A Brief Analysis of the Factors Restricting the Effectiveness of Controlling Cotton Verticillium wilt by Using Biocontrol Bacteria in the Field

doi:10.6048/j.issn.1001-4330.2022.01.018

Abstract

Abstract:

【Objective】 To identify the main factors that restrict the effectiveness of controlling cotton Verticillium wilt by using biocontrol bacteria in the field, and to provide a theoretical basis for improving the techniques for biocontrol of cotton Verticillium wilt in Xinjiang. 【Method】 Cotton Verticillium wilt in the Korla cotton areas was taken as the subject and the occurrence of Verticillium wilt and data about agricultural operations were investigated in the field. The microsclerotia of Verticillium dahlia were separated from the soil using selective media, and gfp-AL7 was used as the marker strain to analyze its colonization capability in soil and its ability to inhibit Verticillium dahlia microsclerotia. 【Result】 Antimicrobial agents with water droplets were typically applied in the Korla cotton area in mid-Junes, which is seriously later than the infection and occurrence periods of cotton Verticillium wilt. More than 90% of Verticillium dahlia microsclerotia in the soil were distributed in the 0-20 cm plough layer, and a small amount existed in the 20-40 cm plough layer. The extracted liquid of the cotton field soil had no inhibitory effect on the growth of the strain AL7, but the marker strain gfp-AL7 had a poor ability to naturally proliferate in the soil. Its number of colonization began declined over time, mainly colonized in the 0-20 cm soil layer. The corrected inhibition rate of microsclerotia in soil by the fermentation broth of biocontrol bacteria AL7 was 70.5%, while that of microsclerotia by the 1∶200 diluted fermentation broth was only 5.5%. The biological agents had no inhibitory effect on the microsclerotia of Verticillium dahliae, instead, they accelerated the increase in the number of microsclerotia. 【Conclusion】 The biocontrol strain AL7 has poor proliferation ability in soil and undesirable inhibition on the microsclerotia of Verticillium dahliae. The application techniques of biocontrol bacteria should be modified according to the occurrence pattern of cotton Verticillium wilt.

Key words: cotton Verticillium wilt; biological control; microsclerotia; colonization; suppression

摘要：

【目的】研究田间利用生防菌防治棉花黄萎病效果的制约因素,为完善新疆棉花黄萎病的生物防治技术提供理论依据。【方法】以库尔勒棉区棉花黄萎病为研究对象,采集田间调查黄萎病发生情况及数据,利用选择性培养基分离土壤中棉花黄萎病菌微菌核,以gfp-AL7为标记菌株分析其在土壤中的定植能力及对黄萎病菌微菌核的抑制能力。【结果】库尔勒棉区随水滴施生防菌剂时间为6月中旬,迟于棉花黄萎病侵染和发生时期。土壤中90%以上的黄萎病菌微菌核分布于0~20 cm耕层,少量存在于20~40 cm耕层中。棉田土壤浸提液对菌株AL7的生长无抑制作用,但是标记菌株gfp-AL7在土壤中自然增殖能力较差,其定植数量随着时间延长开始下降,主要定殖在0~20 cm的土层中。生防菌AL7发酵原液对土壤中微菌核的校正抑制率为70.5%,而200倍发酵液对微菌核的校正抑制率仅为5.5%;生物菌剂对黄萎病菌微菌核没有抑制效果,相反还促进了微菌核数量的增加。【结论】 生防菌株AL7在土壤中的增殖能力以及对棉花黄萎病菌微菌核的抑制能力较差;生防菌施用技术需根据棉花黄萎病发生规律进行调整。

关键词: 棉花黄萎病, 生物防治, 微菌核, 定植, 抑制

CLC Number:

S435.62

LIU Haiyang, WANG Wei, ZHANG Renfu, Wenqiemu Abulizi, YAO Ju. A Brief Analysis of the Factors Restricting the Effectiveness of Controlling Cotton Verticillium wilt by Using Biocontrol Bacteria in the Field[J]. Xinjiang Agricultural Sciences, 2022, 59(1): 155-161.

刘海洋, 王伟, 张仁福, 温切木·阿布列孜, 姚举. 利用生防菌防治棉花黄萎病效果的制约因素[J]. 新疆农业科学, 2022, 59(1): 155-161.

