灵芝多糖对番茄抗灰霉病的诱导效应

doi:10.3864/j.issn.0578-1752.2016.11.007

摘要/Abstract

摘要： 【目的】诱导抗病剂可以诱导寄主植株产生系统抗病性，具有持效性和广谱性的特点。论文以番茄植株为模式植物，番茄灰霉菌（Botrytis cinerea）为目标菌，进行温室盆栽试验，以期明确灵芝多糖诱导剂对番茄灰霉病的诱导抗性。【方法】用一定质量浓度的灵芝多糖溶液（50、100、200和400 mg·L^-1）喷雾处理盆栽番茄植株第1、2片真叶（正反面），每2 d喷施1次，共诱导3次，最后一次诱导2 d后全株喷雾接种供试菌孢子悬浮液（（1—2）×10⁶个孢子/mL），对照组施用等量清水代替灵芝多糖溶液，与各处理平行喷雾接种番茄灰霉菌孢子悬浮液。在接种孢子悬浮液之前用注射器将番茄苗茎基部刺伤，注意伤口不能太大。然后再罩上塑料薄膜保湿24 h，接种孢子悬浮液2 d内植株遮阴处理，并用加湿器提高温室内的相对湿度，相对湿度控制在不低于90%，温度控制在（15±5）℃，接种后第3 天正常光照。通过调查植株病情指数，计算相对防治效果以评价灵芝多糖对番茄抗灰霉病的诱导效果，并且从防御酶活性、丙二醛（MDA）和叶绿素含量的变化角度评价其诱导抗性的作用机制。同时用一定质量浓度的灵芝多糖溶液（50、100、200和400 mg·L^-1）处理番茄种子并育苗，20 d后测定番茄幼苗株高、鲜重等多项生长指标。【结果】与直接施用等量清水后挑战接菌的对照组病情指数49.25相比，灵芝多糖处理组的番茄植株病情指数明显下降，在32.96—43.85。其中经400 mg·L^-1灵芝多糖处理的番茄植株病情指数最低为32.96，相对防效达到33.07%。经灵芝多糖处理的番茄植株，其体内与抗病有关的防御酶活性也发生显著变化，过氧化氢酶（CAT）和多酚氧化酶（PPO）的活性在多糖诱导第3天达到最高，分别达到162和98 U·min^-1·g^-1 FW，是对照组的2.13和1.71倍；过氧化物酶（POD）的活性在诱导后第4天达到最高值434 U·min^-1·g^-1 FW，是对照处理组的3.29倍。灵芝多糖能够系统地诱导番茄体内 CAT、POD 及 PPO 活性，显著抑制了感染灰霉菌后番茄植株叶片叶绿素含量的下降。灵芝多糖处理组的MDA含量与对照组相比有所下降，对照处理组在取样期间内MDA含量持续上升，而灵芝多糖处理组在取样期间内MDA含量呈先上升后趋于平稳的变化趋势。各质量浓度灵芝多糖对番茄种子的发芽率、芽长和幼苗株高、植株的鲜重均有不同程度的促进作用。其中200 mg·L^-1灵芝多糖浸泡的番茄种子发芽率最高，达87.3%，比对照组的发芽率77.3%提高了10.0%；且该处理对番茄幼苗的株高和地上部分鲜重的促进作用也最大，分别较对照增加了12.9%和33.3%；经100 mg·L^-1灵芝多糖处理后番茄幼苗的芽长和地下部分鲜重变化最大，与对照组相比分别提高了0.16 cm和0.33 g。【结论】灵芝多糖能够诱导番茄植株对灰霉病产生系统抗病性，同时对番茄种子发芽和番茄植株幼苗生长具有一定的促进作用。

关键词: 灵芝多糖, 番茄, 灰霉病, 诱导抗性, 防御酶

Abstract: 【Objective】The systemic long-lasting and broad-spectrum resistance was induced with natural or synthetic compounds. The objective of this study is to investigate the induced resistant effect of polysaccharide extracted from the fruiting body of Ganoderma lucidum (GLP) in tomato against gray mold.【Method】The 1-2 true leaf of tomato plants were sprayed with polysaccharides (50, 100, 200 and 400 mg·L^-1)for 3 times once every 2 days and inoculated spore suspension of Botrytis cinerea after 2 days later of the last time GLP treatment ((1-2)×10⁶spores/mL), meanwhile, the plants treated with the same volume of distilled water used as control instead of GLP solution and inoculated spore suspension of B. cinerea. A syringe stabbed the stem of tomato plant before inoculated spore suspension. Covering plastic film in order to moisturize 24 h and keeping humidity the 2 days of inoculating spore suspension of B. cinerea, the greenhouse relative humidity was not less than 90% and temperature was (15±5) ℃, normal light was recovered 3 days later. The disease index and disease reduction against gray mold in tomato plants was investigated. Simultaneously, the tomato seedlings were soaked in the concentration (50, 100, 200 and 400 mg·L^-1) of polysaccharides and then cultivated in pots. The germination rate, root length and plant height, shoot weight, fresh weight of tomato seedlings were measured 20 days after treatment with polysaccharides. 【Result】Compared with the clear water control’s disease index 49.25, the GLP treatment groups’ disease index were between with 32.96-43.85, significantly reduced, it was found that the lowest disease index 32.96 at the concentration of 400 mg·L^-1 GLP treatment and the disease reduction was 33.07%. The activity of protective enzyme in leaves showed different dynamic changes after GLP treatment, catalase (CAT) and polyphenoloxidase (PPO) activities remarkably increased, and the activities reached the highest peak after 3 days of induction. The highest CAT (162 U·min^-1·g^-1 FW) and PPO (98 U·min^-1·g^-1FW) were 2.13 and 1.71 times compared with the control group, respectively. Peroxidase (POD) activity reached the highest peak after 4 days of induction then decreased gradually, significantly higher than the control. The highest POD (434 U·min^-1·g^-1 FW) was 3.29 times compared with the control groups. MDA content after application of GLP showed a down trend compared to that of control. When the seed was treated with polysaccharides, the germination rate, root length and plant height, shoot weight, root weight of tomato seedlings all increased to a certain degree. The treatment at the concentration of 200 mg·L^-1 GLP showed the highest germination rate 87.3%, 10.0% higher than the control group germination rate 77.3%. Simultaneously, the treatment at the concentration of 200 mg·L^-1 GLP showed the highest promotion in the tomato plant height and shoot weight, enhanced 12.9% and 33.3% respectively than the control. The tomato seed germinal length and root weight enhanced 0.16 cm and 0.33 g respectively than the control at the concentration of 100 mg·L^-1 GLP.【Conclusion】GLP at appropriate concentration had the ability to induce systematic resistance against gray mold. In addition, GLP promoted the growth of tomato seedlings.

Key words: Ganoderma lucidum polysaccharides, tomato, gray mold, induced resistance, defensive enzymes

宁玉波，王红艳，乔康，刘秀梅，王开运. 灵芝多糖对番茄抗灰霉病的诱导效应[J]. 中国农业科学, 2016, 49(11): 2103-2112.

NING Yu-bo, WANG Hong-yan, QIAO Kang, LIU Xiu-mei, WANG Kai-yun. Induced Resistance by Polysaccharides Isolated from Ganoderma lucidum in Tomato Against Gray Mold[J]. Scientia Agricultura Sinica, 2016, 49(11): 2103-2112.

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