Team:Kyoto/Human Practices
Human Practices
Prologue
Our project “FLOWEREVER” started with a single proposal of one member who loves flowers. At first, we
questioned, “Why flowers?” However, the more we researched, the more we realized how integral flowers are in
our daily lives, and we decided to promote the project in earnest.
From there, we had a series of meetings. However, since we had little knowledge of the reality of the flower industry, it was hard to make a framework for the project. We brought our ideas to florists, distribution centers, and farmers where the problem is happening and listened to their opinions. After all these interviews and discussions with them, we were able to change our project to be beneficial to the end-users and had great potential to solve the problem of flower loss.
We also shared what we learned from experts about "flowers" with the public. It is said that the demand for flowers has been on the decline in recent years. In such a situation, this project is trying to find a way to make it possible for people to continue to love flowers, which are a natural emotion that we have in common. This time, as one of the activities aimed at promoting the feature of flowers, we would like to introduce the Japanese culture of “Ikebana” to everyone. Fortunately, we were able to learn about “Ikebana” from an expert in this field.
What you are about to see is the process of turning "FLOWERVER" into a wonderful project with the advice of people in the flower industry and experts. We hope you enjoy it.
From there, we had a series of meetings. However, since we had little knowledge of the reality of the flower industry, it was hard to make a framework for the project. We brought our ideas to florists, distribution centers, and farmers where the problem is happening and listened to their opinions. After all these interviews and discussions with them, we were able to change our project to be beneficial to the end-users and had great potential to solve the problem of flower loss.
We also shared what we learned from experts about "flowers" with the public. It is said that the demand for flowers has been on the decline in recent years. In such a situation, this project is trying to find a way to make it possible for people to continue to love flowers, which are a natural emotion that we have in common. This time, as one of the activities aimed at promoting the feature of flowers, we would like to introduce the Japanese culture of “Ikebana” to everyone. Fortunately, we were able to learn about “Ikebana” from an expert in this field.
What you are about to see is the process of turning "FLOWERVER" into a wonderful project with the advice of people in the flower industry and experts. We hope you enjoy it.
Go to Part2
Go to Part1
Part 1
Introduction -Identifying the Problem-
We began researching the problem of flower loss around April of this year. A simple Google search with
the keyword “flower loss”, brought up so many papers, data, photos, and videos. Most of them showed how
many flowers have been discarded in recent years.
We were concerned if the issue was related to the recent COVID-19 pandemic and if it was temporary or permanent. As we all know, flowers are used for big events such as marriages, festivals, and ceremonies. Maybe the flower loss occurred because those events were canceled due to COVID-19. Furthermore, if it is an ongoing problem we needed to consider the cause, and how it could be solved. To start, we interviewed several flower shops in Kyoto City to get a general idea of flower loss.
We were concerned if the issue was related to the recent COVID-19 pandemic and if it was temporary or permanent. As we all know, flowers are used for big events such as marriages, festivals, and ceremonies. Maybe the flower loss occurred because those events were canceled due to COVID-19. Furthermore, if it is an ongoing problem we needed to consider the cause, and how it could be solved. To start, we interviewed several flower shops in Kyoto City to get a general idea of flower loss.
Hanacho Florist
[Main Points]
We confirmed the existence of flowers that are out of standard and are discarded. We also got a hint that if we visit the growers, we might be able to find out more about the actual situation.
Hanacho Florist is a local florist located near Kyoto University. Due to the current situation, we asked questions via email and they provided us with a lot of information.
They say that there are few cases of flower loss that occur in retail stores. Rather flowers that do not meet a particular standard are discarded at the production phase.
If substandard products are distributed and spread in the market, the bad image that "flowers wither quickly" will be established among consumers. So, it is a shame to waste substandard products, but we learned that they are not sold to customers for the sake of a system that allows all three parties (producers, sellers, and consumers) to be happy, and most importantly, for consumers to be able to buy with peace of mind. As you may know, appearance is everything about flowers -You don't like flowers that wither quickly or have insect bites, do you?
Now we learned that substandard flowers were being thrown away, we decided to design a project based on the idea of how to avoid or reduce the number of substandard flowers at the production stage.
We confirmed the existence of flowers that are out of standard and are discarded. We also got a hint that if we visit the growers, we might be able to find out more about the actual situation.
Hanacho Florist is a local florist located near Kyoto University. Due to the current situation, we asked questions via email and they provided us with a lot of information.
They say that there are few cases of flower loss that occur in retail stores. Rather flowers that do not meet a particular standard are discarded at the production phase.
If substandard products are distributed and spread in the market, the bad image that "flowers wither quickly" will be established among consumers. So, it is a shame to waste substandard products, but we learned that they are not sold to customers for the sake of a system that allows all three parties (producers, sellers, and consumers) to be happy, and most importantly, for consumers to be able to buy with peace of mind. As you may know, appearance is everything about flowers -You don't like flowers that wither quickly or have insect bites, do you?
Now we learned that substandard flowers were being thrown away, we decided to design a project based on the idea of how to avoid or reduce the number of substandard flowers at the production stage.
Ginkaen Florist
[Main Points]
Ginkaen is what is called a general florist, doing business not only with B to C (selling flowers to customers) but also B to B(selling flowers and flower arrangements for florists). Mr. Masahiro Inoue, the representative of the florist, said that the florist's policy is to sell the flowers immediately after purchasing and use life-prolonging drugs at a minimum. Also, he insisted that the flowers they buy are always of the highest quality even though they are expensive. Low-quality flowers are cheap but likely to wither quickly so in the long term, they cannot satisfy the consumers which corresponds with the opinion of Hanacho florists.
