杨树溃疡病拮抗链霉菌的筛选及鉴定

doi:10.11707/j.1001-7488.20210710

Abstract

Abstract:

Objective: In order to control the poplar canker disease by biological control agent,the present study aimed to screen and obtain antagonistic strains from Streptomyces that can effectively inhibit poplar canker. Method: A streptomyces strain with good biocontrol effect against the pathogen of poplar canker was obtained from the soil in the larch forest Hengshan Protection Station,Changbaishan Mountain National Nature Reserve,Jilin Province. The number was recorded as HS1. Multiple screening was used to determine the antibacterial activity of the strain and its fermentation broth; morphological,physiological and biochemical characteristics,16S rDNA sequence analysis were used to determine its taxonomic status; artificial inoculation method was used to determine the preventive and therapeutic effects of HS1 fermentation broth on poplar canker. Result: The result showed that strain HS1 and its fermentation broth had inhibitory effect on the tested plant pathogens,and the antibacterial spectrum was relatively wide. Among them,it had the strongest inhibitory effect on Botryosphaeria dothidea. The width of inhibition zone averagely was more than 20.66 mm and the diameter of the bacteriostatic circle of the fermentation broth reached 40.26 mm.The 16S rDNA sequence alignment analysis showed that the strain had the highest homology with Streptomyces scopuliridis; The result of artificial inoculation test showed that the preventive and therapeutic effects of strain HS1 fermentation broth on poplar canker were 60.87% and 71.74% respectirly. Conclusion: The screened strain HS1 and its fermentation broth could effectively prevent and inhibit the occurrence and expansion of poplar canker disease,and also have a good antibacterial effect on other pathogens of many important agricultural and forest diseases. Therefore,it has a good for development and application prospects in biocontrol of forest pathogens.

Key words: poplar canker, Streptomyces, biological control, screening and identification

CLC Number:

S432.1

Yuequ Chen,Qingzhen Liu,Limei Li,Yang Zhang,Jiao Han,Yong'an Zhang. Screening and Identification of Antagonistic Streptomyces for Biocontrol of Poplar Canker[J]. Scientia Silvae Sinicae, 2021, 57(7): 92-100.

