Effects of Penicillium spp.and Trichoderma spp. on Pleurotus ostreatus Growth and Screening of Effective Disinfectants

Peisheng QlN，Qin YANG，Fuchang HUANG，Bin LlU，Jinfeng LlInstitute of Applied Microbiology，College of Agriculture，Guangxi University，Nanning 530005，China

Effects of Penicillium spp.and Trichoderma spp. on Pleurotus ostreatus Growth and Screening of Effective Disinfectants

Peisheng QlN，Qin YANG，Fuchang HUANG，Bin LlU*，Jinfeng Ll
Institute of Applied Microbiology，College of Agriculture，Guangxi University，Nanning 530005，China

［Objective］This study was conducted to investigate the effects of Penicillium spp.and Trichoderma spp.on the growth of Pleurotus ostreatus （oyster mushroom）and to screen effective disinfectants to control them.［Method］Six species of Penicillium and Trichoderma were inoculated into PDA plates growing with P.ostreatus to observe the growth of their mycelia.And then the inhibitory effects of hydrogen peroxide，bromogeramine bromide，84 disinfectant，lysol，potassium permanganate，and 75%medical alcohol on Trichoderma pleuroticola，Trichoderma viride，Trichoderma harzianum，Penicillium citrinum，Penicillium thomii and Penicillium brevicompactum were detected.［Result］Confrontation test revealed that Penicillium spp. and Trichoderma spp.competed for nutrients，antagonized against the growth of P.ostreatus mycelia，invaded and twined around P.ostreatus mycelia，and also produced toxins poisoning P.ostreatus mycelia.The six disinfectants exhibited different inhibitory effects against different species of Penicillium and Trichoderma.Among them the inhibitory effects of bromogeramine bromide against the six species of Penicillium and Trichoderma were significantly different.However，the inhibitory effects of 3%hydrogen peroxide，84 disinfectant，5%lysol，potassium permanganate and 75%medical alcohol were significantly different among Penicillium spp.，but not significantly different among Trichoderma spp.［Conclusion］Bromogeramine bromide can be used to control the bacteria and fungi in mushroom production for it has significant inhibitory effects on the six species of both Penicillium and Trichoderma.

Penicillium；Trichoderma；Pleurotus ostreatus；Disinfectant

Materials and Methods

Pleurotus ostreatus strains，Penicillium spp.and Trichoderma spp. to be tested

Pleurotus ostreatus strains P015，P012，P615，P39，P558 and P650 were provided by Institute of Applied Microbiology，College of Agriculture，Guangxi University. Trichoderma pleuroticola，Trichoderma viride，Trichoderma harzianum，Penicillium citrinum，Penicillium thomii and Penicillium brevicompactum were also isolated by the institute.

Disinfectants

（A）3%Hydrogen peroxide solution was a product of Shandong Lerkam Disinfection Technology Co.，Ltd （voluntary standard Q/3714AL× 29）.（B）Benzalkonium bromide was also produced by Shandong Lerkam Disinfection Technology Co.，Ltd（voluntary standard Q/1400AL×004，original concentration of 2.7-3.3 g/L）and diluted 15 times with sterile water.（C）84 Disinfectant was produced by Nanchang Huaxin Pharmaceutical Chemical Co.，Ltd.（chlorine concentration of 4.0%-7.0%，active ingredient:sodium hypochlorite，voluntary standard:Q/NH×010-2008）and diluted 100 times with sterile water.（D）5%Lysol was produced by Jiangxi Caoshanhu Disinfection Co.，Ltd.（cresol concentration of 5.0%，voluntary standard ISO9001:Q/JCSH006-2010:2008）and diluted 100 times with sterile water.（E）Potassium permanganate was produced by Shantou Xilong Chemical Co.，Ltd.（voluntary standard:GB/T643-2008）and diluted500 times with sterile water.（F）75% alcohol was self-made.

Table 1 Influence of six disinfectants on mycelial growth of T.pleuroticola

Mycelium confrontation test

Pleurotus ostreatus strains were activated and inoculated into PDA plates before being cultivated in an incubator at 25℃ for 2 d.Meanwhile，mycelium discs of six mold species with diameter of 5 were punched and inoculated to above PDA plates，3-5 cm away from P.ostreatus mycelia，cultured at 25℃ to observe the mycelial growth of both mold and mushroom in plates，especially the wilt，dissolution and winding of their mycelia.

Test on inhibitory effects of disinfectants on mycelial growth of Penicillium spp.and Trichoderma spp.

The Penicillium spp.and Trichoderma spp.were activated and inoculated into PDA plates，cultivated in an incubator at 25℃for 1 d.Sterile filter paper discs（5 diameter）were dipped in disinfectant solutions and then placed near Penicillium spp.and Trichoderma spp.colonies in PDA plates［12］.Three repetitions were set for each treatment.

Statistical methods

After 24 h of culture at 25℃，colony diameter of the six mold species was measured by crossing method.Least significant range（LSR）values were calculated for statistical analysis［13］.

