Leyva-Mir Santos Gerardo, García-León Elizabeth, Camacho-Tapia Moises, Villasenor-Mir Hector-Eduardo, Leyva-Madrigal Karla Yeriana, Mora-Romero Guadalupe Arlene, Tovar-Pedraza Juan Manuel
Universidad Autónoma Chapingo, Departamento de Parasitología Agrícola, km 38.5 Carretera Mexico-Texoco, Texcoco, Estado de México, Mexico, 56230;
Instituto Nacional de Investigaciones Agrícolas, Forestales y Pecuarias, Campo Experimental Valle del Fuerte, CARRETERA INTERNACIONAL MEXICO-NOGALES, KM.1609, GUASAVE, SINALOA, MEXICO, Mexico, 81110;
Plant Dis. 2022 Mar 12. doi: 10.1094/PDIS-11-21-2467-PDN.
Fusarium head blight (FHB) is one of the most important diseases affecting wheat production worldwide. In Mexico, and are the dominant species causing FHB of wheat (Cerón-Bustamante et al. 2018). During the 2017 to 2019 surveys, FHB symptoms were observed in wheat fields in the Highlands region of Mexico. Symptomatic spike samples were collected from 19 wheat fields in five states (Tlaxcala, Hidalgo, Puebla, Estado de México, and Morelos). -like colonies were consistently isolated on potato dextrose agar (PDA) and 95 monoconidial isolates were obtained. Morphological features of seven isolates were consistent with the description of the species complex (Xia et al. 2019). On PDA, colonies exhibited white and fluffy aerial mycelia, with diffused pink pigment on the reverse side after 7 days of incubation at 25℃. On carnation leaf agar (CLA), macroconidia (n = 100) were hyaline, falcate, with 3 to 6 septa, measuring 25.2 to 43.1 × 2.8 to 5.1 μm, and foot-shaped basal cell. Chlamydospores were ellipsoidal or subglobose and produced in chains. These seven isolates were selected for multilocus phylogenetic analysis and pathogenicity tests. Isolates were deposited in the Culture Collection of Phytopathogenic Fungi of the Department of Agricultural Parasitology at the Chapingo Autonomous University under acc. nos. UACH428 to UACH434. For molecular identification, genomic DNA was extracted, and the internal transcribed spacer (ITS) region, partial sequences of translation elongation factor 1-alpha (EF1-α) and the second largest subunit of RNA polymerase II (RPB2) genes were amplified, and sequenced with the primer sets ITS5/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), and RBP2-5F/RPB2-7R (Liu et al. 1999), respectively. A phylogenetic tree, including published ITS, EF1-α, and RPB2 sequence data, was constructed for the species complex (FIESC) based on Maximum Likelihood. Three species of the FIESC were identified into (five isolates), (one isolate), and (one isolate). The sequences were deposited in GenBank with accession nos. OL347713 to OL347719 for ITS, OL365078 to OL365084 for EF1-α, and OL365072 to OL365077 for RPB2. The pathogenicity of the isolates was confirmed on wheat cv. Nana F2007 at the flowering stage in a glasshouse assay. The heads of 20 wheat plants were sprayed with a conidial suspension (1 × 10 spores/ml) of each isolate. Ten plants mock-inoculated with sterilized water served as the controls. All plants were placed in a moist chamber for 48 h. At 10 days after inoculation, typical FHB symptoms were visible on the inoculated plants, whereas the control plants remained asymptomatic. The pathogenicity test was repeated twice with similar results. The fungi were reisolated from the infected heads and found to be morphologically identical to the isolates used for inoculation, fulfilling Koch's postulates. Previously, three isolates of sp. belonging to the FIESC, were associated with FHB of wheat in Mexico (Cerón-Bustamante et al. 2018); however, this is the first report of , , and causing FHB of wheat in Mexico and worldwide (Farr and Rossman 2021). Further studies should be focused on determining the distribution, prevalence, and toxigenic potential of the isolates of the FIESC associated with wheat diseases in Mexico.
小麦赤霉病(FHB)是影响全球小麦生产的最重要病害之一。在墨西哥, 和 是引起小麦赤霉病的优势种(Cerón - Bustamante等人,2018年)。在2017年至2019年的调查中,在墨西哥高地地区的小麦田中观察到了赤霉病症状。从五个州(特拉斯卡拉、伊达尔戈、普埃布拉、墨西哥州和莫雷洛斯)的19个小麦田中采集了有症状的穗样本。在马铃薯葡萄糖琼脂(PDA)上始终能分离出类似 的菌落,并获得了95个单孢分离株。七个分离株的形态特征与 种复合体的描述一致(Xia等人,2019年)。在PDA上,菌落呈现白色蓬松的气生菌丝体,在25℃下培养7天后,背面有扩散的粉红色色素。在香石竹叶琼脂(CLA)上,大分生孢子(n = 100)透明,镰刀形,有3至6个隔膜,大小为25.2至43.1×2.8至5.1μm,基部细胞呈足形。厚垣孢子椭圆形或近球形,成链产生。选择这七个分离株进行多位点系统发育分析和致病性测试。分离株保存在查平戈自治大学农业寄生虫学系植物病原真菌培养保藏中心,保藏编号为UACH428至UACH434。为了进行分子鉴定,提取了基因组DNA,并扩增了内部转录间隔区(ITS)区域、翻译延伸因子1 - α(EF1 - α)的部分序列和RNA聚合酶II的第二大亚基(RPB2)基因,分别用引物对ITS5/ITS4(White等人,1990年)、EF1 - 728F/EF1 - 986R(Carbone和Kohn,1999年)以及RBP2 - 5F/RPB2 - 7R(Liu等人)进行测序。基于最大似然法构建了包括已发表的ITS、EF1 - α和RPB2序列数据的 种复合体(FIESC)的系统发育树。FIESC的三个种被鉴定为 (五个分离株)、 (一个分离株)和 (一个分离株)。这些序列已存入GenBank,ITS的登录号为OL347713至OL347719,EF1 - α的登录号为OL365078至OL365084,RPB2的登录号为OL365072至OL365077。在温室试验中,在小麦品种Nana F2007的开花期证实了分离株的致病性。用每个分离株的分生孢子悬浮液(1×10孢子/ml)喷洒20株小麦的穗。用无菌水模拟接种的10株植物作为对照。所有植物都放在潮湿的培养箱中48小时。接种后10天,接种的植物上出现了典型的赤霉病症状,而对照植物仍无症状。致病性测试重复了两次,结果相似。从感染的穗上重新分离出真菌,发现其形态与用于接种的分离株相同,符合柯赫氏法则。此前,属于FIESC的三个 种的分离株与墨西哥的小麦赤霉病有关(Cerón - Bustamante等人,2018年);然而,这是 、 和 在墨西哥和全球引起小麦赤霉病的首次报道(Farr和Rossman,2021年)。进一步的研究应集中在确定与墨西哥小麦病害相关的FIESC分离株的分布、流行情况和产毒潜力上。