娄彻氏链霉菌HM85的鉴定及其对甜菜的防病促生作用

doi:10.13304/j.nykjdb.2024.0144

中国农业科技导报 ›› 2024, Vol. 26 ›› Issue (5): 148-155.DOI: 10.13304/j.nykjdb.2024.0144

• 动植物健康 • 上一篇

娄彻氏链霉菌HM85的鉴定及其对甜菜的防病促生作用

杨智敏¹(), 张慧豪²(), 张园园², 杜红岩², 刘晓东², 侯亚光¹, 王毅¹, 徐道龙¹, 黄金贵², 程晓宁², 随洋¹, 王瑞利¹, 于超¹, 赵玲玲¹, 陈春梅¹, 雅茹¹, 贾丽¹, 张明月², 王宏伟¹, 姚淞耀³, 赵莹²(), 邵科¹()

^1.内蒙古科学技术研究院生物技术研究所，呼和浩特 010000
^2.河南农业大学植物保护学院，郑州 450002
^3.内蒙古自治区乡村振兴研究中心，呼和浩特 010000

收稿日期:2024-02-28 接受日期:2024-03-28 出版日期:2024-05-15 发布日期:2024-05-14
通讯作者: 赵莹,邵科
作者简介:杨智敏 E-mail：yangzhimin0110@126.com
张慧豪 E-mail：zhanghuihao0420@163.com第一联系人：杨智敏和张慧豪为共同第一作者。
基金资助:
内蒙古自治区科技计划项目(2020GG0271);国家糖料产业技术体系建设项目(CARS-170725);内蒙古自治区 “揭榜挂帅”项目(2022JBGS0029);内蒙古自治区科技成果转化专项资金项目(2021CG0042)

Identification of Streptomyces rochei HM85 and Its Biocontrol and Growth-promoting Effects on Sugar Beet

Zhimin YANG¹(), Huihao ZHANG²(), Yuanyuan ZHANG², Hongyan DU², Xiaodong LIU², Yaguang HOU¹, Yi WANG¹, Daolong XU¹, Jingui HUANG², Xiaoning CHENG², Yang SUI¹, Ruili WANG¹, Chao YU¹, Lingling ZHAO¹, Chunmei CHEN¹, Ru YA¹, Li JIA¹, Mingyue ZHANG², Hongwei WANG¹, Songyao YAO³, Ying ZHAO²(), Ke SHAO¹()

^1.Institute of Biotechnology，Inner Mongolia Academy of Science and Technology，Hohhot 010000，China
^2.College of Plant Protection，Henan Agricultural University，Zhengzhou 450002，China
^3.Rural Revitalization Research Center of Inner Mongolia Autonomous Region，Hohhot 010000，China

Received:2024-02-28 Accepted:2024-03-28 Online:2024-05-15 Published:2024-05-14
Contact: Ying ZHAO,Ke SHAO

摘要/Abstract

摘要：

由甜菜尾孢菌（Cercospora beticola）引起的甜菜褐斑病是一种广泛发生、危害严重的真菌性病害，严重制约了我国糖料产业的发展。为挖掘对甜菜褐斑病具有生防应用潜力的拮抗菌株，从内蒙古自治区呼伦贝尔黑土地采集甜菜健康植株根际土壤样本，通过梯度稀释平板法和对峙培养法进行分离、纯化、筛选，获得1株具有良好拮抗效果的菌株HM85。基于其形态学特征和生理生化特征，结合16S rRNA基因序列分析将菌株HM85鉴定为娄彻氏链霉菌（Streptomyces rochei）。平板对峙试验结果显示，菌株HM85对甜菜尾孢菌HB2-2-2菌株的生长有明显抑制作用，抑菌率为58.68%。盆栽试验结果表明，HM85在防治甜菜褐斑病的同时对甜菜植株具有一定的促生作用，株高、株鲜重、根长、根鲜重和根直径与对照组相比分别增加22.73%、20.47%、36.10%、32.52%和75.03%。由此可见，娄彻氏链霉菌HM85具有作为促生菌剂和生防菌剂的研发潜力。

关键词: 娄彻氏链霉菌, 甜菜尾孢菌, 甜菜褐斑病, 拮抗活性, 促生作用

Abstract:

Cercospora leaf sport of beet （CLS）， caused by Cercospora beticola， is the most destructive foliar fungal disease of sugar beet worldwide which seriously restricts the development of China’s sugar industry. In order to screen and identify the antagonistic strains with biocontrol applications against Cercospora beticola， the rhizosphere soil samples of sugar beet were collected from Hulunbuir， Inner Mongolia autonomous region and the antagonistic strain HM85 with stable antimicrobial activity was screened by dilution coating flat plate method and dual culture assay. The strain HM85 was identified as Streptomyces rochei based on its morphological，physiological and biochemical characteristics along with 16S rRNA gene sequence analysis．The results of dual culture test showed that the HM85 strain could significantly inhibit the growth of Cercospora beticola HB2-2-2 with inhibitory rate of 58.68%. In addition， the fermentation broth of S. rochei HM85 had antagonistic activity against Cercospora beticola and could promote the growth of sugar beet in pot tests，which plant height， fresh weight of plant，root length，fresh weight of root and root diameter was increased by 22.73%， 20.47%， 36.10%， 32.52%， 75.03% compared to the control， respectively. In summary， S. rochei HM85 could be exploited as a potential bio-inoculant improving production and biocontrol agent for sugar beet leaf sport disease control.

