嗜碱假单胞菌Ej2对稻瘟病的防治效果及对水稻内源激素的影响

doi:10.3864/j.issn.0578-1752.2022.02.007

摘要/Abstract

摘要：

【目的】明确水稻内生菌嗜碱假单胞菌（Pseudomonas alcaliphila）菌株Ej2对镰孢菌（Fusarium spp.）等病原菌的抑菌效果,重点研究对稻瘟病的防治作用及对水稻内源激素的影响,为稻瘟病菌（Magnaporthe oryzae）杀菌剂的研发提供新的生防菌资源。【方法】嗜碱假单胞菌菌株Ej2分离于水稻叶片,采用对峙培养法测试该菌株对稻瘟病菌以及对镰孢菌、草莓炭疽病菌（Colletotrichum gloeospoioides）、链格孢苹果专化型（Alternaria alternata f. sp. mali）BJ-A5和草莓黑斑病菌（A. alternata) BJ-ST24等病原真菌的抑菌效果,并检测其挥发性物质对稻瘟病菌的抑菌作用以及发酵液对水稻种子萌发的促进作用,盆栽法检测对水稻叶瘟的温室防治效果,通过小区试验验证对叶瘟和穗颈瘟的田间防治效果及对水稻的促生效果,采用基于MRM方法的靶向代谢组学技术检测发酵液喷施3次对胁迫激素、细胞分裂素等水稻内源激素的影响。【结果】嗜碱假单胞菌菌株Ej2及其挥发性物质对稻瘟病菌菌丝生长具有显著拮抗作用,对水稻叶瘟的温室防治效果为78.26%,与75%三环唑可湿性粉剂无显著差异;对叶瘟和穗颈瘟的田间防治效果分别为75.07%—83.67%和62.79%—75.09%,对叶瘟的田间防治效果与稻瘟酰胺有显著差异,对穗颈瘟的田间防治效果与稻瘟酰胺无显著差异。菌株Ej2对西瓜枯萎病菌（F. oxysporum f. sp. niveum）、茄镰孢（F. solani）、串珠镰孢（F. moniliforme）、草莓炭疽病菌、链格孢苹果专化型和草莓黑斑病菌5种植物病原真菌具有抑菌活性。与清水对照相比,Ej2发酵液显著增加水稻种子萌发率、株高和穗重;喷施3次后,植物胁迫激素中的乙烯（ETH）和脱落酸（ABA）含量显著增加,细胞分裂素中的玉米素核苷（czR）、反式玉米素核苷（tZR）和异戊烯基腺嘌呤核苷（iPR）激素含量显著下降,植物生长素（IAA）、茉莉酸（JA）、顺反茉莉酸类激素(cis-OPDA)、茉莉酸-异亮氨酸（JA-LIE）、顺式玉米素（cZ）、反式玉米素（tZ）、异戊烯基腺嘌呤（iP）、水杨酸（SA）等植物内源激素含量无显著变化。【结论】嗜碱假单胞菌菌株Ej2具有防病、促进作物生长及增强水稻植株抗逆性的作用,是一种可以防治稻瘟病等真菌病害的生防微生物。

关键词: 嗜碱假单胞菌, 稻瘟病, 生物防治, 植物病原真菌, 水稻内源激素

Abstract:

【Objective】The objective of this study is to clarify the antagonist activity of Pseudomonas alcaliphila strain Ej2 against various phytopathogens (Fusarium spp., etc.) in vitro, focusing on the control efficacy of P. alcaliphila strain Ej2 against rice blast and influences of endogenous hormones in rice plant. The result will provide new biological control resources for the research and development of fungicides against rice blast. 【Method】P. alcaliphila strain Ej2 was isolated from rice leaf tissue. Antagonistic action of strain Ej2 against Magnaporthe oryzae and other pathogenic fungi such as Fusarium spp., Colletotrichum gloeospoioides, Alternaria alternata which can cause apple spot fallen leaf disease and strawberry black spot, etc, was determined by dual culture assay. In addition, antagonistic activity of volatile substance produced by P. alcaliphila strain Ej2 against M. oryzae and promoting efficacy of fermentation broth increasing the germinating rate of rice seeds were tested in the experiment. The control efficacy of strain Ej2 was determined for the control of rice leaf blast under greenhouse condition by pot-culture method. The plot experiments were conducted to verify the growth promoting effect on rice and control efficacy against leaf blast and neck blast of strain Ej2. Targeted metabonomics based on MRM method was used to detect the effect of strain Ej2 on endogenous hormones in rice plant such as stress hormone and cytokinin after spraying fermentation broth for three times. 【Result】The strain and volatile substance significantly suppressed the hypha growth rate of M. oryzae P131. The control efficacy against rice blast under greenhouse condition was 78.26% and it had no significant difference with 75% tricyclazole wettable powder. The control efficacy against rice leaf blast and neck blast in fields was 75.07%-83.67% and 62.79%-75.09%, respectively. It had a significant difference between the control efficacy of strain Ej2 and 20% fenoxanil suspending agent against rice leaf blast, but it had no significant difference between the control efficacy of strain Ej2 and 20% fenoxanil suspending agent against rice neck blast. P. alcaliphila strain Ej2 showed strong antagonist activity against other five pathogenic fungi, such as F. oxysporum f. sp. niveum, F. solani, F. moniliforme, C. gloeospoioides, A. alternata. The fermentation broth of P. alcaliphila strain Ej2 significantly increased the germinating rate of rice seeds, plant height, and panicle weight compared with water control. After spraying three times, the levels of the stress hormone in plant such as ethylene (ETH) and abscisic acid (ABA) were significantly increased, but the levels of cytokinin such as zeatin riboside (czR), trans-zeatin riboside (tZR) and N6-isopentenyladenine riboside (iPR) were significantly decreased. In addition, the levels of several endogenous hormones in rice plant including IAA, jasmonic acid (JA), cis-OPDA, jasmonoyl-L-isoleucine (JA-LIE), cis-zeatin (cZ), trans-zeatin (tZ), N6-isopentenyladenine (iP) and salicylic acid (SA) had no significant change. 【Conclusion】P. alkalophilis strain Ej2 possesses the application value to control disease, promote crop growth and enhance stress resistance of rice plant, and it is considered to be a biological control microorganism against rice blast and other fungi diseases.

