Direct method in transgenic plant production

1. DIRECT METHOD IN
TRANSGENIC PLANT
PRODUCTION
GROUP MEMBERS-
ASHISH KUMAR – 2019-AJ-26
TRIBENI GOSWAMI – 2018-AJ-112
MUDANG HUTO – 2018-AJ-61
HIROK JYOTI KONWAR – 2019-AJ-43

2. * What is Transgenic plant ?
 Transgenic plants are plants that have had their genomes modified
through genetic engineering techniques either by the addition of a
foreign gene or removal of a certain detrimental gene .
 The first transgenic plant was developed through the insertion of nptII
bacterial antibiotic resistance gene into tobacco in the year 1983 in
USA by Fraley et al .
 The main purpose in the production of transgenic plants is to produce
crops, which have ideal traits, quality, and high yield. Besides being
beneficial to the agriculture sector, the plants are found to be able to
act as the factory for pharmaceutical protein production .

4. *Transformation -
 Transformation is the process by which genetic makeup of
an organism is altered by the insertion of new gene (or
exogenous DNA) into its genome .
 Transformation Was first demonstrated in 1928 by British
bacteriologist Frederick Griffith in Diplococcus
pneumonia.
 In 1944 this "transforming principle" was identified as
being genetic by Oswald Avery , Colin Macleod, and
Maclyn McCarty.

6. * Vectorless or Direct gene transfer
method -
 Introduction of DNA into plant cells without the involvement of
biological agents such as Agrobacterium and leading to stable
transformation is called direct gene transfer.
 The term ‘direct gene transfer’ is used to discriminate between
methods of plant transformation that rely on the use of Agrobacterium
(indirect methods) and those that do not (direct method).
 Direct gene transfer methods rely on the delivery of large amounts of
‘naked’ DNA whilst the plant cell is transiently (rapidly)
permeabilized.

7. * PHYSICAL METHOD
* Electroporation technique -
 Is the process where by electrical impulses of high field strength are used to
reversibly permeabalize cell membrane to facilitates uptake of large
molecules, including DNA.
 Transformation using electroporation was developed in the late 1980s.
 It can be used to deliver DNA into plant cells and protoplasts.
 Plant materials is incubated in a buffer solution containing DNA and subjected
to high-voltage electric pulse.
 The DNA then migrates through highvoltage-induced pores in the plasma
membrane and integrates into the genome .
 It can be used to transform all the major cereals particularly rice, wheat ,
maize .

10. * Advantages and Disadvantages of
Electroporation -
• Advantages –
i. Both intact cells and tissue can be transformed.
ii. The efficiency of transformation depends upon the plant materials,
electroporation and tissue treatment conditions used for
transformation.
• Disadvantage –
i. Only 40 to 50% incubated cells receive DNA.
ii. ~50% of the transformed cells can survive.