Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated 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
Biotechnology, Bharathidasan University

Engineering,

School

of

Applied Biochemistry and biotechnology (2015)

Objetivo
Desenvolver a transformao gentica eficiente de n cotiledonar
de soja cv. PK 416, utilizando sonicao e infiltrao vcuo;

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Soybean
Cultivars
Germinao e preparo
do explante
Caractersticas da construo usada na transformao
LBA4404, EHA101 e EHA105
pCAMBIA 1304 - 6.128 bp
Gene marcador
Gene reprter

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Soybean Cultivars
Infeco por Agrobacterium e co-cultivo de explantes
CMM
Mtodos de transformao
EHA105
0
5
10
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 AgrobacteriumMediated Genetic Transformation in Indian
Meios de regenerao
Soybean Cultivars
Cefotaxima, vancomicina e higromicina B
SEM

SIM

RM

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Soybean Cultivars

no. of GUS-GFP+ plants/total no. of infected explants100

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Soybean Cultivars

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Soybean Cultivars

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Soybean Cultivars
Anlise histoquimica

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Ensaio Visual GFP Soybean Cultivars

ST

AND

ST

AN
T

G
P

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Anlise molcular Soybean Cultivars
Primes especificos gene hpt II - amplificam fragmentos de 407 bp
Presena e integrao do gene
EcoRI

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Soybean Cultivars

Application of Sonication in Combination with

Vacuum Infiltration Enhances the AgrobacteriumMediated Genetic Transformation in Indian
Soybean Cultivars
Concluses
Desenvolvimento de um protocolo de transformao gentica
reprodutvel e altamente eficiente mediada por Agrobacterium
para ns cotiledonares de soja;

Possibilidade de infectar mais nmero de explantes dentro de

um curto perodo de tempo;

A combinao de ultra-sons e infiltrao a vcuo auxilia na

transferncia de T-DNA resultando em melhor eficincia de
transformao
Grande potencial para ser usada em diversas cultivares de soja
para transferir e expressar traos agronomicamente e
economicamente importantes