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Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in Indian Soybean Cultivars
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Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
M. Arun; K. Subramanyam; T. S. Mariashibu; J. Theboral; G. Shivanandhan; M. Manickavasagam ; A. Ganapathi -
Department of Biotechnology & Genetic Engineering, School of Biotechnology, Bharathidasan University
Applied Biochemistry and biotechnology (2015)
Objetivo
•Desenvolver a transformação genética eficiente de nó cotiledonar de soja cv. PK 416, utilizando sonicação e infiltração á vácuo;
•Germinação e preparo do explante•Características da construção usada na transformação
•LBA4404, EHA101 e EHA105
•pCAMBIA 1304 - 6.128 bp
•Gene marcador
•Gene repórter
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
• Infecção por Agrobacterium e co-cultivo de explantesCMM• Métodos de transformaçãoEHA105
0 510 20 30 40 50 s
20s 0, 100, 250, 500, ou 750 (mm Hg)
0, 1, 2, ou 3 min.
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
• Meios de regeneração
Cefotaxima, vancomicina e higromicina B
SIM
SEM
RM
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
no. of GUS-GFP+ plants/total no. of infected explants×100
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
Análise histoquimica
AND ST
ST
GPANT
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean CultivarsEnsaio Visual GFP
Primes especificos gene hpt II - amplificam fragmentos de 407 bpPresença e integração do gene EcoRI
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean CultivarsAnálise molécular
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
• Conclusões
• Desenvolvimento de um protocolo de transformação genética reprodutível e altamente eficiente mediada por Agrobacterium para nós cotiledonares de soja;
• Possibilidade de infectar mais número de explantes dentro de um curto período de tempo;
• A combinação de ultra-sons e infiltração a vácuo auxilia na transferência de T-DNA resultando em melhor eficiência de transformação
• Grande potencial para ser usada em diversas cultivares de soja para transferir e expressar traços agronomicamente e economicamente importantes
Application of Sonication in Combination with Vacuum Infiltration Enhances the Agrobacterium-Mediated Genetic Transformation in
Indian Soybean Cultivars
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