactive oxygen species, but also acted as signaling molecules playing an important role in the disease resistance of ISR. Callose was mainly deposited in sclerenchyma cells, phloem, xylem tissues and epidermis seedling leaves in ISR reaction induced by BDO. Callose deposition was highest in sclerenchyma cell and least in the epidermis. Callose deposition areas had differences in ethylene signal compounds but became less significant with reduced treatment time. Ethylene concentration affected callose deposition in ISR reaction induced by BDO; callose deposition increased significantly with reduced treatment time. Areas of callose deposition occupied 9.916 mm² under 100 μmol·L^-1 ACC treatment after 5 d, and increased to 38.396 mm² after 10 d. However, in the later stages of disease invasioncallose deposition area decreased to 20.052 mm² under 100 μmol·L^-1 ACC after 15 d. These results suggest that the ethylene molecular signalhas a short-term effect on callose deposition and increased the disease-resistance of creeping bentgrass. This research provides a theoretical basis for the exploration of ET signal molecules to control the physiological characteristics of resistance in ISR resistance response in creeping bentgrass.

Key words: creeping bentgrass (Agrostis stolonifera), AsA-GSH cycle, callose, ethylene, butanediol, ISR disease-resistance response, paraffin sectioning

姜寒玉, 王亚峰, 徐明, 甘培文, 张金林, 马晖玲. 乙烯对匍匐翦股颖ISR抗病反应中AsA-GSH循环及胼胝质沉积的影响[J]. 草业学报, 2018, 27(9): 120-131.

JIANG Han-yu, WANG Ya-feng, XU Ming, GAN Pei-wen, ZHANG Jin-lin, MA Hui-ling. Effects of ethylene on the ascorbate-glutathione cycle and callose deposition in ISR disease-resistance reaction in creeping bentgrass[J]. Acta Prataculturae Sinica, 2018, 27(9): 120-131.

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