Pioli R N, Morandi E N, Gosparini C O, Borghi A L
Fisiología Vegetal, Facultad de Ciencias Agrarias.
Micología, Facultad Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, P.O. Box 14, 2123 Zavalla, Santa Fe, Argentina.
Plant Dis. 1999 Nov;83(11):1071. doi: 10.1094/PDIS.1999.83.11.1071B.
The objective of this study was to characterize the pathogenicity of several local isolates of Diaporthe phaseolorum (Cooke & Ellis) Sacc. var. meridionalis Fernández and its anamorph, Phomopsis phaseoli (Desmaz.) Sacc. meridionalis Morgan-Jones, the causal agent of southern stem canker of soybean (Glycine max (L.) Merr.), in soybean lines carrying major resistance genes. Soybean plants with typical stem canker symptoms were collected during the 1996 to 1997 and 1997 to 1998 growing seasons in the central and southern areas of Santa Fe Province, Argentina. The pathogen was isolated from the internal tissues of infected stems, cultured on potato glucose agar acidified with 0.2% lactic acid (APGA), amended with streptomycin at 100 mg/liter, and maintained in the dark at 25 ± 1°C. Isolates were characterized based on the morphology of colonies, perithecia, and pycnidia and measurement of asci, bicellular, biguttulate ascospores, and alpha conidia (1). Soybean cultivars used to assay pathogenicity included Tracy M (Rdc1 and Rdc2 genes), Isoline I (Tracy Misoline with only the Rdc1 gene), Isoline II (Tracy M isoline with only the Rdc2 gene), Crockett (Rdc3 gene), Hutchinson (Rdc4 gene), and RA 702 (susceptible cultivar). Hypocotyls of 14-day-old seedlings grown in the greenhouse were inoculated by the toothpick method. Four replicates of nine seedlings each were used. Seedlings punctured with sterile toothpicks served as controls. The experiment was repeated twice with similar results. The D. phaseolorum var. meridionalis isolates assayed and their collection locations were Dpm1 (Malabrigo), Dpm2 (Los Molinos), Dpm3 (San Justo), Dpm5 (Oliveros), Dpm6 (San Jerónimo), and Dpm7 (Clarke). Twenty-eight days after inoculation, stem canker reactions were measured as the percentage of dead plants. The pathogen was reisolated from stems of randomly chosen symptomatic plants on day 14 after inoculation. These plants were included in the calculation of the percentage of dead plants. In control plants, lesions were not detected, and mycelial growth did not occur from stem portions plated on APGA. Tracy M and RA 702 had 0 to 7% dead plants and 70 to 95% dead plants, respectively, with all assayed isolates. Cultivars with single resistance genes reacted differently to various isolates. Isolates Dpm1 and Dpm3 caused little or no stem canker (<10% dead plants) on all cultivars with resistance genes. Isolates Dpm2 and Dpm6 killed 56 and 52%, respectively, of Isoline II (Rdc2 gene) plants. Isolates Dpm2 and Dpm7 killed 25% of cv. Hutchinson (Rdc4 gene) and Isoline I (Rdc1 gene) plants, respectively. Isolate Dpm5 killed <12% of plants with genes Rdc1, Rdc2, or Rdc3. The reaction of isolate Dpm5 with Hutchinson (Rdc4 gene) was not evaluated. The pathogenic diversity of these isolates of D. phaseolorum var. meridionalis may have been induced by the wide diffusion of resistant host cultivars (2). References: (1) F. A. Fernández and R. T. Hanlin. Mycologia 88:425, 1996. (2) A. W. Zhang et al. Phytopathology 88:1306, 1998.
本研究的目的是鉴定菜豆间座壳(Diaporthe phaseolorum (Cooke & Ellis) Sacc. var. meridionalis Fernández)及其无性型菜豆拟茎点霉(Phomopsis phaseoli (Desmaz.) Sacc. meridionalis Morgan-Jones)的几个本地分离株的致病性,它们是大豆(Glycine max (L.) Merr.)南方茎溃疡病的病原菌,对携带主要抗性基因的大豆品系进行研究。在1996至1997年以及1997至1998年生长季期间,于阿根廷圣菲省中部和南部地区收集了具有典型茎溃疡症状的大豆植株。从受感染茎的内部组织分离病原菌,在添加了100毫克/升链霉素并用0.2%乳酸酸化的马铃薯葡萄糖琼脂(APGA)上培养,并在25±1°C黑暗条件下保存。根据菌落、子囊壳和分生孢子器的形态以及对子囊、双细胞、具双油球的子囊孢子和α分生孢子的测量对分离株进行鉴定(1)。用于测定致病性的大豆品种包括特雷西M(Rdc1和Rdc2基因)、同型系I(仅含Rdc1基因的特雷西M同型系)、同型系II(仅含Rdc2基因的特雷西M同型系)、克罗克特(Rdc3基因)、哈钦森(Rdc4基因)和RA 702(感病品种)。通过牙签接种法对接种在温室中生长14天的幼苗的下胚轴进行接种。每个品种使用9株幼苗,重复4次。用无菌牙签穿刺伤的幼苗作为对照。该实验重复两次,结果相似。所测定的菜豆间座壳变种南方分离株及其采集地点分别为Dpm1(马拉布里戈)、Dpm2(洛斯莫利诺斯)、Dpm3(圣胡斯托)、Dpm5(奥利韦罗斯)、Dpm6(圣赫罗尼莫)和Dpm7(克拉克)。接种28天后,以死株百分比来测定茎溃疡反应。在接种后第14天从随机选择的有症状植株的茎中重新分离病原菌。这些植株被纳入死株百分比的计算。在对照植株中,未检测到病斑,接种在APGA上的茎段也未出现菌丝生长。对于所有测定的分离株,特雷西M和RA 702的死株率分别为0至7%和70至95%。具有单个抗性基因的品种对不同分离株的反应不同。分离株Dpm1和Dpm3对所有具有抗性基因的品种造成的茎溃疡很少或没有(死株率<10%)。分离株Dpm2和Dpm6分别使同型系II(Rdc2基因)植株的56%和52%死亡。分离株Dpm2和Dpm7分别使哈钦森品种(Rdc4基因)和同型系I(Rdc1基因)植株的25%死亡。分离株Dpm5使具有Rdc1、Rdc2或Rdc3基因的植株死亡比例<12%。未评估分离株Dpm5对哈钦森(Rdc4基因)的反应。这些菜豆间座壳变种南方分离株的致病多样性可能是由抗性寄主品种的广泛传播所诱导的(2)。参考文献:(1)F. A. Fernández和R. T. Hanlin。《真菌学》88:425,1996。(2)A. W. Zhang等人。《植物病理学》88:1306,1998。