- The life span of flowers is short and their quality is easily degraded, but there are currently not enough efforts being made to deal with it. Therefore, the idea of reducing the amount of flower loss was found to be worthwhile!
- There are many safety and efficacy issues with the existing chemicals to extend the life of flowers. A safe and simple to use life-extension product would be well received by any florist
Ginkaen is what is called a general florist, doing business not only with B to C (selling flowers to customers) but also B to B(selling flowers and flower arrangements for florists). Mr. Masahiro Inoue, the representative of the florist, said that the florist's policy is to sell the flowers immediately after purchasing and use life-prolonging drugs at a minimum. Also, he insisted that the flowers they buy are always of the highest quality even though they are expensive. Low-quality flowers are cheap but likely to wither quickly so in the long term, they cannot satisfy the consumers which corresponds with the opinion of Hanacho florists.
- Where and Why does flower loss occur?
To keep flowers for a long time, they need to be kept refrigerated and the time between the production site and the consumer has to be short. Since Ginkaen has several stores in Kyoto city and uses high-quality refrigerators, it can store a relatively large quantity of flowers at once. Smaller flower shops, on the other hand, do not have such storage facilities and purchase through various intermediaries, making it easier for the flowers to wither and discolor. Once the flowers are discolored, they cannot be sold as products and therefore discarded. So, one of the reasons for the flower loss is due to the equipment and purchasing conditions of the florist.
- Current countermeasures and their problems
Florists don't just watch the flowers go bad, they make an effort to deal with it. For example, lilies can be used for flower arrangements when they are in the bud stage and for funerals when they have bloomed. They can also sell flowers that are no longer of good quality at a low price at a 1$ shop. There are many ways to sell different stages of flowers.
However, during the "Higan" season (Buddhist services held during the week of the equinox), a large number of flowers need to be stocked for a long period, so a life-prolonging agent is used. These chemicals are costly and have weaknesses such as being sensitive to moisture. If there is an improved life-prolonging agent, Mr. Inoue says he thinks that many florists would find it useful. However, in terms of waste, he thinks that it happens more on the farmer's side than on the retail side. Some flowers can be replanted even though they are withered, but basically, substandard flowers are discarded. Last year, many events were canceled due to COVID-19, and some of the high-quality flowers could not be handled and discarded at the farm.
Summary So Far
Based on our interviews with the above two florists, we discovered that the majority of
flowers are lost in the flower production process. With a kind introduction from Mr. Inoue, we continued
our research at the Kyoto City Flower Local Wholesale Market.
Kyoto City Flower Local Wholesale Market
We visited Kyoto City Flower Local Wholesale Market.
[Main Points]
- The market is making efforts in establishing a good transportation system to avoid discarding flowers on-site. If there is waste, it will probably be substandard flowers that are identified and thrown away by the farmers.
The role of markets is to purchase flowers from farmers and distribute them. Last year (2020), events
were canceled all over Japan, and high-quality flowers could not be shipped and had to be thrown away.
Now that the situation is gradually returning back to normal, they are able to forecast the demand and
control the amount of distribution. Unlike vegetables, flowers are short-life, so they only purchase as
much as they can sell.
The main storage is always air-conditioned to avoid heat and the entire procedure of shipping is refrigerated using a cold chain system. There is also a bucket transportation method (wet transportation method) where the products are always immersed in treated water, so they are kept fresh.
The market has its own standard of purchase, and they are strict in order to deliver good flowers to the consumers. Therefore, farmers are encouraged to throw away flowers that do not meet the standards. Mr. Yoshiyuki Ozasa, a staff of the market, implied that this might be the cause of the flower loss.
The main storage is always air-conditioned to avoid heat and the entire procedure of shipping is refrigerated using a cold chain system. There is also a bucket transportation method (wet transportation method) where the products are always immersed in treated water, so they are kept fresh.
The market has its own standard of purchase, and they are strict in order to deliver good flowers to the consumers. Therefore, farmers are encouraged to throw away flowers that do not meet the standards. Mr. Yoshiyuki Ozasa, a staff of the market, implied that this might be the cause of the flower loss.
Kurokawa Floriculture
So far, we have been looking at various flower industries. Mr. Kurokawa is a flower farmer in Osaka City
who grows flowers on a large scale. Our encounter with this person had a major impact on the design of
the project.
According to Mr. Kurokawa, the cause of flower loss can be divided into two categories: planned and unplanned. The former type of planned loss is not limited to the flower industry but happens in all production industries. They produce more than the demand to maximize profits, so it is difficult to change this system. On the other hand, unplanned loss caused by climate change, viruses, and pests are of concern.
The causes of loss include...
According to Mr. Kurokawa, the cause of flower loss can be divided into two categories: planned and unplanned. The former type of planned loss is not limited to the flower industry but happens in all production industries. They produce more than the demand to maximize profits, so it is difficult to change this system. On the other hand, unplanned loss caused by climate change, viruses, and pests are of concern.
The causes of loss include...
- Climate change
Due to abnormal weather conditions in recent years, it is difficult to calculate how many plants will grow and when they will flower. However, Mr.Kurokawa can estimate the approximate amount through his experience of a long career, and it is not yet that serious. - Diseases (such as viruses, viroids and mildew)
Although prevention and treatment are done based on the type of infection, it is difficult to control fully due to the emergence of resistance.
