BAHIR DAR UNIVERSITY

INSTITUTE OF TECHNOLOGY

FACULTY OF CHEMICAL AND FOOD ENGINEERING

DEPARTMENT OF FOOD TECHNOLOGY

Post Graduate Program (MSc)

Assignment of Advanced Food Biotechnology

Title: - Genetically Improve Soybean to Enhance the Yield of Protein and Oil

By Berhanu Regassa

ID No:-BDU1301020

Submitted to: Dr. Neela Satheesh

December, 2020
Bahir Dar, Ethiopia
1. Introduction

Soybean [Glycine max (L.) Merrill.] is an economically important legume with 2n = 40

chromosomes, whose seeds contain an average of 40% protein and 20% oil, and its plants enrich
the soil by fixing nitrogen in symbiosis with nitrogen-fixing bacteria. Soybean seeds contain
carbohydrates (11%), Water (15%) dietary fibers (9%), vitamins, and minerals as well. For the
last few decades the majority of research laboratories have been investigating genetic traits to
improve the yield of protein or oil in soybean seeds through genetic engineering, thereby
achieving improved quantity and quality of soybean seeds. Until now, most of the transformation
experiments have implemented a single functional gene not multiple genes. Those agronomical
and economically important traits affect the enhancement of grain quantity and quality.
However, the majorities of agronomic and genetic traits such as complex metabolic, biological,
and pharmaceutical pathways are polygenetic traits and are produced in a complex pathway.
Therefore, those traits are encoded and regulated by a number of genes. In an attempt to study
and manipulate those pathways, the transfer of multigene or large inserts into plants have been
developed by multigene engineering technology and have also been involved in metabolic
engineering. Several examples of multigene or large insert transfers have been reported such as
the application of carotenogenic genes in rice, canola, and maize, and of polyunsaturated fatty
acid and vitamin E genes in soybean and Arabidopsis. Therefore, reliable systems for
transforming large DNA fragments into plants make it feasible to introduce a natural gene cluster
or a series of previously unlinked foreign genes into a single locus. (Hyeyoung Lee, 2014).

Genetic improvements of the crop have focused mainly on enhancement of seed and oil yield,
development of varieties suited to different cropping systems and locations, and breeding
resistant/tolerant varieties for various biotic and abiotic stresses.( Besufikad Enideg Getnet,
2019)

Transfer of DNA into plant cells has been achieved by using several methods. In soybeans, the
most frequently employed plant genetic engineering methods are Agrobacterium-mediated
transformation and particle bombardment. Both systems have successfully been used in
genetic transformation of soybean. Since the initial reports of fertile transgenic soybean
production various efforts have been made to improve the transformation efficiency and to
produce transgenic soybean. Particularly, the preferred and reproducible transformation is the
use of the cotyledonary node as a plant material, which is based on Agrobacterium-mediated
gene transfer. Nevertheless, new methods have been developed for more efficient soybean
transformation. There still remain, however, many challenges for genotype- and tissue- specific
independent transformation of soybean. This review provides an overview of historical efforts in
developing and advancing soybean regeneration and transformation systems. (Hyeyoung Lee,
2014)

2. Approaches for Soybean Gene Modification

In soybean transformation, two major methods are now widely utilized: Agrobacterium-
mediated transformation of different explant tissues and particle bombardment. The
Agrobacterium-mediated method usually produces single or low copy numbers of insertions with
relatively rare rearrangement. On the other hand, bombardment technique directly introduces
desired genes into the target plant cell with small tungsten or gold particles. The success of this
approach critically depends upon the ability of the target tissue to proliferate as well as proper
pre-cultures to make a target plant. (Hyeyoung Lee, 2014 and Owens, L. D., & Smigocki, A. C.
1988)

There are three main requirements to establish an efficient transformation system: - (6) (1)

1. A source of totipotent cells that serve as recipients of the delivered DNA,

2. A means of delivering DNA into the target cells (c)

3. A system for selecting or identifying the transformed cells.

2.1. Particle Bombardment (Projectile Acceleration)

The introduction of DNA into intact cells and tissues by accelerated micro particles driven at
high speeds. Accelerated particles are able to cross the cell wall and the cell and nuclear
membranes. In the nucleus, exogenous DNA fragments are liberated and may be integrated into
chromosomal DNA through the processes of illegitimate or homologous recombination, which
depend exclusively on cellular components. The main advantage of particle bombardment lies in
the possibility of transferring genes to any cell or tissue type independently of genotype and
without having to consider the compatibility between the host and bacterium, as required by the
Agrobacterium system. Particle bombardment is also quicker and easier to use. However, this
technique introduces multiple DNA copies that may be fragmented or recombined. Particle
bombardment can be achieved through high-or low-helium pressure gene guns, enabling the
penetration of the target tissue to be controlled very accurately so that the majority of the
particles carrying the DNA can be directed to a specific cell layer. This feature is extremely
important because different explants may require different acceleration conditions for optimum
particle penetration. Meristems (composed of multiple cell layers) require the employment of
high-pressure equipment to reach the inner layers. Zhang, 2014,) ( Hyeyoung Lee, So-Yon Park
and Zhanyuan J.

