Long-term T-DNA insert stability and transgene expression consistency in field propagated sugarcane.

Plant molecular biology 2017 Mar

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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.

Citation

Kerry Hosmer Caffall, Chengkun He, Michele Smith-Jones, Kristin Mayo, Pearl Mai, Shujie Dong, John Ke, Erik Dunder, Michele Yarnall, Rachel Whinna, Joe DeMaio, Weining Gu, Judith Sheldon, Martin Allen, Tricia Costello, Kristin Setliff, Rakesh Jain, Ada Snyder, Clark Lovelady, Eric Rawls, Eric Palmer, Yan Zhang, Nicholas Bate, Liang Shi, Ian Jepson. Long-term T-DNA insert stability and transgene expression consistency in field propagated sugarcane. Plant molecular biology. 2017 Mar;93(4-5):451-463