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Int. J. Mol. Sci. 2018, 19(6), 1692; https://doi.org/10.3390/ijms19061692

Particle Bombardment of the cry2A Gene Cassette Induces Stem Borer Resistance in Sugarcane

Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture and Key Laboratory of Crop Genetics and Breeding and Comprehensive Utilization, College of Crop Science, Fujian Agriculture and Forestry University, Ministry of Education, Fuzhou 350002, China
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Received: 3 May 2018 / Revised: 31 May 2018 / Accepted: 4 June 2018 / Published: 6 June 2018
(This article belongs to the Special Issue Plant Genetics and Molecular Breeding)
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Abstract

Sugarcane borer is the most common and harmful pest in Chinese sugarcane fields, and can cause damage to the whole plant during the entire growing season. To improve borer resistance in sugarcane, we constructed a plant expression vector pGcry2A0229 with the bar gene as the marker and the cry2A gene as the target, and introduced it into embryogenic calli of most widely cultivated sugarcane cultivar ROC22 by particle bombardment. After screening with phosphinothricin in vitro and Basta spray, 21 resistance-regenerated plants were obtained, and 10 positive transgenic lines harboring the cry2A gene were further confirmed by conventional PCR detection. Real-time quantitative PCR (RT-qPCR) analysis showed that the copy number of the cry2A gene varied among different transgenic lines but did not exceed four copies. Quantitative ELISA analysis showed that there was no linear relationship with copy number but negatively correlated with the percentage of borer-infested plants. The analysis of industrial and agronomic traits showed that the theoretical sugar yields of transgenic lines TR-4 and TR-10 were slightly lower than that of the control in both plant cane and ratoon cane; nevertheless, TR-4 and TR-10 lines exhibited markedly lower in frequency of borer-infested plants in plant cane and in the ratoon cane compared to the control. Our results indicate that the introduction of the cry2A gene via bombardment produces transgenic lines with obviously increased stem borer resistance and comparable sugar yield, providing a practical value in direct commercial cultivation and crossbreeding for ROC22 has been used as the most popular elite genitor in various breeding programs in China. View Full-Text
Keywords: sugarcane; cry2A gene; particle bombardment; stem borer; resistance sugarcane; cry2A gene; particle bombardment; stem borer; resistance
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Gao, S.; Yang, Y.; Xu, L.; Guo, J.; Su, Y.; Wu, Q.; Wang, C.; Que, Y. Particle Bombardment of the cry2A Gene Cassette Induces Stem Borer Resistance in Sugarcane. Int. J. Mol. Sci. 2018, 19, 1692.

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