Bienapfl J C, Malvick D K, Percich J A
Department of Plant Pathology, University of Minnesota, St. Paul 55108.
Plant Dis. 2010 Aug;94(8):1069. doi: 10.1094/PDIS-94-8-1069B.
Multiple Fusarium species have been found in association with soybean (Glycine max) plants exhibiting root rot in the United States (3). Soybean plants that lacked apparent foliar symptoms, but exhibited 2- to 5-mm brown, necrotic taproot lesions and lateral root necrosis were observed in Minnesota in one field each in Marshall and Otter Tail counties in July of 2007, as well as in one field in Marshall County in July of 2008. Sampling was conducted as part of a study investigating root rot in major soybean-production areas of Minnesota. Plants were arbitrarily dug up at the R3 growth stage. Root systems were washed, surface disinfested in 0.5% NaOCl for 3 min, rinsed in deionized water, and dried. Fusarium isolates were recovered from root sections with necrotic lesions embedded in modified Nash-Snyder medium (1). One resulting Fusarium colony from one plant per county was transferred to half-strength acidified potato dextrose agar (PDA) and carnation leaf agar (CLA) to examine morphological characteristics (4). Culture morphology on PDA consisted of flat mycelium with sparse white aerial mycelium. On CLA, thick-walled macroconidia with a hooked apical cell and a foot-shaped basal cell were produced in cream-colored sporodochia. Macroconidia ranged from 32.5 to 45.0 μm long. Microconidia were oval to cylindrical with 0 to 1 septa, ranged from 7.5 to 11.25 μm long, and were produced on monophialides. Chlamydospores were produced abundantly in chains that were terminal and intercalary in the hyphae of 4-week-old cultures. Morphological characteristics of the three isolates were consistent with descriptions of F. redolens (2,4). The identity of each isolate was confirmed by sequencing the translation elongation factor 1-α (TEF) locus (4). BLAST analysis of the TEF sequences from each isolate against the FUSARIUM-ID database resulted in a 100% match for 17 accessions of F. redolens (e.g., FD 01103, FD 01369). Each F. redolens isolate was tested for pathogenicity on soybean. Sterile sorghum grain was infested with each isolate and incubated for 2 weeks. Sterile sorghum was used for control plants. Soybean seeds of cv. AG2107 were planted in 11.4-cm pots ~1 cm above a 25-cm layer of infested sorghum or sterile sorghum. Two replicate pots containing four plants each were used per treatment and the experiment was repeated once. Root rot was assessed 28 days after planting. Each F. redolens isolate consistently caused taproot necrosis on inoculated plants, whereas control plants did not exhibit root necrosis. Isolations were made from roots of inoculated and control plants and the isolates recovered from inoculated plants were identified as F. redolens based on morphological characteristics and TEF sequences. Fusarium species were not isolated from control plants. To our knowledge, this is the first report of F. redolens causing root rot of soybean; however, it is possible F. redolens has been found previously and misidentified as F. oxysporum (2,4). Results from inoculations suggest that F. redolens may be an important root rot pathogen in Minnesota soybean fields. References: (1) J. C. Bienapfl et al. Acta Hortic. 668:123, 2004. (2) C. Booth and J. M. Waterston. No. 27 in: CMI Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, England, 1964. (3) G. L. Hartman et al. Compendium of Soybean Diseases. 4th ed. The American Phytopathological Society, St. Paul, MN, 1999. (4) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006.
在美国,已发现多种镰刀菌与表现出根腐病的大豆(Glycine max)植株有关联(3)。2007年7月,在明尼苏达州马歇尔县和奥特塔尔县的一块田地中,以及2008年7月在马歇尔县的一块田地中,观察到了一些大豆植株,这些植株没有明显的叶片症状,但主根出现了2至5毫米的褐色坏死病斑,侧根也发生了坏死。采样是作为一项调查明尼苏达州主要大豆产区根腐病的研究的一部分进行的。在R3生长阶段随机挖掘植株。将根系冲洗干净,在0.5%次氯酸钠中进行表面消毒3分钟,用去离子水冲洗,然后干燥。从嵌入改良Nash-Snyder培养基(1)的坏死病斑根段中分离镰刀菌菌株。从每个县的一株植物上获得的一个镰刀菌菌落转移到半强度酸化马铃薯葡萄糖琼脂(PDA)和香石竹叶琼脂(CLA)上,以检查形态特征(4)。在PDA上的培养形态为扁平菌丝体,有稀疏的白色气生菌丝体。在CLA上,在奶油色的分生孢子座中产生厚壁大分生孢子,顶端细胞呈钩状,基部细胞呈足状。大分生孢子长32.5至45.0μm。小分生孢子椭圆形至圆柱形,有0至1个隔膜,长7.5至11.25μm,在单瓶梗上产生。厚垣孢子在4周龄培养物的菌丝中大量成链产生,位于末端和中间。这三个分离株的形态特征与红镰孢(F. redolens)的描述一致(2,4)。通过对翻译延伸因子1-α(TEF)基因座进行测序,确认了每个分离株的身份(4)。将每个分离株的TEF序列与FUSARIUM-ID数据库进行BLAST分析,结果显示与17个红镰孢登录号(如FD 01103、FD 01369)100%匹配。对每个红镰孢分离株进行了大豆致病性测试。用每个分离株侵染无菌高粱籽粒,并培养2周。无菌高粱用作对照植株。将AG2107品种的大豆种子种植在11.4厘米的花盆中,距离25厘米厚的侵染高粱层或无菌高粱层约1厘米。每个处理使用两个重复花盆,每个花盆种植四株植物,实验重复一次。种植28天后评估根腐病情况。每个红镰孢分离株在接种植株上均持续引起主根坏死,而对照植株未表现出根坏死。从接种和对照植株的根部分离菌株,根据形态特征和TEF序列,从接种植株中分离得到的菌株被鉴定为红镰孢。未从对照植株中分离到镰刀菌属物种。据我们所知,这是关于红镰孢引起大豆根腐病的首次报道;然而,之前有可能已发现红镰孢但被误鉴定为尖孢镰孢(F. oxysporum)(2,4)。接种结果表明,红镰孢可能是明尼苏达州大豆田中的一种重要根腐病病原菌。参考文献:(1)J. C. Bienapfl等人,《园艺学报》668:123,2004年。(2)C. Booth和J. M. Waterston,载于《CMI病原真菌和细菌描述》第27号。CMI,英国邱园,1964年。(3)G. L. Hartman等人,《大豆病害简编》第4版。美国植物病理学会,明尼苏达州圣保罗,1999年。(4)J. F. Leslie和B. A. Summerell,《镰刀菌实验室手册》。布莱克韦尔出版社,爱荷华州艾姆斯,2006年。