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

Peisheng QlN，Qin YANG，F(xiàn)uchang 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，F(xiàn)uchang 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

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青霉、木霉對(duì)平菇菌絲生長(zhǎng)的影響及消毒液的初篩

覃培升，楊琴，黃福常，劉斌*，黎金鋒 （廣西大學(xué)農(nóng)學(xué)院應(yīng)用微生物研究所，廣西南寧530005）

［目的］研究青霉、木霉對(duì)平菇菌絲生長(zhǎng)的影響，并篩選用于防控空氣及環(huán)境中病原菌的有效消毒液。［方法］在平菇生長(zhǎng)PDA培養(yǎng)基上接種霉菌開(kāi)展菌絲對(duì)峙試驗(yàn)，觀察相交時(shí)兩者菌絲生長(zhǎng)狀況。研究過(guò)氧化氫、新潔爾滅、84消毒液、來(lái)蘇爾、高錳酸鉀、75%醫(yī)用酒精對(duì)側(cè)耳木霉、綠色木霉、哈茨木霉、桔青霉、托姆青霉和短密青霉共6種青霉、木霉菌株的生長(zhǎng)抑制作用。［結(jié)果］菌絲對(duì)峙試驗(yàn)表明，青霉、木霉菌對(duì)平菇的危害主要有爭(zhēng)奪營(yíng)養(yǎng)、競(jìng)爭(zhēng)性殺傷菌絲、產(chǎn)生拮抗、侵襲纏繞穿透平菇營(yíng)養(yǎng)菌絲、產(chǎn)生毒素毒害生長(zhǎng)菌絲。6種消毒液對(duì)不同的霉菌抑制作用各有不同，均有一定的抑制，效果強(qiáng)弱與霉菌的不同屬有明顯的區(qū)別。新潔爾滅對(duì)6種霉菌的抑制作用差異顯著，3%過(guò)氧化氫、84消毒液、5%來(lái)蘇爾、高錳酸鉀和75%酒精對(duì)3種木霉作用差異不顯著，對(duì)3種青霉作用差異顯著。［結(jié)論］新潔爾滅對(duì)青霉木霉菌的預(yù)防抑制效果顯著，可作為生產(chǎn)中消毒防雜用藥使用。

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.

側(cè)耳屬食用菌優(yōu)良菌株篩選及標(biāo)準(zhǔn)化繁育技術(shù)研究——廣西側(cè)耳屬食用菌種質(zhì)資源評(píng)價(jià)及抗病蟲(chóng)新品種選育 （桂科攻1222012-1B）。

覃培升（1973-），男，廣西橫縣人，碩士，實(shí)驗(yàn)師，從事農(nóng)業(yè)微生物和食用菌實(shí)驗(yàn)教學(xué)科研工作，E-mail：faxsheng@gxu.edu.cn。*通訊作者，教授，博士生導(dǎo)師，從事菌物系統(tǒng)學(xué)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