Detection of Mycovirus in Sclerotium Rolfsii, A Phytopathogenic Fungus in Bangladesh

M. Faruk, N. Suzuki
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Abstract

A research project was conducted to detect presence of mycovirus, a probable virocontrol agent in Sclerotium rolfsii, a destructive soil-borne plant pathogenic fungus. A total of 500 isolates of the pathogen was isolated from infected wheat, bush bean, tomato, eggplant, lentil, chickpea, eggplant, chilli and okra grown Bangladesh following tissue planting methods and brought to the Virology laboratory of Okayama University, Japan. After brought to Japan, a total of 472 isolates were re-isolated and the fungal isolates were cultured on potato dextrose agar for extraction of DNA and dsRNA. The extracted nucleic acids were tested for the presence of viruses using rolling circle amplification (RCA) (for DNA viruses) and cellulose column chromatography (for RNA viruses). The detected S. rolfsii viruses were further characterized molecularly. For detection of DNA virus, a total of 120 isolates were tested, none were found positive for DNA viruses. For detection of dsRNA virus, a total of 472 isolates were tested, 7 isolates showed possible dsRNA positive band. The partial cDNA sequences of the dsRNA segments isolated from the strain SR336 were obtained. BlastX database search with the partial sequence from SR336 isolates showed similarities with randorna like virus. To screen the presence of randorna like virus in S. rolfsii isolates, 64 isolates were selected from 472 isolates of S. rolfsii. The isolates of S. rolfsii were tested by RT-PCR using randorna virus specific primers. Out of 64 isolates 13, (20.31%) isolates were randorna virus positive strain and rest of the isolates were absent of randorna virus. Bangladesh J. Agril. Res. 46(3): 331-341, September 2021
孟加拉国一种植物病原真菌罗尔夫菌核中分枝病毒的检测
摘要对一种具有破坏性的土传植物病原真菌罗尔夫菌核菌(Sclerotium rolfsii)进行了分枝病毒(mycovirus)的检测研究。从孟加拉国种植的小麦、灌木豆、番茄、茄子、扁豆、鹰嘴豆、茄子、辣椒和秋葵中分离出500株病原菌,并送到日本冈山大学病毒学实验室。带回日本后,对472株真菌进行再分离,并在马铃薯葡萄糖琼脂上培养,提取DNA和dsRNA。提取的核酸采用滚环扩增(RCA)(用于DNA病毒)和纤维素柱层析(用于RNA病毒)检测病毒的存在。对检测到的S. rolfsii病毒进行了进一步的分子表征。在DNA病毒检测方面,共检测了120株分离株,均未发现DNA病毒阳性。在dsRNA病毒检测方面,共检测了472株分离株,7株分离株显示可能的dsRNA阳性带。获得了SR336分离的dsRNA片段的部分cDNA序列。在BlastX数据库中检索SR336分离株的部分序列,发现该病毒与随机样病毒具有相似性。为了筛选罗尔夫氏梭菌分离株中是否存在随机样病毒,从472株罗尔夫氏梭菌中筛选出64株。采用随机病毒特异性引物RT-PCR技术对罗尔夫氏梭菌分离株进行检测。64株分离物中有13株(20.31%)为随机病毒阳性株,其余分离物无随机病毒。孟加拉国J. Agril。Res. 46(3): 331-341, 2021年9月
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