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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

Bioassay in the seedling stage.A: Symptoms of non-transgenic sugarcane plantlets challenged with sugarcane borers; B: Symptoms of transgenic sugarcane plantlets challenged with sugarcane borers; C: Sugarcane borer feeding with non-transgenic sugarcane plantlets; D: Sugarcane borer feeding with transgenic sugarcane plantlets.
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pone.0153929.g005: Bioassay in the seedling stage.A: Symptoms of non-transgenic sugarcane plantlets challenged with sugarcane borers; B: Symptoms of transgenic sugarcane plantlets challenged with sugarcane borers; C: Sugarcane borer feeding with non-transgenic sugarcane plantlets; D: Sugarcane borer feeding with transgenic sugarcane plantlets.

Mentions: The transgenic sugarcane lines were generally more resistant to sugarcane borer compared to the non-transgenic lines. In the seedling stage, 7 d after inoculation, the typical effects of borer attack leading to dead heart were seen on the non-transgenic plants. The same effects were observed after 10 d in several transgenic lines. Plantlets of non-transgenic sugarcane withered slowly 10 d after inoculation. At 15 d, only transgenic lines continued to grow normally (Fig 5B), while non-transgenics withered or died (Fig 5A). In the transgenic lines, only 10%-20% of borers survived but these appeared weak and were small (Fig 5D). In contrast, the borers in non-transgenic control plants had significantly higher survivorship and were larger (Fig 5C). In the field trials, the transgenic lines were also resistant to the sugarcane borer damage to leaves and stems compared to the non-transgenic control (Fig 6). The borer damage rate in all transgenic lines was lower than the control and differences were significant except for lines T-7, T-10, and T-12. There was no significant difference between the lines T-2, T-3, T-5, T-8, T-11, and T-14, all of which had higher protein expression and a medium cry1Ac copy number (Table 2). Although the transgenic sugarcane lines were still attacked by stem borers, the degree of damage was clearly less than the damage done to the non-transgenic control line (Fig 7).


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)

Bioassay in the seedling stage.A: Symptoms of non-transgenic sugarcane plantlets challenged with sugarcane borers; B: Symptoms of transgenic sugarcane plantlets challenged with sugarcane borers; C: Sugarcane borer feeding with non-transgenic sugarcane plantlets; D: Sugarcane borer feeding with transgenic sugarcane plantlets.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153929.g005: Bioassay in the seedling stage.A: Symptoms of non-transgenic sugarcane plantlets challenged with sugarcane borers; B: Symptoms of transgenic sugarcane plantlets challenged with sugarcane borers; C: Sugarcane borer feeding with non-transgenic sugarcane plantlets; D: Sugarcane borer feeding with transgenic sugarcane plantlets.
Mentions: The transgenic sugarcane lines were generally more resistant to sugarcane borer compared to the non-transgenic lines. In the seedling stage, 7 d after inoculation, the typical effects of borer attack leading to dead heart were seen on the non-transgenic plants. The same effects were observed after 10 d in several transgenic lines. Plantlets of non-transgenic sugarcane withered slowly 10 d after inoculation. At 15 d, only transgenic lines continued to grow normally (Fig 5B), while non-transgenics withered or died (Fig 5A). In the transgenic lines, only 10%-20% of borers survived but these appeared weak and were small (Fig 5D). In contrast, the borers in non-transgenic control plants had significantly higher survivorship and were larger (Fig 5C). In the field trials, the transgenic lines were also resistant to the sugarcane borer damage to leaves and stems compared to the non-transgenic control (Fig 6). The borer damage rate in all transgenic lines was lower than the control and differences were significant except for lines T-7, T-10, and T-12. There was no significant difference between the lines T-2, T-3, T-5, T-8, T-11, and T-14, all of which had higher protein expression and a medium cry1Ac copy number (Table 2). Although the transgenic sugarcane lines were still attacked by stem borers, the degree of damage was clearly less than the damage done to the non-transgenic control line (Fig 7).

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