Figures/Tables 12

Table 1

Fig.1

Table 2

Table 3

Fig.2

Fig.3

Table 4

Table 5

Table 6

Fig.4

Fig.5

Fig.6

References 0

	陈欣怡. 2016. 苹果轮纹病生防解淀粉芽胞杆菌(Bacillus amyloliquefaciens)PG12主要生防机制研究. 北京: 中国农业大学博士学位论文.
	Chen X Y. 2016. Study on the main biocontrol mechanism of Bacillus amyloliquefaciens PG12 against apple ring rot disease. Beijing: PhD thesis of China Agricultural University. [in Chinese]
	高芬, 吴元华, 王梦亮. 可可链霉菌182-2活性代谢产物对烟草赤星病菌的作用. 中国农业科学, 2013, 46 (23): 4933- 4940. doi: 10.3864/j.issn.0578-1752.2013.23.009
	Gao F , Wu Y H , Wang M L . Action of the metabolite produced by Streptomyces cacaoi strain 182-2 on Alternaria alternate. Scientia Agricultura Sinica, 2013, 46 (23): 4933- 4940. doi: 10.3864/j.issn.0578-1752.2013.23.009
	贺建武, 陈奇辉, 刘祝祥, 等. 波赛链霉菌合成蒽环类抗肿瘤抗生素研究进展. 中国抗生素杂志, 2011, 36 (3): 161- 164.
	He J W , Chen Q H , Liu Z X , et al. Advances on biosynthesis of anthracycline antibiotics by Streptomyces peucetius. Chinese Journal of Antibiotics, 2011, 36 (3): 161- 164.
	冀瑞卿. 2007. 鳞柄白鹅膏抑菌成分及其对杨树烂皮病菌抑制机理. 哈尔滨: 东北林业大学博士学位论文.
	Ji R Q. 2007. Research on lnhibiting substance of Amanita virosa and its inhibiting mechanism on Cytospora chrysosperma. Harbin: PhD thesis of Northeast Forestry University. [in Chinese]
	康海婷, 凌丹燕, 吴志娟, 等. 蓝莓枝干溃疡病病原鉴定及品种抗病性研究. 浙江农业学报, 2019, 31 (3): 436- 443. doi: 10.3969/j.issn.1004-1524.2019.03.13
	Kang H T , Ling D Y , Wu Z J , et al. Pathogen isolation and identification of blueberry stem canker and screening of disease resistant cultivars. Acta Agriculturae Zhejiangensis, 2019, 31 (3): 436- 443. doi: 10.3969/j.issn.1004-1524.2019.03.13
	李永丽, 王亚红, 周洲, 等. 新疆野苹果内生娄彻氏链霉菌A-m1的鉴定和发酵条件优化及抑菌广谱作用. 林业科学, 2020, 56 (7): 70- 81.
	Li Y L , Wang Y H , Zhou Z , et al. Identification and optimization of fermentation conditions and antibacterial activity of endophytic Streptomyces rochei A-ml from the branch of Malus sieversii. Scientia Silvae Sinicae, 2020, 56 (7): 70- 81.
	刘若寒. 2015. 链霉菌Streptomyces scopuliridis和S. glaucus次生代谢产物的研究. 沈阳: 东北大学硕士学位论文.
	Liu R H. 2015. Study on the secondary metabolites of Streptomyces scopuliridis and S. glaucus. Shenyang: MS thesis of Northeastern University. [in Chinese]
	阮继生, 黄英. 放线菌快速鉴定与系统分类. 北京: 科学出版社, 2011.
	Ruan J S , Huang Y . Rapid identification and systematics of Actinobacteria. Beijing: Science Press, 2011.
	孙鑫垚. 2010. 陕西省苹果轮纹病病原菌鉴定与多样性研究. 杨凌: 西北农林科技大学硕士学位论文.
	Sun X Y. 2010. Pathogen identification and diversity of apple white rot in shaanxi province. Yangling: MS thesis of Northwest A&F University. [in Chinese]
	王琼伟. 2019. 山核桃干腐病菌的基础生物学及其侵染、传播的定量监测技术. 杭州: 浙江农林大学硕士学位论文.
	Wang Q W. 2019. Basic biology of Botryosphaeria dothidea causing Chinese hickory(Carya cathayensis) trunk canker and quantitative monitoring its infection and transmission. Hangzhou: MS thesis of Zhejiang A&F University. [in Chinese]
	王世强. 2014. 链霉菌JD211对水稻的防病促生效应及机制. 南昌: 江西农业大学硕士学位论文.
	Wang S Q. 2014. Research on the effects of Streptomyces D211 on rice growth promotion and disease prevention mechanism. Nanchang: MS thesis of Jiangxi Agricultural University. [in Chinese]
	魏晓丽. 2013. 拮抗与内生链霉菌的定殖及促生作用研究. 杨凌: 西北农林科技大学硕士学位论文.
	Wei X L. 2013. Study on colonization capability and growth promotion of antagonistic and endophytic Streptomyces. Yangling: MS thesis of Northwest A&F University. [in Chinese]
	胥丽娜, 徐亮, 李磊, 等. 枯草芽孢杆菌对杨树水泡溃疡病菌菌丝生长的抑制作用. 菌物学报, 2007, 5 (3): 165- 168.
	Xu L N , Xu L , Li L , et al. Inhibition effect of Bacillus subtilis on the growth of Botryosphaeria dothidea. Mycosystema, 2007, 5 (3): 165- 168.
	于冰, 申屠旭萍, 俞晓平. 丰加霉素对黄瓜立枯丝核菌的拮抗作用. 中国生物防治学报, 2011, 27 (3): 373- 377.
	Yu B , Shen T X P , Yu X P . Antifungal activity of toyocamycin on Rhizoctonia solani. Chinese Journal of Biological Control, 2011, 27 (3): 373- 377.
	杨蕾, 周国英, 梁军. 2种生防菌株对杨树溃疡病原葡萄座腔菌的抑制作用. 林业科学, 2015, 51 (8): 68- 73.
	Yang L , Zhou G Y , Liang J . Inhibitory effects of two biocontrol fungous strains on poplar canker Botryosphaeria dothdea. Scientia Silvae Sinicae, 2015, 51 (8): 68- 73.
	杨瑞, 王岐, 张露, 等. 放线菌扫描电镜样品制备方法比较研究. 电子显微学报, 2014, 33 (1): 84- 89.
	Yang R , Wang Q , Zhang L , et al. Comparison of preparation method for actinomyces scanning electron microscopy sample. Journal of Chinese Electron Microscopy Society, 2014, 33 (1): 84- 89.
	张倩, 李阳, 杨埔, 等. 甜樱桃葡萄座腔菌的生物学特性研究. 食品工业科技, 2020, 41 (10): 101- 106.
	Zhang Q , Li Y , Yang P , et al. Biological characteristics of Botryosphaeria dothoidea from sweet cherry. Science and Technology of Food Industry, 2020, 41 (10): 101- 106.
	中国科学院微生物研究所放线菌分类组. 1975. 链霉菌鉴定手册. 北京: 科学出版社.
	Actinomycetes Classification Group, Institute of Microbiology, Chinese Academy of Sciences. 1975. Streptomyces identification manual. Beijing: Science Press. [in Chinese]
	周德庆. 微生物学教程. 3版北京: 高等教育出版社, 2011.
	Zhou D Q . Essential microbiology. 3rd ed Bejing: Higher Education Press, 2011.
	Ara I , Bukhari N A , Aref N M , et al. Antiviral activities of streptomycetes against tobacco mosaic virus(TMV) in Datura plant: evaluation of different organic compounds in their metabolites. African journal of Biotechnology, 2012, 11 (8): 2130- 2138.
	Baharlouei A , Sharifi-Sirchi G R , Shahidi Bonjar G H . Identification of an antifungal chitinase from a potential biocontrol agent, Streptomyces plicatus strain 101, and its new antagonistic spectrum of activity. The Philippine agricultural scientist, 2010, 93 (4): 439- 445.
	Farris M H , Duffy C , Findlay R H , et al. Streptomyces scopuliridis sp. nov., a bacteriocin-producing soil streptomycete. International Journal of Systematic and Evolutionary Microbiology, 2011, 61 (9): 2112- 2116.
	Hoyt S . Integrated chemical control of insects and biological control of mites on apple in Washington. Journal of Economic Entomology, 1969, 62 (1): 74- 86.
	Li L Y , Liu R H , Han L , et al. Structure determination of two new nerolidol-type sesquiterpenoids from the soil actinomycete Streptomyces scopuliridis. Magnetic Resonance in Chemistry, 2016, 54 (7): 606- 609.
	McEvoy P B . Theoretical contributions to biological control success. BioControl, 2018, 63 (1): 87- 103. doi: 10.1007/s10526-017-9852-6
	Tan L T H , Ser H L , Yin W F , et al. Investigation of antioxidative and anticancer potentials of Streptomyces sp. mum256 isolated from malaysia mangrove soil. Frontiers in Microbiology, 2015, 6 (11): 1- 12.
	Zhou H X , Yu Y , Tan X M , et al. Biological control of insect pests in apple orchards in China. Biological Control, 2014, 68, 47- 56.

