RT-PCR Detection of dsRNA Virus LeV in Lentinula edodes

Abstract

Abstract: Lentinula edodes mycovirus HKB(LeV)is a fungal virus with the potential symptoms and exists widely in Lentinula edodes strains. In order to detect LeV from abundant strains rapidly and accurately, a pair of primers were designed according to the sequence of LeV (AB.429556.2)and amplified by RT-PCR method. In a total of 25 strains, 20 strains were positive for LeV and 5 strains were negative. The result of dsRNA extraction also showed dsRNA were obtained in the positive strains and the negative strains were not. In conclusion, the study proved that RT-PCR is a rapid and sensitive method for detection of LeV in Lentinula edodes strains and can attribute to the control of strains quality.

Key words: dsRNA, Mycovirus, RT-PCR, Lentinula edodes

Guo Jie, Wu Xiaoping. RT-PCR Detection of dsRNA Virus LeV in Lentinula edodes[J]. Biotechnology Bulletin, 2014, 0(1): 147-150.

References

[1] Yumi M. Molecular characterization of a novel mycovirus in the cultivated mushroom, Lentinula edodes[J]. Virology Journal,2012, 9: 60.
[2] 潘迎捷, 陈明杰, 沈雪仁. 香菇病毒的分离、诊断、侵染途径和生物学特性[J]. 上海农业学报, 1992, 8(4): 7-11.
[3] Ohta C, Taguchi T, Takahashi S, et al. Detection of double stranded RNA elements in cultivated Lentinula edodes(in Japanese)[J].Mushroom Sci Biotechnol, 2008, 16: 155-158.
[4] 吴小平, 王丽, 谢宝贵, 陈剑, 香菇菌株的脱毒研究[J]. 中国农学通报, 2009, 25(24): 44-49.
[5] Kim YJ, Kim JY, Kim JH, et al. Identification of a Novel Pleurotus ostreatus dsRNA virus and determination of the distribution of viruses in mushroom spores[J]. The Journal of Microbiology,2008, 46(1): 95-99.
[6] Jea YH, Sun LJ, Joo LN, et al. Identification of three isometric viruses from Pleurotus ostreatus[J]. Journal of Huazhong Agricultural University, 2004, 23(1): 150-156.
[7] Magae Y, Hayashi N. Double-stranded RNA and virus-like particles in the edible basidiomycete Flammulina velutipes(Enokitake)[J].FEMS Microbiol Lett, 1999, 180(2): 331-335.
[8] 姚立, 陈春乐, 张忠信, 等. 一种新香菇病毒基因组部分cDNA 序列及病毒RT-PCR 检测[J]. 微生物学报, 2010, 37(1): 61-70.
[9] Gildow EF, Ballinger ME, Rochow WF. Identification of doublestranded RNAS associated with barley yellow dwarf virus infection in oats[J]. Phytopathology, 1983, 73: 1570-1572.
[10] French RC, Price MA, Derrick KS. Circular double-stranded RNA in potato spindle tuber viroid-infected tomatoes[J]. Nature,1982, 295: 259-260.
[11] Grcgan HM, Aclie BA, Gaze RH, et al. Double-stranded RNA elements associated with the MVX disease of Agaricus bisporus[J].Mycol Res, 2003, 107(2): 147-154.
[12] 曹庆, 易干军, 陈金印, 等. 柑桔裂皮病类病毒RT-PCR 检测体系优化分析[J]. 南昌工程学院学报, 2008, 27(03): 64- 66.
[13] 郭立新, 向本春, 朱水芳. 苹果茎沟病毒实时荧光RT-PCR 反应体系的优化[J]. 北方园艺, 2009(3): 8-11.
[14] 谢志勤, 谢芝勋, 廖敏, 等. RT-PCR 检测猪瘟病毒方法的建立与应用[J]. 畜牧与兽医, 2002, 34(11): 11-14.
[15] Peter AR, Peter JW. RT-PCR detection of dsRNAs associated with La France disease of the cultivated mushroom Agaricus bisporus (Lange)Imbach[J]. Journal of Virological Methods, 1997, 63: 17-26.
[16] Kim YJ, Park S, Yie SW, et al. RT-PCR Detection of dsRNA Mycoviruses Infecting Pleurotus ostreatus and Agaricus blazei Murrill[J]. Plant Pathol, 2005, 21(4): 343-348.