Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus

  • BMC Biotechnol. 2018 Aug 9;18(1):48. doi: 10.1186/s12896-018-0459-5.
Jing Xi  1  2 ,  Minesh Patel  1  3 ,  Shujie Dong  4 ,  Qiudeng Que  4 ,  Rongda Qu  5
Affiliations
  • 1. Department of Crop and Soil Sciences, North Carolina State University, Raleigh, NC, 27695, USA.
  • 2. Present address: Department of Biochemistry, University of California, Riverside, CA, 92521, USA.
  • 3. Present address: BASF Corporation-R&D Center, Durham, NC, 27709, USA.
  • 4. Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709, USA.
  • 5. Department of Crop and Soil Sciences, North Carolina State University, Raleigh, NC, 27695, USA. [email protected].
PMID: 30092808 DOI: 10.1186/s12896-018-0459-5
Abstract

Background: Large T-DNA fragment transfer has long been a problem for Agrobacterium-mediated transformation. Although vector systems, such as the BIBAC series, were successfully developed for the purpose, low transformation efficiencies were consistently observed.

Results: To gain insights of this problem in monocot transformation, we investigated the T-strand accumulation of various size of T-DNA in two kinds of binary vectors (one copy vs. multi-copy) upon acetosyringone (AS) induction and explored ways to improve the efficiency of the large T-DNA fragment transfer in Agrobacterium-mediated rice transformation. By performing immuno-precipitation of VirD2-T-strands and quantitative Real-Time PCR assays, we monitored the accumulation of the T-strands in Agrobacterium tumeficiens after AS induction. We further demonstrated that extension of AS induction time highly significantly improved large-size T-DNA transfer to rice cells.

Conclusions: Our data provide valuable information of the T-strand dynamics and its impact on large T-DNA transfer in monocots, and likely dicots as well.

Keywords
Agrobacterium; BIBAC; Immuno-precipitation; Large T-DNA; Monocot transformation.
Products