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Assessing the bioconfinement potential of a Nicotiana hybrid platform for use in plant molecular farming applications.

Rice JH, Mundell RE, Millwood RJ, Chambers OD, Stewart CN, Davies HM - BMC Biotechnol. (2013)

Bottom Line: The introduction of pharmaceutical traits in tobacco for commercial production could benefit from the utilization of a transgene bioconfinement system.GFP was used as a useful proxy for pharmaceutical transgenes.In two field experiments, one each in Tennessee and Kentucky, we detected outcrossing at only one location (Tennessee) at 1.4%.

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Affiliation: Department of Plant Sciences, University of Tennessee, Knoxville, TN 37996, USA.

ABSTRACT

Background: The introduction of pharmaceutical traits in tobacco for commercial production could benefit from the utilization of a transgene bioconfinement system. It has been observed that interspecific F1Nicotiana hybrids (Nicotiana tabacum × Nicotiana glauca) are sterile and thus proposed that hybrids could be suitable bioconfined hosts for biomanufacturing. We genetically tagged hybrids with green fluorescent protein (GFP), which was used as a visual marker to enable gene flow tracking and quantification for field and greenhouse studies. GFP was used as a useful proxy for pharmaceutical transgenes.

Results: Analysis of DNA content revealed significant genomic downsizing of the hybrid relative to that of N. tabacum. Hybrid pollen was capable of germination in vitro, albeit with a very low frequency and with significant differences between plants. In two field experiments, one each in Tennessee and Kentucky, we detected outcrossing at only one location (Tennessee) at 1.4%. Additionally, from 50 hybrid plants at each field site, formation of 84 and 16 seed was observed, respectively. Similar conclusions about hybrid fertility were drawn from greenhouse crosses. In terms of above-ground biomass, the hybrid yield was not significantly different than that of N. tabacum in the field.

Conclusion: N. tabacum × N. glauca hybrids show potential to contribute to a bioconfinement- and biomanufacturing host system. Hybrids exhibit extremely low fertility with no difference of green biomass yields relative to N. tabacum. In addition, hybrids are morphologically distinguishable from tobacco allowing for identity preservation. This hybrid system for biomanufacturing would optimally be used where N. glauca is not present and in physical isolation of N. tabacum production to provide total bioconfinement.

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Fresh aboveground biomass productivity. (A) Productivity was measured for nontransgenic hybrid plants and nontransgenic TN 90 plants in a field experiment over two years with three measurements per year. (B) A greenhouse study was conducted during one year with two measurements of the hybrid GFP line, the transgenic parental lines of the aforementioned hybrid, and a nontransgenic hybrid line. Mean separation is by Fishers LSD and bars marked by the same letter are not significantly different (p < 0.05). Error bars are the standard error of the means.
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Figure 3: Fresh aboveground biomass productivity. (A) Productivity was measured for nontransgenic hybrid plants and nontransgenic TN 90 plants in a field experiment over two years with three measurements per year. (B) A greenhouse study was conducted during one year with two measurements of the hybrid GFP line, the transgenic parental lines of the aforementioned hybrid, and a nontransgenic hybrid line. Mean separation is by Fishers LSD and bars marked by the same letter are not significantly different (p < 0.05). Error bars are the standard error of the means.

Mentions: For interspecific hybrid Nicotianas to have potential as production-host plants in biomanufacturing they should produce high biomass; e.g., comparable to commercial tobacco. There were not significant differences of biomass between hybrids and TN 90 in the field experiment (p = 0.738; Figure 3A). There were biomass differences among plant types in the greenhouse study (Figure 3B). We conclude that interspecific hybrid tobacco performed comparably to N. tabacum in biomass.


Assessing the bioconfinement potential of a Nicotiana hybrid platform for use in plant molecular farming applications.

Rice JH, Mundell RE, Millwood RJ, Chambers OD, Stewart CN, Davies HM - BMC Biotechnol. (2013)

Fresh aboveground biomass productivity. (A) Productivity was measured for nontransgenic hybrid plants and nontransgenic TN 90 plants in a field experiment over two years with three measurements per year. (B) A greenhouse study was conducted during one year with two measurements of the hybrid GFP line, the transgenic parental lines of the aforementioned hybrid, and a nontransgenic hybrid line. Mean separation is by Fishers LSD and bars marked by the same letter are not significantly different (p < 0.05). Error bars are the standard error of the means.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750662&req=5

Figure 3: Fresh aboveground biomass productivity. (A) Productivity was measured for nontransgenic hybrid plants and nontransgenic TN 90 plants in a field experiment over two years with three measurements per year. (B) A greenhouse study was conducted during one year with two measurements of the hybrid GFP line, the transgenic parental lines of the aforementioned hybrid, and a nontransgenic hybrid line. Mean separation is by Fishers LSD and bars marked by the same letter are not significantly different (p < 0.05). Error bars are the standard error of the means.
Mentions: For interspecific hybrid Nicotianas to have potential as production-host plants in biomanufacturing they should produce high biomass; e.g., comparable to commercial tobacco. There were not significant differences of biomass between hybrids and TN 90 in the field experiment (p = 0.738; Figure 3A). There were biomass differences among plant types in the greenhouse study (Figure 3B). We conclude that interspecific hybrid tobacco performed comparably to N. tabacum in biomass.

Bottom Line: The introduction of pharmaceutical traits in tobacco for commercial production could benefit from the utilization of a transgene bioconfinement system.GFP was used as a useful proxy for pharmaceutical transgenes.In two field experiments, one each in Tennessee and Kentucky, we detected outcrossing at only one location (Tennessee) at 1.4%.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Plant Sciences, University of Tennessee, Knoxville, TN 37996, USA.

ABSTRACT

Background: The introduction of pharmaceutical traits in tobacco for commercial production could benefit from the utilization of a transgene bioconfinement system. It has been observed that interspecific F1Nicotiana hybrids (Nicotiana tabacum × Nicotiana glauca) are sterile and thus proposed that hybrids could be suitable bioconfined hosts for biomanufacturing. We genetically tagged hybrids with green fluorescent protein (GFP), which was used as a visual marker to enable gene flow tracking and quantification for field and greenhouse studies. GFP was used as a useful proxy for pharmaceutical transgenes.

Results: Analysis of DNA content revealed significant genomic downsizing of the hybrid relative to that of N. tabacum. Hybrid pollen was capable of germination in vitro, albeit with a very low frequency and with significant differences between plants. In two field experiments, one each in Tennessee and Kentucky, we detected outcrossing at only one location (Tennessee) at 1.4%. Additionally, from 50 hybrid plants at each field site, formation of 84 and 16 seed was observed, respectively. Similar conclusions about hybrid fertility were drawn from greenhouse crosses. In terms of above-ground biomass, the hybrid yield was not significantly different than that of N. tabacum in the field.

Conclusion: N. tabacum × N. glauca hybrids show potential to contribute to a bioconfinement- and biomanufacturing host system. Hybrids exhibit extremely low fertility with no difference of green biomass yields relative to N. tabacum. In addition, hybrids are morphologically distinguishable from tobacco allowing for identity preservation. This hybrid system for biomanufacturing would optimally be used where N. glauca is not present and in physical isolation of N. tabacum production to provide total bioconfinement.

Show MeSH
Related in: MedlinePlus