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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

Partially regenerated plantlets with PPT resistance in differentiation culture.
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pone.0153929.g001: Partially regenerated plantlets with PPT resistance in differentiation culture.

Mentions: The pGcry1Ac0229 plasmid DNA was used to prepare microprojectiles for bombardment transformation, and the embryonic calli of the sugarcane cultivar FN15 were used as receptors. Based on previously determined screening concentrations of PPT (0.75 mg/L), cultures were screened and the putative transformants that survived the stress of PPT were obtained. Wild type cells without PPT resistance gradually turned brown and then died (Fig 1). Regenerated plantlets were transferred to rooting medium without PPT and the total number of survivors was 189. These plantlets were planted in soil in 72-cave trays and the second round of screening in vitro was conducted by spraying a solution containing 3.0 ‰ Basta on the plantlets. After 15–20 d, most of them gradually became yellow, wilted, and dried (S3 Fig). The final total of surviving plants was 26.


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)

Partially regenerated plantlets with PPT resistance in differentiation culture.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153929.g001: Partially regenerated plantlets with PPT resistance in differentiation culture.
Mentions: The pGcry1Ac0229 plasmid DNA was used to prepare microprojectiles for bombardment transformation, and the embryonic calli of the sugarcane cultivar FN15 were used as receptors. Based on previously determined screening concentrations of PPT (0.75 mg/L), cultures were screened and the putative transformants that survived the stress of PPT were obtained. Wild type cells without PPT resistance gradually turned brown and then died (Fig 1). Regenerated plantlets were transferred to rooting medium without PPT and the total number of survivors was 189. These plantlets were planted in soil in 72-cave trays and the second round of screening in vitro was conducted by spraying a solution containing 3.0 ‰ Basta on the plantlets. After 15–20 d, most of them gradually became yellow, wilted, and dried (S3 Fig). The final total of surviving plants was 26.

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