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| Cloning and Analysis of Reverse Transcriptase Gene (RT) of Ty1-copia Retrotransposons in Lycoris radiata |
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Abstract At present, the evolution mechanism of Lycoris radiata genome is unclear and retrotransposons serve as the main source of plant genome evolution. Ty1-copia retrotransposon reverse transcriptase gene (RT) fragments were cloned from genomic DNA of diploid (2n=2x=22) and triploid (2n=3x=33) L. radiata, respectively, using universal primers of Ty1-copia retrotransposon RT, and then analyzed by the relevant bioinformatics softwares in this study. Sixteen (2RA1~2RA16, GenBank accession: KP275626~KP275641) and 17 (3RA1~3RA17, GenBank accession: KP275642~KP275658) RT sequences were cloned from diploid and triploid L. radiata, respectively. The length of these sequences ranged from 248 to 266 bp. The similarity of RT nucleotide sequences from diploid L. radiata ranged from 49.05% to 87.83%, while that from triploid L. radiata ranged from 40.75% to 89.73%. And the similarity of RT inferred amino acid sequences from diploid L. radiata ranged from 18.60% to 91.95%, while that from triploid L. radiata ranged from 18.07% to 88.51%. The results showed that 33 RT sequences obtained from L. radiata had polymorphism and high heterogeneity, and the difference of RT sequences among triploid L. radiata was larger than that among diploid L. radiata. Phylogenetic trees, which were constructed by MEGA 4.0 using the Neighbor-Joining method, indicated that Ty1-copia retrotransposons in L. radiata happened not only horizontal transmission, but also vertical transmission. In addition, there was no direct correlation between classification and ploidy. Compared with other plants, it was found that they had higher homology with herbaceous plants: Volvox carteri from algae, Dicranum scoparium from bryophytes, Osmunda cinnamomea and Equisetum scirpoides from fern, and monocotyledonous plants (Zea mays, Triticum aestivum, Equisetum scirpoides and Dendrobium officinale), and dicotyledonous plants (Arabidopsis thaliana, Lycopersicon esculentum, Populus ciliate, Picea abies, Beta procumbens and Glycine max). Moreover, 2RA16 from diploid L. radiata was the most ancient Ty1-copia retrotransposon RT sequences. Diploid L. radiata was likely to be the primitive group in L. radiata complexes, and triploid L. radiata was proved to be homologous triploid. This study is helpful for understanding intra- specific evolution of L. radiata and the origin of triploid L. radiata, and it provides scientific evidences for further revealling karyotype evolutionary mechanism and theoretical foundation for polyploid breeding of L. radiata in the future.
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Received: 08 October 2014
Published: 01 March 2015
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