Conventionally, agricultural chemicals are used to do deal with but experienced farmers can easily tell by intuition that plants have a disease. However, it should even help experienced farmers to avoid mistakes and identify problems early and with confidence. - Insect damage (damage caused by thrips)
Currently, farmers use pesticides but it is difficult to adjust the amount because unlike vegetables, flower petals are more delicate and easily cause stains. Also they have to spray pesticides at least once a week, becoming a burden on the labor force.
Fig. This figure diagrams the journey of our human practice work.
Interlude - The Cultures of Flowers in Japan -
Normally, how do you interact with flowers? You might decorate your table with flowers blooming in your
garden. You might buy the flowers displayed at the storefront and decorate your tasteless room with
them. You might sow seeds of your favorite flower and cultivate them.
There will be different relationships between you and flowers.
“Ikebana” can be counted as one of the ways to enjoy flowers. Ikebana is a Japanese traditional style of flower arrangement, but this is not enough to explain it. It has more interesting features. This time, Mr. Shimadu, who is a teacher at Ikenobo, Kyoto's oldest Ikebana school, taught us about Ikebana. Before we move on to next human practice work, please stop by and enjoy learning the history and artistry of flowers.
There will be different relationships between you and flowers.
“Ikebana” can be counted as one of the ways to enjoy flowers. Ikebana is a Japanese traditional style of flower arrangement, but this is not enough to explain it. It has more interesting features. This time, Mr. Shimadu, who is a teacher at Ikenobo, Kyoto's oldest Ikebana school, taught us about Ikebana. Before we move on to next human practice work, please stop by and enjoy learning the history and artistry of flowers.
The History of Ikebana
While Ikebana has a lot of schools, Ikenobo is the head family (iemoto, or original) of
Ikebana itself and we do not call Ikenobo a “derivative” school. It has a long history of more
than 560 years as the oldest record of Ikenobo can be found in "Tofukuji Hekizan Nichiroku", which is a
valuable document that provides an insight into the Muromachi period (approx. 1336 - 1573).
Ikenobo is said to originate from arranging flowers as a Buddhist ceremony. Since then, Ikenobo has undergone many transitions in its policies and their style has been handed down to the present day as it has become a part of our daily lives.
For example, until the middle of the Edo period (approx. 1603 - 1868), Ikebana was a pastime of the masters machiya, traditional (merchant's) houses. In that time, they faithfully followed the rules (etiquette) of Ikebana to create large works of art.
Then, in the early Meiji period (approx. 1868 - 1912), people in Japan began to realize the need for the opportunity of education for women as well in order to compete with other countries. Because of this movement, Ikebana was incorporated into schools as one of the education for female students. This time, the style was not regulated by rules and done freely as opposed to Ikebana in the Edo period.
In the Showa period (1926-1989), Ikebana as well as tea ceremony boomed as Hanayome Syugyou (lessons for women to learn various manners before marriage) and became very familiar to the general public. In those days it was said that every home had Suiban and Kenzan, special tools used in flower arrangement.
However, today, the number of Ikebana students is declining due to the diversification of lessons such as piano and dance. We also associated Ikebana with a difficult lesson to start casually until this interview, although it is one of the most fascinating cultures for everyday life in Japan. This will be discussed later.
What Is Important in Ikebana
-Ikebana is for a “person”
Originally, Ikebana began as a way of omotenashi (hospitality) for hotokesama, the Buddha, and ancestors. Over time, it changed to be practiced when people do omotenashi for house guests. As you can see, Japanese people consistently have entertained and are entertaining a person with Ikebana based on the idea of welcoming “someone else.”
-” Even more beautiful”
Flowers are beautiful enough as they are, and almost everyone will be happy to receive them. When we see flowers in the entrance or reception room of a house we are invited to, we will feel the hospitality of the destination and it will make us happy. Even if you are handed a dandelion from the roadside in a clumsy manner on the way home together with your friend whom you fought with, you will feel happy too.
Unlike music, painting, and haiku, which are created from a blank sheet, or what is called "nothing," and then "something" appears, Ikebana is trying to create something “more beautiful” from a flower that is already beautiful. A desire for making something more beautiful is the rules and spirituality of Ikebana.
However, the most important thing in Ikebana is to be exposed to flowers and enjoy from your heart.
Ikenobo is said to originate from arranging flowers as a Buddhist ceremony. Since then, Ikenobo has undergone many transitions in its policies and their style has been handed down to the present day as it has become a part of our daily lives.
For example, until the middle of the Edo period (approx. 1603 - 1868), Ikebana was a pastime of the masters machiya, traditional (merchant's) houses. In that time, they faithfully followed the rules (etiquette) of Ikebana to create large works of art.
Then, in the early Meiji period (approx. 1868 - 1912), people in Japan began to realize the need for the opportunity of education for women as well in order to compete with other countries. Because of this movement, Ikebana was incorporated into schools as one of the education for female students. This time, the style was not regulated by rules and done freely as opposed to Ikebana in the Edo period.
In the Showa period (1926-1989), Ikebana as well as tea ceremony boomed as Hanayome Syugyou (lessons for women to learn various manners before marriage) and became very familiar to the general public. In those days it was said that every home had Suiban and Kenzan, special tools used in flower arrangement.
However, today, the number of Ikebana students is declining due to the diversification of lessons such as piano and dance. We also associated Ikebana with a difficult lesson to start casually until this interview, although it is one of the most fascinating cultures for everyday life in Japan. This will be discussed later.
