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Transgenic Sugarcane with a cry1Ac Gene Exhibited Better Phenotypic Traits and Enhanced Resistance against Sugarcane Borer.

Gao S, Yang Y, Wang C, Guo J, Zhou D, Wu Q, Su Y, Xu L, Que Y - PLoS ONE (2016)

Bottom Line: We developed sugarcane plants with improved resistance to the sugarcane borer, Diatraea saccharalis (F).Compared to the non-transgenic control plants, all transgenic lines with medium copy numbers had relatively equal or lower sucrose yield and significantly improved sugarcane borer resistance, which lowered susceptibility to damage by insects.This suggests that the transgenic sugarcane lines harboring medium copy numbers of the cry1Ac gene may have significantly higher resistance to sugarcane borer but the sugarcane yield in these lines is similar to the non-transgenic control thus making them superior to the control lines.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.

ABSTRACT
We developed sugarcane plants with improved resistance to the sugarcane borer, Diatraea saccharalis (F). An expression vector pGcry1Ac0229, harboring the cry1Ac gene and the selectable marker gene, bar, was constructed. This construct was introduced into the sugarcane cultivar FN15 by particle bombardment. Transformed plantlets were identified after selection with Phosphinothricin (PPT) and Basta. Plantlets were then screened by PCR based on the presence of cry1Ac and 14 cry1Ac positive plantlets were identified. Real-time quantitative PCR (RT-qPCR) revealed that the copy number of cry1Ac gene in the transgenic lines varied from 1 to 148. ELISA analysis showed that Cry1Ac protein levels in 7 transgenic lines ranged from 0.85 μg/FWg to 70.92 μg/FWg in leaves and 0.04 μg/FWg to 7.22 μg/FWg in stems, and negatively correlated to the rate of insect damage that ranged from 36.67% to 13.33%, respectively. Agronomic traits of six transgenic sugarcane lines with medium copy numbers were similar to the non-transgenic parental line. However, phenotype was poor in lines with high or low copy numbers. Compared to the non-transgenic control plants, all transgenic lines with medium copy numbers had relatively equal or lower sucrose yield and significantly improved sugarcane borer resistance, which lowered susceptibility to damage by insects. This suggests that the transgenic sugarcane lines harboring medium copy numbers of the cry1Ac gene may have significantly higher resistance to sugarcane borer but the sugarcane yield in these lines is similar to the non-transgenic control thus making them superior to the control lines.

No MeSH data available.


Related in: MedlinePlus

Electrophoresis of PCR amplified products of the candidate cry1Ac transgenic sugarcane plants.M, DNA Marker; 1–9, Basta-resistant plants; 10, Positive control (plasmid pGcry1Ac0229); 11, Negative control (non-transgenic sugarcane without bombardment); 12, Blank control.
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pone.0153929.g002: Electrophoresis of PCR amplified products of the candidate cry1Ac transgenic sugarcane plants.M, DNA Marker; 1–9, Basta-resistant plants; 10, Positive control (plasmid pGcry1Ac0229); 11, Negative control (non-transgenic sugarcane without bombardment); 12, Blank control.

Mentions: PCR validation was performed on the Basta resistant regenerated plants. A total of 14 independent plants amplified the expected size DNA consistent with the positive control (pGcry1Ac0229), while the negative non-transgenic line and the blank control did not amplify any band (Fig 2). Further sequencing showed that this band was precisely the 211 bp partial sequence of cry1Ac, suggesting that the cry1Ac gene was successfully inserted into the sugarcane genome. A total of 14 among the 26 Basta-resistant regenerated plants tested positive by PCR.


Transgenic Sugarcane with a cry1Ac Gene Exhibited Better Phenotypic Traits and Enhanced Resistance against Sugarcane Borer.

Gao S, Yang Y, Wang C, Guo J, Zhou D, Wu Q, Su Y, Xu L, Que Y - PLoS ONE (2016)

Electrophoresis of PCR amplified products of the candidate cry1Ac transgenic sugarcane plants.M, DNA Marker; 1–9, Basta-resistant plants; 10, Positive control (plasmid pGcry1Ac0229); 11, Negative control (non-transgenic sugarcane without bombardment); 12, Blank control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836700&req=5

pone.0153929.g002: Electrophoresis of PCR amplified products of the candidate cry1Ac transgenic sugarcane plants.M, DNA Marker; 1–9, Basta-resistant plants; 10, Positive control (plasmid pGcry1Ac0229); 11, Negative control (non-transgenic sugarcane without bombardment); 12, Blank control.
Mentions: PCR validation was performed on the Basta resistant regenerated plants. A total of 14 independent plants amplified the expected size DNA consistent with the positive control (pGcry1Ac0229), while the negative non-transgenic line and the blank control did not amplify any band (Fig 2). Further sequencing showed that this band was precisely the 211 bp partial sequence of cry1Ac, suggesting that the cry1Ac gene was successfully inserted into the sugarcane genome. A total of 14 among the 26 Basta-resistant regenerated plants tested positive by PCR.

Bottom Line: We developed sugarcane plants with improved resistance to the sugarcane borer, Diatraea saccharalis (F).Compared to the non-transgenic control plants, all transgenic lines with medium copy numbers had relatively equal or lower sucrose yield and significantly improved sugarcane borer resistance, which lowered susceptibility to damage by insects.This suggests that the transgenic sugarcane lines harboring medium copy numbers of the cry1Ac gene may have significantly higher resistance to sugarcane borer but the sugarcane yield in these lines is similar to the non-transgenic control thus making them superior to the control lines.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.

ABSTRACT
We developed sugarcane plants with improved resistance to the sugarcane borer, Diatraea saccharalis (F). An expression vector pGcry1Ac0229, harboring the cry1Ac gene and the selectable marker gene, bar, was constructed. This construct was introduced into the sugarcane cultivar FN15 by particle bombardment. Transformed plantlets were identified after selection with Phosphinothricin (PPT) and Basta. Plantlets were then screened by PCR based on the presence of cry1Ac and 14 cry1Ac positive plantlets were identified. Real-time quantitative PCR (RT-qPCR) revealed that the copy number of cry1Ac gene in the transgenic lines varied from 1 to 148. ELISA analysis showed that Cry1Ac protein levels in 7 transgenic lines ranged from 0.85 μg/FWg to 70.92 μg/FWg in leaves and 0.04 μg/FWg to 7.22 μg/FWg in stems, and negatively correlated to the rate of insect damage that ranged from 36.67% to 13.33%, respectively. Agronomic traits of six transgenic sugarcane lines with medium copy numbers were similar to the non-transgenic parental line. However, phenotype was poor in lines with high or low copy numbers. Compared to the non-transgenic control plants, all transgenic lines with medium copy numbers had relatively equal or lower sucrose yield and significantly improved sugarcane borer resistance, which lowered susceptibility to damage by insects. This suggests that the transgenic sugarcane lines harboring medium copy numbers of the cry1Ac gene may have significantly higher resistance to sugarcane borer but the sugarcane yield in these lines is similar to the non-transgenic control thus making them superior to the control lines.

No MeSH data available.


Related in: MedlinePlus