High School Students Publish a Paper on Plant Physiology in a Notable Journal

— Written by Tim Marzullo —

In an article we previously published in June 2022 about our scientific paper that dealt with play behavior in fish, I concluded at the end of the article:

“I think it is possible for novices and high school students to publish papers (and it is the dream and goal of our team)… That is why we are planning an experiment. We want to publish with a school in Santiago, Chile, collaborating with second and third year high school students. We are collecting data on electrical signals in plants… If it works, we will tell you…”

Dear readers, 21 months after writing this, the day has arrived. We did it! Our paper recently appeared in the academic journal “Plant Signaling and Behavior” about our experiments in electrophysiology in plants, with 5 high school students as the first authors. You can read the paper here.

Electrical signals in plants? What? Yes, it is understudied and often misunderstood, but plants do have signals similar to the electrical signals we have in our hearts, muscles, and brain. However, they are much slower (1,000-15,000 times slower). But what are they for? In the famous examples of the venus flytrap and the sensitive mimosa, the electrical signals coordinate their fast movements, but electrical signals also exist in plants that do not move quickly, such as tomatoes, chili peppers, basil, etc.

One of the functions of electrical signals in plants is as an alarm signal. For example, if a herbivore is eating a plant, an electrical signal passes through the branches saying “we are under attack” and the plant can synthesize bitter compounds so that the leaves taste bitter. A plant cannot escape when under attack, and it has the problem that it is “stuck in place forever” (i.e., it cannot run away from a threat, or fight physically), but there are protection systems and defenses (thorns, poisons, production of bitter compounds, etc.).

As electrophysiology in plants is understudied, we wanted to further investigate electrical signals in plants that do not necessarily move rapidly. And with that idea, we began to work on an ambitious project with the (high school) Colegio Alberto Blest Gana (CABG) in San Ramón, Santiago.

But before discussing the results, we must give a little more context about the scientific publication process.

The Value of Publication in Science

In the scientific world, the most important thing is publishing your results in academic journals. In the entrepreneurship world, the most important things for a business are its sales and its impact on society (what problem it is solving), but for the science world the most important things for a research team are 1) the amount of the papers published, and 2) the quality/impact of the papers. The two most famous scientific magazines in the world are “Science” from the USA and “Nature” from the United Kingdom, (the discovery of the structure of DNA was described in Nature and the discovery of CRISPR was published in Science, for example). There are literally thousands of scientific journals in all areas of science, and of course, some more prestigious than others (in biology, “Cell” is considered very good, in neuroscience “Neuron,” and in medicine “Lancet”).

During a scientist’s training, some publish their first publication in the last years of their undergraduate studies (not so common), but the majority of scientists in training publish their first papers during their postgraduate studies, working with their principal investigator (head of research). Normally, graduate students do the experiments, prepare the figures, and their principal investigator helps with the writing. For the first paper, the principal investigators do a majority of the writing, but over time the graduate students begin to write their entire papers in the process of learning how to write in precise and technical language.

Recently, we at Backyard Brains wanted to try something very novel – involving high school students in the process of contributing knowledge to the scientific record. There are some examples of grammar school students who have published (Blackthorton School) or examples of scientists who have published with their children (including in Nature). However, to publish a paper, professional scientists need to be very involved due to technical writing and literature knowledge, which is why it is uncommon for high school students to publish original scientific research.

The Alliance Between Colegio (High School) Alberto Blest Gana and Backyard Brains

Our team has worked with Colegio Alberto Blest Gana, in San Ramón, Santiago for 10 years (our first appearance there was in 2014), and each year we have tried something more ambitious. When we started, we did standard Backyard Brains talks (experiments on cockroaches, human physiology, etc.), and after that we started doing workshops, where the students learned how to assemble homemade capacitors and batteries, how to use breadboards, and do basic Arduino programming, etc. After that, we did individual projects (sports physiology, learning in insects, playing video games with muscle electrical signals, for example).

This last series of classes with the high school was in 2018, and in 2019 I went to Seoul, Korea to open a branch of Backyard Brains there. For a while we paused in the activities with Colegio Alberto Blest Gana. In March of 2020, while I was still participating in the Korea Start-Up Grand Challenge program, an entrepreneurship program of the Korean government, the first case of COVID-19 outside of China arrived in Korea, and shortly after that COVID-19 reached the whole world. We know all too well what happened afterward.

One result of COVID-19 was that the educational world transformed and became more accommodating to video classes (Google Video, Zoom, Microsoft Teams, etc.). Before 2018, all Backyard Brains´ classes at Colegio Alberto Blest Gana were in person, but during COVID-19, the director of the high school, Ricard Román, invited me to give some talks to the faculty and students remotely. After the peak of the pandemia passed I returned to Chile to work with colleagues and in the fall of 2022 I did in-person classes with the school.

