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1 Chinese Journal of Applied Entomology 2016, 53(1): DOI: /j.issn 柑桔全爪螨高致病力菌株芽枝状枝 * 孢霉的生物学特性研究王 ** 娟 *** 胡军华龙艳玲周娜彭凤格姚廷山 / 摘要目的 Panonychus citri McGregor Cladosporium cladosporioides CQBBW3) 方法 ph CQBBW3 结果 CQBBW3 LC /ml LT d ph ph 7.0 结论 CQBBW3 25 关键词 Biological characteristics of a strain of Cladosporium cladosporioides that is highly pathogenic to Panonychus citri (McGregor) WANG Juan ** HU Jun-Hua *** LONG Yan-Ling ZHOU Na PENG Feng-Ge YAO Ting-Shan (Citrus Research Institute, Southwest University, National Engineering Research Center for Citrus, Scientific Observing and Experimental Station of Fruit Tree Science(Southwest Region), Ministry of Agriculture, Chongqing , China) Abstract [Objectives] To explore the biological characteristics of Cladosporium cladosporioides, a newly isolated fungal strain that is highly toxic to Panonychus citri. [Methods] The effects of different factors, such as culture medium, carbon sources, nitrogen sources, temperature, and ph, on the hyphal growth and sporulation of C. cladosporioides, were tested using single factor tests. [Results] The results indicate that the LC 50 of the CQBBW3 strain was spores/ml and the LT 50 was d. The optimum culture medium for growing the CQBBW3 strain was Czapek's Medium. Mannitol and Glycine were the best carbon and nitrogen sources for hyphal growth, and Lactose and Lysine were the best carbon and nitrogen sources for sporulation. The most suitable culture temperature for hyphal growth was 25, and the most suitable temperatures for sporulation were 15 and 30.The optimum ph for hyphal growth and sporulation were and 7.0, respectively. Although ph had no significant effect on fungal growth rate it had a marked effect on sporulation. [Conclusion] The CQBBW3 strain was toxic to P. citri, and grew best under conditions of 25, neutral ph and high organic nutrition. This study provides a scientific basis for both the rapid large-scale culture of the CQBBW3 strain and its effective utilization in the field. Key words Cladosporium cladosporioides, Panonychus citri, biological characteristic * Supported projects cstc2014fazktjcsf BAD16B ** First author wangnvkouyue@163.com *** Corresponding author hujunhua@cric.cn Received Accepted

2 1 : 23 Panonychus citri McGregor Chandler et al Fawcett Cladosporium cladosporioides Fres de Vries 10% 80% Eken Hayat 2009 Tetranychus urticae 50.95% 74.76% Cladosporium cladosporioides CQBBW3 ph CQBBW3 1 材料与方法 1.1 材料 CQBBW3 PDA 25 7 d 1.2 CQBBW3 菌株杀螨活性测定 CQBBW3 PDA d (12L.. 12D) 0.02% /ml % ml 10 ml 0.02% L.. 12D RH95% 7 d CQBBW3 PBT LC 50 LT 不同培养基对 CQBBW3 菌株生长和产孢的影响 CQBBW3 5 28±1 12L.. 12D RH95% 14 d SPSS20 5 PDA 200 g 20 g 20 g ml SYA 40 g 5 g 20 g ml Czapek's 3 g 1 g 0.5 g 0.5 g 0.01 g 30 g 20 g ml SDAY 40 g 10 g 10 g 20 g ml SEA 40 g 10 g 10 g 20 g 5

3 24 Chinese Journal of Applied Entomology ml L-broth 10 g NaCl 5 g 3 g 20 g 20 g ml 1.4 不同碳源氮源对 CQBBW3 菌株生长及产孢的影响 7 30 g d 6 3 g d 1.5 不同温度和 ph 值对 CQBBW3 菌株生长和产孢的影响 CQBBW PDA 14 d ph CQBBW3 1 mol L 1 HCl 1 mol L 1 NaOH PDA ph ph d 1.6 环境相容性农药的筛选 % WP 40% EC 80% WP 10% DP 43% SC[ ( ) ] 20% SC 50% WP 70% WP ( ) 25% SC 80% WP 50% WDG 1 g 1 ml 100 ml 100 ml PDA CQBBW d PBT EC 50 r 2 结果与分析 2.1 CQBBW3 菌株的杀螨活性 CQBBW /ml 7 50% 48% 7 LC /ml 95% Y= X r /ml LT d 95% Y= X r 不同培养基对 CQBBW3 菌株生长和产孢的影响 1 CQBBW3 SEA mm/d PDA L-broth SDAY 1.32 mm/d /mm 2 PDA SEA /mm 2

