UNIVERSIDAD SAN FRANCISCO DE QUITO

FOOD BIOTECHNOLOGY

REVIEW QUESTIONS, CHAPTER 20: Engieering plant quality and proteins

Samanta Rojas (214353) December, 7, 2023

1. How can plants be genetically manipulated to produce flowers with unusual colors?

Altering the color of flowers by manipulating plants involves modifying the activity of
genes responsible for pigmentation. Techniques like transgene insertion, CRISPR/Cas9
gene editing, and RNAi can be employed for this purpose. The objective is to amplify or
suppress the synthesis of pigments like anthocyanins, carotenoids, and betaines to
attain the preferred color. Furthermore, adjusting environmental elements such as
light intensity, temperature, and nutrient availability can also impact the coloration of
flowers.

2. How can soybeans be genetically manipulated to increase their lysine content?

Soybeans can undergo genetic modification to enhance their lysine levels by

introducing an external gene encoding a protein abundant in lysine into the soybean
genetic makeup. This introduced gene can be controlled using a seed-specific
promoter to guarantee that the lysine-rich protein is exclusively produced in the seeds.
Selection of the transformed soybeans can be done through selectable markers, and
the lysine content can be validated through chemical analysis. Another strategy
involves diminishing the expression of genes associated with lysine breakdown,
fostering the buildup of lysine in the seeds.

3. How can vitamins be overproduced in rice plants?

Genetic engineering can lead to the excessive production of vitamins in rice plants.
One method involves incorporating genes responsible for vitamin biosynthesis into the
rice genome. For instance, introducing the phytoene synthase gene can boost the
synthesis of vitamin A. Alternatively, genetic modification can be used to augment the
expression of native genes associated with vitamin biosynthesis. For example,
elevating the expression of the GSA (glutamate-1-semialdehyde aminotransferase)
gene can enhance folate production in rice plants.

4. How would you engineer soybean plants to overproduce long-chain omega-3 and
omega-6 fatty acids?

Through genetic manipulation of soybean genes, it becomes feasible to transform

linoleic acid and alpha-linolenic acid into long-chain omega-3 and omega-6 fatty acids,
with the potential for further conversion into arachidonic acid and eicosapentaenoic
acid. Additionally, it is achievable to modify pathways within soybeans to generate
long-chain polyunsaturated fatty acids.
5. What is the effect of increasing the level of oxidized glutathione within a plant? How
would you genetically manipulate a plant to do this?

To diminish oxidative stress in a plant, the process entails extracting cDNA regulated
by the 35S promoter and integrating it into the plant using a binary Ti plasmid. This
action results in an elevation of oxidized glutathione levels.

6. How can the level of bioavailable iron in plants be increased?

The bioavailable iron content in plants can be heightened through various means, such
as overexpressing iron transporters, manipulating genes related to iron metabolism,
and improving the plant's capacity to convert ferric iron into ferrous iron. Additional
strategies involve incorporating iron chelators into the soil or employing plant growth-
promoting rhizobacteria to enhance iron solubility in the soil. Furthermore, genetic
engineering allows for the introduction of genes from other organisms that generate
iron-binding peptides or proteins.

7. How would you increase the amount of amylose compared to amylopectin in potato
starch? How would you increase the amount of amylopectin compared to amylose?
What is the advantage of performing these manipulations?

Increasing amylose concentration in plants can be accomplished by introducing

antisense versions of the starch branching enzyme using Agrobacterium tumefaciens
and the 35S promoter. Similarly, antisense versions of three starch synthase genes can
be employed to impact amylopectin concentration. These alterations offer the
advantage of creating a potato better suited for frozen and other industrial products
by decreasing amylopectin content.

8. How does an antigen express in a transgenic plant act as an edible vaccine?

When an antigen is expressed in a transgenic plant, it serves as an edible vaccine by

eliciting an immune response in the human body upon consumption. The antigen,
being a foreign substance, prompts the immune system to generate antibodies against
it. Ingesting a transgenic plant expressing the antigen leads to the recognition of the
antigen by the immune system, triggering the production of antibodies. This
establishes immunity to the disease associated with the antigen, eliminating the need
for conventional vaccine administration methods like injections. The edible vaccine
approach is considered a promising solution for vaccinating large populations,
particularly in developing countries where conventional vaccine delivery systems may
be insufficient or unavailable.

9. How would you engineer an edible vaccine directed against V. cholerae-caused

diarrhea?

