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Summary Integration ofAgrobacterium tumefacienstransferred 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.We conducted quantitative transformation assays of wild‐type andpolQmutantArabidopsisand rice, analyzed T‐DNA/plant DNA junction sequences, and (forArabidopsis) measured the amount of integrated T‐DNA in mutant and wild‐type tissue.Unexpectedly, we were able to generate stable transformants of all tested lines, although the transformation frequency ofpolQmutants was c.20% that of wild‐type plants. T‐DNA/plant DNA junctions from these transformed rice andArabidopsis polQmutants closely resembled those from wild‐type plants, indicating that loss of PolQ activity does not alter the characteristics of T‐DNA integration events.polQmutant plants show growth and developmental defects, perhaps explaining previous unsuccessful attempts at their stable transformation.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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Mutations in the predicted DNA polymerase subunit POLD3 result in more rapid flowering of Brachypodium distachyon
Summary The timing of reproduction is a critical developmental decision in the life cycle of many plant species.Fine mapping of a rapid‐flowering mutant was done using whole‐genome sequence data from bulked DNA from a segregating F2 mapping populations. The causative mutation maps to a gene orthologous with the third subunit of DNA polymerase δ (POLD3), a previously uncharacterized gene in plants. Expression analyses of POLD3 were conducted via real time qPCR to determine when and in what tissues the gene is expressed.To better understand the molecular basis of the rapid‐flowering phenotype, transcriptomic analyses were conducted in the mutant vs wild‐type. Consistent with the rapid‐flowering mutant phenotype, a range of genes involved in floral induction and flower development are upregulated in the mutant.Our results provide the first characterization of the developmental and gene expression phenotypes that result from a lesion inPOLD3in plants.
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- Award ID(s):
- 1755224
- PAR ID:
- 10456176
- Publisher / Repository:
- Wiley-Blackwell
- Date Published:
- Journal Name:
- New Phytologist
- Volume:
- 227
- Issue:
- 6
- ISSN:
- 0028-646X
- Page Range / eLocation ID:
- p. 1725-1735
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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