Key words: Pseudomonas alcaliphila, rice blast, biological control, plant pathogenic fungi, endogenous hormones in rice

沙月霞, 黄泽阳, 马瑞. 嗜碱假单胞菌Ej2对稻瘟病的防治效果及对水稻内源激素的影响[J]. 中国农业科学, 2022, 55(2): 320-328.

SHA YueXia, HUANG ZeYang, MA Rui. Control Efficacy of Pseudomonas alcaliphila Strain Ej2 Against Rice Blast and Its Effect on Endogenous Hormones in Rice[J]. Scientia Agricultura Sinica, 2022, 55(2): 320-328.

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处理 Treatment	病情指数 Disease index	防治效果 Control efficacy (%)
嗜碱假单胞菌Ej2 P. alcaliphila Ej2	6.81±1.32b	78.26±1.22a
清水对照Water control	31.33±1.53a	/
三环唑Tricyclazole	6.92±1.14b	77.91±1.43a

处理 Treatment	叶瘟Leaf blast		穗颈瘟Panicle blast
处理 Treatment	病情指数 Disease index	防治效果 Control efficacy (%)	病情指数 Disease index	防治效果 Control efficacy (%)
嗜碱假单胞菌Ej2 P. alcaliphila Ej2	6.17±1.32b	75.07±1.71a	8.03±0.64bc	75.09±1.62a
稻瘟酰胺Fenoxanil	7.22±1.19b	64.05±1.00b	8.72±0.51bc	74.60±2.08a
芽孢杆菌Bacillus	6.44±1.74b	78.04±0.98a	9.11±0.87b	72.76±1.02a
清水对照Water control	24.72±1.25a	/	34.32±2.06a	/

处理 Treatment	叶瘟Leaf blast		穗颈瘟Panicle blast
处理 Treatment	病情指数 Disease index	防治效果 Control efficacy (%)	病情指数 Disease index	防治效果 Control efficacy (%)
嗜碱假单胞菌Ej2 P. alcaliphila Ej2	1.33±1.32b	83.67±0.71a	8.77±0.64b	62.79±1.22a
稻瘟酰胺Fenoxanil	1.78±1.19b	81.28±1.81a	9.69±0.51b	58.87±1.88ab
芽孢杆菌Bacillus	1.62±1.21b	82.88±1.03a	8.14±1.74b	65.46±1.58a
清水对照Water control	9.78±1.25a	/	23.56±1.76a	/

病原菌 Target pathogen	抑菌效果 Antagonist activity (%)
茄镰孢F. solani N18-1-2	43.42±1.51
串珠镰孢F. moniliforme N19-2-2	42.11±1.21
西瓜枯萎病菌F. oxysporum f. sp. niveum M8	55.74±2.10
草莓炭疽病菌C. gloeospoioides ZDP21	50.37±1.44
链格孢苹果专化型A. alternata f. sp. mali BJ-A5	54.18±1.72
草莓黑斑病菌A. alternata BJ-ST24	13.00±0.62

处理 Treatment	株高 Plant height (cm)	穗长 Panicle length (cm)	穗重 Panicle weight (g)	千粒重 Thousand kernel weight (g)
嗜碱假单胞菌Ej2 P. alcaliphila Ej2	90.88±2.03a	18.18±1.05a	3.32±1.01a	25.85±0.93ab
芽孢杆菌Bacillus	90.82±0.70a	18.82±1.95a	3.56±0.78a	26.56±0.46a
稻瘟酰胺Fenoxanil	86.72±1.72b	18.12±1.08a	2.77±1.34b	26.01±0.03ab
清水对照Water control	86.20±1.72b	18.10±2.57a	2.86±1.15b	24.63±1.31b