Xu Jing, Kong Fanxin, Zhang Haijin, Zhang Wenfei, Wu Hongsheng, Chen Guoqiu
Dongling Road No.84Shenyang, China, 110161;
Chaoyang, China;
Plant Dis. 2022 Nov 22. doi: 10.1094/PDIS-06-22-1388-PDN.
Foxtail millet [ (L.) P. Beauv.] is one of the most important nutritious food crops in China. In August 2020, plants of the foxtail millet cultivar Xiao Huang Miao were found that were wilted and root rot symptoms of 25-75% incidence in a field production area of about 3000 m near Tongliao of Inner Mongolia and Chaoyang cities of Liaonning province. The wilted plants showed yellowing, stunting, and the lower stalk became straw colored, softened, with gray-white mould on the surface of the stem nodes. The root system was poorly developed, brown and rotted. Symptomatic roots were surface-disinfested with 70% ethanol for 1 min and in 2% sodium hypochlorite (NaOCl) for 3 min, rinsed with sterilized water three times, and placed on potato dextrose agar (PDA) and incubated at 26ºC for 5 days. Ten pure cultures were obtained from single conidia with an inoculation needle under stereomicroscope. The cultures were transferred to carnation leaf agar (CLA) medium and incubated two weeks in the dark at 26ºC for microscopic observation. Macroconidia had one to four septa (three septa dominated), and were slender and straight with curved apical cell and foot-shaped basal cell, 25.5 - 30.5 × 2.5 - 4.5 μm (n=50). Microconidia were non-septate, oval, and were formed in short chains or false heads on monophialides, 2.5 - 15 × 2.75 - 4.0 μm (n=50). Chlamydospores were singly or in chains, circular or subcircular, 5.25 - 11.5 μm in diameter (n=50). Morphologically, the fungus was identified as Burgess & Trimboli (Klaasen and Nelson,1998; Leslie and Summerell, 2006,). To validate this identification, rDNA internal transcribed spacer (ITS), partial translation elongation factor 1 alpha (TEF-á) gene, and RNA polymerase II second largest subunit (rpb2) of the ten isolates were amplified and sequenced (White et al.1990;O'Donnell K. et al. 2015,2010). Identical sequences were obtained and the sequence of the isolate GZGF23 was submitted to GenBank. BLASTn analysis of the ITS (OL964384), TEF-á (OL961517) and RPB2(ON756204) sequence of isolate GZGF23 revealed 99.86% (MH862671, 557/565bp), 100% (MT011009, 713/1770bp) and 100% (MT010976, 1002/3907bp) sequence similarity respectively with (CBS749.97). Pathogenicity studies were conducted on outdoor potted ground and with the foxtail millet cultivar "Xiao Huang miao". Five 12-L pots were filled with sterilized field soil mixed with 300ml conidial suspension at 3 × 10 spores/ml. Another five 12-L pots were filled with sterilized field soil mixed with 300ml sterilized water that served as controls. About twenty seeds per pot were surface disinfected in 2% NaOCl for 3 min, and rinsed with sterilized water. The foxtail millet seeds were sown the same day as soil inoculation and 6 plants were left in each pot when seedling emerged. Five weeks after seedling emergence, all inoculated plants exhibited symptoms similar to the syptoms observed in the field but control plants had no symptoms. The same results were obtained when pathogenicity tests were repeated two times in the same manner. was reisolated from inoculated plants and its morphological and molecular characteristics matched the original isolate, but the fungus was not reisolated from control plants. This is the first report of root rot caused by on foxtail millet in China. The disease might bring a threat to foxtail millet production and effective control measures should be identified to reduce losses. References: Klaasen J. A. and Nelson P. E. 1998. Mycopathologia 140: 171-176. Leslie J. F. and Summerell B. A. 2006. Blackwell Publishing, Oxford, U.K. O'Donnell K., et al. 2015. Phytoparasitica 43:583-595. White T. J., et al. 1990. Academic Press, San Diego, CA, pp 315-322. O'Donnell K et al. 2010. J.Clin.Microbiol. 48:3708.
