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Human practice

• 1) Abstract
• 2) Discussion with forestry association and forest survey
• 3) Screening of "Genetic Roulette"
• 4) Events

1) Abstract

Nightingale once said as follows

'Regardless to any work, it is only in the field is to be able to learn in practice.'

This is a very suggestive idea for those who work greatly for society. Instead of conducting research on the laboratory alone, unless it is in accordance with the actual circumstances of the problem, it becomes meaningless even though you use cutting-edge technology. The important thing is to reflect the voices and realities of the site. We tried to make our project suitable for the reality both by visiting the Forestry Association and asked their opinions and by going out to the mountain by ourselves.

2) Discussion with forestry association and forest survey

Discussion with Forestry Association

In addition to gathering information on pine-wilt disease on the Internet, we went to two Forestry Associations managing the forest in Kyoto city where we live and discussed them in order to know the current situation such as how widely pine-wilt disease spread and what kind of measures are taken in Kyoto city. Through discussion we got the following.

• Drug spreading that we found on the Internet is not used now.
• pine trees in Kyoto city are now in a state of being destroyed.
• They use an injection to save remaining pine trees every year, but the results have been decreasing year by year due to the expansion of the damage of pine-wilt disease.
• Such measures against pine-wilt disease are accompanied by a physical burden.

If we come up with great measure against pine-wilt disease, the measure does not make any sense when pines are not left. As we felt strongly so, we decided to breed RNAi pine using genetic engineering techniques to regain the beautiful pine forest in Kyoto city.

Forest survey

In summer, we visited the forest behind Kiyomizudera temple (Kodaiji mountain national forest), which is said to have a lot of damage from pine wood nematode in Kyoto city, and carried out field survey of pine-wilt disease and collection of pine wood nematode. It is said that this mountain was once forested mainly by red pine, but now it was forested mainly by chinquapin like a picture. Eventually we could only find 5 pines in a 2 hours survey. (2 of 5 pine trees seem to be infected by pine-wilt disease) In order to take wild pine wood nematodes from those 5 pines, we collected wood chips and carried out extraction of pine wood nematodes. Then, 4 nematodes which seemed to be pine wood nematodes were obtained. We attempted to cultivate those 4 to perform the feeding RNAi experiment using the obtained wild type nematode, but unfortunately failed. So we went to Yoshidayama mountain near the Kyoto university and again conducted a forest survey and collection of pine wood nematodes. Yoshidayama mountain was once a mountain where many red pine trees grew, but here too, many pines withered due to pine-wilt disease, and now it is forested mainly by chinquapin. In addition, there was a trace of injection in all of the remaining pine and we recognized again how serious pine-wilt disease caused by pine wood nematode is. We collected branches of pines that are seemed to be affected by pine-wilt disease and tried to extract pine wood nematodes. However, although many nematodes were obtained, no nematodes could be obtained that seemed to be a pine wood nematode. The following hypotheses can be considered as the reason why pine wood nematode was not obtained.

• Long horn beetles decreased drastically as withered pines were cut down every year, and pine wood nematodes were unable to move to new trees and decreased.
• Since the season in which the survey was conducted was around winter, nematodes moved to the pupal chamber of long horn beetles in a tree, and pine wood nematodes had disappeared in the branches.
• Pine wood nematodes could not infect pine trees by the drug of injection of last year.
• The remaining pine trees of Yoshidayama Mountain have resistance to pine wood nematodes.

3) Screening of "Genetic Roulette"

4) Events