Articles
MANNOSE SELECTION SYSTEM TO SCREEN VGB TRANSGENIC CHRYSANTHEMUM PLANT
Article number
953_31
Pages
223 – 230
Language
English
Abstract
Chrysanthemum morifolium is an important crop in the flower market.
It is an historic flower in China.
There are more than 3000 species domestically, which are rich in both pattern and color.
Ground-cover chrysanthemum is an important landscape plant in China but it cannot be resistant to waterlogging.
The traditional breeding method cannot solve this problem.
Therefore, the advantages about targeted improvement and short breeding cycle of transgenic technology are particularly important in Chrysanthemum breeding.
Furthermore, the safety problem of molecular breeding is the focus in society.
This concern could be settled if a non-antibiotic mannose selection system using the phosphomannose-isomerase (Pmi) gene is used.
The Vgb gene is isolated from the bacterium Vitreoscilla. The strict aerobic bacterium Vitreoscilla stercoraria expresses hemoglobin-like protein, Vhb, which facilitates oxygen transportation and efficiently dissociates the oxygen for aerobiosis.
It can improve the plants tolerance ability under water.
Its function has been proven in many plants.
In this study, the new selectable marker – Pmi gene was used.
This selection system utilizes the Pmi gene encoding for phosphomannose-isomerase that convert mannose-6-phosphate to fructose-6- phosphate.
Only transformed cells are capable of utilizing mannose as a carbon source.
A vector, pCAMBIA1301-Pmi-Vgb, with Vgb gene and Pmi gene was constructed.
With this vector the genes Pmi and Vgb have been transformed into ground-cover chrysanthemum Fenditan by Agrobacterium-mediated method.
The antibiotics-resistant seedlings were respectively detected by PCR with primers depending on Vgb. The PCR result showed that the Vgb gene has been transmitted into chrysanthemum successfully.
It is an historic flower in China.
There are more than 3000 species domestically, which are rich in both pattern and color.
Ground-cover chrysanthemum is an important landscape plant in China but it cannot be resistant to waterlogging.
The traditional breeding method cannot solve this problem.
Therefore, the advantages about targeted improvement and short breeding cycle of transgenic technology are particularly important in Chrysanthemum breeding.
Furthermore, the safety problem of molecular breeding is the focus in society.
This concern could be settled if a non-antibiotic mannose selection system using the phosphomannose-isomerase (Pmi) gene is used.
The Vgb gene is isolated from the bacterium Vitreoscilla. The strict aerobic bacterium Vitreoscilla stercoraria expresses hemoglobin-like protein, Vhb, which facilitates oxygen transportation and efficiently dissociates the oxygen for aerobiosis.
It can improve the plants tolerance ability under water.
Its function has been proven in many plants.
In this study, the new selectable marker – Pmi gene was used.
This selection system utilizes the Pmi gene encoding for phosphomannose-isomerase that convert mannose-6-phosphate to fructose-6- phosphate.
Only transformed cells are capable of utilizing mannose as a carbon source.
A vector, pCAMBIA1301-Pmi-Vgb, with Vgb gene and Pmi gene was constructed.
With this vector the genes Pmi and Vgb have been transformed into ground-cover chrysanthemum Fenditan by Agrobacterium-mediated method.
The antibiotics-resistant seedlings were respectively detected by PCR with primers depending on Vgb. The PCR result showed that the Vgb gene has been transmitted into chrysanthemum successfully.
Publication
Authors
Y. Wang, S. Shi, X. Li, Y. Gao, Q. Zhang
Keywords
safety marker, Pmi gene, Vgb gene, chrysanthemum
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