Study on Control Effect of Streptomyces phaeoluteichromatogenes HEBRC45958 Strain on Corynespora Leaf Spot of Tomato

doi:10.13304/j.nykjdb.2023.0416

Abstract

Abstract:

In order to clarify the control effect of Streptomyces phaeoluteichromatogenes strain HEBRC45958 against Corynespora leaf spot of tomato， the antifungal activity was determined in vitro by using dual culture assay combined with microscope and scanning electron microscope observation. The siderophore production and the activities of hydrolases were also determined. The control effect of the fermentation broth of HEBRC45958 strain on Corynespora leaf spot of tomato was determined in pots. The results showed that the inhibition rate HEBRC45958 on Corynespora cassiicola was 52.65% in the dual culture. Compared with control， the mycelium treated with HEBRC45958 strain was distorted and cracked by scanning microscopy， and the space between the cell wall and cell membrane was widened by transmission electron microscopy. The HEBRC45958 strain could produce amylase， cellulase， chitinase and siderophore. The control effect of fermentation broth against Corynespora leaf spot of tomato was 100% in pot experiment， which showed that HEBRC45958 strain had a good prospect of biocontrol application for Corynespora leaf spot of tomato.

Key words: Streptomyces phaeoluteichromatogenes, tomato, Corynespora cassiicola, control effect

摘要：

为明确产褐黄色链霉菌HEBRC45958菌株对番茄棒孢叶斑病的防控效果，采用平板对峙培养结合光学显微镜和扫描电镜观察确定HEBRC45958离体抑菌活性，并测定其产铁载体能力和水解酶活性。同时采用盆栽试验确定HEBRC45958发酵液对番茄棒孢叶斑病防控效果。结果表明，对峙培养条件下 HEBRC45958对多主棒孢的抑菌率为52.65%。扫描电镜下观察，与对照相比，HEBRC45958处理后多主棒孢菌丝发生扭曲和破裂，透射电镜下观察，多主棒孢细胞膜和细胞壁距离增加。HEBRC45958菌株能产生淀粉酶、纤维素酶、几丁质酶和嗜铁素。盆栽试验结果表明，HEBRC45958发酵液对番茄棒孢叶斑病的防效为100%，具有较好的生防应用前景。

关键词: 产褐黄色链霉菌, 番茄, 多主棒孢, 防效

CLC Number:

S476

Daye HUANG, Zhibin YU, Zhongyi WAN, Dan YANG, Jinping LI, Chunxia CAO. Study on Control Effect of Streptomyces phaeoluteichromatogenes HEBRC45958 Strain on Corynespora Leaf Spot of Tomato[J]. Journal of Agricultural Science and Technology, 2024, 26(11): 136-142.

黄大野, 余志斌, 万中义, 杨丹, 李金萍, 曹春霞. 产褐黄色链霉菌HEBRC45958菌株防控番茄棒孢叶斑病研究[J]. 中国农业科技导报, 2024, 26(11): 136-142.

Figures/Tables 7

Table 1 In vitro inhibition on plant pathogens with HEBRC45958 strain

植物病原菌Plant pathogen	抑菌率Inhibition rate/%
瓜果腐霉Pythiumaphanidermatum	30.49±1.20 c
辣椒疫霉Phytophthora capsici	0.00±0.00 d
灰葡萄孢Botrytis cinerea	59.63±4.40 a
多主棒孢Corynespora cassiicola	52.65±2.50 a
尖孢镰刀菌Fusarium oxysporum	0.00±0.00 d
胶孢炭疽菌Colletotrichum gloeosporioides	36.67±3.80 b
核盘菌Sclerotinia sclerotiorum	53.33±2.10 a

Fig. 1 Inhibitory effect of HEBRC45958 strain on C. cassiicolaA：Control；B：HEBRC45958 treatment

Fig. 2 Scanning electron microscopy of the effect of HEBRC45958 strain on mycelia of C. cassiicolaA：Control；B：HEBRC45958 treatment

Fig. 3 Transmission electron microscopy observations of C. cassiicola hyphae treated with HEBRC45958 strainA：Control；B：HEBRC45958 treatment

