烟草米根霉对6种杀菌剂的敏感性及菌株保存条件筛选
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摘要
采用孢子萌发法测定了烟草上米根霉Rhizopus oryzae对6种杀菌剂(代森锰锌、多菌灵、嘧霉胺、嘧菌酯、啶酰菌胺及氟啶胺)的敏感性,并就其适宜的保存条件进行了筛选,同时采用离体叶片法测定了上述6种杀菌剂对烟叶霉烂病的防治效果。结果表明:6种杀菌剂对米根霉孢子萌发和烟叶霉烂病均表现出了不同的抑制活性。其中,抑制孢子萌发活性最为明显的是氟啶胺和啶酰菌胺,其EC_(90)值分别为0.67和1.53 mg/L;其次为代森锰锌和嘧菌酯,15.16和17.66 mg/L;最弱为嘧霉胺和多菌灵,71.87和81.96 mg/L。对烟叶霉烂病防效最好的为嘧菌酯,50 mg/L处理的防效为85%;其次为啶酰菌胺,200 mg/L处理时防效为83%;氟啶胺的防效较差,1 000 mg/L处理时仅为48%;而代森锰锌、多菌灵和嘧霉胺在最高使用剂量(分别为4 000、800和800 mg/L)时防效均低于20%。病原菌保存方法筛选结果表明,保存后米根霉的孢子萌发率均发生了不同程度降低。其中,4℃保存于20%甘油组的孢子悬浮液萌发率为60%;4℃保存的孢子干样萌发率为36%;4℃保存的孢子悬浮液和–20℃保存于20%甘油的孢子悬浮液萌发率均低于20%;20℃保存的孢子干样萌发率为11%;–20℃保存的孢子悬浮液萌发率为6%。研究结果可为烘烤期烟叶霉烂病防治和米根霉孢子的保存提供参考依据。
The sensitivity of Rhizopus oryzae to six fungicides(mancozeb, carbendazim, pyrimethanil,azoxystrobin, boscalid, fluazinam) and the preservation conditions of this pathogen were determined using the conidia germination method. Efficacy of these fungicides against tobacco pole rot disease was also evaluated on the detached leaves. Results showed that all the six fungicides presented different inhibitory activities against both conidia germination and pole rot. For the conidia germination, the highest inhibitions were found in the cases with fluazinam and boscalid, with EC_(90) values of 0.67 mg/L and 1.53 mg/L, respectively. Lower inhibition rates were detected in the cases with mancozeb and azoxystrobin(15.16 and 17.66 mg/L). And the lowest inhibition rates were observed in the cases with pyrimethanil and carbendazim(71.87 and 81.96 mg/L). For the disease control efficacy, the highest inhibition rate was observed in the group treated by azoxystrobin at 50 mg/L with the efficacy of 85%.Lower inhibition rate was detected after the boscalid treatment, which showed 83% efficacy at200 mg/L, while a poor inhibition rate(48%) was observed by fluazinam treatment at the concentration of 1 000 mg/L. And mancozeb, carbendazim, pyrimethanil all exhibited efficacy of < 20% at their respectively highest dosages of 4 000, 800 and 800 mg/L. The screening results of six different pathogen preservation conditions showed that conidia germination ratio of R. oryzae decreased variously by using different methods. The conidia germination ratio of the conidial suspension in 20%glycerol was 60% at 4 ℃, and, for dry conidia samples, that was 36% at 4 ℃. In terms of the conidial suspension stored at 4 ℃ and the conidial suspension stored at –20 ℃, the germination ratios in 20%glycerol were both < 20%. With regard to the dry conidia samples stored at –20 ℃, the conidia germination ratio was 11%, and, for the conidial suspension stored at –20 ℃, that was 6%. Results of this study have provided reference and basis for both the chemical control of tobacco pole rot and the storage of R. oryzae.