Results and Analysis

Influence of six species of Penicillium and Trichoderma on the growth of P.ostreatus mycelia

As shown in Fig.1，T.pleuroticola grew rapidly，produced a small amount of spores，directly twined and covered P.ostreatus mycelia；no antagonistic streak was observed from the bottom of plate.Meanwhile，P.ostreatus mycelia could not resist the invasion of T.pleuroticola，thus they were absorbed，melted and finally disappeared.T.viride also had strong potential to invade，grew rapidly and produced a large amount of spores and finally covered P.ostreatus mycelia；no antagonistic streak was observed from the bottom of plate.P.ostreatus mycelia could not resist the invasion of T.viride，so they were absorbed，melted and finally disappeared.T.harzianum also had strong potential to invade，grew rapidly and produced a large amount of spores and finally covered P.ostreatus mycelia；pale yellow antagonistic streak was observed from the bottom of plate.P.ostreatus mycelia had certain resistance to T.viride，but the wrapped P.ostreatus mycelia wilted，died and were finally absorbed.P.citrinum mycelia grew slowly，spread through spores，and had a strong smell.White antagonistic streak formed between the mycelia of P.citrinum and P.ostreatus，where the growth of P.ostreatus mycelia was inhibited.P.ostreatus mycelia had resistance to P.citrinum.P.thomii mycelia grew slowly，and produced a large amount of spores. White antagonistic streak was observed between the mycelia of P. thomii and P.ostreatus，where the growth of P.ostreatus mycelia was inhibited.From the bottom of plate，it was observed that the edge of P. thomii mycelia was pale yellow and covered P.ostreatus mycelia.P.brevicompactum mycelia grew slowly and produced a large amount of spores，while the growth of P.ostreatus mycelia was reduced.White antagonistic streak was observed between the mycelia of P.drevicompactum and P.ostreatus，where the mycelia of both P.brevicompactum and P.ostreatus was hindered.

To sum up，Penicillium spp.and Trichoderma spp.competed for nutrients with P.ostreatus，antagonized against the growth of P.ostreatus mycelia，invaded and twined around P.ostreatus mycelia，and also produced toxins poisoning P.ostreatus.

Table 5 Influence of six disinfectants on mycelial growth of P.thomii

Table 6 Influence of six disinfectants on mycelial growth of P.brevicompactum

Influence of six disinfectants on mycelial growth of Penicillium spp. and Trichoderma spp.

It could be concluded from Table 1-Table 6，the six disinfectants exhibited different inhibitory effects against different species of Penicillium and Trichoderma.Among them the inhibitory effects of bromogeramine bromide against the six species of Penicillium and Trichoderma were significantly different；3%hydrogen peroxide，84 disinfectant，5%Lysol，potassium permanganate and 75%medical alcohol showed inhibitory effects on all of the six mold species，but their inhibitory effects were significantly different among Penicillium spp.，but not significantly different among Trichoderma spp.

Discussion

The dual culture of the six coon P. ost reatus strains and the six mold species revealed that Penicillium spp. and Trichoderma spp.had significantly different pathogenicity against P.ostreatus.The three species of genus Trichoderma:T.pleuroticola，T.viride and T. harzianum were highly pathogenic to P.ostreatus，as they inhibited the mycelial growth of P.ostreatus at early stage and then almost covered P.ostreatus mycelia，so that P.ostreatus mycelia wilted and finally died.The three species of genus Penicillium:P.citrinum，P.thomii and P.brevicompactum did not show strong pathogenicity because of their mycelia growth limitations，but they also inhibited the mycelia growth of P.ostreatus.Obvious antagonistic streak was observed between the mycelia of Penicillium spp.and P.ostreatus，where their mycelia wrinkled and wilted.Above results prove that Trichoderma spp.have serious harm and may cause large economic loss to oyster mushroom production.

Among the six disinfectants we tested，dromogeramine bromide has significantly different inhibitory effects on the six species of Penicillium and Trichoderma，and thus can be used for the sterilization of environment，mushroom strains，tools and labors in the production of edible mushroom.

References

［1］HUANG NL（黄年来）.Mushroom pest and disease control:a color handbook［食用菌病虫害诊治 （彩色）手册］［M］. Beijing:China Agriculture Press（北京:中国农业出版社），2001.

［2］QIN PS（覃培升），LIU B（刘斌），LI JF（黎金锋），et al.Isolation，identification and characteristics of Penicillium and Trichoderna from polluted oyster mushroom（平菇培养料青、绿霉菌的分离鉴定及特性）［J］.Guangdong Agricultural Sciences（广东农业科学），2013，4:57-60.

［3］WUXP（吴小平）.Identification，pathogenic mechanism and control of Trichoderma spp.（食用菌致病木霉的鉴定、致病机理及防治研究）［D］.Fujian:Fujian Agriculture and Forestry University（福建:福建农林大学），2008.