Fig. 4 Hydrolase activities and the ability to produce siderophoreA： Amylase medium； B： Chitin medium； C： CAS medium； D： Carboxyl methyl cellulose medium

Table 2 Control effect of the fermentation broth of HEBRC45958 strain on Corynespora leaf spot of tomato

处理Treatment	病情指数Disease index	防效Control effect/%
发酵原液Fermentation broth	0.00±0.00 c	100.00±0.00 a
25倍发酵液Fermentation broth 25×	6.90±5.00 b	92.88±5.16 b
50倍发酵液Fermentation broth 50×	11.11±0.35 b	88.53±0.36 b
啶酰菌胺Bascalid	12.34±3.51 b	87.26±3.63 b
对照Control	96.88±5.40 a	—

Fig. 5 Control effect of HEBRC45958 strain on Corynespora leaf spot of tomato

References 24

1	李宝聚,高苇,石延霞,等.多主棒孢和棒孢叶斑病的研究进展[J].植物保护学报,2012,39(2):171-176.
	LI B J, GAO W, SHI Y X, et al.. Progress in researches on Corynespora leaf spot [J]. Acta Phytophylacica Sin., 2012, 39(2):171-176.
2	田守波,朱为民,朱龙英,等.番茄棒孢叶斑病病原鉴定及生物学特性研究[J].植物保护,2020, 46(3):157-162.
	TIAN S B, ZHU W M, ZHU L Y, et al.. Identification of the pathogen from Corynespora leaf spot of tomato and research on its biological characteristics [J]. Plant Prot., 2020, 46(3):157-162.
3	李明远.李明远断病手迹(六十一)北京发现番茄棒孢叶斑病[J].农业工程技术,2015(10):84-85.
4	DUAN Y B, XIN W J, LU F, et al.. Benzimidazole- and QoI-resistance in Corynespora cassiicola populations from greenhouse-cultivated cucumber: an emerging problem in China [J]. Pesticide Biochem. Physiol., 2019, 153: 95-105.
5	ZHU F D, SHI Y X, XIE X W, et al.. Occurrence distribution and characteristics of boscalid-resistant Corynespora cassiicola in China [J]. Plant Dis., 2019, 103(1) :69-76.
6	SHI Y, ZHU F, SUN B, et al.. Two adjacent mutations in the conserved domain of SdhB confer various resistance phenotypes to fluopyram in Corynespora cassiicola [J]. Pest Manage. Sci., 2021, 77(9):3980-3989.
7	SHI Y, SUN B, XIE X, et al.. Site-directed mutagenesis of the succinate dehydrogenase subunits B and D from Corynespora cassiicola reveals different fitness costs and sensitivities to succinate dehydrogenase inhibitors [J]. Environ. Microbiol., 2021, 23(10):5769-5783.
8	SUN B X, ZHU G X, XIE X W, et al.. Double mutations on succinate dehydrogenase are involved in SDHI resistance in Corynespora cassiicola [J/OL]. Microorganisms, 2022, 10(1):132 [2023-04-30]. .
9	赵昱榕,李磊,谢学文,等.贝莱斯芽胞杆菌ZF2对多主棒孢病菌防治效果[J].中国生物防治学报,2019,35(2):217-225.
	ZHAO Y R, LI L, XIE X W, et al.. Biocontrol effect of Bacillus velezensis strain ZF2 against Corynespora cassiicola [J]. J. Chin. Biol. Control., 2019, 35(2):217-225.
10	李新宇,李磊,石延霞,等.黄瓜棒孢叶斑病拮抗细菌的筛选,鉴定及防治效果[J].植物保护学报,2020,47(3):620-627.
	LI X Y, LI L, SHI Y X, et al.. Screening, identification and control effects of antagonistic bacteria against cucumber Corynespora leaf spot [J]. J. Plant Prot., 2020, 47(3):620-627.
11	黄大野,贲海燕,曹春霞,等.死亡谷芽胞杆菌NBIF-001防治番茄棒孢叶斑病研究[J].植物保护,2017,43(6):192-195.
	HUANG D Y, BEN H Y, CAO C X, et al.. Control effect of Bacillus vallismortis NBIF001 on Corynespora leaf spot on tomato [J]. Plant Prot., 2017, 43(6):192-195.
