Scientia Agricultura Sinica ›› 2016, Vol. 49 ›› Issue (15): 2977-2987.doi: 10.3864/j.issn.0578-1752.2016.15.012

;

• PLANT PROTECTION • Previous Articles     Next Articles

Influence of Subculture on Virulence to Frankliniella occidentalis and Conidial Production of the Entomopathogenic Fungus Beauveria bassiana

ZHANG Hui, WU Sheng-yong, LI Juan, ZHANG Lu-lu, ZHANG Lin-ya, LEI Zhong-ren   

  1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193
  • Received:2016-03-28 Online:2016-08-01 Published:2016-08-01

Abstract: 【Objective】The objective of this study is to assess the capability of the Beauveria bassiana against Frankliniella occidentalisin the field, and explore the influence of subculture on virulence and conidial production of the B. bassiana, propose an effective method for preventing the retrogression during successive subculture, and provide a basis for the large-scale production technology of B. bassiana. 【Method】 The virulence of 28 fungal strains to adult F. occidentalis was evaluated by soaking method with B. bassiana concentration of 1×107 conidia/mL in the laboratory. The adults were dipped for 5 s in the conidial suspension. Conidial production of the 4 high virulent strains was measured on corn meal agar (CMA) medium and sabouraud dextrose agar yeast (SDAY) medium, screening high conidial production strain and spore production medium by comparing conidial production of different strains on different media. Subsequently, strain DZDC-9 was subcultured on three media (CMA medium, CMA medium with cicada exuviae, and CMA medium with F. occidentalis cadaver powder) to compare its virulence and conidial production over five subcultures. The relationship between conidial production and virulence was analyzed. 【Result】Among 28 strains of entomopathogenic fungi B. bassiana tested for virulence against F. occidentalis in laboratory bioassays, strains DZDC-9, GZGY-5, WLMQ1-8 and SZ-26 were found as the most potent. The corrected cumulative mortalities were over 90% after 5 days, and LT50<3 d. Conidial production obtained from the CMA medium of each strain was higher than from SDAY medium, and strain DZDC-9 had the higher conidial production among the four strains. The results demonstrated that the virulence of strain DZDC-9 declined with successive subculture. However, when the strain was subcultured on the CMA medium with 0.5% cicada exuviae or 0.5% F. occidentalis cadaver powder as an additive, the virulence increased with successive subculture. Conidial production was not significantly different between the same generation subculture on CMA medium with cicada exuviae and CMA medium with F. occidentalis cadaver powder, but they were significantly greater than conidial production obtained from CMA medium. A positive correlation was found between sporulation and virulence of B. bassiana conidia under different media and culture conditions.【Conclusion】The potential of B. bassiana strain DZDC-9 for the control of F. occidentalis was studied, and proposed an effective method for stabilizing the virulence and sporulation of B. bassiana. Conidial production could be used as an indicator for virulence evaluation.

Key words: Frankliniella occidentalis, Beauveria bassiana, subculture, virulence, conidial production

[1]    Kirk W D, Terry L I. The spread of the western flower thrips Frankliniella occidentalis (Pergande). Agricultural and Forest Entomology, 2003, 5(4): 301-310.
[2]    Reitz S R, Gao Y L, Lei Z R. Thrips: pests of concern to China and the United States. Agricultural Sciences in China, 2011, 10(6): 867-892.
[3]    郑长英, 刘云虹, 张乃芹, 赵希丽. 山东省发现外来入侵有害生物——西花蓟马. 青岛农业大学学报(自然科学版), 2007, 24(3): 172-174.
Zheng C Y, Liu Y H, Zhang N Q, Zhao X L. Invaded insect pest-Frankliniella occidentalis first reported in Shandong Province. Journal of Qingdao Agricultural University (Natural Science), 2007, 24(3): 172-174. (in Chinese)
[4]    吕要斌, 张治军, 吴青君, 杜予洲, 张宏瑞, 于毅, 王恩东, 王铭华, 王满囷, 童晓立, 吕立华, 谭新球, 付卫东. 外来入侵害虫西花蓟马防控技术研究与示范. 应用昆虫学报, 2011, 48(3): 488-496.
