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Title: Generation of a selectable marker free, highly expressed single copy locus as landing pad for transgene stacking in sugarcane

Journal Article · · Plant Molecular Biology
 [1];  [2];  [1];  [3];  [1];  [1];  [4];  [4];  [5];  [5];  [5];  [6]; ORCiD logo [4]
  1. Univ. of Florida, Gainesville, FL (United States). Agronomy Dept. Plant Molecular and Cellular Biology Program. Genetics Inst.
  2. Univ. of Florida, Gainesville, FL (United States). Agronomy Dept. Plant Molecular and Cellular Biology Program. Genetics Inst.; Kongju National Univ., Yesan (Korea, Republic of). Dept. of Plant Resources. College of Industrial Science
  3. Univ. of Florida, Gainesville, FL (United States). Agronomy Dept. Plant Molecular and Cellular Biology Program. Genetics Inst.; Korea Inst. of Science and Technology (KIST), Gangwon (Korea, Republic of). Smart Farm Research Center. Inst. of Natural Products
  4. Univ. of Florida, Gainesville, FL (United States). Agronomy Dept. Plant Molecular and Cellular Biology Program. Genetics Inst. DOE Center for Advanced Bioenergy and Bioproducts Innovation
  5. Syngenta Crop Protection, LLC, Research Triangle Park, NC (United States)
  6. Univ. of Arkansas, Fayetteville, AR (United States). Crop, Soil and Environmental Sciences
+ Show Author Affiliations

Sugarcane, a tropical C4 grass in the genus Saccharum (Poaceae), accounts for nearly 80% of sugar produced worldwide and is also an important feedstock for biofuel production. Generating transgenic sugarcane with predictable and stable transgene expression is critical for crop improvement. We generated a highly expressed single copy locus as landing pad for transgene stacking. Transgenic sugarcane lines with stable integration of a single copy nptII expression cassette flanked by insulators supported higher transgene expression along with reduced line to line variation when compared to single copy events without insulators by NPTII ELISA analysis. Subsequently, the nptII selectable marker gene was efficiently excised from the sugarcane genome by the FLPe/FRT site-specific recombination system to create selectable marker free plants. This study provides valuable resources for future gene stacking using site-specific recombination or genome editing tools.

Research Organization:
Univ. of Florida, Gainesville, FL (United States); Syngenta Crop Protection, LLC, Research Triangle Park, NC (United States); Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
SC0018420
OSTI ID:
1505510
Journal Information:
Plant Molecular Biology, Vol. 100, Issue 3; ISSN 0167-4412
Publisher:
SpringerCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 6 works
Citation information provided by
Web of Science

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59 BASIC BIOLOGICAL SCIENCES
sugarcane
selection marker removal
site-specific recombination
insulator
FLP
FLPe