| In this study,16 strains belonging to 16 species of the genus Pleurotus were used. Primer CO332F and CO332R were designed by the CO1 sequence of the genus Pleurotus which have been obtained in lab. This primer used to RT-PCR amplification the coding sequences, amplification productions were clone and seqenced. Make use of the DNAMAN and ClustalX software to analysis the coding sequences of CO1. The results indicated that the coding sequence of CO1 is conservation in the genus Pleurotus, seqence similarities between species ranged from 0.888 to 1.000.Through analyzing the seqences we found that the difference of spiecies CO1 in the genus Pleurotus mainly from intron. Meanwhile one intron was found in P. ostreatus, P. citrinopileatus, P. cornucopiae and P. eryngii var. tuoliensis; two introns in P. eryngii, P pulmonarius, P. abieticola, P. tuber-regium and P. rattenburyi; three intrion in P. australis and P. smithii. and none intron in P. calyptratus, P. cystidiosu, P. djamor, P. dryinus and P. eryngii var. ferulae.In order to rapid identify the Pleurotus species, In this study we used primers CO332F and CO332R to conduct the first amplification. Results of the first-step PCR amplification showed that all the strains only get one band. And then the 16 strains were divided into four groups by the single band size. Group one including P. calyptratus, P. cystidiosu, P. djamor, P. dryinus and P. eryngii var. ferulae; Group two including P. ostreatus, P. citrinopileatus, P. cornucopiae and P. eryngii var. tuoliensis; Group three including P. eryngii, P. pulmonarius, P. abieticola and P. tuber-regium; Group four including P. rattenburyi, P. australis and P. smithii, Species-specific primers were designed accordingly. Results of the second-step PCR amplification showed that each species can get one band by the species-specific primers. In conclusion, the 16 species of Pleurotus can be identified by two-step PCR in accordance with the band size and the presence or absence of the band. |
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