What Is Important in Ikebana
-Ikebana is for a “person”
Originally, Ikebana began as a way of omotenashi (hospitality) for hotokesama, the Buddha, and ancestors. Over time, it changed to be practiced when people do omotenashi for house guests. As you can see, Japanese people consistently have entertained and are entertaining a person with Ikebana based on the idea of welcoming “someone else.”
-” Even more beautiful”
Flowers are beautiful enough as they are, and almost everyone will be happy to receive them. When we see flowers in the entrance or reception room of a house we are invited to, we will feel the hospitality of the destination and it will make us happy. Even if you are handed a dandelion from the roadside in a clumsy manner on the way home together with your friend whom you fought with, you will feel happy too.
Unlike music, painting, and haiku, which are created from a blank sheet, or what is called "nothing," and then "something" appears, Ikebana is trying to create something “more beautiful” from a flower that is already beautiful. A desire for making something more beautiful is the rules and spirituality of Ikebana.
However, the most important thing in Ikebana is to be exposed to flowers and enjoy from your heart.
The Techniques and Uniqueness of Ikebana
Ikebana is a three-dimensional art form in which you use plants to express yourself, and this is
the feature that foreign flower arrangement has in common with Ikebana. However, as you know, it
is very difficult to express yourself and requires various techniques. Here are some examples:
1. Arranging and composing “flowers”
Ikebana artists arrange and compose flowers in the same way that composers arrange notes, painters arrange colors, and haiku poets arrange words. In addition, of course, plants are living things, so artists cannot force them too strongly, but they sometimes bend them a little bit to form the right contour to express themselves.
2. Removing "ma (間)" - feeling what is not there -
What is “ma (間) ?” “Ma” can be translated into English as “opening,” “empty space,” or “space.” Please look at the figure below. As you can see, when the large leaf spreads out on only the left side to create a semicircle, the object naturally creates a "ma" on the right side. The beautiful silhouette appearing in this way deepens in taste. Moreover, for example, this kind of methodology makes it possible to create Ikebana that makes you feel some wind in a windless room.
1. Arranging and composing “flowers”
Ikebana artists arrange and compose flowers in the same way that composers arrange notes, painters arrange colors, and haiku poets arrange words. In addition, of course, plants are living things, so artists cannot force them too strongly, but they sometimes bend them a little bit to form the right contour to express themselves.
2. Removing "ma (間)" - feeling what is not there -
What is “ma (間) ?” “Ma” can be translated into English as “opening,” “empty space,” or “space.” Please look at the figure below. As you can see, when the large leaf spreads out on only the left side to create a semicircle, the object naturally creates a "ma" on the right side. The beautiful silhouette appearing in this way deepens in taste. Moreover, for example, this kind of methodology makes it possible to create Ikebana that makes you feel some wind in a windless room.
Fig. Rikka formal style (courtesy of 井上太市池坊いけばな教室)
As a Consumer
As a consumer of cut flowers, Mr. Shimadzu also gave us some advice on our project.
Now, we are trying to make the flowers last longer. Regarding this, he remarked as the following.
“I believe that flowers are beautiful only because they wither. However, in exhibitions, the work needs to remain intact while it is seen by the audience for a day. In such cases, I put a lot of effort into it. For example, when arranging bamboo, I pull out the knots and fill it with water, or apply hand cream to prevent evaporation from the leaves.”
“Besides, once I have arranged flowers, I have an attachment to it, and come to want it to live as long as possible.” He went on and answered about another goal we are aiming for, which is to make flowers last longer “in an environmentally friendly way”. “Environmental considerations are a more and more major concern for many people today. Therefore, who would not be happy to be able to love their favorite flowers more or less longer while still saving the environment?”
Not only this, but Mr. Shimazu also provided us with some interesting ideas. “If you want consumers to use it, it should be solid like a capsule. It would be hard to be accepted because it would be a hassle to measure, add, and mix liquids. On the other hand, the flower sellers may accept your products in liquid types.” Thus, we found that we need to reconsider "solid or liquid" depending on "who will use it” (See the Engineering page and the Implementation page for details).
Now, we are trying to make the flowers last longer. Regarding this, he remarked as the following.
“I believe that flowers are beautiful only because they wither. However, in exhibitions, the work needs to remain intact while it is seen by the audience for a day. In such cases, I put a lot of effort into it. For example, when arranging bamboo, I pull out the knots and fill it with water, or apply hand cream to prevent evaporation from the leaves.”
“Besides, once I have arranged flowers, I have an attachment to it, and come to want it to live as long as possible.” He went on and answered about another goal we are aiming for, which is to make flowers last longer “in an environmentally friendly way”. “Environmental considerations are a more and more major concern for many people today. Therefore, who would not be happy to be able to love their favorite flowers more or less longer while still saving the environment?”
Not only this, but Mr. Shimazu also provided us with some interesting ideas. “If you want consumers to use it, it should be solid like a capsule. It would be hard to be accepted because it would be a hassle to measure, add, and mix liquids. On the other hand, the flower sellers may accept your products in liquid types.” Thus, we found that we need to reconsider "solid or liquid" depending on "who will use it” (See the Engineering page and the Implementation page for details).
Sense of Values
Where do you think has the greatest demand for cut flowers in Japan? The answer is Kagoshima. Kagoshima
is one of the prefectures located in the southwest part of Japan. Then, why Kagoshima? Because Kagoshima
has a culture that values ancestors so much. Most Japanese people leave flowers at graves only during
certain times of the year such as Obon and Higan , but people in Kagoshima leave flowers
all year round. The custom of offering flowers for ancestors during a walk is completely integrated into
their daily lives. The reason for this is exactly the sense of value we place on our ancestors.
