title: Long-term T-DNA insert stability and transgene expression consistency in field propagated sugarcane.
abstract: This study addresses T-DNA insert stability and transgene expression consistency in multiple cycles of field propagated sugarcane. T-DNA inserts are stable; no transgene rearrangements were observed. AmCYAN1 and PMI protein accumulation levels were maintained. There was no evidence that production of either protein declined across generations and no transgene silencing was observed in three commercial sugarcane varieties through commercially relevant ratooning, propagation-by-setts, and micro-propagation generation processes over 4 years of field testing. Long term transgene expression consistency and T-DNA insert stability can be achieved in sugarcane, suggesting that it is highly probable that transgenic sugarcane can be successfully commercialized. This study addresses T-DNA insert stability and transgene expression consistency in multiple cycles of field propagated sugarcane. These data are critical supporting information needed for successful commercialization of GM sugarcane. Here seventeen transgenic events, containing the AmCYAN1 gene driven by a CMP promoter and the E. coli PMI gene driven by either a CMP or Ubi promoter, were used to monitor T-DNA insert stability and consistency of transgene encoded protein accumulation through commercially relevant ratooning, propagation-by-setts, and micro-propagation generation processes. The experiments were conducted in three commercial sugarcane varieties over 4 years of field testing. DNA gel blot analysis showed that the T-DNA inserts are stable; no transgene rearrangements were observed. Quantitative ELISA showed no evidence of decreasing AmCYAN1 and PMI protein levels across generations and no transgene silencing was observed. These results indicate that long term transgene expression consistency and T-DNA insert stability can be achieved in sugarcane, suggesting that it is highly probable that transgenic sugarcane can be successfully commercialized.
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Kerry Hosmer
authors.initials: KH
authors.lastname: Caffall
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA. chengkun.he@syngenta.com.
authors.forename: Chengkun
authors.initials: C
authors.lastname: He
authors.affiliation: Syngenta Seeds Incorporated, 2369 330th Street, Slater, IA, 50244, USA.
authors.forename: Michele
authors.initials: M
authors.lastname: Smith-Jones
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Kristin
authors.initials: K
authors.lastname: Mayo
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Pearl
authors.initials: P
authors.lastname: Mai
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Shujie
authors.initials: S
authors.lastname: Dong
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: John
authors.initials: J
authors.lastname: Ke
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Erik
authors.initials: E
authors.lastname: Dunder
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Michele
authors.initials: M
authors.lastname: Yarnall
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Whinna
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Joe
authors.initials: J
authors.lastname: DeMaio
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Weining
authors.initials: W
authors.lastname: Gu
authors.affiliation: Syngenta Jealott's Hill Research Center, Bracknell, BRK, RG42 6EY, UK.
authors.forename: Judith
authors.initials: J
authors.lastname: Sheldon
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Allen
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Tricia
authors.initials: T
authors.lastname: Costello
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Kristin
authors.initials: K
authors.lastname: Setliff
authors.affiliation: Vero Beach Research Center, Syngenta Crop Protection, LLC, 7145 58th Avenue, Vero Beach, FL, 32967, USA.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Jain
authors.affiliation: Vero Beach Research Center, Syngenta Crop Protection, LLC, 7145 58th Avenue, Vero Beach, FL, 32967, USA.
authors.forename: Ada
authors.initials: A
authors.lastname: Snyder
authors.affiliation: Vero Beach Research Center, Syngenta Crop Protection, LLC, 7145 58th Avenue, Vero Beach, FL, 32967, USA.
authors.forename: Clark
authors.initials: C
authors.lastname: Lovelady
authors.affiliation: Vero Beach Research Center, Syngenta Crop Protection, LLC, 7145 58th Avenue, Vero Beach, FL, 32967, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Rawls
authors.affiliation: Vero Beach Research Center, Syngenta Crop Protection, LLC, 7145 58th Avenue, Vero Beach, FL, 32967, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Palmer
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Bate
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Liang
authors.initials: L
authors.lastname: Shi
authors.affiliation: Syngenta Crop Protection, LLC, 9 Davis Drive, Research Triangle Park, Durham, NC, 27709-2257, USA.
authors.forename: Ian
authors.initials: I
authors.lastname: Jepson
mesh_terms: D000818:Animals; D004269:DNA, Bacterial / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008359:Mannose-6-Phosphate Isomerase / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015203:Reproducibility of Results; D031786:Saccharum / Q000235:genetics* / Q000254:growth & development; D012615:Sea Anemones / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D019076:Transgenes / Q000235:genetics*
chemical_list: C485182:Cyan Fluorescent Protein; D004269:DNA, Bacterial; C036483:T-DNA; D049452:Green Fluorescent Proteins; D008359:Mannose-6-Phosphate Isomerase
keywords: AmCYAN1; Expression consistency; PMI; T-DNA insert stability; Transgenic sugarcane