谷子[Setaria italica (L.) P. Beauv.]是中国最重要的营养粮食作物之一。2020年8月,在内蒙古通辽市和辽宁省朝阳市附近约3000平方米的田间生产区,发现谷子品种小黄苗的植株出现萎蔫和根腐症状,发病率为25%-75%。萎蔫植株表现为叶片发黄、生长受阻,下部茎秆变为稻草色,变软,茎节表面有灰白色霉层。根系发育不良,呈褐色并腐烂。将有症状的根用70%乙醇进行表面消毒1分钟,再放入2%次氯酸钠(NaOCl)中消毒3分钟,用无菌水冲洗三次,然后置于马铃薯葡萄糖琼脂(PDA)上,在26℃下培养5天。在体视显微镜下用接种针从单个分生孢子中获得了10个纯培养物。将培养物转移到香石竹叶琼脂(CLA)培养基上,在26℃黑暗条件下培养两周用于显微镜观察。大型分生孢子有1-4个隔膜(以3个隔膜为主),细长且直,顶端细胞弯曲,基部细胞呈足形,25.5 - 30.5×2.5 - 4.5μm(n = 50)。小型分生孢子无隔膜,椭圆形,在单瓶梗上形成短链或假头状,2.5 - 15×2.75 - 4.0μm(n = 50)。厚垣孢子单个或成链状,圆形或近圆形,直径5.25 - 11.5μm(n = 50)。从形态学上,该真菌被鉴定为Burgess & Trimboli(Klaasen和Nelson,1998;Leslie和Summerell,2006)。为验证该鉴定结果,对10个分离株的核糖体DNA内转录间隔区(ITS)、部分翻译延伸因子1α(TEF-α)基因和RNA聚合酶II第二大亚基(rpb2)进行了扩增和测序(White等人,1990;O'Donnell K.等人,2015,2010)。获得了相同的序列,并将分离株GZGF23的序列提交到GenBank。对分离株GZGF23的ITS(OL964384)、TEF-α(OL961517)和RPB2(ON756204)序列进行BLASTn分析,结果显示分别与[Fusarium proliferatum (Matsushima) Nirenberg](CBS749.97)的序列相似度为99.86%(MH862671,557/565bp)、100%(MT011009,713/1770bp)和100%(MT010976,1002/3907bp)。在室外盆栽地上对谷子品种“小黄苗”进行了致病性研究。五个12升的花盆装满灭菌的田间土壤,并与300毫升浓度为3×10⁶个孢子/毫升的分生孢子悬浮液混合。另外五个12升的花盆装满灭菌的田间土壤,并与300毫升灭菌水混合作为对照。每盆约20粒种子在2% NaOCl中进行表面消毒3分钟,并用无菌水冲洗。谷子种子在土壤接种当天播种,出苗后每盆保留6株植株。出苗后五周,所有接种植株均表现出与田间观察到的症状相似的症状,但对照植株无症状。以同样的方式重复进行两次致病性试验,得到了相同的结果。从接种植株中重新分离出了[Fusarium proliferatum (Matsushima) Nirenberg],其形态和分子特征与原始分离株相符,但未从对照植株中重新分离出该真菌。这是中国关于谷子根腐病由[Fusarium proliferatum (Matsushima) Nirenberg]引起的首次报道。该病可能对谷子生产构成威胁,应确定有效的防治措施以减少损失。参考文献:Klaasen J. A.和Nelson P. E. 1998. Mycopathologia 140: 171 - 176。Leslie J. F.和Summerell B. A. 2006. Blackwell Publishing, Oxford, U.K.。O'Donnell K.,等人,2015. Phytoparasitica 43:583 - 595。White T. J.,等人,1990. Academic Press, San Diego, CA, pp 315 - 322。O'Donnell K等人,2010. J.Clin.Microbiol. 48:3708。