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Pollen-mediated flow of bar gene in transgenic herbicide-resistant turf grass Zoysia japonica

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Abstract

Weed control can be most effectively achieved through the use of herbicide-resistance transgene. A preliminary study bar-transgenic Zoysia japonica posed no serious risk on the unintended escape of the transgene from its cultivation site. The present follow-up investigation of the dispersion of pollen and its short-distance escape outside of the bar-transgenic Zoysia japonica habitats were ascertained in terms of environmental factors affecting anthesis and pollen viability. In a 24-h day cycle, zoysiagrass pollen was released predominantly between 08:00 and 10:00, and the pollen was most viable during the same time interval. Optimal temperature and humidity for pollen viability was 15–20 °C and 80–90%, respectively. The pollen germinated in 120 min after anthesis, but under cloudy conditions germination time doubled. No differences in pollen viability/longevity between the transgenic and non-transgenic plants were observed. The pollen-mediated gene flow of transgenic Zoysia japonica to wild-type non-transgenic zoysiagrass species was monitored by measuring the cross-over rate of the bar gene in the context of three different models. At distances within 5 m, the rate of gene flow ranged from 3 to 5.7% according to the models used. The greater the distance from the transgenic plant site, the lower the gene flow rate. The furthest transgene detected was 38 m away and exhibited a 0.25% gene flow rate. The radial model yielded a 3.7% escape rate within a 3 m radius and was wind direction dependent. The distance- and direction-dependent gene flow events were influenced by wind direction and velocity during flowering season.

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Acknowledgments

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) of the Ministry of Education (2019R1A6A1A11052070), Republic of Korea.

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Correspondence to Hong-Gyu Kang or Hyo-Yeon Lee.

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11816_2021_667_MOESM1_ESM.jpg

Supplementary Fig. S1. Monitoring of long distance cross fertilization. Pool detection limit of the LL strip test (A) and PCR (B). Lane 1 and 2 are WT and JG21, respectively. Lane 3 to 7 mixed WT to JG21 at a certain ratio. Lane 3, 1:1; lane 4, 3:1; lane 5, 10:1; lane 6, 20:1; lane 7, 50:1. Arrow heads in Supplementary Fig. S1A and 1B indicate the PAT protein band and the targeted PCR product (810 bp), respectively. C, Each strip was tested with samples pooled from 9 sites and 107 strips were used for samples of total 963 sites. D, Each PCR was tested with samples pooled from 9 sites and 107 PCRs were used for samples of total 963 sites. The right agarose gel image shows the result of the first 50 out of 107 PCRs. The 0 lane is the result from the mixture of WT to JG21 (10:1). As a PCR control marker, an actin gene (900 bp) of zoysiagrass was amplified with 5′-GACATGGTAC TGGAATGGTC-3′ and 5′-AGGATTCAAA GGAGCTTCAG-3′ primer set (the left image)

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Kang, HG., Chung, OC., Bae, TW. et al. Pollen-mediated flow of bar gene in transgenic herbicide-resistant turf grass Zoysia japonica. Plant Biotechnol Rep 15, 241–250 (2021). https://doi.org/10.1007/s11816-021-00667-4

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