Lü Y B, Zhang Z J, Wu Q J, Du Y Z, Zhang H R, Yu Y, Wang E D, Wang M H, Wang M Q, Tong X L, Lü L H, Tan X Q, Fu W D. Research progress of the monitoring, forecast and sustainable management of invasive alien pest Frankliniella occidentalis in China. Chinese Journal of Applied Entomology, 2011, 48(3): 488-496. (in Chinese)
[5]    Reitz S R, Yearby E L, Funderburk J E, STAVISKY J, MOMOL M T, OLSON S M. Integrated management tactics for Frankliniella thrips (Thysanoptera: Thripidae) in field-grown pepper. Journal of Economic Entomology, 2003, 96(4): 1201-1214.
[6]    雷仲仁, 问锦曾, 王音. 危险性外来入侵害虫——西花蓟马的鉴别、危害及防治. 植物保护, 2004, 30(3): 63-66.   
Lei Z R, Wen J Z, Wang Y. The identification, damage and control of an invasive western flower thrips (Frankliniella occidentalis). Plant Protection, 2004, 30(3): 63-66. (in Chinese)
[7]    Helyer N L, Brobyn P J. Chemical control of western flower thrips (Frankliniella occidentalis Pergande). Annals of Applied Biology, 1992, 121(2): 219-231.
[8]    Niassy S, Maniania N K, Subramanian S, Gitonga L M, Mburu D M, Masiga D, Ekesi S. Selection of promising fungal biological control agent of the western flower thrips Frankliniella occidentalis (Pergande). Letters in Applied Microbiology, 2012, 54(6): 487-493.
[9]    Shah F A, Allen N, Wright C J, Butt T M. Repeated in vitro subculturing alters spore surface properties and virulence of Metarhizium anisopliae. FEMS Microbiology Letters, 2007, 276(1): 60-66.
[10]   Wang C S, Butt T M, St Leger R J. Colony sectorization of Metarhizium anisopliae is a sign of ageing. Microbiology, 2005, 151(10): 3223-3236.
[11]   徐金柱, 黄勃, 李增智. 球孢白僵菌单孢子分离株在继代培养过程中菌落局变的遗传分析. 菌物学报, 2010, 29(2): 279-284.  
Xu J Z, Huang B, Li Z Z. Genetic analysis of colony sectorization of Beauveria bassiana single-spore isolates during subculturing. Mycosystema, 2010, 29(2): 279-284. (in Chinese) 
[12]   Jacobson R J, Chandler D, Fenlon J, Russell K M. Compatibility of Beauveria bassiana (Balsamo) Vuillemin with Amblyseius cucumeris Oudemans (Acarina: Phytoseiidae) to control Frankliniella occidentalis Pergande (Thysanoptera: Thripidae) on cucumber plants. Biocontrol Science and Technology, 2001, 11(3): 391-400.
[13]   Ugine T A, Wraight S P, Brownbridge M, Sanderson J P. Development of a novel bioassay for estimation of median lethal concentrations (LC50) and doses (LD50) of the entomopathogenic fungus Beauveria bassiana, against western flower thrips, Frankliniella occidentalis. Journal of invertebrate pathology, 2005, 89(3): 210-218.
[14]   Safavi S A, Shah F A, Pakdel A K, Rasoulian G R, Bandani A R, Butt T M. Effect of nutrition on growth and virulence of the entomopathogenic fungus Beauveria bassiana. FEMS Microbiology Letters, 2007, 270(1): 116-123.
[15]   唐晓庆, 樊美珍, 李增智. 球孢白僵菌继代培养中菌落局变现象及环境影响因素的研究. 真菌学报, 1996, 15(3): 188-196.
Tang X Q, Fan M Z, Li Z Z. Study on saltation of Beauveria bassiana, an important entomogenous fungus subcultured in different conditions. Acta Mycologica Sinica, 1996, 15(3): 188-196. (in Chinese)
[16]   XU J Z, HUANG B, QIN C S, LI Z Z. Sequence and phylogenetic analysis of Beauveria bassiana with mitochondrial genome. Mycosystema, 2009, 28(5): 718-723.
[17]   汪黎明, 陈雪, 徐延平, 胡晓磊, 何灵敏, 黄勃, 李增智. 一株球孢白僵菌转基因工程菌在继代培养中的变异. 菌物学报, 2011, 30(5): 738-743.
