Joy Josna, Mahadevakumar S, Mamatha Bhanu L S, Niranjan Raj S, Chandranayaka S, Lakshmidevi N
University of Mysore, 29243, Department of Studies in Microbiology, Mysore, Karnataka, India;
University of Mysore, Department of Studies in Botany, Mycology and Phytopathology Laboratory, Department of Studies in Botany, Manasagangotri, Mysore, Karnataka, Mysore, Karnataka, India, 570006;
Plant Dis. 2022 Feb 8. doi: 10.1094/PDIS-12-21-2834-PDN.
Salvia hispanica L. (Lamiaceae) commonly called 'chia' is an important food crop that has gained significance in recent times globally due to its nutritive value. During a field survey (Mysore district, Karnataka, October, 2021), chia fields were found associated with a characteristic foot rot disease. Further, the presence of mycelial structures along with sclerotial bodies was recorded near the stem-soil interface on the infected plants. The disease incidence ranged 15-21% in an area of approximately 15 hectares of chia fields. The symptoms initially appeared as tan lesions near the stem soil interface and the lesions were colonized by the fast growing mycelium. As the disease progressed, the plants toppled due to death of the stem-root interface region. Infected plants from KM Halli (12º20'90"N; 76º37'68"E) and DMG Halli (12º28'50"N; 76º51'66"E) (n=30) were sampled and associated fungal pathogen isolated on potato dextrose agar (PDA; HiMedia Lab, Mumbai). Fungal mycelia developing from the infected tissues were inoculated on to fresh PDA plates to obtained pure cultures for further identification. Fungal colonies with dense, aerial whitish-cottony mycelia with uniformly globoid sclerotia (0.52.9 mm) were observed after 1012 days of incubation at room temperature. Sclerotia were white at first and turned brown with age. The average number of sclerotia produced per plate ranged from 150 to >280 (n = 10). To further to confirm the identity of the isolates, three representative isolates (SrSh1, SrSh5 and SrSh10) was subjected to molecular identification based on ITS-rDNA sequences. Briefly, genomic DNA was isolated from 12 day old cultures using the CTAB method and ITS-rDNA was amplified using ITS1-ITS4 primers (White et al., 1990). An expected amplicon of >650 bp was obtained and later sequenced from both the directions. The consensus sequences were analysed through nBLAST search which revealed that 100% (643/643 bp) sequence similarity with reference sequences of Athelia rolfsii (S. rolfsii) from GenBank database (KY640622 and AB075298). A phylogenetic tree obtained by the neighbor-joining method using MEGAX shared a common clade with the reference sequences retrieved and computed, thus confirming the identification based on sequence analysis and molecular phylogeny. The representative sequence of A. rolfsii isolates SrSh1, SrSh4 and SrSh7 isolates deposited in GenBank with Accession no OM021878-OM021880. Based on etiology, morphological, cultural and molecular data the pathogen was identified as Athelia rolfsii (Curzi) Tu & Kimbrough (Syn: Sclerotium rolfsii Sacc.) (Mordue, 1974; Mahadevakumar et al., 2016, 2018). Pathogenicity tests were conducted by inoculating the sclerotial bodies near stem soil interface of chia plants grown under green house (at 28 ± 2°C and 70% relative humidity). Briefly, a total of 60 healthy plants were inoculated with sclerotia and covered with polythene bags for 2 days and removed later. Plants (n=20) inoculated without any sclerotia were treated as controls. The development of characteristic foot rot disease was observed after 6-8 days post inoculation. A total of 38 plants showed the foot rot symptoms while control plants remained healthy. The identity of the fungus was confirmed by morphology and molecular sequence analysis after re-isolation. Chia is an important food crop and in recent times has been regarded as super food. Although S. rolfsii is known to be associated with many crops, this is the first report in chia. Therefore, to the best of our knowledge, this is the first report of foot rot disease caused by Sclerotium rolfsii on chia in India. Early diagnosis of this disease will help the farmers to adopt suitable management practices to avoid loss.
西班牙鼠尾草(唇形科)通常被称为“奇亚籽”,是一种重要的粮食作物,近年来因其营养价值在全球范围内受到重视。在一次实地调查(2021年10月,卡纳塔克邦迈索尔区)中,发现奇亚籽田与一种典型的根腐病有关。此外,在受感染植株的茎-土界面附近记录到了菌丝结构和菌核体。在约15公顷的奇亚籽田中,该病发病率为15%-21%。症状最初表现为茎-土界面附近的棕褐色病斑,病斑被快速生长的菌丝定殖。随着病情发展,植株因茎-根界面区域死亡而倒伏。从KM Halli(北纬12°20′90″;东经76°37′68″)和DMG Halli(北纬12°28′50″;东经76°51′66″)采集了受感染植株(n = 30),并在马铃薯葡萄糖琼脂(PDA;孟买HiMedia实验室)上分离相关真菌病原体。将从受感染组织长出的真菌菌丝接种到新鲜的PDA平板上,以获得纯培养物用于进一步鉴定。在室温下培养10-12天后,观察到真菌菌落具有密集的气生白色棉絮状菌丝体和均匀的球状菌核(0.5-2.9毫米)。菌核最初为白色,随着年龄增长变为褐色。每平板产生的菌核平均数量在150至>280之间(n = 10)。为进一步确认分离株的身份,对三个代表性分离株(SrSh1、SrSh5和SrSh10)基于ITS-rDNA序列进行分子鉴定。简要地说,使用CTAB方法从12日龄培养物中分离基因组DNA,并使用ITS1-ITS4引物(White等人,1990)扩增ITS-rDNA。获得了预期的>650 bp扩增子,随后从两个方向进行测序。通过nBLAST搜索分析一致序列,结果显示与GenBank数据库中罗氏阿太菌(S. rolfsii)的参考序列(KY640622和AB075298)有100%(643/643 bp)的序列相似性。使用MEGAX通过邻接法获得的系统发育树与检索和计算的参考序列共享一个共同分支,从而基于序列分析和分子系统发育确认了鉴定结果。罗氏阿太菌分离株SrSh1、SrSh4和SrSh7的代表性序列保存在GenBank中,登录号为OM021878-OM021880。基于病因、形态、培养和分子数据,该病原体被鉴定为罗氏阿太菌(Curzi)Tu & Kimbrough(同义词:齐整小核菌Sacc.)(Mordue,1974;Mahadevakumar等人,2016年,2018年)。通过在温室(28±2°C和70%相对湿度)中生长的奇亚籽植株的茎-土界面附近接种菌核体进行致病性测试。简要地说,总共60株健康植株接种了菌核体,并用聚乙烯袋覆盖2天,随后移除。未接种任何菌核体的植株(n = 20)作为对照。接种后6-8天观察到典型根腐病的发展。共有38株植株表现出根腐症状,而对照植株保持健康。重新分离后通过形态学和分子序列分析确认了真菌的身份。奇亚籽是一种重要的粮食作物,近年来被视为超级食物。虽然已知齐整小核菌与许多作物有关,但这是奇亚籽中的首次报道。因此,据我们所知,这是印度首次关于齐整小核菌引起奇亚籽根腐病的报道。该病的早期诊断将有助于农民采取适当的管理措施以避免损失。