Mr. Shimadzu continued his explanation. The above are just some of the differences in values within Japan, but of course, there are many differences in values when comparing Japan and the West. For example, most people prefer flowers in full bloom, but in Japan, especially in Ikebana , we also love budding flowers. At the root of this is the motivation to see the incompleteness in the bud and to cherish the future contained in it.
Mr. Shimadzu continued his explanation. The above are just some of the differences in values within Japan, but of course, there are many differences in values when comparing Japan and the West. For example, most people prefer flowers in full bloom, but in Japan, especially in Ikebana , we also love budding flowers. At the root of this is the motivation to see the incompleteness in the bud and to cherish the future contained in it.
Ikebana is not art, but culture for everyday life.
It is culture, it changes.
It is culture that has survived because it is part of our daily lives.
It is culture, it changes.
It is culture that has survived because it is part of our daily lives.
to be continued…
Part 2
Now that we have a better understanding of the issues facing the floral industry through
human practice work, we started to think about how we can solve these problems practically.
The target for our project is farmers, distribution center employees, and florists. We kept coming up with new solutions and soon improved our solutions based on their feedback. Thus, we completed the project called FLOWEREVER.
The target for our project is farmers, distribution center employees, and florists. We kept coming up with new solutions and soon improved our solutions based on their feedback. Thus, we completed the project called FLOWEREVER.
Fig. Illustration of our Human Practice
Infectious Disease Detection
Main Points
[Issues to be addressed]
- When infected with viruses, flowers can become oddly shaped.
- As a result, the flowers do not meet consumer standards, and are discarded.
- Development of a simple detection method that does not require expensive equipment and specialized facilities.
- Minimize the damage caused by the disease through early detection.
Understanding the Problem
When we visited Kurokawa floriculture, we were given a farmers' point of view. We learned about the
significant loss of flowers that cannot be shipped due to defects caused by viruses and viroids. Mr.
Kurokawa explained that they are taking preventive measures against the diseases by studying their
characteristics, but it is difficult to deal with them because resistant strains appear quickly. In
addition, he said that accurately determining whether a flower is infected is something that can only be
cultivated through years of experience.
Adapting the Approach
Therefore, we decided to switch our perspective from “solving a problem” to “preventing a problem”.
First, we considered a vaccine-like method to immunize flowering plants with the pathogen as a
preventative measure. However, there are very few practical vaccines that can be used to immunize
plants, and considering reproducibility, we decided that prevention of the virus was not realistic. So,
as a more practical measure, we came up with the idea of detecting viruses and viroids to prevent more
damage. By detecting virus-infected flowering plants early, we could prevent the spread of the virus. We
reasoned that screening which depends on experience could be replicated with a smart device. Designing a
device-based system that can be used in the field would be beneficial to farmers who are having
difficulties with accurate disease detection, or experienced farmers who require more efficient
detection methods.
With this idea as a starting point, we decided to investigate the method of detection in more detail and spoke with Dr. Shunsuke Asano of the Nara Prefectural Agricultural Research and Development Center, which is researching virus detection in dahlias. According to Dr. Asano, in flower production, prefectural officials inspect seedlings about once a year to see if they are infected with the virus.
However, since the producers are also increasing the number of seedlings, there is a risk of infection there, so they rely on visual inspection to identify infected products. This makes it difficult to distinguish the presence of the virus in small seedlings. Therefore, they wanted a cheaper and easier method for everyone.
It would also become more accurate in dealing with virus diseases if farmers could do this detection on their own rather than relying on the nursery companies. Even if a farmer receives an uninfected plant from a nursery, there is a possibility that the plant may become infected during production.
With this idea as a starting point, we decided to investigate the method of detection in more detail and spoke with Dr. Shunsuke Asano of the Nara Prefectural Agricultural Research and Development Center, which is researching virus detection in dahlias. According to Dr. Asano, in flower production, prefectural officials inspect seedlings about once a year to see if they are infected with the virus.
However, since the producers are also increasing the number of seedlings, there is a risk of infection there, so they rely on visual inspection to identify infected products. This makes it difficult to distinguish the presence of the virus in small seedlings. Therefore, they wanted a cheaper and easier method for everyone.
It would also become more accurate in dealing with virus diseases if farmers could do this detection on their own rather than relying on the nursery companies. Even if a farmer receives an uninfected plant from a nursery, there is a possibility that the plant may become infected during production.
Fig. How we came up with Detection.
Search for a Specific Detection Method
We decided to expand the conventional system to make it faster, more specific, and easier
to handle, so that farmers could perform detection daily.
Our research led us to the RT-LAMP method, which does not require a thermal cycler and allows faster detection. However, the RT-LAMP method produces a large number of false positives. The original purpose of detection was to reduce the economic loss caused by viral infection as much as possible. As such a large number of false positives would increase the number of unnecessarily disposed flowers, which is contrary to the original purpose. Then we found out from the paper (https://www.sciencedirect.com/science/article/pii/S0168170220310364) that false positives can be reduced by combining the RT-LAMP method with Cas12a.
Through this process, we decided to detect viruses infecting dahlia using the RT-LAMP method and DETECTR
(using Cas12a), which is a test method based on CRISPR technology.
BLOOM system was established to produce a biomolecular product that supports this CRISPR and RT-LAMP technology. The producer of this technology might use this BLOOM system.