4 1 : 25 Table 1 Medium 表 1 CQBBW3 菌株在不同培养基上的生长速率和产孢量 The growth rates and spore production of CQBBW3 strain on different medium Growth rate mm/d 10 4 /mm 2 Spore production 10 4 spores/mm 2 SDAY 1.32±0.09d 1.61±0.07c L-broth 3.71±0.24b 0.33±0.00cd SEA 4.36±0.08a 0.13±0.00d Czapek s 2.32±0.38c 23.88±0.08a PDA 3.98±0.16b 14.83±0.11b SYA 4.37±0.07a 14.64±0.13b ± 0.05 Duncan s Data are means±se, and followed by same letters in the same column mean no significant difference at 0.05 levels by Duncan s multiple range test. The same below. 2.3 不同碳源氮源对 CQBBW3 菌株生长及产孢的影响 CQBBW mm/d /mm /mm 2 CQBBW mm/d 0.96 mm/d /mm /mm /mm 温度和 ph 对 CQBBW3 菌株生长和产孢的影响 CQBBW3 Table 2 Carbon source 表 2 不同碳源对 CQBBW3 菌株生长和产孢的影响 Effects of carbon sources on growth rates and spore production of CQBBW3 strain Growth rate (mm/d) 10 4 /mm 2 Spore production 10 4 spores/mm 2 Sucrose 2.30±0.26de 23.80±0.25b Mannite 2.92±0.10a 9.96±0.38d Maltose 2.80±0.06ab 10.71±0.06d Glycerol 2.61±0.02bc 16.31±0.05c Lactose 2.51±0.13cd 36.48±0.25a Glucose 2.21±0.05e 34.12±0.07a Starch 2.73±0.15abc 5.47±0.06e

5 26 Chinese Journal of Applied Entomology 53 Table 3 Nitrogen source 表 3 不同氮源对 CQBBW3 菌株生长和产孢的影响 Effects of nitrogen sources on growth rates and spore production of CQBBW3 strain Growth rate (mm/d) 10 4 /mm 2 Spore production (10 4 spores/mm 2 ) Ammonium sulphate 1.45±0.14c 12.84±0.02b Ammonium nitrate 1.46±0.08c 12.46±0.01c Glycine 2.73±0.18a 4.23±0.04e Urea 0.96±0.12d 7.22±0.07d Lysine 2.24±0.09b 17.20±0.01a Potassium nitrate 2.23±0.21b 2.46±0.02f CQBBW mm/d mm/d /mm /mm CQBBW3 ph ph ph mm/d ph ph /mm 2 ph ph CQBBW3 菌株与杀菌剂的相容性筛选 CQBBW % WP CQBBW3 EC mg/ml 50% WDG EC mg/ml 10% DP EC mg/ml 3 结论与讨论 Eken Hayat /ml Table 4 Temperature ( ) 表 4 温度对 CQBBW3 菌株生长和产孢的影响 Effects of temperature on growth rates and spore production of CQBBW3 strain Growth rate (mm/d) 10 5 /mm 2 Spore production (10 5 spores/mm 2 ) ±0.20d 2.04±0.12bc ±0.40cd 4.56±0.35a ±0.35b 1.78±0.09c ±0.22a 2.22±0.06b ±0.05c 4.31±0.13a ±0.00e 0.00±0.00d ±0.00e 0.00±0.00d

6 1 : 27 Table 5 ph ph value 表 5 ph 值对 CQBBW3 菌株生长和产孢的影响 Effects of ph value on the growth rate and spore production of CQBBW3 strain Growth rate (mm/d) 10 4 /mm 2 Spore production (10 5 spores/mm 2 ) ±0.03ab 7.98±0.50c ±0.12a 7.83±0.47c ±0.20a 17.39±0.12a ±0.05a 14.79±0.05b ±0.27b 2.21±0.37d ±0.21b 14.22±0.14b ±0.20b 3.40±0.38d Fungicide 表 6 11 种杀菌剂与 CQBBW3 菌株的相容性 Table 6 Compatibility of 11 fungicides with CQBBW3 strain Toxic regression equation EC 50 (mg/ml) r Correlation coefficient 10% DP Difenoconazole Y= X % EC Flusilazole Y= X % SC Tebuconazole Y= X % WP Oxine-copper Y= X % WP Mancozeb Y= X % WP Zineb Y= X % SC Thiophonate-methyl Y= X % WP Propineb Y= X % SC Azoxystrobin Y= X % WDG Kresoxim-methyl Y= X % WP Cuprous oxide Y= X WP WDG EW SC EC DP AS WP: Wettable powder; WDG: Water dispersible granules; EW: Emulsion in water; SC: Suspending concentrate; EC: Emulsifiable concentrate; DP: Powder; AS: Aqua % 74.76% LT d Jeyarani 2011 LC /ml LT d 50.8% LC 50 LT /ml d CQBBW

