Abstract
Agrobacterium tumefaciens-mediated transformation (ATMT) has been proven to be a powerful strategy for gene disruption in plants and fungi. Patterns associated with transferred DNA (T-DNA) integration in plants and yeast have been studied comprehensively, whereas no detailed analysis of T-DNA integration has been reported yet in filamentous fungi. Here, we reported the T-DNA insertion patterns in the genome of filamentous fungus Magnaporthe oryzae. Using ATMT, a T-DNA tagged population consisting of 6,179 transformants of M. oryzae was constructed. With thermal asymmetric interlaced-PCR (TAIL-PCR), 623 right border (RB) flanking sequences and 124 left border (LB) flanking sequences were generated. Analysis of these flanking sequences indicated a significant integration bias toward non-coding sequences, suggesting distribution of T-DNAs was not random. Comparing to T-DNA RB, LB was nicked inaccurately and truncated frequently during integration. Chromosomal rearrangements, such as deletion, inversion, and translocation, were associated with T-DNA integration in some transformants. Our data suggest that, comparing with plant cells, T-DNA integrates into this filamentous fungus with more precise and simpler patterns. Some phenotypic mutants were observed in our T-DNA tagged population, and these transformants will be very useful for functional genomics research of M. oryzae.
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This project was supported by the Ministry of Science and Technology of China (Grant 2002BA711A15).
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Communicated by K. Borkovich.
Guihua Li and Zhuangzhi Zhou contributed equally to the work.
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Li, G., Zhou, Z., Liu, G. et al. Characterization of T-DNA insertion patterns in the genome of rice blast fungus Magnaporthe oryzae . Curr Genet 51, 233–243 (2007). https://doi.org/10.1007/s00294-007-0122-5
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DOI: https://doi.org/10.1007/s00294-007-0122-5