水稻稻瘟病病原菌分离纯化及分子鉴定

Isolation,purification and molecular identification of the pathogen of rice blast

从牡丹江市不同水稻栽培地采集疑似感染稻瘟病的水稻叶片,用PDA培养基进行病原菌分离纯化培养,通过观察菌落黑色素及分生孢子的形态进行初筛鉴别;采用试剂盒提取菌丝DNA,利用稻瘟病菌特异引物进行PCR扩增,对扩增产物进行测序及Nucleotide Blast比对。结果表明,6个栽培区采集病样经初筛确定5株有黑色素分泌且观察到其分生孢子,测序证实其中2个样品PCR产物序列相同,且与NCBI网站公布的半知菌亚门的灰梨孢(Pyricularia grisea)高度同源,确定获得稻瘟病菌纯菌株。

The leaves of rice infected with rice blast disease were collectted from different rice cultivation areas.Pathogen isolation and purification culture were carried out by PDA medium,and the morphology of colony melanin and conidia were identified by prelim⁃inary screening.Mycelia DNA was extracted with kit,PCR amplification was performed using specific primers of Blast,and the ampli⁃fied products were sequenced and Nucleotide Blast comparisons were performed.The results showed that the disease sample was taken six cultivation areas after the initial screening to determine 5 strains of secretion of melanin and observed its conidium,PCR product sequencing confirmed that the two sample sequence was the same,and the imperfect fungi of NCBI website door of grey pear spore(Pyricularia grisea)highly homologous,determine to obtain pure strains of rice blast.