First report of basal stem rot on sugarcane (var. Badila) caused by Sclerotium rolfsii in China.

IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Plant disease Pub Date : 2024-10-11 DOI:10.1094/PDIS-07-24-1460-PDN
Su-Chan Lao, Su-Juan Lao, Ji-Hua Huang, Zhu-Gui Zhou, Shan-Hai Lin
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Abstract

Badila (Saccharum officinarum) is one of the important chewing cane in south China. During the year 2019-2020, as much as 60.2%-87.5% of sugarcane plants stem showed red rot developments were observed in the fields of Yongning District, Nanning city, Guangxi province. Symptomatic plants showed red rot at basal stem nodes and sheath, when the disease serious, the epidermis and aerial roots decomposed and exfoliated, then formed sclerotiums, the upper stem also occurred the symptom. Infected plant tissues were dissected into small pieces with 0.1 × 0.1cm in size and surface sterilized in 0.1% HCl2 for 2 min, followed by 75% ethanol for 30 s, rinsed three times with sterile distilled water. Then the tissues were placed onto potato dextrose agar (PDA) plates and incubated at 25 °C for 3 days. Numerous white globoid sclerotia were formed on PDA after 5 days of growth. The sclerotia (2 to 3 mm in diameter) were white at first and then gradually turned dark brown. Aerial mycelia usually formed many narrow hyphal strands 4 to 9 μm wide. Five uniform isolates were obtained from diseased sugarcane plants. Pathogenicity of representative strain W1 was confirmed by inoculating 120-day-old Badila plants grown in field. Five plants were inoculated with colonized agar discs (6mm in diameter) by applying toothpick tips to the lower part of the stem. Five non-inoculated plants served as control. The inoculated and non-inoculated plants were sprayed sterile water then incubated with plastic film for maintained high moisture. All the plants were placed inside of a growth chamber at 26 ± 2°C with a 14-h photoperiod and 80% relative humidity. All inoculated plants showed red rot at stem and sheath after 2 weeks, whereas the control plants were symptomless. By the third week, mycelium and sclerotia developed on the crown on the inoculated plants. The fungus was re-isolated from the artificially inoculated plants. To confirm the species-level identification, partial of the ribosomal DNA internal transcribed spacer (ITS), mitocondrial small subunit (SSU), and nuclear ribosomal large subunit (LSU) regions of representative strain W1 were amplified and sequenced using the primers pairs ITS1/ITS4 (White et al. 1990), ITS-Fu-F /ITS-Fu-R and SRLSU1//SRLSU2 (Kumar et al., 2016), respectively. The resulting ITS, SSU and LSU sequences were deposited in GenBank (GenBank accession no. MW620994, MW617878, and MW617872) and shared 99.42%, 100% and 100% sequence identity with Athelia rolfsii isolate (JN017199, OM319631, and MT225781). Phylogenetic analysis conducted with neighbor-joining (NJ) method using MEGA6.0 revealed that the isolate share a common clade with reference sequence of A. rolfsii in GenBank Data Library. Based on morphological and molecular characteristics, the fungus was identified as A. rolfsii (anamorph: Sclerotium rolfsii) (Paul et al. 2017; Paparu et al. 2020). Although S. rolfsii has been reported causing sugarcane sett rot in Australia (Bhuiyan et al., 2019) and seedlings of sugarcane in Indian (Gopi et al., 2023), as we know, this is the first report of sugarcane basal stem rot disease caused by this fungus in China. This study will be helpful for the prevention and control sugarcane basal stem rot in the future.

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在中国首次报道由 Sclerotium rolfsii 引起的甘蔗(变种 Badila)基部茎腐病。
巴迪拉(Saccharum officinarum)是中国南方重要的咀嚼蔗之一。2019-2020年度,广西南宁市邕宁区田间观察到高达60.2%-87.5%的甘蔗植株茎部出现红腐病。发病植株基部茎节和鞘出现红腐病,病情严重时,表皮和气生根腐烂脱落,形成硬腐子囊,茎上部也出现该症状。将感染的植物组织切成 0.1 × 0.1 厘米大小的小块,在 0.1% HCl2 中表面消毒 2 分钟,然后用 75% 乙醇消毒 30 秒,再用无菌蒸馏水冲洗三次。然后将组织放在马铃薯葡萄糖琼脂(PDA)平板上,在 25 °C 下培养 3 天。生长 5 天后,在 PDA 上形成了许多白色球状的硬菌。硬菌(直径 2 至 3 毫米)最初为白色,然后逐渐变成深褐色。气生菌丝通常形成许多宽 4 至 9 μm 的狭窄菌丝。从染病的甘蔗植株中获得了 5 个一致的分离株。通过接种田间生长 120 天的巴迪拉植株,证实了代表性菌株 W1 的致病性。用牙签尖插入茎干下部,将定植琼脂盘(直径 6 毫米)接种到五株植物上。五株未接种的植物作为对照。将接种和未接种的植物喷洒无菌水,然后用塑料薄膜培养以保持高湿度。所有植株均置于生长室中,温度为 26 ± 2°C,光周期为 14 小时,相对湿度为 80%。2 周后,所有接种植物的茎和鞘都出现了红腐病,而对照植物则没有症状。第三周时,接种植株的树冠上长出了菌丝和菌核。从人工接种的植株上重新分离出了真菌。为确认菌种级别的鉴定,使用引物对 ITS1/ITS4 (White 等,1990 年)、ITS-Fu-F /ITS-Fu-R 和 SRLSU1//SRLSU2 (Kumar 等,2016 年)分别对代表性菌株 W1 的部分核糖体 DNA 内部转录间隔区(ITS)、有丝分裂小亚基(SSU)和核糖体大亚基(LSU)区域进行了扩增和测序。得到的 ITS、SSU 和 LSU 序列存入 GenBank(GenBank 编号:MW620994、MW617878 和 MW617872),与 Athelia rolfsii 分离物(JN017199、OM319631 和 MT225781)的序列同一性分别为 99.42%、100% 和 100%。利用 MEGA6.0 进行的邻接法(NJ)系统进化分析表明,该分离物与 GenBank 数据库中的 A. rolfsii 参考序列具有共同的支系。根据形态和分子特征,该真菌被鉴定为 A. rolfsii(拟态:Sclerotium rolfsii)(Paul 等人,2017 年;Paparu 等人,2020 年)。虽然 S. rolfsii 已被报道在澳大利亚(Bhuiyan 等人,2019 年)和印度(Gopi 等人,2023 年)引起甘蔗定植腐烂病和甘蔗幼苗腐烂病,但据我们所知,这是中国首次报道由该真菌引起的甘蔗基部茎腐病。这项研究将有助于今后甘蔗基部茎腐病的防控。
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来源期刊
Plant disease
Plant disease 农林科学-植物科学
CiteScore
5.10
自引率
13.30%
发文量
1993
审稿时长
2 months
期刊介绍: Plant Disease is the leading international journal for rapid reporting of research on new, emerging, and established plant diseases. The journal publishes papers that describe basic and applied research focusing on practical aspects of disease diagnosis, development, and management.
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