Repeated, steady cycles of tissue culture

Because this bacterium has the ability to carry its own genes into plant cells, it is used in the gene transfer of many plants. To select only cells that have received the genes after transfer and regenerate them into the plant body of roses, it is necessary to optimize plant hormones as well as kinds and concentrations of nutrients. The work to perform this procedure in sterilized conditions, using a plant in a test tube, is called “tissue culture.”

In the case of roses, it takes about one year from the introduction of genes to the blooming. How effectively the introduced genes function differs from one genetically modified rose to another, so we have to produce as many genetically modified roses as possible. Efficiency of gene introduction also differs substantially by rose variety. Therefore, we continued tissue culturing, learning through trial and error.

The mystery of why the roses didn’t turn blue—even after we inserted the blue genes

Finally, we were able to introduce genes into red roses. For the first time in 1994, roses to which two kinds of blue genes of petunia opened. However, the color of the flowers was still red and no blue pigment was detected although the genes were introduced without a doubt.

Color changes or delphinidin were not observed even after many promoters (DNA sequence regulation gene expression) were used to drive petunia blue genes. Thus, we decided to introduce blue genes other than from petunias to roses. We isolated blue genes from various plants with blue flowers, such as gentian, butterfly pea, and torenia, and introduced them into roses. However, no matter how many times we conducted experiments, only “roses with blue genes but without blue pigments” bloomed.

We even suspected that roses might decompose blue pigments….（Senior General Manager Yoshikazu Tanaka Ph.D.）

Worked hard to help raise funds in Australia

Around that time, I was busy helping raise funds at the Florigene in Australia. This was a 4-year project originally, but it took a much longer time to produce blue pigments in roses. There was financial support from Suntory during the first two years, but financial difficulties unique to venture companies continued subsequently, posing various barriers.

From the viewpoint of investors, it must have been hard to make an investment in the blue rose project which was taking several years, unlike software development that could be completed in half a year. Finally, we obtained funds, using a taxation system encouraging research and development in Australia.

Florogene could not make use of the system without the approval of Suntory, so I served as a negotiator between Suntory and Florigene.

Steady efforts of repeating trials and errors

It took much trial and error to introduce the heterologous blue genes which had been once regarded as unnecessary by the ancestors of roses into rose cells and regenerate transgenic roses by tissue culture.

We had to use the power of a microorganism to introduce these genes into cells, and needed to conduct experiments to determine the strain of the microorganism to be chosen. At the same time, we also had to actually try to introduce the genes into a stalk and a leaf in order to determine which was better.

Introducing genes into plants and regenerating them required much more patience than isolating blue genes because steady efforts were necessary to test various factors repeatedly.

Tissue culture requiring preciseness and patience

Roses did not turn blue even after blue genes were introduced–We repeated experiments, and at one point, I even suspected that roses might have mechanisms to decompose blue pigments. Because I would have been more stressed out by thinking about this matter too seriously, I kept telling myself that “research and development does not advance smoothly in most cases.”

Under these circumstances, researchers engaged in tissue culture devoted themselves to the experiments of introducing blue genes of various blue flowers into rose cells. Only those with precise and patient personality could be engaged in this aspect of the research. The desks of those engaged in tissue culture were organized tidily, too. I, who left my desk messy, belonged to the molecular biology team to isolate blue genes (Smile).

We had many failures, but each of them taught us a useful lesson for the future (Principal Researcher Yukihisa Katsumoto, Ph.D.)

I demonstrated my aspiration to become a project member

I engaged in plant science at university, and joined the company, longing to conduct research on blue roses in 1991. During the early days, partly as my training, I was given opportunities to conduct various studies including experiments on genes and enzymes. However, I had a strong desire to deal with living plants themselves. So I conducted tissue culture experiments after working hours, produced some results, and negotiated directly with my boss then. As I desired, I became a member of the blue rose project. My action was really in line with the spirit of “Yatte Minahare”

Introduction of blue genes into rose cells was a laborious and time-consuming series of steady work. However, I myself did not mind it at all because that was the work I wanted to do so much.

Teamwork promoted upon mutual respect

We had a series of failures in the development of blue roses, but each failure gave us a useful lesson for future research. Each time we examined flowers that had not turned blue, we found some hint that might be useful. It was enjoyable to find such hints and use them in the next stage. Because of this enjoyment, all the members, including myself, were able to continue daily research.

Each member of the team had his or her own specialty. Some were good at molecular biology, others at tissue culture, and others at chemical analysis. Nobody was able to conduct this research alone, so we depended on each other’s specialties and cherished the sense of cooperation to make the dreams come true.

Regardless of our age and sex, the team members respected each other, recognizing the ability of the other members as professional researchers. Such mutual respect enabled all of us to continue to be positive.

I am thankful to the company, which allowed us to continue even when we could not produce fruitful results

Even when no fruitful results were obtained, the team members did not give up and continued the research, believing the success. I again would like to express my appreciation to the company, which allowed us to continue this project for such a long time.

We knew that creation of something good would take time, as in the case of the aging of whisky. As signified by the term “Yatte Minahare” the challenging corporate spirit is unique to Suntory.

Without the spirit of “Yatte Minahare” the company would not have assigned me to the research team in my first year of employment or would not have allowed us to continue the blue rose project for 14 years in spite of the fact that the prospect for commercialization remained unclear for a long time. Perhaps the project would have been discontinued a long time ago at an ordinary company.

• Research papers on flower
• Conference presentations on flower