菌株 Strains	抑菌带宽 Inhibition zone/mm	菌株Strains	抑菌带宽 Inhibition zone/mm	菌株Strains	抑菌带宽 Inhibition zone/mm
HS1	20.66±0.647 15	HS6	17.70±1.453 41	A-1-14	12.08±0.962 70
HS2	19.92±2.305 91	CX1	13.18±0.870 17	F12	14.92±0.472 86
HS3	19.96±1.833 79	CX2	13.94±1.107 61	F15	10.48±0.468 61
HS4	17.30±1.487 14	CX3	14.60±0.603 99
HS5	15.06±0.554 62	CX4	14.66±1.177 96

病原菌 Pathogens	抑菌带宽 Inhibition zone/mm	抑菌直径 Diameter of inhibition zone/mm
蓝莓枯枝病菌小新壳梭孢Neofusicoccum parvum	12.28±1.36f	20.69±1.15f
杨树烂皮病菌金黄壳囊孢菌Cytospora chrysosperma	20.30±2.36bc	40.02±0.59a
烟草赤星病菌链格孢Alternaria alternata	17.96±0.78d	32.80±0.78c
玉米大斑病菌Exserohilum turcicum	18.46±0.76cd	39.77±0.79a
甜瓜枯萎病菌尖孢镰刀菌Fusarium oxysporium	15.40±0.93e	21.28±1.17f
大豆炭疽病菌Colletotrichum truncatum	21.20±1.10b	37.61±1.55b
蓝莓溃疡病菌葡萄座腔菌Botryosphaeria dothidea	21.72±1.86b	39.84±0.67a
瓜果腐霉病菌Pythium aphanidermatum	14.94±1.07e	17.12±0.87g
水稻恶苗病菌串株镰刀菌Fusarium moniliforme	14.32±0.38e	29.15±0.59d
云杉立枯病菌尖孢镰刀菌Fusarium oxysporium	20.84±1.87b	29.90±1.05d
辣椒炭疽病菌Colletotrichum capsici	15.64±0.71e	33.70±0.44c
茄子褐纹病菌Phomopsis vexans	24.80±0.89a	36.28±0.63b
杨树溃疡病菌葡萄座腔菌Botryosphaeria dothidea	20.66±0.65b	40.26±0.51a

