<br>印度特里普拉邦咖喱叶树 (Murraya koenigii) 首次报告由 Athelia rolfsii 引起的茎腐病。,Plant Disease

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印度特里普拉邦咖喱叶树 (Murraya koenigii) 首次报告由 Athelia rolfsii 引起的茎腐病。
Plant Disease ( IF 4.4 ) Pub Date : 2020-08-17 , DOI: 10.1094/pdis-05-20-1152-pdn
Deeba Kamil ₁ , Amar Bahadur _{2,

3} , Presenjit Debnath ₄ , Anjali Kumari ₅ , Shiv Pratap Choudhary ₆ , Krishna Kumar ₇ , Prameeladevi Thokala ₇

Affiliation

九里香是印度重要的药用植物，俗称咖喱叶树，生长于热带和亚热带地区。咖喱树的叶子被用作草药，因为存在以下重要的活性成分 bismahanine、murrayanine、murrayafoline-A、bi-koeniquinone-A、murrayazolidine 等（Jain 等人，2017）。 2019 年 7 月中旬，特里普拉邦伦布切拉农业学院（印度）的咖喱叶树上观察到茎腐病症状。病害表现为腐烂、萎蔫、枯萎等，发病率8%～10%。最初，受感染的植株逐渐枯萎，在土线处的下部茎表面出现白色菌丝垫。收集受感染的茎样品，用0.25%次氯酸钠表面消毒1分钟，用灭菌蒸馏水洗涤三次，置于含有2%水琼脂的培养皿中。在 26°C 培养三天后，将植物碎片产生的菌丝转移到含有马铃薯葡萄糖琼脂的培养皿中。从疾病样本中收集了十个分离株。获得的纯培养物为丰富的气生白色菌丝体，具有直径为0.8至1.5毫米的圆形至不规则菌核。菌核最初是白色的，但后来变成棕色。根据形态学，该病原体被鉴定为 Athelia rolfsii (Aycock 1966)。为了确认鉴定，使用 ZR 真菌/细菌 DNA 小量制备试剂盒（加利福尼亚州欧文市）从分离株的菌丝体中提取基因组 DNA，并使用通用引物 ITS1 和 ITS1 扩增 rDNA 的内转录间隔区 (ITS) 区域。 ITS4（White 等人，1990）。对 550 bp PCR 产物进行测序，结果显示与 Athelia rolfsii 分离株（GenBank 登录号 MH854711）有 99% 的相似性。生成的序列提交给 GenBank（登录号 MT535585）。鉴定出病原菌后，进行盆栽实验以确认其致病性。土盆（直径29厘米）装满消毒过的土壤并保存在温室中。十株咖喱叶植物（50 天大）从单独盆中的种子生长，并用在马铃薯葡萄糖肉汤中制备的 15 天大菌丝体垫进行接种。在无菌刀片的帮助下，对每株咖喱植物的茎进行人为损伤，并用含有菌丝垫的湿无菌棉花包裹。以人工损伤并接种无菌蒸馏水的五株咖喱叶植物作为对照。将装有植物的土盆单独用塑料袋覆盖，并在26°C的温室中保存约15天。接种后的植物在接种六天后开始出现茎腐烂的症状，并开始干燥。接种植物的茎腐病症状与田间观察到的相似。从接种的植物中重新分离真菌，并根据形态学确认罗尔氏菌的鉴定。在对照植物上没有观察到任何症状。获得的培养物保藏于印度典型培养物保藏中心，植物病理学部，ICAR-印度农业研究所，新德里，印度(ITC-8666)。据我们所知，这是印度和世界范围内首次报道由 A. rolfsii 引起的咖喱叶植物茎腐病。由于其药用、风味和香气特性，它在印度经常使用。咖喱叶植物在印度经常被用作药草，因此这种疾病对生产构成重大风险。

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First Report of Stem Rot Caused by Athelia rolfsii on Curry Leaf Tree (Murraya koenigii) in Tripura, India.

Murraya koenigii is an important medicinal plant of India and commonly known as curry leaf tree grown in tropical and subtropical regions. The leaves of curry tree are used as a herb due to the presence of following important active constituent bismahanine, murrayanine, murrayafoline-A, bi-koeniquinone-A, murrayazolidine etc. (Jain et al. 2017). During mid-July 2019, stem rot disease symptoms were observed on curry leaf trees at the College of Agriculture, Lembucherra, Tripura (India). The disease symptoms consisted of rotting, wilting and blighting with disease incidence ranging from 8 to 10%. Initially, infected plants gradually withered and white mycelia mats appeared on the surface of the lower stem at the soil line. Infected stem samples were collected and surface was sterilized with 0.25% sodium hypochlorite for 1 min, washed thrice with sterilized distilled water and placed in Petri plates containing 2% water agar. After three days of incubation at 26°C, hyphae produced from plant bits were transferred into Petri plates containing potato dextrose agar. Ten isolates were collected from the diseases samples. Pure cultures were obtained as abundant, aerial and white mycelia with round to irregular sclerotia of 0.8 to 1.5 mm in diam. The sclerotia were initially white in color but later turned into brown color. The pathogen was identified as Athelia rolfsii based on morphology (Aycock 1966). To confirm the identification, the genomic DNA was extracted from a mycelia mat of the isolates using ZR fungal/Bacterial DNA miniprep kit (Irvine, CA) and the internal transcribed spacer (ITS) region of rDNA was amplified using the universal primers, ITS1 and ITS4 (White et al. 1990). A 550 bp PCR product was sequenced and showed 99% similarity with Athelia rolfsii isolate (GenBank accession MH854711).The generated sequence was submitted to GenBank (Accession MT535585). After identification of the pathogen a pot experiment was conducted to confirm the pathogenicity. Earthen pots (29 cm. diam.) were filled with sterilized soil and kept in a green house. Ten curry leaf plants (50 days old) were grown from seed in the separate pot were inoculated with 15-day old mycelia mats prepared in potato dextrose broth. The stem of each curry plant were artificially injured with the help of sterile blade and wrapped with moistened sterilized cotton containing the mycelial mat. Five curry leaf plants artificially injured and inoculated with sterilized distilled water were used as control. The Earthen pots with plants were individually covered with plastic bags and kept in the green house at 26°C for approximately 15 days. The inoculated plants started showing symptoms of stem rot six days after inoculation and started drying onward. The symptoms of stem rot on the inoculated plants were similar to those observed in the field. The fungus was re-isolated from the inoculated plants and A. rolfsii identification was confirmed based on morphology. No symptoms were observed on the control plants. The obtained culture was deposited in the Indian Type Culture Collection, Division of Plant Pathology, ICAR - Indian Agricultural Research Institute, New Delhi, India (ITC-8666). To the best of our knowledge this is the first report of stem rot disease of curry leaf plant caused by A. rolfsii in India and worldwide. Due to medicinal, flavour and aroma properties, it is regularly used in India. Curry leaf plant is regularly used as a medical herb in India and therefore this disease poses a significant risk to production.

更新日期：2020-08-17

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