In addition, we have started to develop software and hardware (pest control kit) that enables even beginners to easily detect and judge viruses and viroids, which was our initial goal.
The sample to be examined produces fluorescence that can be quantitatively analyzed for the presence of viruses. The fluorescence is then captured and photographed by the hardware. Finally, we developed software to quantitatively analyze and distinguish between positive and negative results for the presence of viruses and viroids.
Our research led us to the RT-LAMP method, which does not require a thermal cycler and allows faster detection. However, the RT-LAMP method produces a large number of false positives. The original purpose of detection was to reduce the economic loss caused by viral infection as much as possible. As such a large number of false positives would increase the number of unnecessarily disposed flowers, which is contrary to the original purpose. Then we found out from the paper (https://www.sciencedirect.com/science/article/pii/S0168170220310364) that false positives can be reduced by combining the RT-LAMP method with Cas12a.
Fig. RT-LAMP method with Cas12a.
BLOOM system was established to produce a biomolecular product that supports this CRISPR and RT-LAMP technology. The producer of this technology might use this BLOOM system.
In addition, we have started to develop software and hardware (pest control kit) that enables even beginners to easily detect and judge viruses and viroids, which was our initial goal.
The sample to be examined produces fluorescence that can be quantitatively analyzed for the presence of viruses. The fluorescence is then captured and photographed by the hardware. Finally, we developed software to quantitatively analyze and distinguish between positive and negative results for the presence of viruses and viroids.
Fig. Several approaches to help flower farmers
Antimicrobial Peptides
Main Points
[Issues to be addressed]
- When the flowers are soaked in water, bacteria grow and the flowers wither quickly.
- The treatment agents used are expensive, sensitive to moisture, and environmentally hazardous.
- “Synthesis of antimicrobial peptides”
It kills bacteria inside the vase and preventing inhibition of water absorption by blocking plant canals.
Fig. Human Practice at florists
Fig. Human Practice at distribution centers
Current Problems
When we first visited Mr. Masahiro Inoue and Mr. Yoshiyuki Ozasa, we learned that flowers
are put in a bucket with water to preserve. Although the water for flowers is changed daily, different
bacteria proliferate in different flowers when they are kept in water. The growth of bacteria leads to
the development of mold, blockage of the ducts, etc., causing early wilting. STS agent is currently used
to prolong flower ages but it is expensive, environmentally harmful, and can only be used in special
cases. In addition, this treatment is sensitive to humidity and keeps long since the backside of the
flower shop is very humid (in fact, there is a lot of water on the ground).
Fig. Cutflower Problem
The Cause of the Problems
Therefore, we explored the actual situation of treatment agents to solve these problems.
Treatments can be broadly divided into ethylene inhibitors and antibacterial agents. However,
ethylene-induced senescence is quite different from different varieties and cannot be tackled
comprehensively. We thought that a method to inhibit the growth of the bacteria would apply to many
varieties. We investigated the relationship between bacterial growth and flower aging and found that
biofilm (BF) formation was the cause. We went to interview Dr. Nomura at Tsukuba University, who is an
expert on BF, who gave us a very interesting talk.
Fig. Solution for a creation of the biofilm
Developments
~ We had developed a solution using antimicrobial peptides. ~
1. Controlling BF is important for the longevity of flowers
He showed us the actual BF of Pseudomonas aeruginosa and showed us that it is easy to visualize and observe over time using a confocal microscope. Pathogenic fungi create BF in plants, and we were able to visualize the BF inside by making the fungus glow with GFP and enter through the pores.
It was clear that microbial control = BF control. Therefore, the importance of controlling wilting flowers by decomposing the BF produced by the bacteria growth inside the vase and preventing inhibition of water absorption by blocking the plant's ducts was confirmed, which led to the decision to synthesize peptides with antibacterial properties.
2. Determination of the production method of antimicrobial peptides
Antimicrobial peptides can be produced by E. coli culture, but we were not sure how can we detect the peptide and specific assays by using the peptide, so we talked to Dr. Ogawa, Takeuchi, and Watanabe at Kyoto University, who are experts of biomolecular production. In the initial discussion, several methods were proposed for detecting antimicrobial peptides (SDS-PAGE and blotting, HPLC enrichment and amino acid sequencing, liquid-phase chromatography, MS, ultrafiltration, and combination of monolithic column). However, liquid-phase chromatography proved to be the best choice for this experiment. The reason why we chose it was that it was “not excessive”
3. How to check the effect of AMP
In addition, when we asked for advice on how to conduct an assay to check the effect of AMP, we were told that we should isolate bacteria from flower arrangement water and then conduct a disk diffusion method. The goal was to collect bacteria and check the growth under different conditions by simulating by assigning different numerical values to the concentration of AMP or combining multiple AMPs with several enzymes to derive the optimal concentration and combination of antimicrobial substances. In addition, microbiome analysis was also proposed, but at that time, we didn’t know if it was financially feasible. Considering all these facts above, Dr. Ogawa suggested two approaches.
The first method is to build a model of flower arrangement.
We put flowers in the water combined with the ratio of bacteria in the actual water of flower arrangement and test the conditions of AMP concentration. To make it get closer to the actual ratio, analysis of the bacteria is essential.
The second method is to use the actual water from the flower arrangement as it is and put the flowers in it to test the concentration and combination of AMPs. Again, since the water from the flower arrangement was the actual sample, bacterial analysis was required to validate the results (the ratio of bacteria may vary from place to place).
