ST FRANCIS METHODIST SCHOOL

ATAR BIOLOGY
TASK 2
GENETIC ENGINEERING – CREATION OF
BLACK ROSES

NAME: LOURDES KUSUMADI

CLASS: PU2A
Part A
Roses have been cultivated even before the Middle Ages until today roses are one
of the most commonly cultivated ornamental plants in the world. Nowadays, more
than 30,000 cultivated rose varieties are said to exist and they are usually referred to
as Rosa x hybrida. Rosa x hybrida is a description used for most cultivated rose
cultivars. These cultivated rises have arisen over the centuries through complex
crosses involving a number of species of roses.

Traditionally, new varieties by selective breeding (artificial selection) is the process

used by humans to selectively develop a particular characteristic. Selective breeding
of rose focus on of various characteristics to enhance their ornamental value such as
colour, size, form, quality of the bloom, and adaptation towards the environment. For
example, breeding out ‘gemini’ rose, a dual blushing to deep coral colour and ‘sunset
celebration’, apricot burnished with cream rose. This two rose will produce a new
rose that has both blush and apricot petals.

Sunset Celebration Gemini

Gemini x Sunset Celebration

Fig 1 – Selective Breeding

The second method is grafting. Grafting
involves joining together parts of two
plants to function as one plant. It’s an
efficient method of propagation that
produces offspring genetically identical
to the original plant. To graft one plants
provides the lower trunk and root
system (stock/rootstock), while the
other plant provides the upper portion
(steams, leaves, flowers, and fruit) that Fig 2 – Grafting
has the wanted characteristics for the
offspring called the scion. Grafting allows us to create exact genetic duplicates of
superior individual plants. The way grafting work is when it is combined properly, the
cells in their cambium layers will start to form called callus. Eventually, callus will
allow the circulatory system of the stock and scion to heal together, this results in a
functioning plant composed of two genetically different parts.

In order to produce the numbers needed for commercial sale plants are reproduced
asexually. This method does not require the investment required to produce flower
but to push up the production scale, horticulturist will usually do grafting and artificial
selection. Asexual reproduction produces plants that are genetically identical to the
parent plant because no mixing of male and female gametes takes place.
Traditionally, these plants survive well under stable environmental conditions when
compared with plants produced from sexual reproduction because they carry genes
identical to those of their parents.

There are several important rose traits that are the cause for new varieties to rise.
These traits include ease of reproduction, resistance to biotic and abiotic stresses,
plant architecture, and flower development.

Natural evolution is the process by which species adapt to their environment. Natural
evolution leads to evolutionary change when individuals with certain characteristics
have a greater survival, in other words it is the survival of the fittest. It is one of the
process that is responsible for the evolution of adaptations of organisms to their
environment. The process includes to elimination of the non-advantageous
characteristics in the species and addition of characteristics that would create an
upperhand for the species in the environment, and increase their survival rate.
Fig 3 – Kennedia nigricans

Part B

Flower colour is mainly determined by anthocyanins.

Rosa hybrida is not able to produce a black flower. As
Kennedia nigricans is the one of the only plant that
produces black flower naturally. One way to obtain black
rose is by transgenesis. Transgenesis is the process of
introducing a new gene into a living organism so that the
organism will exhibit a new trait and transmit that
property to its offspring. The steps will be as follow:
1. Isolating the desired gene
The mRNA carrying the code for making black pigment is extracted. This
mRNA is incubated with reverse transcriptase. As a result, cDNA is formed.
This DNa carries the code responsible for the black pigment in Kennedia
nigricans. Now, this DNA is cleaved with the help of restriction enzyme.

Fig 4 – Sticky ends vs Blunt ends

As what Fig 4 shows, BamHI cut at covalent phospodiester bonds of both

DNA strands, while Hpal make a simple cut across both strands at a single
point. Any one of these enzyme could be used to cut the DNA.

2. Formation of recombinant DNA

Plasmids are the vector which are used in
genetic engineering. They are obtained by
digesting the bacterial cell enzymatically.
Plasmids are then cut by using a restriction
enzyme. The fragment produced will be
having the complimentary base pair. These
extension will form hydrogen-bonded base
pairs with the DNA strand on other DNA
molecules. These DNA fusions will be made
permanent by DNA ligase which seals the
strand by catalyzing the formation of
phosphodiester bond.

Fig 5 – Recombinant DNA formation

 3. Inserting recombinant DNA to a bacteria host
Once the appropriate vector is made and transferred to Agrobacterium
tumefaciens, it must be integrated into the
plant genome.

4. Growing Rose hybrida in the bacteria

medium
Figure 6 show how the transformation is
carried out. First of all, the rose leaf disks are
punched out and incubated overnight with a
culture of vector-containing A. tumefaciens in
Luria broth. The disks are then paced in
culture plates containing a medium that
induces the leaf disks to start growing shoots.
After a couple of days, these disks are
transferred to plates, which contain antibiotics
corresponding to the antibiotic-resistant
marker genes placed in the vector. Only those Fig 6 – Growing the plant in a
plant cells that have integrated T-DNA from cultured bacteria medium
the vector will have antibiotic resistance. The
medium therefore selects for the transformed plant cells by killing those that
do not contain the vector. Once the transformed cells are selected, the
growing shoots are transferred to a root-inducing medium, where they grow
roots, and then to soil to grow into transgenic plants.

This procedure should be produced in a large scale, to prove the confirmation of

successful gene transfer. To know if its successful or not, time will be taken to wait
for the plant to grow and if its successful, a black rose will be produced. When this
happens, the black rose could asexually reproduce and be available for commercial
or research purposes.