12	黄大野,曹春霞,张亚妮,等.枯草芽孢杆菌NBF809防治番茄棒孢叶斑病研究[J].中国蔬菜, 2018(12):40-44.
	HUANG D Y, CAO C X, ZHANG Y N, et al.. Studies on controlling tomato Corynespora leaf spot by Bacillus subtilis NBF809 [J]. China Veget., 2018(12):40-44.
13	LI Q, NING P, ZHENG L, et al.. Effects of volatile substances of Streptomyces globisporus JK-1 on control of Botrytis cinerea on tomato fruit [J]. Biol. Cont., 2012, 61(2):113-120.
14	王超,申成美,郑丽,等.烟草青枯病生防细菌的筛选与生防效果研究[J].植物保护,2014,40(2):43-47.
15	武志江,王亚军,杨柳,等.百合枯萎病拮抗细菌的筛选,鉴定及其抑菌物质研究[J].微生物学通报,2015,42(7):1307-1320.
	WU Z J, WANG Y J, YANG L, et al.. Isolation and identification of an antagonistic bacterium against Fusarium on lily and its antifungal substances [J]. Microbiol. China, 2015, 42(7):1307-1320.
16	MUDYTWA R M, CHAIBVA P, TAKAWIRA M, et al.. Evaluation of Trichoderma harzianum in controlling damping-off (Pythium spp.) on tomato (Solanum lycopersicum) seedling varieties [J]. Ann. Biolo. Res., 2016, 7(6):6-11.
17	MIYAMOTO T, ISHII H, SEKOT, et al.. Occurrence of Corynespora cassiicola isolates resistant to boscalid on cucumber in Ibaraki Prefecture, Japan [J]. Plant Pathol., 2009, 58(6):1144-1151.
18	ISHII H, YANO K, DATE H, et al.. Molecular characterization and diagnosis of QoI resistance in cucumber and eggplant fungal pathogens [J]. Phytopathology, 2007, 97(11):1458-1466.
19	金丽颖.抗黄瓜棒孢叶斑病放线菌的筛选及Streptomyces inhibens多相分类研究[D].哈尔滨:东北农业大学,2019.
	JIN L Y. Screening of actinomycetes against cucumber target leaf spot disease and polyphasic taxonomy study of Streptomyces inhibens [D]. Harbin: Northeast Agricultural University, 2019.
20	XIONG Z Q, TU X R, WEI S J, et al.. The mechanism of antifungal action of a new polyene macrolide antibiotic antifungalmycin 702 from Streptomyces padanus JAU4234 on the rice sheath blight pathogen Rhizoctonia solani [J/OL]. PLoS One, 2013, 8(8):e73884 [2023-04-30]. https:doi.‍org/10.1371/journal.pone.0073884.
21	董章勇,王振中.植物病原真菌细胞壁降解酶的研究进展[J].湖北农业科学, 2012,51(21): 4697-4700.
	DONG Z Y, WANG Z Z. Research progress of fungal cell wall-degrading enzyme [J]. Hubei Agric. Sci., 2012, 51(21):4697-4700.
22	臧超群,白元俊,张海东,等.暗黑链霉菌PY-1活性产物分析及其对葡萄霜霉病田间防效评价[J].植物保护学报, 2018, 45(4): 864-870.
	ZANG C Q, BAI Y J, ZHANG H D, et al.. Study on bioactive metabolite of Streptomyces atratus PY-1 and the field control efficiency against grapevine downy mildew [J]. J. Plant Prot., 2018, 45(4): 864-870.
23	荣良燕,姚拓,赵桂琴,等.产铁载体PGPR菌筛选及其对病原菌的拮抗作用[J].植物保护, 2011, 37(1):59-64.
	RONG L Y, YAO T, ZHAO G Q, et al.. Screening of siderophore-producing PGPR bacteria and their antagonism against the pathogens [J]. Plant Prot., 2011, 37(1):59-64.
24	MA D C, ZHU J M, JIANG J G, et al.. Evaluation of bioactivity and control efficacy of tetramycin against Corynespora cassiicola [J]. Pest. Biochem. Physiol., 2018, 152(12):106-113.

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