We were also advised that whichever method we use, we should use water that is dirty enough to wither so that we can easily see the effect.
The experimental system we constructed was very useful, and we were able to stably produce and evaluate the target peptides (See the Experiment page for details).
1. Controlling BF is important for the longevity of flowers
He showed us the actual BF of Pseudomonas aeruginosa and showed us that it is easy to visualize and observe over time using a confocal microscope. Pathogenic fungi create BF in plants, and we were able to visualize the BF inside by making the fungus glow with GFP and enter through the pores.
It was clear that microbial control = BF control. Therefore, the importance of controlling wilting flowers by decomposing the BF produced by the bacteria growth inside the vase and preventing inhibition of water absorption by blocking the plant's ducts was confirmed, which led to the decision to synthesize peptides with antibacterial properties.
Fig. Anti-bacteria peptide
2. Determination of the production method of antimicrobial peptides
Antimicrobial peptides can be produced by E. coli culture, but we were not sure how can we detect the peptide and specific assays by using the peptide, so we talked to Dr. Ogawa, Takeuchi, and Watanabe at Kyoto University, who are experts of biomolecular production. In the initial discussion, several methods were proposed for detecting antimicrobial peptides (SDS-PAGE and blotting, HPLC enrichment and amino acid sequencing, liquid-phase chromatography, MS, ultrafiltration, and combination of monolithic column). However, liquid-phase chromatography proved to be the best choice for this experiment. The reason why we chose it was that it was “not excessive”
3. How to check the effect of AMP
In addition, when we asked for advice on how to conduct an assay to check the effect of AMP, we were told that we should isolate bacteria from flower arrangement water and then conduct a disk diffusion method. The goal was to collect bacteria and check the growth under different conditions by simulating by assigning different numerical values to the concentration of AMP or combining multiple AMPs with several enzymes to derive the optimal concentration and combination of antimicrobial substances. In addition, microbiome analysis was also proposed, but at that time, we didn’t know if it was financially feasible. Considering all these facts above, Dr. Ogawa suggested two approaches.
The first method is to build a model of flower arrangement.
We put flowers in the water combined with the ratio of bacteria in the actual water of flower arrangement and test the conditions of AMP concentration. To make it get closer to the actual ratio, analysis of the bacteria is essential.
The second method is to use the actual water from the flower arrangement as it is and put the flowers in it to test the concentration and combination of AMPs. Again, since the water from the flower arrangement was the actual sample, bacterial analysis was required to validate the results (the ratio of bacteria may vary from place to place).
We were also advised that whichever method we use, we should use water that is dirty enough to wither so that we can easily see the effect.
The experimental system we constructed was very useful, and we were able to stably produce and evaluate the target peptides (See the Experiment page for details).
Feedback from Ginkaen
Although the antimicrobial peptides were developed in this way, we eventually went back
to the Ginkaen and asked them if they could put them to practical use. They then commented that the
peptides were resistant to moisture, cheaper than conventional products, and easier to handle.
Killing Pests
Main Points
Fig. Conversation with farmers
- The opinion of the farmer made it possible for us to narrow down a target to thrips among many kinds of agricultural pests. In addition, the exchange of ideas with experts changed the way of dsRNA works, which was the key to the success of the experiments.
Understanding of the problem
When we visited Mr. Kurokawa, we learned how serious the damage caused by thrips is at
the production site. Our team had just started noticing thistles in several papers we read, and we were
not sure whether to target them for insect control. However, thanks to Mr. Kurokawa's voice as a flower
producer, our policy was decided.
Path for Solution
Then, we had many online meetings using Zoom with Dr. Shimada and Dr. Nakano of Kyoto
University, Dr. Nakao of Kyoto Prefectural University, and Prof. Sonoda of Utsunomiya University. Based
on the findings we learned there, we developed a more optimal experimental method.
For example, in the early stages of our project, we had various ideas about what methods would be appropriate for killing thrips. One of the ideas was to create transgenic plants. This idea sounded interesting. However, Dr. Shimada pointed out the length of time it would take to complete the experiment, and we found it difficult to complete it before the iGEM deadline. Therefore, based on the advice by Dr. Shimada, we gave up on actually trying it out for this project.
After many discussions, we decided to use RNAi. Then, we started to plan the details of the experimental method.
For example, in the early stages of our project, we had various ideas about what methods would be appropriate for killing thrips. One of the ideas was to create transgenic plants. This idea sounded interesting. However, Dr. Shimada pointed out the length of time it would take to complete the experiment, and we found it difficult to complete it before the iGEM deadline. Therefore, based on the advice by Dr. Shimada, we gave up on actually trying it out for this project.
After many discussions, we decided to use RNAi. Then, we started to plan the details of the experimental method.
Advice from Experts
With an introduction from Dr. Sonoda, we met Dr. Tokumaru of the Agriculture and Forestry
Center of the Kyoto Prefectural Agriculture, Forestry and Fisheries Technology Center. Dr. Tokumaru
taught us various practical matters necessary to prepare for conducting experiments with thistles.
For instance, we used the primary leaves of green beans because he recommended them to us. Those leaves have fewer hairs, so the percentage of thrips that die when they get their legs caught in the hairs is reduced compared to normal leaves.
In addition, in the assay, we cut the leaves into 1 cm squares instead of using whole leaves because Dr. Tokumaru advised that thistles are too small to see them on whole leaves.
These made it easier for us to see if our dsRNA was really effective in eliminating thrips.
