The No.601 watermelon (citrullus lanatus) seeds were treated with 25 keV N+ implantation at the dosage of 7.8 ×1016 ions/cm2. After treatment, watermelon seeds were incubated with 380μg/μl pumpkin (Cucubita, ma...The No.601 watermelon (citrullus lanatus) seeds were treated with 25 keV N+ implantation at the dosage of 7.8 ×1016 ions/cm2. After treatment, watermelon seeds were incubated with 380μg/μl pumpkin (Cucubita, maxima Duch) DNA solution at 35 ℃ for 5 hours. By two-generations of selection and resistance screening at seedling stage, one transformed material was selected out, whose rind color is similar to that of the donor pumpkin and whose size of seeds is between that of the donor and the receptor. Using AFLP (amplified fragment length polymorphism) technique, two polymorphic DNA fragments were amplified. This primarily testified that the donor DNA fragments/gene were introduced into the receptor cell and integrated into the genomic DNA of the receptor.展开更多
Retrotransposon-like elements are major constituents of most eukaryotic genomes. For example, they account for roughly 90% of the wheat （Triticum aestivum） genome, Previous study on a wheat strain treated by low-ene...Retrotransposon-like elements are major constituents of most eukaryotic genomes. For example, they account for roughly 90% of the wheat （Triticum aestivum） genome, Previous study on a wheat strain treated by low-energy N^＋ ions indicated the variations in AFLP （Amplified Fragment Length Polymorphism ） markers, One such variation was caused by the re-activation of Tyl-copia-like retrotransposons, implying that the mutagenic effects of lowenergy ions might work through elevated activation of retrotransposons, In this paper an expression profile of Tyl-copia-like retrotransposons in wheat treated by low-energy N^＋ ions is reported, The reverse transcriptase （RT） domains of these retrotransposons were amplified by reverse-transcriptional polymerase chain reaction （RT-PCR） and sequentially cloned, 42 and 65 clones were obtained from the treated （CL） and control materials （CK）, respectively, Sequence analysis of each clone was performed by software. Phylogeny and classification were calculated responding to the sequences of the RT domains. All the results show that there is much difference in the RT domain between the control sample and the treated sample, Especially, the RT domains from the treated group encode significantly more functional ORF （open reading frames） than those from the control sample, This observation suggests that the treated sample has higher activation of retrotransposons, possibly as a consequence of low-energy ion beam irradiation, It also suggests that retrotransposons in the two groups impact the host gene expression in two different ways and carry out different functions in wheat cells.展开更多
基金This work was supported by National Key Project (96-538-01-01).
文摘The No.601 watermelon (citrullus lanatus) seeds were treated with 25 keV N+ implantation at the dosage of 7.8 ×1016 ions/cm2. After treatment, watermelon seeds were incubated with 380μg/μl pumpkin (Cucubita, maxima Duch) DNA solution at 35 ℃ for 5 hours. By two-generations of selection and resistance screening at seedling stage, one transformed material was selected out, whose rind color is similar to that of the donor pumpkin and whose size of seeds is between that of the donor and the receptor. Using AFLP (amplified fragment length polymorphism) technique, two polymorphic DNA fragments were amplified. This primarily testified that the donor DNA fragments/gene were introduced into the receptor cell and integrated into the genomic DNA of the receptor.
基金supported by National Natural Science Foundation of China (No.10505018)Natural Science Foundation of Henan Province (No.511030400)
文摘Retrotransposon-like elements are major constituents of most eukaryotic genomes. For example, they account for roughly 90% of the wheat （Triticum aestivum） genome, Previous study on a wheat strain treated by low-energy N^＋ ions indicated the variations in AFLP （Amplified Fragment Length Polymorphism ） markers, One such variation was caused by the re-activation of Tyl-copia-like retrotransposons, implying that the mutagenic effects of lowenergy ions might work through elevated activation of retrotransposons, In this paper an expression profile of Tyl-copia-like retrotransposons in wheat treated by low-energy N^＋ ions is reported, The reverse transcriptase （RT） domains of these retrotransposons were amplified by reverse-transcriptional polymerase chain reaction （RT-PCR） and sequentially cloned, 42 and 65 clones were obtained from the treated （CL） and control materials （CK）, respectively, Sequence analysis of each clone was performed by software. Phylogeny and classification were calculated responding to the sequences of the RT domains. All the results show that there is much difference in the RT domain between the control sample and the treated sample, Especially, the RT domains from the treated group encode significantly more functional ORF （open reading frames） than those from the control sample, This observation suggests that the treated sample has higher activation of retrotransposons, possibly as a consequence of low-energy ion beam irradiation, It also suggests that retrotransposons in the two groups impact the host gene expression in two different ways and carry out different functions in wheat cells.