Key words: Streptomyces rochei, Cercospora beticola, Cercospora leaf sport of beet （CLS）, antagonistic activity, growth-promoting effect

中图分类号:

S182

杨智敏, 张慧豪, 张园园, 杜红岩, 刘晓东, 侯亚光, 王毅, 徐道龙, 黄金贵, 程晓宁, 随洋, 王瑞利, 于超, 赵玲玲, 陈春梅, 雅茹, 贾丽, 张明月, 王宏伟, 姚淞耀, 赵莹, 邵科. 娄彻氏链霉菌HM85的鉴定及其对甜菜的防病促生作用[J]. 中国农业科技导报, 2024, 26(5): 148-155.

Zhimin YANG, Huihao ZHANG, Yuanyuan ZHANG, Hongyan DU, Xiaodong LIU, Yaguang HOU, Yi WANG, Daolong XU, Jingui HUANG, Xiaoning CHENG, Yang SUI, Ruili WANG, Chao YU, Lingling ZHAO, Chunmei CHEN, Ru YA, Li JIA, Mingyue ZHANG, Hongwei WANG, Songyao YAO, Ying ZHAO, Ke SHAO. Identification of Streptomyces rochei HM85 and Its Biocontrol and Growth-promoting Effects on Sugar Beet[J]. Journal of Agricultural Science and Technology, 2024, 26(5): 148-155.

图/表 9

图1 部分菌株对甜菜生尾孢菌HB2-2-2的拮抗作用

Fig. 1 Antagonistic effects of partial strains on C. beticola Sacc. HB2-2-2

图2 部分供试菌株与甜菜生尾孢菌的平板对峙结果A：HB2-2-2；B：HM13 vs HB2-2-2；C：HM47 vs HB2-2-2；D：HM41 vs HB2-2-2；E：HM44 vs HB2-2-2；F：HM85 vs HB2-2-2

Fig. 2 Plate confrontation result between partial strains and C. beticola Sacc.

表1 不同菌株对甜菜尾孢菌的拮抗效果

Table 1 Antagonistic effect of strains on C. beticola Sacc. HB2-2-2

菌株 Strain	抑菌半径 Inhibiting radius/cm	抑菌率 Inhibiting ratio/%
HM13	2.06±0.12 d	50.93±3.08 e
HM47	1.71±0.05 c	42.22±1.35 d
HM41	1.22±0.11 b	30.17±2.82 c
HM44	2.23±0.11 d	55.10±2.81 f
HM85	2.37±0.06 d	58.68±1.60 f
HM74	0.71±0.03 a	17.56±0.70 a
HM45	1.01±0.07 c	25.08±1.68 b
HM66	0.94±0.02 a	23.35±0.57 b

图3 不同处理组植株的生长表型

Fig. 3 Plant growth of different treatment groups

图4 不同处理组甜菜生理指标A：株高；B：株鲜重；C：根长；D：根鲜重；E：根直径。ns表示差异不显著（P>0.05）；*表示差异显著（P<0.05）。

Fig. 4 Physiological indices of Beta vulgaris in different treatment groupsA：Plant height； B：Plant fresh weight； C：Root length； D：Root fresh weight； E：Root diameter. ns indicates no significant difference （P>0.05）； * indicates significant difference （P<0.05）.

表2 不同处理甜菜植株的生理指标和促生效率

Table 2 Physiological indices and growth promotion efficiency of Beta vulgaris plants under different treatments

性状 Trait	对照 CK	HB2-2-2	HM58+HB2-2-2	促生效率 Efficiency of growth promotion/%
株高/mm Plant height	126.41±6.64 b	104.57±4.89 a	128.34±4.52 b	22.73
株鲜重/g Plant fresh weight	1.80±0.38 b	1.62±0.24 a	1.95±0.26 b	20.47
根长/mm Root length	146.60±8.76 a	176.37±9.14 a	240.05±6.92 a	36.10
根鲜重/g Root fresh weight	0.40±0.03 b	0.34±0.03 a	0.46±0.02 b	32.52
根直径/mm Root diameter	3.22±0.04 a	2.96±0.06 a	5.19±0.03 b	75.03

图5 HM85在高氏1号培养基上的菌落形态

Fig. 5 Colony of strain HM85 on GS-1 plate

表3 菌株HM85生理特征

Table 3 Physiological and biochemical characteristics of strain HM85

理化特性 Physicochemical property	HM85
葡萄糖Glucose	+
果糖Fructose	+
木糖Xylose	+
棉子糖Raffinose	+
L-鼠李糖L-rhamnose	+
L-阿拉伯糖L-arabinose	+
甘露醇Mannitol	+
水杨苷Salicin	+
肌醇Inositol	-
蔗糖Cane sugar	-
产H₂S Produce H₂S	NA
产黑色素Produce melanin	NA
水解淀粉Hydrolyzed starch	+
分解纤维素Break down cellulose	+
牛奶凝固Milk coagulated	+
胨化Peptoneization	+
明胶液化Gel atin liquefaction	NA

图6 基于菌株HM85 16S rRNA序列构建的进化树

Fig. 6 Phylogenetic tree based on 16S rRNA sequences of strain HM85

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娄彻氏链霉菌HM85的鉴定及其对甜菜的防病促生作用

Identification of Streptomyces rochei HM85 and Its Biocontrol and Growth-promoting Effects on Sugar Beet

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