处理 Treatment	预防作用Prevention			治疗作用Curation
处理 Treatment	发病率 Rate of disease(%)	病情指数 Disease indexes	预防效果 Effect of Prevention (%)	发病率 Rate of disease(%)	病情指数 Disease indexes	预防效果 Effect of Prevention (%)
HS1	50.00±8.33bc	24.99±3.61b	60.87±5.66a	41.65±8.35c	18.05±2.40c	71.74±3.76a
多菌灵Carbendazim	61.31±9.64b	31.94±2.40b	50.00±3.76b	58.34±8.35bc	33.33±2.08b	47.83±3.26b
对照组CK	94.44±9.62a	63.89±3.19a	—	94.44±9.62a	63.89±3.19a	—

培养基 Medium	气生菌丝体 Aerial mycelium	基质菌丝体 Substrate mycelium	可溶性色素 Soluble pigment	丰茂度 Abundance
高氏一号琼脂 Gause No.1 agar	蚌肉白 Clam white	酪黄 Buttery yellow	-	+++
葡萄糖天门冬素 Glucose asparagine agar	荷花白 Lotus white	蚌肉白 Clam white	-	+++
蔗糖察氏 Czaper’s agar	粉白 Powder white	Ⅰ-3-1′	-	+
克氏合成一号琼脂 Carrot No.1 agar	蚌肉白 Clam white	豆汁黄 Soy sauce yellow	-	+++
马铃薯葡萄糖琼脂 Potato glucose agar	淡红灰 Light red gray	风帆黄 Sail yellow	鲛青 Cyan	+++
酵母膏麦芽膏琼脂 Yeast extract malt extract agar	Ⅳ-1-2′	豆汁黄 Soy sauce yellow	-	+++
淀粉铵琼脂 starch ammonium salt medium	Ⅰ-2-2′	莲子白 Lotus seed white	-	+++
贝奈特琼脂 Bennett’s agar	鹿角棕 Antler brown	鹅掌黄 Liriodendron yellow	-	++
葡萄糖酵母膏琼脂 Glucose yeast extract agar	荷花白 Lotus white	软木黄 Cork yellow	-	+++
甘油天冬素培养基 Glycerol asparagine agar	鲛青 Cyan	珠母灰 Pearlescent gray	Ⅶ-7-1′	++
甘油精氨酸培养基 Glycerine arginine medium	荷花白 Lotus white	粉白 Powder white	-	+++
黄豆粉蛋白胨培养基 Soybean powder peptone medium	Ⅳ-2-1′	粉白 Powder white	-	+++
燕麦粉琼脂 Oatemeal agar	Ⅲ-3-3′	香水玫瑰黄 Perfume rose yellow	-	+++
淀粉琼脂 Soluble starch agar	乳白 Milky	淡蜜黄 Light honey yellow	鹅掌黄 Liriodendron yellow	++
黄豆粉琼脂培养基 Bean agar medium	Ⅳ-3-2′	蚌肉白 Clam white	-	+++

pH	丰茂度 Abundance	NaCl(%)	丰茂度 Abundance	温度 Temperature/℃	丰茂度 Abundance
2	-	0	+	4	+
5	++	10	++	15	++
7	+++	20	++	28	+++
9	++	30	+++	37	-
12	+	40	++	50	-

Screening and Identification of Antagonistic Streptomyces for Biocontrol of Poplar Canker

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 12

References 0

Related Articles 15

Recommended Articles

Metrics

Comments

碳源利用 Utilization of carbon sources	丰茂度 Abundance	碳源利用 Utilization of carbon sources	丰茂度 Abundance
D-木糖D-xylose	+++	麦芽糖Amylomaltose	++
D-果糖D-Fructose	+++	棉子糖Raffinose	-
甘露醇D-mannitol	-	鸟嘌呤Guanine	+
L-鼠李糖L-Rhamnose	-	甘氨酸Glycine	+
α-乳糖α-lactose	+	L-酪氨酸L-tyrosine	++
葡萄糖Glucose	+++	L-阿拉伯糖L-Arabinose	+

