The "Everyone's Euglena Project" (MinMido PJ) Initiated in 2019

Seeking new strains of euglena, we launched a citizen-participation research project where people from all over Japan sent us water samples. The collection of water samples was conducted over three years, from 2019 to 2021. This article presents a comprehensive summary of the samples collected so far!

Participants and Total Number of Samples in the MinMido PJ

Table 1: Summary of MinMido PJ from 2019-2021

Year	Participants	Applications	Arrived Samples	Total Samples
2019	High School Students	17 schools	13 schools	207
2020	Families	80 groups	58 groups	295
2021	Middle & High School Students	18 schools	11 schools	190
	Families	97 groups	70 groups	353
Total	Schools	35 schools*	24 schools*	1045
	Families	177 groups*	128 groups*

Figure 1: Sample Collection Locations

As shown in Table 1, over the past three years, 35 schools and 177 families (including duplicates) participated in the MinMido PJ. Due to the impact of the COVID-19 pandemic, some who applied couldn't unfortunately collect water samples. Nevertheless, we received water samples from 24 schools and 128 families. A total of 1045 samples were collected from all over Japan, from Hokkaido in the north to Okinawa in the south.

About the Obtained Euglena Strains

Table 2: Euglena Strains and Their ITS2 Sequences

Year	Total Samples	Euglena Present	Established Strains	Types of ITS2 Sequences	New ITS2 Sequences
2019	207	14	13	5	1
2020	295	31	34	15	6
2021	543	33	33	12	2
Duplicates	–	–	–	13	2
Total	1045	78	80	19	7

Out of the 1045 samples collected, 78 samples contained euglena. 80 strains of euglena were successfully isolated and cultured. By examining the ITS2 region of DNA, 19 different strains of euglena were identified, including the commonly used Euglena gracilis Z. Notably, 7 of these were new ITS2 sequences not identified before.

While the ITS2 sequence alone doesn't determine the characteristics of a euglena strain, among these new strains, there might be some with unique functionalities.

Origin of the Euglena Strains

Figure 2: Phylogenetic Tree of Euglena Strains (ITS2) and Their Origin Regions

Lastly, the phylogenetic tree on the left of Figure 2 arranges the obtained euglena strains based on their ITS2 sequence differences. The branches of the tree indicate the sample ID and collection location. The origin regions of the euglena strains are shown in green on the map of Japan on the right. Strains were obtained from a wide range of regions, from Hokkaido in the north to Kagoshima in the south. We will analyze whether any of these strains have unique characteristics, such as resistance to heat or cold.

Furthermore, we have the opportunity to gather academic data on whether there's a relationship between the origin region and the DNA sequence or cellular characteristics of the euglena. Understanding the evolution and distribution of euglena in Japan can provide valuable insights.

In conclusion, over the three years since 2019, thanks to your cooperation, we've been able to obtain a wide variety of euglena strains from various locations across Japan, even in challenging times. We sincerely thank everyone for their contributions!

Moving forward, we aim to investigate whether there are euglena strains with new features and to explore the relationship between their origin regions and their characteristics.