The sensitivity of Rhizopus oryzae to six fungicides(mancozeb, carbendazim, pyrimethanil,azoxystrobin, boscalid, fluazinam) and the preservation conditions of this pathogen were determined using the conidia germination method. Efficacy of these fungicides against tobacco pole rot disease was also evaluated on the detached leaves. Results showed that all the six fungicides presented different inhibitory activities against both conidia germination and pole rot. For the conidia germination, the highest inhibitions were found in the cases with fluazinam and boscalid, with EC_(90) values of 0.67 mg/L and 1.53 mg/L, respectively. Lower inhibition rates were detected in the cases with mancozeb and azoxystrobin(15.16 and 17.66 mg/L). And the lowest inhibition rates were observed in the cases with pyrimethanil and carbendazim(71.87 and 81.96 mg/L). For the disease control efficacy, the highest inhibition rate was observed in the group treated by azoxystrobin at 50 mg/L with the efficacy of 85%.Lower inhibition rate was detected after the boscalid treatment, which showed 83% efficacy at200 mg/L, while a poor inhibition rate(48%) was observed by fluazinam treatment at the concentration of 1 000 mg/L. And mancozeb, carbendazim, pyrimethanil all exhibited efficacy of < 20% at their respectively highest dosages of 4 000, 800 and 800 mg/L. The screening results of six different pathogen preservation conditions showed that conidia germination ratio of R. oryzae decreased variously by using different methods. The conidia germination ratio of the conidial suspension in 20%glycerol was 60% at 4 ℃, and, for dry conidia samples, that was 36% at 4 ℃. In terms of the conidial suspension stored at 4 ℃ and the conidial suspension stored at –20 ℃, the germination ratios in 20%glycerol were both < 20%. With regard to the dry conidia samples stored at –20 ℃, the conidia germination ratio was 11%, and, for the conidial suspension stored at –20 ℃, that was 6%. Results of this study have provided reference and basis for both the chemical control of tobacco pole rot and the storage of R. oryzae.
引文
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[9]MIURA Y.Ten-year preservation of mucor,Rhizopus and Aspergillus cultures under liquid paraffim[J].J Agric Chem Soc Japan,1983,57:535-541.
[10]汪汉成,周明国,张艳军,等,戊唑醇对立枯丝核菌的抑制作用及在水稻上的应用[J].农药学学报,2007,9(4):357-362.WANG H C,ZHOU M G,ZHANG Y J,et al.Fungicidal activity of tebuconazole against Rhizoctonia solani and its application to rice[J].Chin J Pestic Sci,2007,9(4):357-362.
[11]WANG H C,WANG J,LI L C,et al.Metabolic activities of five botryticides against Botrytis cinerea examined using the Biolog FF MicroPlate[J].Sci Rep,2016,6:31025.
[12]SUN H Y, WANG H C,CHEN Y,et al.Multiple resistance of Botrytis cinerea from vegetable crops to carbendazim,diethofencarb,procymidone,and pyrimethanil in China[J].Plant Dis,2010,94(5):551-556.
[13]WANG H C,ZHOU M G,WANG J X,et al.Biological mode of action of dimethomorph on Pseudoperonospora cubensis and its systemic activity in cucumber[J].Agric Sci in China,2009,8(2):172-181.
[14]张凤英,吴国平,林丽萍,等.一种长期保存酿造醪糟用米根霉菌种的简易方法:CN 105886402A[P].2016.ZHANG F Y,WU G P, LIN L P,et al.A simple method for long term preservation of Rhizopus oryzae strains used for brewing fermention:CN 105886402A[P].2016.
[15]张玉琴,李青山,王传义,等.烤烟烟叶成熟过程中的颜色参数与烘烤特性研究[J].西南农业学报,2018(1):62-67.ZHANG Y Q,LI Q S,WANG C Y,et al.Study on color parameters and curing characteristics of flue-cured tobacco leaves in process of maturity[J].Southwest China J Agric Sci,2018(1):62-67.
[16]李润根,艾芬婷.龙牙百合软腐病病菌:米根霉的分离鉴定及杀菌剂室内毒力测定初报[J].中国植保导刊,2017,37(11):10-14.LI R G,AI F T.Isolation,identification and inhibition test of Rhizopus oryzae from soft rot disease on Longya lily[J].China Plant Prot,2017,37(11):10-14.
[17]刘艳祥.新疆红枣果实两种真菌病害病原鉴定及关键防治技术研究[D].乌鲁木齐:新疆农业大学,2016:1-52.LIU Y X.Research between pathogens identification and key control techniques of two jujube fruit fungal diseases in Xinjiang[D].Wulumuqi:Xinjiang Agric Univ,2016:1-52.
[2]曾婷英,顾钢,张绍升.烘烤期烟叶霉烂病的病原鉴定[J].中国烟草学报,2014,20(4):65-68.ZENG T Y,GU G,ZHANG S S.Pathogen identification of tobacco leaf mildew rot during flue-curing[J].Acta Tabacaria Sinica,2014,20(4):65-68.