To accomplish this, one can employ Agrobacterium tumefaciens along with the cholera
toxin subunit B gene. This method allows for the genetic modification of a potato,
facilitating the binding of the cholera toxin subunit B to the intestinal receptor.

10. How can a plant’s oxygen levels be manipulated? How does the manipulation of a
plant’s oxygen levels affect the yield of that plant?
 An approach to accomplish this involves the introduction of the C4 pathway of
photosynthesis into plants that conventionally follow the C3 pathway. This leads to a
heightened oxygen supply to the plant cells, consequently promoting improved plant
growth.

11. How would you engineer rice plants to overproduce tetrahydrofolate? Why would
you do this?

By introducing foreign genes sourced from Arabidopsis thaliana, which serve as rate-
limiting enzymes in the biosynthetic pathway, it becomes feasible to augment the flux
and enhance the nutritional content of rice. Enhancing the plant to generate
tetrahydrofolate and tetrahydrofolate polyglutamates can further elevate its
nutritional value.

12. Why is it necessary to genetically engineer soybean plants to have a small amount of
phytate in their seed when low-phytate mutants may be selected following
conventional mutagenesis?

Because it has disadvantages in the plant because high levels of phytates inhibit iron,
hinder germination, and stress, harming efficient and reliable processes compared to
other methods.

13. Briefly describe a vector system that may be used to engineer plants to produce
large amounts of full-size IgG molecules.

One approach for engineering plants to yield substantial quantities of full-size IgG
molecules involves the utilization of the plant viral expression system. In this system,
the heavy and light chain genes of the IgG molecule are incorporated into a viral
vector, which is subsequently introduced into plant cells. The viral vector is designed to
replicate within these cells, leading to elevated expression levels of the IgG genes.
Consequently, the plant cells produce and release significant amounts of full-size IgG
molecules that can be extracted and purified from the plant tissue. This system holds
promise as a cost-effective method for the large-scale production of IgG molecules.

14. Why are plants an attractive host system, compared to bacteria and animal cells in
culture, to produce human therapeutic proteins?

Plants present an appealing platform to produce human therapeutic proteins due to

several advantageous features. Firstly, their cultivation and maintenance are relatively
cost-effective. Additionally, they possess the capability to perform post-translational
modifications, including glycosylation, phosphorylation, and proteolytic processing,
crucial for the correct folding and functioning of numerous therapeutic proteins.
Moreover, they are generally regarded as safer compared to bacterial and animal cell
culture systems as they do not harbor human pathogens. Furthermore, their scalability
for large-scale production is easily achievable. Lastly, plant-based systems also hold
the potential to generate proteins with distinct glycosylation patterns that may prove
beneficial for specific therapeutic applications.

15. What is phytate? How can the levels of phytate in seed be modulated? Why would
you want to do this?
 For the bioavailability of iron in plants, there are compounds such as phytates that do
not allow iron absorption. For this, phytase is used, which regulates and progressively
turns off the ATP binding case transporter in the seeds.

16. Describe a strategy for developing a plant vaccine against type 2 Shiga toxin?

To develop a plant vaccine against Shiga toxin type 2, the gene that encodes the B
subunit of the toxin must be inserted into a plant vector (Ti plasmid from
agrobacterium tumefaciens) so that the plant expresses the B subunit protein of the
toxin and is collected to be purified as a vaccine.

17. How can the lignin content of trees be decreased? What is the benefit of this type of
genetic manipulation?

The decrease in lignin in trees can occur through genetic manipulation that reduces
enzymatic activity during lignin biosynthesis. It can be achieved with RNAi by silencing
genes that encode enzymes or even by introducing genes that encode enzymes and
are the precursors of lignin. lignin

18. How can manipulation of the level of brassinosteroids be used to increase plant
yield?

The level of brassinosteroids can increase plant performance when manipulation

stimulates cell elongation and division to improve stress tolerance, increase in
size and seeds, and optimize photosynthesis. The overexpression of biosynthetic
genes or the application of brassinosteroids are methods.

19. How can plants be genetically modified to increase their effectiveness in

phytoremediation?

Genetic modification in plants is harmful to the environment when metals

accumulate, the solution is to introduce the YCF1 gene in Aravidopsis thaliana
plants. Yeast protein expression detoxifies metals for transport to vacuoles in
plant cells

References

Molecular Biotechnology, principles, and applications of recombinant DNA.

Glick and Pasternak. Library code TP248.2.658.2003