贝莱斯芽孢杆菌E69预防稻瘟病等多种真菌病害的潜力

doi:10.3864/j.issn.0578-1752.2019.11.006

Abstract

Abstract:

【Objective】 The objective of this study is to clarify the antagonistic effect of Bacillus velezensis strain E69 isolated from rice endophytic bacteria against various fungal plant pathogens in vitro, especially the control efficacy of B. velezensis E69 on rice blast, and to reduce the use of chemical pesticides.【Method】 B. velezensis E69 and B. subtilis (most commonly used microorganisms in biological control of rice blast) E66 strains were isolated from endophytic bacteria of rice leaf tissue. The antagonistic effect of strains E69 and E66 and their fermented liquid, sterile supernatant against Magnaporthe oryzae was tested by confrontation culture method. Also, the antagonistic effect of strains E69 and E66 against other 11 pathogenic fungi, such as Rhizoctonia solani, Fusarium spp., Botrytis cinerea, Colletotrichum gloeospoioides, Phytophthora parasitica var. nicotianae, Alternaria alternate, F. oxysporum, et al., was determined. The preventive efficacy of E69 and E66 was tested for the control of rice leaf blast under greenhouse condition. The field experiments were conducted to evaluate the preventive efficacy of E69 and E66 against rice leaf blast and neck blast. The inhibitory effect of E69 and E66 against conidial germination and appressorial formation of M. oryzae was assessed in laboratory. The colonization of GFP-marked strain E69 in rice stem was observed by laser scanning confocal microscope.【Result】 Strains E69 and E66 significantly suppressed the mycelia growth rate of M. oryzae P131, the preventive efficacy against rice blast under greenhouse condition was 83.24% and 76.57%, respectively. The preventive efficacy of E69, E66 against rice leaf blast and neck blast in field was 85.97%, 79.76% and 69.67%, 68.82%, respectively. The preventive effect of E69 on leaf blast was significantly higher than that of 75% tricyclozole wettable powder, but there was no significant difference between the preventive efficacy of strain E69 and 75% tricyclazole powder against rice neck blast. E69 and E66 had significant antagonistic effects against 11 plant pathogens, such as R. solani, Fusarium spp., B. cinerea, C. gloeospoioides, P. parasitica var. nicotianae, A. alternate, F. oxysporum, et al. The antagonistic effect of E69 was higher than that of E66. E69 and E66 could strongly inhibit the conidial germination and appressorial formation of M. oryzae P131, the inhibitory effect of E69 fermented liquid was 95.28% and 94.16%, respectively, and the inhibitory effect of E69 sterile supernatant was 85.36% and 84.31%, respectively. The inhibitory effect of E66 fermented liquid was 89.15% and 87.38%, respectively, and the inhibitory effect of E66 sterile supernatant were 79.65% and 72.45%, respectively. The GFP-marked strain E69 showed good colonization ability in rice stem, and could be stably colonized in rice stem epidermis, parenchyma and vascular bundles. 【Conclusion】 B. velezensis strain E69 is a potential biocontrol strain with obvious preventive effects, which has the application potential of preventing rice blast, sheath blight and other fungal diseases.

Key words: Bacillus velezensis, rice blast, plant pathogenic fungi, biological control, colonization ability

SHA YueXia,SUI ShuTing,ZENG QingChao,SHEN RuiQing. Biocontrol Potential of Bacillus velezensis Strain E69 Against Rice Blast and Other Fungal Diseases[J].Scientia Agricultura Sinica, 2019, 52(11): 1908-1917.

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References 43

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处理Treatment	病情指数Disease index	预防效果Preventive efficacy (%)
E69	3.71±1.11b	83.24±1.41a
E66	5.18±1.05b	76.57±1.15b
75%三环唑Tricyclazole	4.88±1.14b	77.91±1.43b
绿地康Lvdikang	3.42±0.52b	84.51±0.69a
清水对照Water control	22.09±2.43a	—

处理 Treatment	叶瘟Rice leaf blast		穗颈瘟Rice neck blast
处理 Treatment	病情指数 Disease index	预防效果 Preventive efficacy (%)	病情指数 Disease index	预防效果 Preventive efficacy (%)
E69	4.66±0.53b	85.97±0.96a	16.56±2.62b	69.67±2.41a
E66	8.79±2.53b	79.76±2.96b	16.48±4.66b	68.82±4.41a
绿地康Lvdikang	4.04±1.29b	88.74±2.65a	16.03±1.77b	69.67±1.54a
75%三环唑Tricyclazole	10.06±2.19b	77.01±2.01b	15.76±1.15b	70.18±1.08a
清水对照Water control	36.15±2.25a	—	52.86±3.06a	—

病原菌 Target fungal pathogen	抑菌效果 Antagonistic effect
病原菌 Target fungal pathogen	E66	E69
Fusarium oxysporum N16-2-1	++	++++
Fusarium solani N18-1-2	+	++++
Fusarium moniliforme N19-2-2	+	++++
Rhizoctonia solani RS8	+++	+++
Botrytis cinerea ZDP4	+	+++
Colletotrichum gloeospoioides ZDP21	+	++
Alternaria alternate BJ-A5	++	+++
Alternaria alternate BJ-ST24	+	+
Alternaria alternate BJ-H9	++	+++
Fusarium oxysporum f. sp. niveum M8	++	++
Phytophthora parasitica var. nicotianae T15	+	++

处理 Treatment	分生孢子Conidium		附着胞Appressorium
处理 Treatment	萌发率 Germination rate (%)	抑制率 Inhibition rate (%)	形成率 Formation rate (%)	抑制率 Inhibition rate (%)
E69发酵液E69 fermented liquid	4.67±0.82d	95.28±0.83a	5.02±1.14d	94.16±0.15a
E69无菌上清液E69 sterile supernatant	14.49±0.19c	85.36±0.19bc	13.49±0.48bc	84.31±10.55b
E66发酵液E66 fermented liquid	10.74±1.73c	89.15±1.23ab	10.85±0.36c	87.38±0.41b
E66无菌上清液E66 sterile supernatant	20.15±3.44b	79.65±3.47c	23.69±1.59b	72.45±1.86c
清水对照Water control	99.00±0.76a	—	86.00±2.67a	—

Biocontrol Potential of Bacillus velezensis Strain E69 Against Rice Blast and Other Fungal Diseases

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