[3]KORTEKAMP A,SCHMIDTKE M,SERR A.Infection and decay of tobacco caused by Rhizopus oryzae/Die Infektion und Faulnis von Tabak verursacht durch Rhizopus oryzae[J].Zeitschrift Fur Pflanzenkrankheiten Und Pflanzenschutz,2003,1 10(6):535-543.
[4]SURHONE L M,TIMPLEDON M T,MARSEKEN S F.Rhizopus oryzae[M].Betascript Publishing,2010:1-116.
[5]汪汉成,张敏,张长青,等.烟叶米根霉检测、碳源代谢表型及其对7种杀菌剂的敏感性[J].农药学学报,2018,20(6):743-748.WANG H C,ZHANG M,ZHANG C Q,et al.Pathogen detection,carbon metabolic phenotype analysis of Rhizopus oryzae from tobacco and its sensitivity to seven fungicides[J].Chin J Pestic Sci,2018,20(6):743-748.
[6]TURNER T L,KIM E,HWANG C,et al.Short communication:conversion of lactose and whey into lactic acid by engineered yeast[J].J Dairy Sci,2017,100(1):124-128.
[7]HUANG D,WANG R,DU W J,et al.Activation of glycerol metabolic pathway by evolutionary engineering of Rhizopus oryzae to strengthen the fumaric acid biosynthesis from crude glycerol[J].Bioresour Technol,2015,196(11):263-272.
[8]LEZHNEVA M L,PETRASH I P,KOVAL'SKAIA L P.Effect of the incubation temperature on the growth and spore formation of Penicillium italicum and Rhizopus nigricans treated with ionizing radiations[J].Prikl Biokhim Mikrobiol,1973,9(5):664-670.
[9]MIURA Y.Ten-year preservation of mucor,Rhizopus and Aspergillus cultures under liquid paraffim[J].J Agric Chem Soc Japan,1983,57:535-541.
[10]汪汉成,周明国,张艳军,等,戊唑醇对立枯丝核菌的抑制作用及在水稻上的应用[J].农药学学报,2007,9(4):357-362.WANG H C,ZHOU M G,ZHANG Y J,et al.Fungicidal activity of tebuconazole against Rhizoctonia solani and its application to rice[J].Chin J Pestic Sci,2007,9(4):357-362.
[11]WANG H C,WANG J,LI L C,et al.Metabolic activities of five botryticides against Botrytis cinerea examined using the Biolog FF MicroPlate[J].Sci Rep,2016,6:31025.
[12]SUN H Y, WANG H C,CHEN Y,et al.Multiple resistance of Botrytis cinerea from vegetable crops to carbendazim,diethofencarb,procymidone,and pyrimethanil in China[J].Plant Dis,2010,94(5):551-556.
[13]WANG H C,ZHOU M G,WANG J X,et al.Biological mode of action of dimethomorph on Pseudoperonospora cubensis and its systemic activity in cucumber[J].Agric Sci in China,2009,8(2):172-181.
[14]张凤英,吴国平,林丽萍,等.一种长期保存酿造醪糟用米根霉菌种的简易方法:CN 105886402A[P].2016.ZHANG F Y,WU G P, LIN L P,et al.A simple method for long term preservation of Rhizopus oryzae strains used for brewing fermention:CN 105886402A[P].2016.
[15]张玉琴,李青山,王传义,等.烤烟烟叶成熟过程中的颜色参数与烘烤特性研究[J].西南农业学报,2018(1):62-67.ZHANG Y Q,LI Q S,WANG C Y,et al.Study on color parameters and curing characteristics of flue-cured tobacco leaves in process of maturity[J].Southwest China J Agric Sci,2018(1):62-67.
[16]李润根,艾芬婷.龙牙百合软腐病病菌:米根霉的分离鉴定及杀菌剂室内毒力测定初报[J].中国植保导刊,2017,37(11):10-14.LI R G,AI F T.Isolation,identification and inhibition test of Rhizopus oryzae from soft rot disease on Longya lily[J].China Plant Prot,2017,37(11):10-14.
[17]刘艳祥.新疆红枣果实两种真菌病害病原鉴定及关键防治技术研究[D].乌鲁木齐:新疆农业大学,2016:1-52.LIU Y X.Research between pathogens identification and key control techniques of two jujube fruit fungal diseases in Xinjiang[D].Wulumuqi:Xinjiang Agric Univ,2016:1-52.