Figures/Tables 6

Fig.1 Occurrence dynamics of Verticillium wilt in different cotton varieties

Fig.2 Distribution of microsclerotia in different depths of soil Note:Different lowercase letters indicate significant differences (P<0.05), The error bars indicate standard error of means (n=4)

Fig.3 Growth status of microsclerotia isolated from soil on selective medium

Fig.4 The colonization of bio-control bacteria AL7 in soil at different depths

Fig.5 The effect of soil extract on the growth of bio-control bacteria

Table 1 Inhibitory effect of bio-control bacteria(agent)on the microsclerotia of Verticillium dahliae (per/g soil)

处理 Treatment group	施用前 Before applying	施用后10 d 10 d After application	校正抑制率 Correction inhibition rate(%)	2次施用后10 d 10 d After second application	校正抑制率 Correction inhibition rate(%)
清水对照Water control	393±39.7	205±79.5	-	158±13.5	-
AL7发酵原液AL7 fermentation stoste	253±20.3	44±23.0	66.7	10±1.2	70.5
AL7发酵液(200 X)AL7 fermentation(200 X)	327±41.2	162±19.4	5.0	118±12.1	5.5
生物菌剂(200 X)Microbial agents(200 X)	264±37.5	181±98.1	-	224±15.6	-

References 20

[1]	刘海洋, 王琦, 王伟, 等. 新疆棉花黄萎病的发生及其病原菌的分子鉴定与ISSR分析[J]. 植物保护学报, 2018, 45(6):1194-1203.
	LIU Haiyang, WANG Qi, WANG Wei, et al. Molecular identification and ISSR analysis of Verticillium dahliae and the current status of cotton Verticillium wilt in Xinjiang[J]. Journal of Plant Protection, 2018, 45(6):1194-1203.
[2]	刘海洋, 王伟, 张仁福, 等. 新疆主要棉区棉花黄萎病发生概况. 植物保护, 2015, 41(3):138-142.
	LIU Haiyang, WANG Wei, ZHANG Renfu, et al. Occurrence overviews of cotton Verticillium wilt in Xinjiang[J]. Plant Protection, 2015, 41(3):138-142.
[3]	朱荷琴, 冯自力, 赵丽红, 等. 我国棉花黄萎病菌的致病力分化及落叶型菌株的分布[C]. 中国植物病理学会2010年学术年会论文集, 2010:147-148.
	ZHU Heqin, FENG Zili, ZHO Lihong, et al. Pathogenicity differentiation of Verticillium dahliae and the distribution of deciduous strains in China[C]. Proceedings of the 2010 Annual Meeting of the Chinese Society of Plant Pathology, 2010:147-148.
[4]	刘海洋, 姚举, 张仁福, 等. 黄萎病不同发生程度棉田中土壤微生物多样性[J]. 生态学报, 2018, 38(5):1619-1629.
	LIU Haiyang, YAO Ju, ZHANG Renfu, et al. Analysis of soil microbial diversity in cotton fields differing in occurrence of cottonVerticilliumwilt in Xinjiang[J]. Acta Ecologica Sinica, 2018, 38(5):1619-1629.
[5]	商文静, 陈婷, 白应文, 等. 大丽轮枝菌微菌核的萌发条件及致死温度[J]. 菌物学报, 2013. 32(6):986-992.
	SHANG Wenjing, CHEN Ting, BAI Yingwen, et al. Germination condition and lethal temperature for microsclerotia of Verticillium dahliae[J]. Mycosystema, 2013, 32(6):986-992.
[6]	Lópezescudero F J, Mwanza C, Blancolópez M A. Reduction of Verticilliumd ahliae microsclerotia viability in soil by dried plant residues[J]. Crop Protection, 2007, 26(2):127-133. DOI URL
[7]	吴蔼民, 顾本康, 傅正擎, 等. 内生菌对棉花黄萎病的田间防效及增产作用[J]. 江苏农业科学, 2000, (5) :38-39.
	WU Aimin, GU Benkang, FU Zhengqing, et al. The controlling effect of cotton wilt (Verticilliumdahliae) by endophytic bacteria in the fields and its effect of yield increase[J]. Jiangsu Agricultural Sciences, 2000,(5):38-39.
[8]	林玲, 金中时, 马长文, 等. 棉花黄萎病生防内生细菌Jaascd的鉴定及田间防效[J]. 江苏农业学报, 2010, 26(1):65-69.