Wang L M, Chen X, Xu Y P, Hu X L, He L M, Huang B, Li Z Z. Variation of a transgenic strain of Beauveria bassiana during subculturing. Mycosystema, 2011, 30(5): 738-743. (in Chinese)
[18]   蔡国贵, 林庆源, 徐耀昌, 黄芙蓉, 陈金标. 白僵菌菌株退化与培养条件关系及其控制技术. 福建林学院学报, 2001, 21(1): 76-79.  
Cai G G, Lin Q Y, Xu Y C, Huang F R, Chen J B. The relationship between culture condition and strain degeneration of Beauveria bassiana and its control technique. Journal of Fujian College of Forestry, 2001, 21(1): 76-79. (in Chinese)
[19]   樊美珍, 李增智, 唐晓庆. 白僵菌菌种退化及其控制. 安徽农业大学学报, 1996, 23(3): 239-245.                
Fan M Z, Li Z Z, Tang X Q. Strain degeneration and its control of Beauveria bassiana. Journal of Anhui Agricultural University, 1996, 23(3): 239-245. (in Chinese)
[20]   James R R. Effects of exogenous nutrients on conidial germination and virulence against the silverleaf whitefly for two Hyphomycetes. Journal of Invertebrate Pathology, 2001, 77(2): 99-107.
[21]   Shah F A, Butt T M. Influence of nutrition on the production and physiology of sectors produced by the insect pathogenic fungus Metarhizium anisopliae. FEMS microbiology letters, 2005, 250(2): 201-207.
[22]   Furlaneto M C, Paião F G, Pinto F G, Fungaro M H. Transformation of the entomopathogenic fungus Metarhizium flavoviride to high resistance to benomyl. Canadian Journal of Microbiology, 1999, 45(10): 875-878.
[23]   Ansari M A, Butt T M. Effects of successive subculturing on stability, virulence, conidial yield, germination and shelf-life of entomopathogenic fungi. Journal of Applied Microbiology, 2011, 110(6): 1460-1469.
[24]   李会平, 黄大庄, 杜邵华, 高洁. 白僵菌经不同基质传代后对桑天牛幼虫的侵染力比较. 蚕业科学, 2008, 34(2): 339-341.
Li H P, Huang D Z, Du S H, Gao J. Comparative infectivity of different passage reisolates of Beauveria bassiana to Apriona germari larvae. Science of Sericulture, 2008, 34(2): 339-341. (in Chinese)
[25]   关兵兵, 王冰, 刘艳微, 孟鑫睿, 李晓慧, 张佳诗, 徐文静, 李启云, 冯树丹, 张正坤. 蝉蜕诱导对球孢白僵菌生物学特性及其毒力影响的研究. 中国农学通报, 2014, 30(34): 51-55.
Guan B B, Wang B, Liu Y W, Meng X R, Li X H, Zhang J S, Xu W J, Li Q Y, Feng S D, Zhang Z K. Research on effect on biological characteristics and virulence of Beauveria bassiana induced by Periostracum cicadae. Chinese Agricultural Science Bulletin, 2014, 30(34): 51-55. (in Chinese)
[26]   李娟, 吴圣勇, 王晓青, 王帅宇, 雷仲仁. 防治西花蓟马的球孢白僵菌菌株筛选及耐热性测定. 中国生物防治学报, 2015, 31(6): 845-852.   
Li J, Wu S Y, Wang X Q, Wang S Y, Lei Z R. Screening of Beauveria bassiana strains against Frankliniella occidentalis and their conidial thermotolerance. Chinese Journal of Biological Control, 2015, 31(6): 845-852. (in Chinese)
[27]   Wu S Y, Gao Y L, Xu X N, Goettel M S, Lei Z R. Compatibility of Beauveria bassiana with Neoseiulus barkeri for control of Frankliniella occidentalis. Journal of Integrative Agriculture, 2015, 14(1): 98-105.
[28]   张璐璐, 吴圣勇, 王帅宇, 李娟, 雷仲仁. 防治蓟马的球孢白僵菌SDDZ-9菌株液体发酵工艺优化. 中国农业科学, 2015, 48(15): 2985-2994.     