True breeding only produces offspring with the same traits. True-breeding organisms
are genetically identical and have identical alleles for specified traits. The alleles for
these type of organisms are homozygous. True-breeding plants and organisms may
express phenotypes that are either homozygous dominant or homozygous
recessive. In complete dominance inheritance, dominant phenotypes are expressed
and recessive phenotypes are masked in heterozygous individuals. To determine
whether a plant is true breeding or not, simply cross a plant with itself. If all the
offspring have the same phenotype as the parental plant, the plant is homozygous,
and is true-breeding. If a plant produces offspring with varying phenotypes, the plant
is likely heterozygous and is not true-breeding. Gregor Mendel or also known as the
“Father of Genetics” finds out the process by which genes for particular traits are
transmitted.

For example, if we are about to cross a pink colour flower with a white colour flower,
and if all of the offspring are all pink means that the pink colour flower is dominant to
the white colour flower. A true-breeding plant with pink colour would have a genotype
of (AA) for the trait and a true-breeding plant with white colour flower would have a
genotype (aa).

Cross pollination between a true-breeding plant with pink colour and a true-breeding
plant with white colour (AA X aa) results in offspring (F1 generation) that are
heterozygeous dominant for pink colour (Rr). Self-pollination in F1 generation plants
(Rr X Rr) results in offspring (F2 generation) with a 3 to 1 ratio of pink colour to white
colour flower. Half of these plants would be heterozygous for pink colour flower, ¼
would be homozygous dominant for pink colour flower (RR), and ¼ would be
homozygous recessive for white colour flower (rr). For illustration purposes, the
example of this experiment would be in like in Fig 7.

Fig 7 – True-breeding

However, there are some of the consideration that should be taken to the probvlems
that might occur with transgene, such as there might be mutation or contamination
that might occur. For example, the black rose may not 100% be produced
successfully because of some unidentified mutation that could happen during the
recombinant DNA production process or any contamination that happens when the
black rose is planted to the soil, and also the black rose may not be able to produce
any offspring because of a mutation. Since black rose is a new invention, no
problems could be identified just yet.

Part C
The benefits of a production of black rose could range from the producer until the
environment. The benefit could effect the producer of the black rose, growers,
customers, the biodiversity of the rose population, and the environment. The
producer of the black rose will legally provide them with intellectual property (IP)
right, such as a patent or a trade mark to the producer company. The IP will grant
them with an exclusive right to decide how or whether the invention could be used by
others. Additionally, this will increase the market share, and profit for the producer.
Developing a black rose that is not available in the market yet can lead to profit
growth and enable economies of scale and increase the producer market power.

The next benefit will effect the growers of the black rose and customers. The
growers of rose will be able to increase their profit because they could sell the rose
in a higher price, learning new skills, and to open up a new market for them.
Moreover, rose has always been the best selling flower ornaments, and gift in the
market, therefore black rose may be accepted well by the customers with the
addition of a unique variety to the rose population.

The last benefit is for the biodiversity and the environment. The black rose will
increase biodiversity to the rose population, and biodiversity boosts ecosystem
productivity where each species has a role to play. Greater species diversity ensures
natural sustainability for all life forms, and increase the aesthetic of a place where
the black rose is being grown, and it may provide an example for an extensive
research to be done in the particular place.

The risks that must be accounted for the producer of black rose is that innovation is
costly, companies can expect to spend around 3.5% to more than 20% of their total
revenue in research and development. Moreover, it also wasted time and resources
because the black rose may not accepted well into the market, and there are no
buyer for it or the demand is not there. The producer may also suffer a loss if they
need to sell the black rose in a cheaper price to attract customers while a huge
amount of money have been consumed for the research.

Since black rose is a new species that is newly introduced to the biodiversity and the
environment. The growers of black rose may not know the long-term effect that it
may cause because it may harm the environment in an unexpected way such as it
may attract weed or parasites, and change to an invasive species, such as being
harmful to rose population because there may a mutation that happens that turns the
black rose into a cancerous species. It disturbs the sustainability of the environment
and may also cause some extinction to other rose population. Lastly, there may be
some mutation that could endanger customers who buys it the black rose, such as
they may be carrying a bacteria known as Pseudomonas aeruginosa which can
cause infection in humans and can invade nearly any tissue in the human body.
Symptoms vary widely, from urinary tract infections to dermatitis, gastrointestinal
infections and even systemic illness.

Part D
Patent is an exclusive right granted for an investor, in this case is a black rose is the
product that needs to be patented. The principle benefit claimed for patents is that
rewarding an inventor creates a positive environment for progress of research. The
ethical arguments that are expressed when supporting patenting of biotechnology
inventions are:
 Patenting promises useful consequences (e.g. new products/research)
 Patenting rewards innovation
 If patenting is not permitted, useful information will become trade secrets
  Patent law regulations inventiveness, not commercial uses of invention.

However, the arguments against patenting include:

 Patenting promotes inappropriate human control over information that is
common heritage
 Patenting produces excessive burdens on customers (increased costs,
payment of royalties for succeeding generations)

This issue is closely related to the commercialisation of genetic engineering, but

some sort of information protection is already accepted as an incentive to invest in
research of benefit to society. In other words the benefits should be in terms of
general medical or agricultural development, rather than economic prosperity of one
company or country over another. Since the patent that the company will be granted
is on the black rose, and black rose does not have any other purpose except for
aesthetics and for trades. The company should be taking to the consideration where
the research may upset public order and morality; likewise, making the black rose
available on the market may be ethically problematic (e.g. harmful effects), but the
actual patent protection, which only gives an exclusive right to use, ca n hardly be
said to upset the public perception of morality. When discussing patent ethics, it
therefore seems appropriate to isolate the particular issue that is ethically
problematic in respect of the actual patent issuance, rather than the research or
general use.
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