Ellis M L, Arias M M Díaz, Jimenez D R Cruz, Munkvold G P, Leandro L F
Department of Plant Pathology and Microbiology, Iowa State University, Ames 50011.
Plant Dis. 2013 Feb;97(2):284. doi: 10.1094/PDIS-07-12-0644-PDN.
During 2007 to 2009, symptomatic and asymptomatic soybean plants were collected from fields in 18 Iowa counties. Fusarium isolates were recovered from surface-sterilized root tissue on peptone PCNB agar (2). Single-spore isolates were transferred to synthetic low nutrient agar (SNA) overlain with pieces (1 × 2 cm) of sterile filter paper, and to potato dextrose agar (PDA), and placed in the dark for 10 to 14 days for morphological identification (4). Twenty-three isolates were identified as Fusarium commune K. Skovg., O'Donnell & Nirenberg, previously in the F. oxysporum species complex (4). Colonies on PDA had white, fluffy, aerial mycelium with magenta to violet pigmentation in the medium. On SNA, macroconidia, chlamydospores, and microconidia on monophialides and polyphialides were consistent with the species description (4). Identification of all 23 isolates was confirmed by DNA sequencing of the translation elongation factor (EF1-α) gene, using ef1 and ef2 primers, and the mitochondrial small subunit (mtSSU), using primers MS1 and MS2 (4) [GenBank accessions for two representative isolates: EF1-α (JX289892 and JX289893), and mtSSU (JX289894 and, JX289895)]. Pathogenicity of two representative isolates of F. commune was tested on soybean (cv. AG2403) in a greenhouse, in water baths set at 18°C, using autoclaved soil mixed with infested sand-cornmeal inoculum (3). The experiment entailed a completely randomized design (CRD) with five replications (single plant/150 ml cone) per treatment, and was conducted three times. Dry root and shoot weights, and root rot severity (visual estimate of percent root rot on the entire root system) were evaluated after 6 weeks. Mean seedling emergence in soil infested with F. commune was 47 and 40% for the two isolates; in contrast, non-inoculated control plants had 100% emergence. There were significant differences in root (P < 0.0001) and shoot (P < 0.0001) weights, and root rot severity (P < 0.0001), between inoculated and non-inoculated plants. Seedlings that emerged were severely stunted and had dark brown lesions. F. commune was reisolated from infected roots of inoculated plants, but not from non-inoculated plants. Pathogenicity of both isolates to soybean (cv. MN1805) was also tested using a petri dish assay, in which eight seeds were placed on a plate with a 4-day-old culture growing on 2% water agar (1). Plates were rated 7 days later for seed germination, seed rot, and lesion development, using an ordinal scale (1). The experiment entailed a CRD with three replicate plates/treatment, and was conducted three times. Germination of inoculated seeds ranged from 37.5 to 75.0%, and germinated seedlings had dark brown lesions on the taproots. There was a significant difference between isolates in the petri dish assay (P = 0.0030); one isolate was less aggressive, but both isolates resulted in significantly more disease than on the non-inoculated control plants, which had 100% germination and no symptoms (P < 0.0001). F. oxysporum is a known soybean pathogen (1), but isolates of F. commune may have been misidentified as F. oxysporum in previous studies. To our knowledge, this is the first report of F. commune as a pathogen on soybean in the U.S.A. References: (1) K. E. Broders et al. Plant Dis. 91:727, 2007. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, UK, 2006. (3) G. P. Munkvold and J. K. O'Mara. Plant Dis. 86:143, 2002. (4) K. Skovgaard et al. Mycologia. 94:630, 2003.
2007年至2009年期间,从爱荷华州18个县的农田中采集了有症状和无症状的大豆植株。在蛋白胨PCNB琼脂培养基(2)上,从表面消毒的根组织中分离出镰刀菌。将单孢分离物转移到覆盖有无菌滤纸碎片(1×2厘米)的合成低营养琼脂(SNA)上,并转移到马铃薯葡萄糖琼脂(PDA)上,置于黑暗中10至14天进行形态学鉴定(4)。23个分离物被鉴定为镰刀菌属(Fusarium commune K. Skovg.),O'Donnell & Nirenberg,以前属于尖孢镰刀菌(F. oxysporum)复合种(4)。PDA上的菌落有白色、蓬松的气生菌丝体,培养基中有紫红色至紫色色素沉着。在SNA上,单瓶梗和多瓶梗上的大分生孢子、厚垣孢子和小分生孢子与该物种的描述一致(4)。使用ef1和ef2引物对翻译延伸因子(EF1-α)基因进行DNA测序,并使用引物MS1和MS2对线粒体小亚基(mtSSU)进行DNA测序,确认了所有23个分离物的鉴定结果(4)[两个代表性分离物的GenBank登录号:EF1-α(JX289892和JX289893),以及mtSSU(JX289894和JX289895)]。在温室中,在18°C的水浴中,使用与受侵染的沙-玉米粉接种物混合的高压灭菌土壤,对两种代表性的镰刀菌属分离物在大豆(品种AG2403)上进行致病性测试(3)。该实验采用完全随机设计(CRD),每个处理有五个重复(单株/150毫升圆锥体),并进行了三次。6周后评估干根和地上部重量以及根腐病严重程度(对整个根系根腐病百分比的目测估计)。两种分离物侵染的土壤中,大豆幼苗的平均出苗率分别为47%和40%;相比之下,未接种的对照植株出苗率为100%。接种和未接种植株在根重(P < 0.0001)、地上部重量(P < 0.0001)和根腐病严重程度(P < ......