	LIN Ling, JIN Zhongshi, MA Changwen, et al. Identificationand field control efficacyofendophytic bacterial strain Jaascd against cotton Verticillium wilt[J]. Jiangsu Agricultural Sciences, 2010, 26(1):65-69.
[9]	石磊, 吕宁, 李全胜, 等. 生物农药随水滴施防治棉花黄萎病用量筛选研究[J]. 西南农业学报, 2019, 32(4):829-836.
	SHI Lei, LÜ Ning, LI Quansheng, et al. Amount screening of different biological agents application through drip irrigation to prevent cotton Vertillium wilt[J]. Southwest China Journal of Agricultural Sciences, 2019, 32(4):829-836.
[10]	喻树迅, 范术丽. 我国棉花遗传育种进展与展望[J]. 棉花学报, 2003, 15(2):120-124.
	YU Shuxun, FAN Shuli. The evolutions and prospect of cotton genetics and breeding in China[J]. Cotton Science, 2003, 15(2):120-124.
[11]	刘海洋, 姚举, 张仁福, 等. 甲基营养型芽孢杆菌AL7对棉花黄萎病的盆栽防治效果及定植能力[J]. 中国生物防治学报, 2017, 33(3):378-384.
	LIU Haiyang, YAO Ju, ZHANG Renfu, et al. Effect of Bacillus methylotrophicusAL7 against cotton Verticillium Wilt and its colonization in rhizosphere of cotton[J]. Chinese Journal of Biological Control, 2017, 33(3):378-384.
[12]	冯争光, 鹿秀云, 彭巧慧, 等. 一种新的检测土壤棉花黄萎菌微菌核的选择性培养基[J]. 华北农学报, 2004,(19):44-47.
	FENG Zhengguang, LU Xiuyun, PENG Qiaohui, et al. A novel selective medium for the detection of the microsclerotia[J]. Acta Agriculturae Boreali-Sinica, 2004,(19):44-47.
[13]	罗佳, 赵爽, 袁玉娟, 等. 施用微生物有机肥对棉花抗病性相关酶活性的影响[J]. 南京农业大学学报, 2011, 34(3):89-93.
	LUO Jia, ZHAO Shuang, YUAN Yujuan, et al. Effects of a bio-organic fertilizer on the disease resistance-related enzymes activities of cotton[J]. Journal of Nanjing Agricultural University, 2011, 34(3):89-93.
[14]	赵卫松, 李社增, 鹿秀云, 等. 西兰花植株残体还田对棉花黄萎病的防治效果及其安全性评价[J]. 中国生物防治学报, 2019, 35(3):449-455.
	ZHAO Weisong, LI Shezeng, LU Xiuyun, et al. Safety evaluation of broccoli residues returning on cotton growth and its control effect on Verticillium wilt[J]. Chinese Journal of Biological, 2019, 35(3):449-455.
[15]	Inderbitzin P, Ward J, Barbella A, et al. Soil microbiomes associated with Verticillium wilt suppressive broccoli and chitin amendments are enriched with potential biocontrol agents[J]. Phytopathology, 2017, 108(1):31-43. DOI URL
[16]	罗燕娜, 杜娟, 李俊华. 滴灌条件下枯草芽胞杆菌S37和S44 对棉花黄萎病的防治效果[J]. 植物保护, 2011, 37(2):174-176.
	LUO Yanna, DU Juan, LI Junhua. Control efficacy of Bacillus subtilis S37 and S44 against cotton Verticillium wilt by under-mulch-drip irrigation[J]. Plant Protection, 2011, 37(2):174-176.
[17]	Marzano S Y L, Villamil M B, Wander M M, et al. Organic Transition Effects on Soilborne Diseases of Soybean and Populations of Pseudomonadaceae[J]. Agronomy Journal, 2015, 388(3).
[18]	Olanya O M, Lambert D H, Porter G A. Effects of pest and soil management systems on potato diseases[J]. American Journal of Potato Research, 2006, 83(5):397-408. DOI URL
[19]	Eo J, Park K C.) Effects of manure composts on soil biota and root-rot disease incidence of ginseng (Panax ginseng)[J]. Applied Soil Ecology, 2013, 71:58-64. DOI URL
[20]	Compant S, Duffy B, Nowak J, et al. Use of Plant growth-promoting bacteria for biocontrol of plant disease: principles, mechanisms of action, and future prospects[J]. Applied and Environmental Microbiology, 2005, 71(9):4951-4959. DOI URL