7 28 Chinese Journal of Applied Entomology % Czapek s 2% 15% 10~31 24~25 35 ph 5.0~ ph 2015 杧 PDA 20 ph ph 6.0 L ph ph 6.0~8.0 ph ph ph 7.0 CQBBW3 11 CQBBW3 CQBBW Hemiberiesia pitysophila ( 1991) Oracella acuta 2007 Aleurocanthus spiniferus ( ) CQBBW3 参考文献 (References) Chandler D, Davidson G, Pell JK, Ball BV, Shaw K, Sunderland KD, Fungal biocontrol of acari. Biocontrol Science & Technology, 10(4): Chen QS, Zhao S, Zou J, Shi L, He L, Monitoring of acaricide resistance in Tetranychus cinnabarinus. Chinese Journal of Applied Entomology, 49(2): [,,,,, , 49(2): ] Eken C, Hayat R, Preliminary evaluation of Cladosporium cladosporioides (Fresen.) de Vries in laboratory conditions, as a potential candidate for biocontrol of Tetranychus urticae Koch. World Journal of Microbiology & Biotechnology, 25(3): Fawcett HS, Biological control of citrus insects by parasitic fungi and bacteria. Citrus Industry, 2: Han BY, Li ZZ, Studies on infectivity of Pleurodesmospora coccorum to Aleurocanthus spiniferus. Journal of Anhui Agricultural University, 19(4): [,, , 19(4): ] Han BY, Zhang HH, Cui L, Epizootic infection and spatial pattern within epizootic peak period of Cladosporium sp. to the population of Aleurocanthus spiniferus. Entomological Journal of East China, 40(1): [,,, , 40(1): ]

8 1 : 29 He L, Yang Y, Fu JZ, Wang JJ, Zhao ZM, Resistance selection and relative fitness of Tetranychus cinnabarinus (Boisduval) to abamectin. Journal of Plant Protection, 31(4): [,,,,, , 31(4): ] Jeyarani S, Banu JG, Ramaraju K, First record of natural occurrence of Cladosporium cladosporioides (Fresenius) de Vries and Beauveria bassiana (Bals. -Criv. ) Vuill on two spotted spider mite, Tetranychus urticae Koch from India. Journal of Entomology, 8(3): Liang ZC, Zeng ZT, Parasitization of Cladospirium cladosporioides (Fres) de Vries on citrus red mites. Journal of Microbiology, 20(1): [,, , 20(1): ] Li SQ, Zheng JR, Zhang Y, Identification and biological characterisation of a Cladosporium cladosporioides strain. Hangzhou Agricultural Science and Technology, (4): [,,, , (4): ] Li YY, Research on control of Tetranychus cinnabarinus experimental population with Beauveria bassiana Bb025. Master Dissertation. Chongqing: Southwest University. [, Bb025.. :.] MA XP, Gu Y, Liu L, Liu XM, Ling SS, Hou JF, Wang CD, Biological characterisation of Cladosporium cladosporioides isolated from Giant Panda. Chinese Veterinary Science, 43(1): 1 8. [,,,,,,, , 43(1): 1 8.] Pan WY, Chen SL, Application of Cladosprium cladosporioides against Chinese chestnut red scale, Kermococcus nawae. Chinese Entomogenous Research and Application, (3): [,, , (3): ] Shi WB, Microbial control of spider mites based on fungal biocontrol agents. Doctoral Dissertation. Hangzhou: Zhejiang University. [, :.] Wang J, Influence of Acremonium hansfordii infecting Tetranychus urticae Koch and related enzymes activity of its epidermis. Master Dissertation. Lanzhou: Gansu Agricultural University. [, :.] Xie QM, Liu JY, A preliminary study on the mildew shellac (Laccifer lacca). Journal of Fujian College of Forestry, 6(1): [,, , 6(1): ] Xie QM, Huang JH, A preliminary study on the Cladosporin. Journal of Fujian College of Forestry, 8(1): [,, , 8(1): ] Yang YL, Pu JJ, Zhang H, Wei YX, Li ZP, Liu XM, Biological characteristics of Cladosporium cladosporioides causing mango sooty blotch disease. Journal of Fruit Science, 32(2): [,,,,,, 杧., 32(2): ] Zhao YM, Research on control of wetland pine mealybug with Cladosporium cladosporioides. Plant Protection and Modern Agriculture-China Society of Plant Protection 2007 Annual Conference Proceedings. Guilin [, ]

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文章(copy) 农学学报 016,6(5):18- Journal of Agriculture www.caaj.org 5 种杀菌剂防治番茄茎腐病的室内筛选和田间使用效果任凤山 1, 高亮 1, 张博 ( 1 山东省农业科学院植物保护研究所 / 山东省植物病毒学重点实验室, 济南 50; 山东农