Zhang L L, WU S Y, WANG S Y, LI J, Lei Z R. Optimization of fermentation process of Beauveria bassiana of SDDZ-9 against Frankliniella occidentalis. Scientia Agricultura Sinica, 2015, 48(15): 2985-2994. (in Chinese)
[29]   张志光, 李建宗, 欧笑兰, 徐正平, 刘敏颂. 白僵菌生活史的研究——(Ⅱ)异核体的形成和准性生殖的显微摄影观察. 湖南师范大学自然科学学报, 1981(2): 38-44.
Zhang Z G, Li J Z, Ou X L, Xu Z P, Liu M S. Life history research of Beauveria bassiana-(Ⅱ) Photomicrography observation of forming the nucleus and associate different sexual reproduction. Journal of Natural Science of Hunan Normal University, 1981(2): 38-44. (in Chinese)
[30]   李会平, 黄大庄, 王志刚, 郑建伟, 黄秋娴. 不同基质传代白僵菌菌株胞外蛋白酶和几丁质酶产生水平与菌株对桑天牛幼虫毒力的关系. 蚕业科学, 2008, 34(3): 521-524.
Li H P, Huang D Z, Wang Z G, Zheng J W, Huang Q X. Relationships between extracellular protease and chitinase activity level of different successive reisolates of Beauveria bassiana and virulence against the Apriona germari larvae. Science of Sericulture, 2008, 34(3): 521-524. (in Chinese)
[31]   Fang W, Leng B, Xiao Y, Jin K, Ma J, Fan Y, Feng J, Yang X, Zhang Y, Pei Y. Cloning of Beauveria bassiana chitinase gene Bbchit1 and its application to improve fungal strain virulence. Applied & Environmental Microbiology, 2005, 71(1): 363-370.
[32]   Lohmann U, Sikora R A, Höfer M. Influence of phospholipids on growth, sporulation and virulence of the endoparasitic fungi Drechmeria coniospora, Verticillium balanoides, and Harposporium anguillulae, in liquid culture. Journal of Phytopathology, 1989, 125(2): 139-147.
[33]   胡景江, 樊美珍. 球孢白僵菌胞外蛋白酶与其毒力的关系. 安徽农业大学学报, 1996, 23(3): 273-278.          
Hu J J, Fan M Z. Relation between extracellaluar protease of Beauveria bassiana and its virulence to Dendrolimus punctatus. Journal of Anhui Agricultural University, 1996, 23(3): 273-278. (in Chinese)
[34]   李农昌, 樊美珍, 李春如, 李华, 高智辉, 朱润德. 白僵菌有关培养条件及其与毒力关系的研究. 安徽农业大学学报, 1996, 23(3): 254-259.
Li N C, Fan M Z, Li C R, Li H, Gao Z H, Zhu R D. relationship between culture condition and virulence of Beauveria bassiana. Journal of Anhui Agricultural University, 1996, 23(3): 254-259. (in Chinese)
[1] LIU RUI, ZHAO YuHan, FU ZhongJu, GU XinYi, WANG YanXia, JIN XueHui, YANG Ying, WU WeiHuai, ZHANG YaLing. Distribution and Variation of PWL Gene Family in Rice Magnaporthe oryzae from Heilongjiang Province and Hainan Province [J]. Scientia Agricultura Sinica, 2023, 56(2): 264-274.
[2] WANG WenJuan,SU Jing,CHEN Shen,YANG JianYuan,CHEN KaiLing,FENG AiQing,WANG CongYing,FENG JinQi,CHEN Bing,ZHU XiaoYuan. Pathogenicity and Avirulence Genes Variation of Magnaporthe oryzae from a Rice Variety Meixiangzhan 2 in Guangdong Province [J]. Scientia Agricultura Sinica, 2022, 55(7): 1346-1358.
[3] TANG ZiYun,HU JianXin,CHEN Jin,LU YiXing,KONG LingLi,DIAO Lu,ZHANG FaFu,XIONG WenGuang,ZENG ZhenLing. Relationship Between Biofilm Formation and Molecular Typing of Staphylococcus aureus from Animal Origin [J]. Scientia Agricultura Sinica, 2022, 55(3): 602-612.
