中国农业科学 ›› 2015, Vol. 48 ›› Issue (19): 3848-3856.doi: 10.3864/j.issn.0578-1752.2015.19.007

• 植物保护 • 上一篇    下一篇

氟唑活化酯对黄瓜抗枯萎病的诱导作用

石延霞1,徐玉芳2,谢学文1,柴阿丽1,王微微1,李宝聚1   

  1. 1中国农业科学院蔬菜花卉研究所,北京 100081
    2华东理工大学药学院,上海 200237
  • 收稿日期:2015-03-17 出版日期:2015-10-01 发布日期:2015-10-01
  • 通讯作者: 李宝聚,E-mail:libaoju@caas.cn
  • 作者简介:石延霞,E-mail:shiyanxia@caas.cn
  • 基金资助:
    “十二五”国家科技支撑计划(2010CB126101)、中国农业科学院科技创新工程、农业部园艺作物生物学与种质创制综合性重点实验室项目

Effects of FBT on Induction of Systemic Resistance in Cucumber Against Cucumber Fusarium Wilt Caused by Fusarium oxysporum f. sp. cucumerinum Owen

SHI Yan-xia1, XU Yu-fang2, XIE Xue-wen1, CHAI A-li1, WANG Wei-wei1, LI Bao-ju1   

  1. 1Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081
    2School of Pharmacy, East China University of Science and Technology, Shanghai 200237
  • Received:2015-03-17 Online:2015-10-01 Published:2015-10-01

摘要: 【目的】诱导抗病剂可以诱导寄主植物的系统抗病性作用,具有持效性和广谱性的特点,研究旨在明确新型诱导抗病剂氟唑活化酯(fluoro-substituted benzothiadiazole derivatives,FBT)对黄瓜抗枯萎病的诱导作用,为该化合物的诱导抗病性机理的深入研究提供参考。【方法】利用50 mg·L-1 FBT在黄瓜苗期移栽前3—4片真叶时喷雾诱导1次,3 d后移栽定植于含有黄瓜枯萎病菌(Fusarium oxysporum f. sp. cucumerinum Owen)的土壤中,缓苗后以叶面喷雾的方式进行第2次诱导,之后每隔7 d喷雾诱导1次,共诱导3次,对照诱导抗病剂苯并噻二唑(BTH)也采用同样的诱导方法,对照杀菌剂70%甲基硫菌灵可湿性粉剂1 500倍液则采用灌根施药,通过调查病情指数以评价其对枯萎病的抗病作用;研究FBT对枯萎病菌侵染黄瓜的影响,将供试黄瓜催芽至0.5 cm胚根后,浸泡于50 mg·L-1 FBT中胚根诱导1次,待播种后黄瓜生长至2片子叶期开始第2次诱导,以后每隔7 d诱导1次,共诱导3次,第3次诱导后24 h灌根接种黄瓜枯萎病菌,并于接种后1、3、5、7、9、11、14、16、20、24、29 d分别取黄瓜根组织,于冰水中清洗,利用酸性品红染色技术评价FBT诱导黄瓜后对枯萎病菌侵染的影响,利用Maule反应和甲苯胺蓝染色技术评价FBT对黄瓜根组织中木质素和酚类物质的沉积情况的影响,利用分光光度-比色法测定FBT诱导黄瓜后根组织中富含羟脯氨酸糖蛋白(HRGP)及β-1,3-葡聚糖酶活性,以分析FBT诱导黄瓜后根组织抗病性变化情况。【结果】诱导抗病性表达发现,50 mg·L-1的FBT对黄瓜抗枯萎病的诱导效果可达62.01%,高于对照诱导抗病剂BTH和杀菌剂70%甲基硫菌灵可湿性粉剂的防治效果。FBT诱导后的黄瓜在接种后7 d开始有枯萎病菌的侵染,此时未诱导只接种的黄瓜根组织因受枯萎病菌侵染,已经出现大量菌丝及孢子,FBT的诱导增强了寄主的抗病性反应,从而抑制了病原菌的侵染。同时经FBT诱导后4 d未接种及诱导后接种两个处理的根组织中具有大量褐色木质素沉积,尤其是根表皮细胞壁及韧皮部木质素沉降量明显,表现深褐色。对酚类物质的观察发现,在FBT诱导后2 d开始出现酚类物质沉积,诱导后4和6 d时,根组织中荧光信号较强,酚类物质沉积量较大,诱导后8 d时,根组织中荧光信号消失,酚类无明显沉积,木质素和酚类物质积累量的增加,以达到增强细胞壁的抗性,抵御病原菌的侵染的作用。在FBT诱导后1 d抗病相关蛋白HRGP和β-1,3-葡聚糖酶活性也明显增强,并一直保持在较高水平,为抵御病原菌侵染做准备。【结论】FBT诱导黄瓜产生了对枯萎病的抗病作用,同时黄瓜根组织通过次生代谢物质沉积量的增加及抗病相关蛋白活性增强,达到增强寄主根组织细胞壁的抗性,从而抵御病原菌的侵染,是具有发展前途的新型诱导抗病剂。