The actual results can be seen at the Experiment page in details.
However, we have come up against a challenge here. It is a question, whether what we are trying to
create is actually adaptable to the flower production stage. To answer this question, we revisited the
journey of cut flowers from production to consumption.
When we interviewed the people again, we found that damage caused by thistles and other insects occurs not only in the production stage, but also during transportation, sales, and consumption, and solutions are needed. Therefore, we proceeded to think about how we could use what we were trying to create, and whether we could verify it with our current experimental design.
After several interviews with Ginkaen, we found many cut flowers are transported in containers filled with water from production to distribution. So one possible application is to add a solution containing our dsRNA to the water used for harvested flowers. In the wet transportation process, in the florist's store, or in the vase of the consumer's home, this will work and flowers will last longer.
Considering this usage, our method of immersing the petiole directly in water containing dsRNA was exactly practical. However, to make what we have created more practical, we will need additional experiments with dsRNA absorption from the stem as well, which is more basal than the petiole.
Meanwhile, Mr. Kurokawa, a flower farmer, said, "Considering that we usually apply pesticides in the form of spraying, it will be difficult to conduct treatment for each leaf individually because it requires a lot of labor.”
Hence, we decided to plan the future development of our experiment.
According to Mr. Kurokawa, it seems that there is a need in the production field to be able to induce RNAi in insects by absorbing dsRNA from their roots. If we continue this study in the future, we would extend our work on experiments in which Arabidopsis thaliana absorbs dsRNA from its roots. However, we do not expect these experiments to directly show the effects on thrips. Since Arabidopsis thaliana has small leaves and is very difficult to assay observing insects, we would like to confirm that it is possible to spread dsRNA throughout the plant and induce pests RNAi by checking whether anti-aging RNAi is triggered in the flowers.
Experiments in this project have already shown that allowing plants to absorb dsRNA from the petiole is also effective in killing thrips to some extent. So, if this planned experiment goes well, we will see more clearly that the effectiveness of our dsRNA as an insecticide is excellent.
For instance, we used the primary leaves of green beans because he recommended them to us. Those leaves have fewer hairs, so the percentage of thrips that die when they get their legs caught in the hairs is reduced compared to normal leaves.
In addition, in the assay, we cut the leaves into 1 cm squares instead of using whole leaves because Dr. Tokumaru advised that thistles are too small to see them on whole leaves.
These made it easier for us to see if our dsRNA was really effective in eliminating thrips.
The actual results can be seen at the Experiment page in details.
Fig. Zoom with Dr. Tokumaru
When we interviewed the people again, we found that damage caused by thistles and other insects occurs not only in the production stage, but also during transportation, sales, and consumption, and solutions are needed. Therefore, we proceeded to think about how we could use what we were trying to create, and whether we could verify it with our current experimental design.
After several interviews with Ginkaen, we found many cut flowers are transported in containers filled with water from production to distribution. So one possible application is to add a solution containing our dsRNA to the water used for harvested flowers. In the wet transportation process, in the florist's store, or in the vase of the consumer's home, this will work and flowers will last longer.
Considering this usage, our method of immersing the petiole directly in water containing dsRNA was exactly practical. However, to make what we have created more practical, we will need additional experiments with dsRNA absorption from the stem as well, which is more basal than the petiole.
Meanwhile, Mr. Kurokawa, a flower farmer, said, "Considering that we usually apply pesticides in the form of spraying, it will be difficult to conduct treatment for each leaf individually because it requires a lot of labor.”
Hence, we decided to plan the future development of our experiment.
According to Mr. Kurokawa, it seems that there is a need in the production field to be able to induce RNAi in insects by absorbing dsRNA from their roots. If we continue this study in the future, we would extend our work on experiments in which Arabidopsis thaliana absorbs dsRNA from its roots. However, we do not expect these experiments to directly show the effects on thrips. Since Arabidopsis thaliana has small leaves and is very difficult to assay observing insects, we would like to confirm that it is possible to spread dsRNA throughout the plant and induce pests RNAi by checking whether anti-aging RNAi is triggered in the flowers.
Experiments in this project have already shown that allowing plants to absorb dsRNA from the petiole is also effective in killing thrips to some extent. So, if this planned experiment goes well, we will see more clearly that the effectiveness of our dsRNA as an insecticide is excellent.
Future Application
Furthermore, if this future experiment is successful, we can apply this dsRNA pesticide
to hydroponic cultivation, which has already started with tomatoes. If we can approach not only flowers
but also vegetables, the products we are trying to produce will be more in compliance with the
principles of the SDGs.
At the same time, we tried to come up with another method. We ended up with a thistle trap. The design is as follows. First, we apply our dsRNA to green beans, a favorite food of thistles. Second, we put it around the field. Then, we can keep thistles away from flowering plants and avoid exposing the products to feeding damage.
We also came up with a transgenic plant based on the same principle to induce RNAi in thistles. This was one we gave up on once in the early stages of our project, but decided to review it as the need for solutions in the production phase became clear through Human Practice (See the Implementation page for details).
At the same time, we tried to come up with another method. We ended up with a thistle trap. The design is as follows. First, we apply our dsRNA to green beans, a favorite food of thistles. Second, we put it around the field. Then, we can keep thistles away from flowering plants and avoid exposing the products to feeding damage.
We also came up with a transgenic plant based on the same principle to induce RNAi in thistles. This was one we gave up on once in the early stages of our project, but decided to review it as the need for solutions in the production phase became clear through Human Practice (See the Implementation page for details).