[4] CHEN ChaoXi,LI YuHan,TAN Min,WANG Lu,HUANG ZhiHong. Biofilm-Forming Phenotype, Antibacterial Resistance Genes, Integrase Genes and Virulence Genes Detection of Escherichia coli Isolated from Yaks and Tibetan Pigs in Northwest Sichuan Plateau [J]. Scientia Agricultura Sinica, 2021, 54(23): 5144-5162.
[5] CAI Ni,YAN DuoZi,NONG XiangQun,WANG GuangJun,TU XiongBing,ZHANG ZeHua. Adhesin Gene mad2 Knockout and Functional Effects on Biological Characteristics and Inducing Plant Responses in Metarhizium anisopliae [J]. Scientia Agricultura Sinica, 2021, 54(22): 4800-4812.
[6] WANG ChengLi,YIN ZhiYuan,NIE JiaJun,LIN YongHui,HUANG LiLi. Identification and Virulence Analysis of CAP Superfamily Genes in Valsa mali [J]. Scientia Agricultura Sinica, 2021, 54(16): 3440-3450.
[7] GuangZhi ZHANG,MingYan WANG,Yi LUO,ShiHang XU,ShunDong HE,ShangJin CUI. The Zoonotic Bacterium-Helicobacter suis and Its Research Progress [J]. Scientia Agricultura Sinica, 2020, 53(6): 1278-1286.
[8] ZHANG Lei,JIA Qi,WU Wei,ZHAO LuPing,XUE Bing,LIU HuanHuan,SHANG Jing,YONG TaiWen,LI Qing,YANG WenYu. Species Identification and Virulence Determination of Beauveria bassiana Strain BEdy1 from Ergania doriae yunnanus [J]. Scientia Agricultura Sinica, 2020, 53(14): 2974-2982.
[9] YANG ZhiYuan,DUAN HuiJuan,WANG XiaoLei,LIU LiXin,ZHAO JiCheng,PAN Jie,LIU YueHuan,LIN Jian. Virulence, E Gene Sequence and Antigenic Difference of 4 Duck Tembusu Virus Isolations [J]. Scientia Agricultura Sinica, 2019, 52(23): 4406-4414.
[10] MENG Feng,ZHANG YaLing,JIN XueHui,ZHANG XiaoYu,JIANG Jun. Detection and Analysis of Magnaporthe oryzae Avirulence Genes AVR-Pib, AVR-Pik and AvrPiz-t in Heilongjiang Province [J]. Scientia Agricultura Sinica, 2019, 52(23): 4262-4273.
[11] SUN Ke,ZHU HaoDan,HE KongWang,WANG DanDan,ZHOU JunMing,YU ZhengYu,Lü LiXin,NI YanXiu. Biological Characteristics of Transcriptional Regulator GalR in Streptococcus suis Serotype 4 [J]. Scientia Agricultura Sinica, 2019, 52(19): 3485-3494.
[12] LIU XiaoChen, WU ShengYong, LEI ZhongRen, WANG HaiHong. Growth Kinetics and Virulence of Two Beauveria bassiana Strains in Frankliniella occidentalis Under Different Temperatures [J]. Scientia Agricultura Sinica, 2018, 51(8): 1484-1492.
[13] REN ShiLong, BAI Hui, WANG yongFang, QUAN JianZhang, DONG ZhiPing, LI ZhiYong, XING JiHong. Identification and Analysis of Magnaporthe oryzae of Foxtail Millet Avirulence Genes [J]. Scientia Agricultura Sinica, 2018, 51(6): 1079-1088.
[14] YIN Xin, MA ShuJie, LI Mei, DENG GuoHua, HOU YuJie, CUI PengFei, SHI JianZhong, CHEN HuaLan. Amino Acid Substitutions of E627V in Polymerase Basic Protein 2 Gene Increases the Pathogenicity of the H7N9 Influenza Virus in Mice [J]. Scientia Agricultura Sinica, 2018, 51(17): 3379-3388.
[15] DU JiGe, XUE Qi, ZHU Zhen, LI QiHong, YIN ChunSheng, YAO WenSheng, KANG Kai, CHEN XiaoYun. Expression and Evaluation of Protective Efficacy of No-toxic Clostridium perfringens ε Toxin Derivative [J]. Scientia Agricultura Sinica, 2018, 51(11): 2206-2215.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!