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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

Cry1Ac protein expression in the leaves and stems of 14 different transgenic sugarcane lines detected by ELISA.
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pone.0153929.g004: Cry1Ac protein expression in the leaves and stems of 14 different transgenic sugarcane lines detected by ELISA.

Mentions: Double-antibody sandwich ELISA was performed to determine the Cry1Ac protein content in mature stage leaves and stems of 14 transgenic sugarcane lines. Standard Bt samples were used to construct a standard curve for quantitative protein detection (Fig 3). The slope was Y = 0.766X+0.017, R2 = 0.999, where Y is absorbance at OD450, and X is the concentration of standard Bt Cry1Ac protein samples. Significant correlation was found between the absorbance and the concentration of the standard protein (R2 = 0.999). Thus, the protein expression of the tested samples was measured based on the linear equation (Fig 4). The results showed that Cry1Ac protein expression was only detected in leaves and stems of seven transgenic lines. Expression in leaves was variable and ranged from a minimum of 0.85μg/FWg in T-1 to a maximum of 70.92 μg/FWg in T-2. In stems, the expression was also variable and ranged from a minimum of 0.04 μg/FWg in T-1 to a maximum of 7.22 μg/FWg in T-2. It should be noted that although there was no Cry1Ac protein expression in the remaining seven transgenic lines, the expression of cry1Ac gene was detected in all these lines.


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)

Cry1Ac protein expression in the leaves and stems of 14 different transgenic sugarcane lines detected by ELISA.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153929.g004: Cry1Ac protein expression in the leaves and stems of 14 different transgenic sugarcane lines detected by ELISA.
Mentions: Double-antibody sandwich ELISA was performed to determine the Cry1Ac protein content in mature stage leaves and stems of 14 transgenic sugarcane lines. Standard Bt samples were used to construct a standard curve for quantitative protein detection (Fig 3). The slope was Y = 0.766X+0.017, R2 = 0.999, where Y is absorbance at OD450, and X is the concentration of standard Bt Cry1Ac protein samples. Significant correlation was found between the absorbance and the concentration of the standard protein (R2 = 0.999). Thus, the protein expression of the tested samples was measured based on the linear equation (Fig 4). The results showed that Cry1Ac protein expression was only detected in leaves and stems of seven transgenic lines. Expression in leaves was variable and ranged from a minimum of 0.85μg/FWg in T-1 to a maximum of 70.92 μg/FWg in T-2. In stems, the expression was also variable and ranged from a minimum of 0.04 μg/FWg in T-1 to a maximum of 7.22 μg/FWg in T-2. It should be noted that although there was no Cry1Ac protein expression in the remaining seven transgenic lines, the expression of cry1Ac gene was detected in all these lines.

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