| This study is designed to conduct molecular identification of tomato yellow leaf curlvirus (TYLCV) arised in Hebei Province, and analyze their variation and phylogeneticrelations. In the study, a pair of specific primers PA1-F/R was designed according toknown genome sequence of TYLCV to conduct PCR detection. The result showed that allspecific segments attained from the4virus samples by PA1-F/R primers wereapproximate645bp. And then the generated specific segments were collected, cloned andsequenced. Sequence analizing showed that the4samples show more than99%ofhomology with TYLCV, which suggests that they were surely infected by TYLCV.Next, a new pair of primers PA2-F/R was redesigned according to the sequence tomplify the near whole sequence of virus sample, and conducting homology comparisonand molecular evolution analyzing. Also, by sequencing and analyzing on DNA-A, itdiscovered that total length of the4virus samples were2781-2782nucleotides, coding6ORFs in all. The four ORFs AC1(nucleotides1542-2615), AC2(nucleotides1226-1633),AC3(nucleotides1081-1485), AC4(nucleotides2171-2464) were located on thecomplemntary sense strand. On the sense strand two ORFs, AV1(nucleotides308-1084)and AV2(nucleotides148-498) were found, and the AV2gene coded for the coat proteincontaining256amino acids. Through comparison of genomes, it found that the4sampleDNA-A had the highest probability in homology with TYLCV reported in Shandong,Shanghai, Zhejiang, Anhui, Japan and Australia at above99.0%, and belonged to the samelarge branch with TYLCV reported in America, Afric and Europe. This studydemonstrated that the4samples were infected with TYLCV, and it is highly probable thatthey were isolates of TYLCV-Israel strain.Finally, in TYLCV-Handanquzhou sample for example, we found out a set of TYLCVinfection cloning preparation methods. This method is to make the contribution forTYLCV disease resistance breeding and TYLCV virus research in the future. |
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