Integration of We conducted quantitative transformation assays of wild‐type and Unexpectedly, we were able to generate stable transformants of all tested lines, although the transformation frequency of We suggest that either multiple redundant pathways function in T‐DNA integration, and/or that integration requires some yet unknown pathway.
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Summary Agrobacterium tumefaciens transferred DNA (T‐DNA) into the plant genome is the last step required for stable plant genetic transformation. The mechanism of T‐DNA integration remains controversial, although scientists have proposed the participation of various nonhomologous end‐joining (NHEJ) pathways. Recent evidence suggests that inArabidopsis , DNA polymerase θ (PolQ) may be a crucial enzyme involved in T‐DNA integration.polQ mutantArabidopsis and rice, analyzed T‐DNA/plant DNA junction sequences, and (forArabidopsis ) measured the amount of integrated T‐DNA in mutant and wild‐type tissue.polQ mutants wasc. 20% that of wild‐type plants. T‐DNA/plant DNA junctions from these transformed rice andArabidopsis polQ mutants closely resembled those from wild‐type plants, indicating that loss of PolQ activity does not alter the characteristics of T‐DNA integration events.polQ mutant plants show growth and developmental defects, perhaps explaining previous unsuccessful attempts at their stable transformation.