Sallato B V, Latorre B A, Aylwin G
Facultad de Agronomía e Ingeniería Forestal, Pontificia Universidad Católica de Chile, Casilla 306-22, Santiago Chile.
Frutera San Fernando (FRUSAN), San Fernando, Chile.
Plant Dis. 2006 Mar;90(3):375. doi: 10.1094/PD-90-0375A.
Resistance to the QoI fungicide (kresoxim-methyl) was detected in Venturia inaequalis (Cooke) Winter isolates from apple scab lesions collected from apple orchards in Chile after 5 to 6 years of commercial use of QoI fungicides. To reduce or prevent resistance development, QoI fungicides have been applied in blocks of two or three consecutive applications followed by an application of a fungicide with a different mode of action. However, the efficacy of kresoxim-methyl declined dramatically, providing almost no disease control in several orchards under high scab pressure in southern Chile, and resistance to the QoI fungicides was suspected. Conidia from at least five sporulating leaf scab lesions were collected from each of 10 and 14 orchards in 2003 and 2004, respectively. Fungicide sensitivity was determined by using a conidial germination test on potato dextrose agar acidified with 0.5 ml/liter of 95% lactic acid (APDA) containing kresoxim-methyl (Stroby 50 SC) at 0.0, 0.01, 0.1, and 1.0 μg/ml. The kresoxim-methyl concentration (0.01 μg/ml) has been used as a discriminatory dose in previous resistance studies (2). Germination was assessed for at least 100 conidia at each inhibitor concentration after 18 to 24 h of incubation at 20°C (1,2). Sensitivities were expressed as the percentage of germination relative to that on unamended APDA (1,2). These assays were repeated and all isolates with 50% effective dose (ED) at >0.1 μg/ml were considered resistant. Isolates from eight of 10 and two of 14 orchards were resistant to kresoxim-methyl in 2003 and 2004, respectively. Relative germination varied from 42.4 to 100% for isolates from resistant orchards on 0.1 μg/ml of kresoxim-methyl, with eight isolates having ED at >0.1 μg/mL. Conidia from isolates obtained in 2003 also germinated on APDA amended with 1.0 μg/ml of kresoxim-methyl, with two isolates having ED at >1.0 μg/mL. Relative to the mean ED of 0.02 μg/ml of kresoxim-methyl determined for relative germination in North America baseline population of V. inaequalis (2), resistance factors for these Chilean isolates were estimated to range from 5 to 50. Conidia of resistant isolates continued to germinate in the presence of 0.1 μg/ml of kresoxim-methyl when 100 μg/ml of salicylhydroxamic acid was added to the amended APDA, suggesting that an alternative oxidase pathway was not responsible for the in vitro results obtained (2). A G143A target mutation has been detected in a V. inaequalis isolate from Chile, although no details concerning this finding were reported (Fungicide Resistance Action Committee, online publication, 2004 [ http://www.frac.info ]). The poor control of apple scab with QoI fungicides in Chilean orchards appears to be due to the development of resistant strains of V. inaequalis that occurred despite the limited use of these materials in blocks of two or three consecutive applications, as initially suggested for resistance management. Farmers have been advised to make only one application of QoI fungicide per year in a tank mixture with another type of fungicide or even to abandon the use of QoI fungicides for apple scab control. To our knowledge, this is the first report of a severe outbreak of apple scab due to the presence of populations of V. inaequalis resistant to QoI fungicides under commercial conditions in Chile. References: (1) R. B. Küng Färber et al. Pest Manage. Sci. 58:261, 2002. (2) G. Olaya and W. Köller. Plant Dis. 83:274, 1999.
在智利苹果园,醌类(QoI)杀菌剂(醚菌酯)商业化使用5至6年后,从苹果黑星病病斑分离得到的苹果黑星病菌(Venturia inaequalis (Cooke) Winter)菌株中检测到了对该杀菌剂的抗性。为减少或预防抗性的产生,QoI杀菌剂采用连续两到三次施用的方式,随后施用一种具有不同作用模式的杀菌剂。然而,醚菌酯的药效急剧下降,在智利南部黑星病发病压力高的几个果园中几乎无法控制病害,怀疑存在对QoI杀菌剂的抗性。2003年和2004年分别从10个和14个果园中,从至少5个产生孢子的叶斑病斑采集分生孢子。通过在添加了0.5毫升/升95%乳酸酸化的马铃薯葡萄糖琼脂(APDA)上进行分生孢子萌发试验来测定杀菌剂敏感性,APDA中含有浓度为0.0、0.01、0.1和1.0微克/毫升的醚菌酯(翠贝50 SC)。醚菌酯浓度(0.01微克/毫升)在之前的抗性研究中被用作鉴别剂量(2)。在20°C下培养18至24小时后,在每个抑制剂浓度下对至少100个分生孢子的萌发情况进行评估(1,2)。敏感性以相对于未添加药剂的APDA上的萌发率的百分比表示(1,2)。重复这些试验,所有在>0.1微克/毫升时有效剂量(ED)为50%的分离株被视为抗性株。2003年和2004年,分别从10个果园中的8个和14个果园中的2个果园分离得到的菌株对醚菌酯具有抗性。对于来自抗性果园的分离株,在0.1微克/毫升醚菌酯上的相对萌发率在42.4%至100%之间,有8个分离株的ED>0.1微克/毫升。2003年获得的分离株的分生孢子在添加了1.0微克/毫升醚菌酯的APDA上也能萌发,有2个分离株的ED>1.0微克/毫升。相对于在北美苹果黑星病菌基线群体中测定的醚菌酯相对萌发的平均ED为0.02微克/毫升(2),这些智利分离株的抗性因子估计在5至50之间。当向添加药剂的APDA中添加100微克/毫升水杨羟肟酸时,抗性分离株的分生孢子在0.1微克/毫升醚菌酯存在下仍能继续萌发,这表明交替氧化酶途径与体外试验结果无关(2)。在一株来自智利的苹果黑星病菌分离株中检测到了G143A靶标突变,尽管未报告该发现的详细情况(杀菌剂抗性行动委员会,在线出版物,2004 [http://www.frac.info])。智利果园中QoI杀菌剂对苹果黑星病防治效果不佳似乎是由于苹果黑星病菌抗性菌株的产生,尽管如最初为抗性管理所建议的那样,这些药剂以连续两到三次施用的方式使用有限。已建议果农每年仅将QoI杀菌剂与另一种类型的杀菌剂混合使用一次,甚至放弃使用QoI杀菌剂来防治苹果黑星病。据我们所知,这是智利商业条件下因存在对QoI杀菌剂具有抗性的苹果黑星病菌群体而导致苹果黑星病严重爆发的首次报道。参考文献:(1)R. B. Küng Färber等人,《有害生物管理科学》58:261,2002。(2)G. Olaya和W. Köller,《植物病害》83:274, 1999。