[1]	Ran He,Ruizhen Wang,Yue Ying,Liangjian Qu,Yong'an Zhang. Phenotypic and Virulence Gene Expression Difference of Esteya vermicola, A Biocontrol Fungus for Pine Wood Nematode under Carbon and Nitrogen Conditions [J]. Scientia Silvae Sinicae, 2021, 57(4): 107-115.
[2]	Ran He,Xia Cui,Yue Ying,Liangjian Qu,Ruizhen Wang,Yong Zhang. Screening and Identification of Beauveria bassiana Strains for Biocontrol of Monochamus alternatus Adults (Coleoptera: Cerambycidae) [J]. Scientia Silvae Sinicae, 2020, 56(10): 129-134.
[3]	Yuting Mao,Tao Ma,Laijiao Lan,Xiujun Wen. Advances in Esteya vermicola, A Potential Biocontrol Fungus for Pine Wood Nematode [J]. Scientia Silvae Sinicae, 2020, 56(1): 180-190.
[4]	Wang Hao, Shi Juan, Luo Youqing, Sheng Maoling, Zhou Feng, Zheng Zijin. Survey of Natural Enemies of Sirex noctilio(Hymenoptera: Siricidae)and Biology of the Dominant Parasitoids in China [J]. Scientia Silvae Sinicae, 2018, 54(3): 91-96.
[5]	Huang Cuiqin. Control Potential of the Predator Scymnus (Pullus) posticalis (Coleoptera: Coccinellidae) against Pseudoregma bambusicola (Homoptera: Hormaphididae) [J]. Scientia Silvae Sinicae, 2016, 52(11): 157-164.
[6]	Li Huiping, Huang Qiuxian, Wang Jing, Li Wen. Potential of Entomopathogen Beauveria bassiana for Controlling Fall Webworm Hyphantria cunea [J]. Scientia Silvae Sinicae, 2015, 51(9): 65-70.
[7]	Yang Lei, Zhou Guoying, Liang Jun. Inhibitory Effects of Two Biocontrol Fungous Strains on Poplar Canker Botryosphaeria dothdea [J]. Scientia Silvae Sinicae, 2015, 51(8): 67-73.
[8]	Yang Lei, Liang Jun, Zhou Guoying, Ni Yang, Lü Quan, Zhang Xingyao. Isolation and Identification of Bio-control Microorganisms in Soil to Poplar Canker [J]. Scientia Silvae Sinicae, 2015, 51(4): 116-125.
[9]	Gao Fen, Li Jinghong, Wang Junhong, Wang Mengliang. Screening in vitro of Antagonistic Streptomyces against the Black Rot Disease (Alternaria tenuissima) of Zanhuang Jujube Fruit and Optimization of Fermentation Conditions [J]. Scientia Silvae Sinicae, 2015, 51(12): 132-140.
[10]	Wang Zhiyong, Yang Zhongqi, Zhang Yanlong, Wang Xiaoyi, Tang Yanlong, Sun Qingjiang, Xu Zheng. Biological Control of Agrilus mali (Coleoptera：Buprestidae) by Applying Four Species of Bethylid Wasp(Hymenoptera：Bethylidae) on Malus sieversii in Xinjiang [J]. Scientia Silvae Sinicae, 2014, 50(8): 97-101.
[11]	Wang Xiaohong, Yang Zhongqi, Wang Xiaoyi, Tang Yanlong, Zhang Yanlong. Biological Control of Apriona swainsoni (Coleoptera：Cerambycidae) by Applying Three Parasitoid Species [J]. Scientia Silvae Sinicae, 2014, 50(1): 103-108.
[12]	Feng Xiaohui;Zhang Xingyao;Yan Donghui. Detection to Pathogens of Poplar Cankers by the Multiplex PCR Technique [J]. Scientia Silvae Sinicae, 2012, 48(5): 72-77.
[13]	Zhang Yanlong;Yang Zhongqi;Wang Xiaoyi;Wu Chengjin;Ma Shengfu;Lu Zhigui. Laboratory Test on Control of Japanese Pine Sawyer by Two Entomopathogenic Nematode Species [J]. Scientia Silvae Sinicae, 2012, 48(12): 64-70.
[14]	Zhang Yabo;Wu Panpan;Wang Peng;Liu Jian;Xu Tiansen;Wang Haojie;Shu Jinping. Identification and Biological Characteristics of A Metarhizium pingshaense Strain Isolated from Melanotus cribricollis Larva [J]. Scientia Silvae Sinicae, 2012, 48(12): 134-140.
[15]	Yang Zhongqi;Li Jianqing;Mei Zengxia;Wang Xiaoyi;Jing Zhigao;Meng Xiangdong. Biocontrol of Batocera horsfieldi (Coleoptera: Cerambycidae)Attacking Ash Trees by Releasing the Parasitoid Dastarcus helophoroides (Coleoptera: Bothrideridae) [J]. Scientia Silvae Sinicae, 2011, 47(12): 78-84.