Transformation of Arabidopsis plants through Agrobacterium mediated transformation and selective screening of putative transgenic plants through PCR analysis

Running head: GENE CLONING EXPRESSION AND ANALYSIS
Title: Transformation of Arabidopsis plants through Agrobacterium mediated
transformation and selective screening of putative transgenic plants through PCR analysis
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1GENE CLONING EXPRESSION AND ANALYSIS
Abstract
Over the years experiments based on plant transformations have been in demand due to the
burden on the food industry. Transformation of desirable genes into a host plant has proved
effective in production for genetically modified crops. Moreover these transfusion
experiments have also helped in the constitution of genetic makeup in the upcoming
generations. The following lab report would focus on the successful transformation of
Arabidopsis plants though the growth of seedlings. This lab report further focuses on the
screening of the putative Arabidopsis gene where Transformation has taken place due to
Agrobacterium mediated transformation. Thus the successful transfer of foreign genes
through plant transformation can prove to be a fruitful method of dealing with the increasing
crop demand and the use of biofuels in the upcoming years.

2GENE CLONING EXPRESSION AND ANALYSIS
1. Aim:
The main aim of the given lab report is to
identify transformed Arabidopsis plants
through Agrobacterium mediated
transformation. It also involves subsequent
screening of such putative transgenic
plants through molecular methods. In the
experiment, seeds represent the T1
generation which have been obtained from
the Arabidopsis plants and will undergo
the process of Agrobacterium mediated
transformation. This will be followed by
extraction of genomic DNA for molecular
analysis from the plantlets which will use
the polymerase chain reaction or PCR for
screening of putative transgenic plants.
2. Introduction:
2.1. General Background:
Plant transformation is defined as a
scientific method of inserting foreign DNA
of superior quality into the genome of the
species of interest .Such plants termed as
transgenic are important for both
agriculture as well as research. Tradition
techniques of plant breeding along with e
consideration of the intragenic
crosses ,chemicals s well as physical
mutagenesis along with different breeding
methods have been non-specific have been
the reason behind transgenic have been
chosen over traditional methods.
Moreover, random mutation of large
segments along with excess time
consumption of backcrosses for the
segregation of the undesired and
unrequired changes involved in the
offspring (Ran, Liang & Gao, 2017 ).
Transgenic technology is also involved in
improving the man characteristics of crops
which are economically important. Traits
involve diseases resistance, abiotic stress
tolerance as well as nutrient imbalance.
Arabidopsis is considered as an ideal
model for transformation procedures.
Development of highly efficient as well as
simple protocols without the
implementation of plant regeneration
procedures are the reasons behind
Arabidopsis being the primary model for

3GENE CLONING EXPRESSION AND ANALYSIS
conducting transformation experiments
(Rivero et al.2014). Zimenowoicz (2014)
states that agrobacterium is the best choice
for controlling such experiments as they
are the most commonly used vectors for
the creation of transgenic plants.
Moreover, this technique helps in the
enhancement of crop productivity,
resistance towards pests especially through
agrobacterium.
2.2. Similar research done in the field:
Similar kind of agrobacterium mediated
transformation has been done in Sorghum
plants where the application of an
improvised protocol has led to large scale
sorghum transformation experiments.
Through the implementation of modified
experiments, the frequency of
transformation have increased to almost
33% .Through the varieties of TX430, 848
independent calluses have been formed.
Quantitative PCR analysis has been done
for molecular analysis followed by
statistical quantitative studies using chi
square tests. Change in medium has
significantly improved transformation
frequencies. Strain and size of DNA have
also been important factors for this
experiment (Wu et.al 2014).
Agrobacterium mediated transformation is
basically a type of transient transformation
and often fluorescent fusion proteins are
used in this method for the transient
transformation for following the in vivo
behavior of the included proteins. Similar
studies of such agrobacterium mediated
transformation has been done in onion
epidermis which have ultimately helped in
determination of the protein subcellular
localization of large scales (Xu et
al.,2014).
2.3. Gap in literature
Transformation has been done before
using various plat species. However,
extraction of genomic DNA from various
plantlets has not been done on wide basis.
Gene analysis like that of TAIR has been
done for the transformation studies
involved in transformation. Moreover,
molecular techniques used for extraction