［4］MA L（马林），SONG JD（宋金俤），QU SX（曲绍轩）.Effects of six biocides on growth of mushroom and bacteria（六种生物杀菌剂对平菇及杂菌生长的影响）［J］.Edible Fungi（食用菌），2010（3）:58-59.

［5］MA L（马林），SONG JD（宋金俤），QU SX（曲绍轩），et al.Inhibitory effects of six biocides on growth of oyster mushroom and mildew（不同生物杀菌剂在袋栽平菇中防治竞争性霉菌试验）［J］.Edible Fungi（食用菌），2010（1）:52-53.

［6］SONG JD（宋金俤），HUA XH（华秀红），LIU C（刘超），et al.Screening of bactericides to edible fugus Trichoderma viride and Mycogone perniciosa Magn（食用菌绿色木霉及湿泡病高效杀菌剂配方筛选）［J］.Edible Fungi of China（中国食用菌），2007，26（6）:47-50.

［7］WANG HY（王慧阳）.Comparative Study on the effect of inhibitory mould in seven disinfectants of edible mushrooms（七种食用菌消毒剂抑制霉菌效果的比较研究）［J］.Edible Fungi of China（中国食用菌），2006，25（3）:29-31.

［8］WANG ZS（王宗善），WU MG（武摸戈）. Effect of 4 fungicide on Trichoderma vider and the growth of Flaulina velutiper mycelium （四种杀菌剂对绿霉及金

（Continued on page 473）

青霉、木霉对平菇菌丝生长的影响及消毒液的初筛

覃培升，杨琴，黄福常，刘斌*，黎金锋 （广西大学农学院应用微生物研究所，广西南宁530005）

［目的］研究青霉、木霉对平菇菌丝生长的影响，并筛选用于防控空气及环境中病原菌的有效消毒液。［方法］在平菇生长PDA培养基上接种霉菌开展菌丝对峙试验，观察相交时两者菌丝生长状况。研究过氧化氢、新洁尔灭、84消毒液、来苏尔、高锰酸钾、75%医用酒精对侧耳木霉、绿色木霉、哈茨木霉、桔青霉、托姆青霉和短密青霉共6种青霉、木霉菌株的生长抑制作用。［结果］菌丝对峙试验表明，青霉、木霉菌对平菇的危害主要有争夺营养、竞争性杀伤菌丝、产生拮抗、侵袭缠绕穿透平菇营养菌丝、产生毒素毒害生长菌丝。6种消毒液对不同的霉菌抑制作用各有不同，均有一定的抑制，效果强弱与霉菌的不同属有明显的区别。新洁尔灭对6种霉菌的抑制作用差异显著，3%过氧化氢、84消毒液、5%来苏尔、高锰酸钾和75%酒精对3种木霉作用差异不显著，对3种青霉作用差异显著。［结论］新洁尔灭对青霉木霉菌的预防抑制效果显著，可作为生产中消毒防杂用药使用。

青霉、木霉；平菇；消毒液

D uring the growth of Pleurotus ostreatus（oyster mushroom），the synthetic nutrients are suitable for growing not only mushroom，but also Penicillium spp.and Trichoderma spp.，both of which are the main fungi causing damage to the growth of mushroom［1］.The fungi grow rapidly，compete for nutrients and place with mushroom，and secrete various toxins which inhibit the development of mycelia and fruiting bodies，reducing mushroom quantity and quality.The critical step is to control the pollution source of Penicillium spp. and Trichoderma spp.，as well as pathogens in air and environment.In the present study，confrontation tests between six species of Penicillium and Trichoderma isolated in mushroom production with six P.ostreatus strain coonly grown by local farmers were carried out［2］to investigate their pathogenicity to P.ostreatus and P.ostreatus mycelia growth after their attack. According to previous study，fungicides were usually used for the control of Penicillium spp.and Trichoderma spp.［3］.In present study，the inhibitory effects of six effective and environmentally friendly disinfectants hydrogen peroxide，bromogeramine bromide，84 disinfectant，lysol，potassium permanganate 75%medical alcohol on Penicillium spp.and Trichoderma spp.were detected to screen a disin-fectant for the prevention and control of pathogens in air and environment in mushroom production.

侧耳属食用菌优良菌株筛选及标准化繁育技术研究——广西侧耳属食用菌种质资源评价及抗病虫新品种选育 （桂科攻1222012-1B）。

覃培升（1973-），男，广西横县人，硕士，实验师，从事农业微生物和食用菌实验教学科研工作，E-mail：faxsheng@gxu.edu.cn。*通讯作者，教授，博士生导师，从事菌物系统学研究，E-mail：liubin@gxu.edu.cn。

2014-12-10

2015-02-03

Supported by Standardized Breeding Technology Research for Screening Excellent Edible Fungus Species of Pleurotus-Evaluation of Guangxi Provincial Edible Fungus Germplasms of Pleurotus and Breeding of New Cultivars Resistant to Diseases and Pests（1222012-1B）.

.E-mail:liubin@gxu.edu.cn

December 10，2014Accepted:February 3，2015