Zhang H, Luo W, Pan Y, Xu J, Xu J S, Chen W Q, Feng J
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences (CAAS), Beijing 100193, China. Funding provided by National Natural Science Foundation (No. 31201477).
Plant Dis. 2014 Sep;98(9):1273. doi: 10.1094/PDIS-02-14-0124-PDN.
In China, several diseases of maize (Zea mays L.) including ear rot are caused by Fusarium spp., leading to significant yield losses and potential risk of mycotoxin contamination (2,3). In 2013, a survey was conducted to determine the population composition of Fusarium species on maize ears in Jilin Province. Symptomatic maize ears with pink or white mold were collected and surface disinfested with 70% ethanol and 10% sodium hypochlorite, followed by three rinses with sterile distilled water and placed onto potato dextrose agar (PDA). After 3 days of incubation at 25°C in the dark, newly grown-out mycelia were transferred onto fresh medium and purified by the single-spore isolation method (4). Fusarium spp. were identified by morphological characteristics (2) and sequence analysis of translation elongation factor-1α (TEF) gene (1). A large number of Fusarium spp. were found including F. graminearum species complex and F. verticillioides. In addition, a new species, F. temperatum, recently described in Belgium (2), was also identified. F. temperatum was originally described as F. subglutinans, but a robust polyphasic approach proved it to be a new biological species closely related to F. subglutinans (2). Previous studies had reported ~15% of Fusarium maize ear rot in Jilin was F. subglutinans. In this study, we found both F. subglutinans s. str. and F. temperatum in the proportion of 16.3% and 9.2%, respectively. Similar to previous studies (2), colonies of our strains on PDA were initially white cottony mycelium that become pinkish white. Conidiophores formed abundantly on SNA that were erect, branched, and terminated in 1 to 3 phialides. Microconidia were abundant, hyaline, 0 to 2 septa, obovoid to oval, and not produced in chains. Chlamydospores were absent. Typically macroconidia were falcate, 3 to 5 septate (mostly 4 septate), hyaline with a curved and blunt apical cell and a distinct foot-shaped basal cell. In order to validate this result, partial translation elongation factor (TEF-1α, 629 bp) gene sequences of isolates were generated (GenBank Accession No. KJ137018) (1). BLASTn analysis revealed 100% sequence identity to F. temperatum (HM067690). A pathogenicity test was performed on maize cv. Zhengdan958. Four days after silk emergence, 2 ml conidial suspension (10 macroconidia/ml) of each isolate was injected into each of 10 maize ears through silk channel. Control plants were inoculated with sterile distilled water. Twenty days after inoculation, typical Fusarium ear rot symptoms (reddish-white mold) was observed on inoculated ears and no symptoms were observed on water controls. Koch's postulates were fulfilled by re-isolating the same fungus from the infected seeds. Although F. temperatum was reported to attack maize kernels in southern China where the annual average temperatures are moderately high (3), to our knowledge, this is the first report of F. temperatum causing Fusarium ear rot in northern China, where the winter is long and very cold, the annual average temperature is 4 to 5°C, and the lowest temperature is lower than -35°C. This indicated that F. temperatum was widely distributed in different ecological regions in China. Furthermore, the northeast spring corn region that includes Jinlin is the most important corn belt, with corn production of this region accounting for 42% of the total corn production in China. Therefore, we should pay more attention to the new species in this region and consider them in the development of maize cultivars with broad-based resistance to the pathogens. References: (1) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (2) J. Scauflaire et al. Mycologia 103:586, 2011. (3) J. H. Wang et al. J. Phytopathol. 162:147, 2014. (4) L. Yang et al. Phytopathology 98:719, 2008.
在中国,包括穗腐病在内的几种玉米病害是由镰刀菌属引起的,导致产量大幅损失以及存在霉菌毒素污染的潜在风险(2,3)。2013年,开展了一项调查以确定吉林省玉米穗上镰刀菌属的种群构成。采集了带有粉色或白色霉层的有症状玉米穗,先用70%乙醇和10%次氯酸钠进行表面消毒,然后用无菌蒸馏水冲洗三次,再置于马铃薯葡萄糖琼脂(PDA)上。在25°C黑暗条件下培养3天后,将新长出的菌丝转移到新鲜培养基上,并通过单孢分离法进行纯化(4)。通过形态特征(2)和翻译延伸因子-1α(TEF)基因的序列分析(1)对镰刀菌属进行鉴定。发现了大量镰刀菌属,包括禾谷镰刀菌复合种和轮枝镰孢菌。此外,还鉴定出了一个最近在比利时描述的新物种——温和镰刀菌(2)。温和镰刀菌最初被描述为拟胶孢镰刀菌,但一种可靠的多相分类方法证明它是一个与拟胶孢镰刀菌密切相关的新生物种(2)。先前的研究报道吉林约15%的玉米穗腐病由拟胶孢镰刀菌引起。在本研究中,我们分别发现了16.3%的拟胶孢镰刀菌狭义种和9.2%的温和镰刀菌。与先前的研究(2)相似,我们菌株在PDA上的菌落最初是白色棉絮状菌丝体,之后变为粉白色。在燕麦片琼脂(SNA)上大量形成分生孢子梗,分生孢子梗直立、分枝,末端有1至3个瓶梗。小型分生孢子丰富,透明,0至2个隔膜,倒卵形至椭圆形,不成链状产生。无厚垣孢子。典型的大型分生孢子呈镰刀形,3至5个隔膜(大多为4个隔膜),透明,顶端细胞弯曲且钝圆,基部细胞明显呈足形。为了验证这一结果,生成了分离株的部分翻译延伸因子(TEF-1α,629 bp)基因序列(GenBank登录号:KJ137018)(1)。BLASTn分析显示与温和镰刀菌(HM067690)的序列同一性为100%。对玉米品种郑单958进行了致病性测试。吐丝4天后,通过花丝通道将每个分离株的2 ml分生孢子悬浮液(10个大型分生孢子/ml)注射到10个玉米穗中的每个穗上。对照植株接种无菌蒸馏水。接种20天后,在接种的玉米穗上观察到典型的镰刀菌穗腐病症状(红白霉层),而在水对照上未观察到症状。通过从感染种子中重新分离出相同真菌,柯赫氏法则得到了验证。尽管据报道温和镰刀菌在中国南方年平均温度适中的地区会侵害玉米籽粒(3),但据我们所知,这是温和镰刀菌在冬季漫长且非常寒冷、年平均温度为4至5°C且最低温度低于-35°C的中国北方引起镰刀菌穗腐病的首次报道。这表明温和镰刀菌在中国不同生态区域广泛分布。此外,包括吉林在内的东北春玉米区是最重要的玉米带,该地区的玉米产量占中国玉米总产量的42%。因此,我们应更加关注该地区的新物种,并在培育对病原体具有广泛抗性的玉米品种时加以考虑。参考文献:(1)D. M. Geiser等人,《欧洲植物病理学杂志》110:473,2004年。(2)J. Scauflaire等人,《真菌学》第103卷:586页,2011年。(3)J. H. Wang等人,《植物病理学杂志》162:147,2014年。(4)L. Yang等人,《植物病理学》98:719,2008年。