关键词: 氟唑活化酯, 黄瓜枯萎病菌, 诱导抗病性, 组织结构, 土传病害

Abstract: 【Objective】The systemic long-lasting and broad-spectrum resistance of cucumber seedlings was induced with natural or synthetic compounds. The objective of this study is to shed light on the resistance induced by fluoro-substituted benzothiadiazole derivatives (FBT) to cucumber Fusarium wilt and provide data for elucidating its mechanism.【Method】Before transplanting, 50 mg·L-1 of FBT was used to spray so as to induce the cucumber seedlings at 3-4 true leaf stage. Three days later, the cucumber seedlings were transplanted into the soil with Fusarium oxysporum f. sp. cucumerinum Owen, and sprayed with FBT so as to induce resistance for the second time. Sequently, the cucumber seedlings were sprayed every 7 days and for 3 times continuously. Meanwhile, contrast agent BTH also was used with the same induction method and 70% thiophanate-methyl at 1 500 times was applied through root-irrigation. The efficiency on the blight disease resistance was assessed through investigating disease index. For studying the influence of FBT on F. oxysporum f. sp. cucumerinum invasion of cucumber, the cucumber’s radicle length was sprouted to 0.5 cm and then the radicle immersed in 50 mg·L-1 of FBT. The resistance of cucumber seedlings was induced another time at the stage of 2 pieces of leaves. Since then, the cucumber seedlings were sprayed in order to induce the resistance every 7 days and total for 3 times. Twenty-four hours after the last induction, cucumber was inoculated with F. oxysporum f. sp. cucumerinum by the root-drenching method. Treated roots and the controls were harvested at 1, 3, 5, 7, 9, 11, 14, 16, 20, 24 and 29 days after inoculation and then cleaned in ice water. After induction, acid fuchsin dyeing technology was used to evaluate the FBT impacts on fusarium infection. In addition, the Maule reaction and toluidine blue staining technique were used to evaluate the change of lignin and phenolic substances in cucumber root tissue deposition. Moreover, metabolic alterations were investigated and enzyme activities of HRGP and β-1,3-glucanase were determined to analyze the physiological changes in BTH-treated cucumber by speetropho tometer-chromatometry.【Result】Because of the expression of induced resistance, it was found that the control efficiency of FBT was 62.01% at concentration of 50 mg·L-1, higher than BTH treatment and thiophanate-methyl treatment at the same conditions. Seven days after inoculation, colonization of F. oxysporium f. sp. cucumerinum began to appear in the cucumber root tissue induced by FBT. Whereas a large number of hyphae and spores had appeared on the cucumbers not induced with FBT, it indicated that cucumber induced with FBT could gain resistance so as to surpresse the colonization of pathogens. The brown lignin was accumulated in the cucumber roots induced by FBT, especially in epidermal cell walls and phloem xylem. Observation of phenolic substances showed that, two days after FBT induction, phenolic compounds appeared in 4 and 6 days, fluorescence signal in root tissue stronger, phenolic accumulated. Eight days after induction, the phenomena that root tissue fluorescence signal disappeared, phenolic no deposition, lignin and phenolic compounds accumulation increased, resulted in the enhancement of the resistance of the cell wall.【Conclusion】It was found that the resistance induced in cucumber seedlings by FBT includes the accumulation of secondary metabolites in the root and the activation of disease-resistant proteins, suggesting FBT is a promising novel inducer.

Key words: fluoro-substituted benzothiadiazole derivatives, Fusarium oxysporum f. sp. cucumerinum Owen, induced resistance, ultrastructure, soil-born disease