2.2. Agrobacterium system

Agrobacterium is a soil-borne Gram-negative phytopathogenic bacterium that naturally infects

different plants. These phytopathogens cause a variety of neoplasms, including crown gall
disease (A. tumefaciens and A. vitis), hairy root disease (A. rhizogenes), and cane gall disease (A.
rubi) on numerous plant species. The origin of these sicknesses is interkingdom horizontal gene
transfer. When virulent strains of Agrobacterium infect plant cells, they transfer one or more
segments of DNA (transferred DNA or T-DNA) from the Ti (Tumor inducing) or Ri (Root
inducing) plasmids into the host plant cells. To obtain engineered (binary) vectors derived from
Ti or Ri plasmids, genes present in the T-DNA region are replaced by foreign DNA. The
advantages of Agrobacterium-mediated gene transfer over particle bombardment include the
possibility of transferring relatively large segments of DNA, a lower number of transgene copies
integrated into the plant genome, rare transgene rearrangement, a lower frequency of genomic
DNA interspersion and reduced abnormal transgene expression. This system involves a low
operating cost and simple transformation protocols. However, plants differ greatly in their
susceptibility to Agrobacterium-mediated transformation. These differences occur among
species, cultivars or tissues. (Vergunst, A. C., et al. 2000)

2.2.1. Agrobacterium tumefaciens and target tissues

Agrobacterium was initially considered to be nonpathogenic to soybean. However soybean may

be susceptible to this bacterium. The addition of acetosyringone during bacterial infection,
selection of the most appropriate A. tumefaciens strain and soybean cultivar, and development of
super-virulent plasmids have contributed to the increased efficiency of soybean transformation.
Recovery of the first transgenic soybean plants using A. tumefaciens-mediated transformation
using cotyledonary nodes as the target tissue. Subsequently, advances in transformation
techniques were achieved, and several research teams have reported the generation of transgenic
plants using different target tissues, such as cotyledonary nodes, hypocotyls, half-, organogenic
callus, and immature zygotic cotyledons.( Hyeyoung Lee, So-Yon Park and Zhanyuan J. Zhang,
2014,)

Figure 1. Scheme for genetic transformation of soybean cotyledonary nodes.

3. Advantages of Soybean Modification

Genetic improvement in soybean has resulted from the continual development and release of
new cultivars with greater genetic yield potential than their predecessors. Genetically modified
soybean has great yield potential, wide range of adaptability, high nutritional value for both food
and feed, and a great importance in cropping systems. Modified soybean has undergone rapid
expansion herbicide-tolerant crops. Characteristics that have been the focus of traditional crop
breeding include lodging resistance, reduced plant height, seed size, seed quality, oil quality,
shattering resistance and resistance to insects, diseases and nematodes. Genetically modified
soybean has generally had different advantages.( Leonard P. Gianessi, Janet E. Carpenter 2000)

1. Enhancement of quality and quantity of the crop, especially the improvement of Oil and
protein yield (Nutrition benefits to serve malnutrition)
 2. Pest resistance: - soybean losses from insect pests can be staggering, resulting in
devastating financial loss for producers.
3. Herbicide tolerance: - It is not cost-effective to remove weeds by physical means such as
tilling so farmers will often spray large quantities of different herbicides (weed killer) to
destroy weeds, a time-consuming and expensive process that requires care so that the
herbicide harms the crop plant or the environment. Crop plants genetically-engineered to
be resistant to one very powerful herbicide could help prevent environment damage by
reducing the amount of herbicides needed. Soybeans genetically modified to be not
affected by their herbicide product Roundup.
4. Disease resistance
5. Drought tolerance/salinity tolerance

4. References
1. Hyeyoung Lee, So-Yon Park and Zhanyuan J. Zhang, 2014, An Overview of Genetic
Transformation of Soybean
2. Leonard P. Gianessi, Janet E. Carpenter 2000, Agricultural Biotechnology: Benefits of
Transgenic Soybeans
3. Besufikad Enideg Getnet, 2019, Soybean (Glycine Max L. Merill) Genetic Improvement
In Ethiopia: A Review Gambela University, volume 7
4. Vergunst, A. C., et al. (2000). VirB/D4 -dependent protein translocation from
Agrobacterium into plant cells. (Translated from eng). Science, 290(5493), 979-982, (in
eng).
5. Owens, L. D., & Smigocki, A. C. (1988). Transformation of Soybean Cells Using Mixed
Strains of Agrobacterium tumefaciens and Phenolic Compounds. (Translated from eng).
Plant Physiol, 88(3), 570-573, (in eng).

6. Milena Schenkel Homorich, 2012  998–1010 Soybean genetic transformation: A valuable

tool for the functional study of genes and the production of agronomically improved plants
7. Karimi M, Inze D, Depicker A. GATEWAY vectors for Agrobacterium-mediated plant
transformation. Trends Plant Sci. 2002;7:193–195. 
8. Paula P. Chee*, Krystal A. Fober, and Jerry L. Slightom Molecular Biology-Unit 7242, The
Upjohn Company, Kalamazoo, Michigan 49007, 1989