A High School Accepting the Risk to Try to Publish a Paper

On April 9, 2022, I presented an in-person talk to the students where we made the first electrical recording on a non-rapid movement – basil. After the talk, I had lunch with the faculty to discuss future projects at the school (and, of course, gossip about our personal lives). At the excellent Chilean restaurant Del Beto, in San Miguel, Ricardo Ramón, the director of the school, and the technology coordinator, Fabién Ovalle, said to me and the other faculty present – “We have done very ambitious things together for 8 years. What shall we do for this year?” I thought, and said, “We should try to do original research, with the goal of publishing an academic paper, with the high school students as the first authors. It would be very novel, and I think we can organize something with the experiments in plant physiology.”

Ricardo, Fabián, and the faculty were intrigued by the idea, but they wanted a hybrid model: that we could work with a group of 4-6 students to do original experiments, and also, at the same time, do general Backyard Brains classes with a larger group to have more inclusion with the students at the high school.

With the word and informal agreement said and done, it was time for action and planning. Thus, we spent a few months preparing the budgets, the schedule, the shipment of equipment, the curriculum, and the organization of the faculty and students to begin after the winter holidays – a project of the second semester of the school year. For the original experiments with plants, and due to the distance because I was in Korea, I could not do the work alone. Thus we also had a biology teacher faculty, Angélica Romero, working with the students, Fabián, the coordinator, and finally, my German colleague and dear friend, the neuroscientist Étienne Serbe-Kamp from the Max Planck Institute for Biological Intelligence and the LMU in München. As a teenager, Étienne spent a few months in Santiago as an exchange student, and he is trilingual in German, Spanish, and English. With the team of teachers assembled, we were going to work with five third and fourth year high school students – Danae Madariaga, Derek Arro, Catalina Irarrázaval, Alejandro Soto, and Felipe Guerra.

For the CABG faculty, the idea of working with a global team with the mission of publishing an academic paper was an intriguing idea that was worth a shot. Risky, sure, but intriguing.

Why risky? Because everyone who has worked on academic publications knows that it is a long process with many possibilities of failure: 1) the research results may not be interesting enough for publication, or 2) the paper could be rejected many times with many revisions, and when that occurs, it is very difficult to maintain morale of continuing to try to publish, even for professional scientists. But Backyard Brains’ mission is to make science more open, with more movement between high schools, scientists, universities, companies, and institutes, and we thank the enthusiasm of the CABG faculty and students for taking the risk with us.

We will now return to discussing the results of the project.

A Cross-Continental Lab: From Data Collection to Publication

On August 9, 2022, after winter break, we started classes. Classes typically occurred at 10 a.m. Chile time, once a week. It was a little complicated to coordinate everything, because we were doing video classes on three different continents, in Chile (the high school students), Germany (Étienne), and Korea (me). Also due to the time differences, while in Chile it was 10:00 in the morning, it was 16:00 in the afternoon in Germany, and 23:00 at night in Korea. It was also complicated with the equipment and plant inventory needs. We sent everything to the home of coordinator Fabián, whose apartment building in La Florida had a concierge, where it was more convenient to receive shipments. Fabián brought everything to the school personally.

During the first two classes, we discussed the scientific process of converting an idea into a scientific paper, using as an example our recently published paper, stated above, on play in fish (it had been published only 2 months before at the time). To publish something with young scientists with the goal of being released within 1-2 years after the start of the experiments, it had to be an experiment where it is possible to record data quickly, repeatedly, and without a requirement for expensive equipment that is fragile and difficult to use. Thus we discussed that experiments with plant electrophysiology seemed a good candidate, as it is understudied and there is a lot of room for novel research (and novel research has fewer barriers for publication, as there are not many other similar papers).

With this goal, during the third week and the third class, we began the first experiments, using the ornamental plants the school already had in the teacher’s lounge: the Argentine dollar. A few weeks before we had sent two amplifiers they needed for the experiments: the Plant SpikerBox, which we could hook up to the school laptops and begin the process of collecting data. Below is a photo of the first experiment they did.

The Methodology and Configuration of the Experiments

Thus, every week, from August 9 to December 16, 19 classes in total, each class lasting between 60 – 75 minutes, we made recordings from many different plants, 16 species in total:

1. Araucaria
2. Argentine Dollar
3. Basil
4. a type of fern
5. a type of ivy
6. Lemon Balm
7. Mint
8. Oregano
9. Papyrus
10. a type of ornamental plant of the genus Peperomia
11. Rosemary
12. Ruda
13. Sensitive Mimosa
14. Drosera (Sundew)
15. Tomato
16. Venus Flytrap

With each experiment, we put a silver wire around the branches, a pin in the ground for the electrical ground (engineering joke), with an amplifier and software recording the electrical signal. The students held a flame with a long-necked lighter for a few seconds to the leaf closest to the electrodes to measure “the wound response.”

We observed electrical signals in response to the flame in 9 of the 16 plant species we studied. Unfortunately we did not observe any in Araucaria, which would have been notable because it is very likely that we would have been the first researchers in the world to electrophysiologically study this sacred plant, but we continue. It is very likely that all plants in the world use some form of electrical signals, but the electrodes and interfaces must be designed in a way that is ideal for each plant. The plants that did show electrical signals in response to a flame were:

1. Basil
2. Argentine Dollar
3. Drosera
4. Mint
5. Mimosa Púdica
6. Rosemary
7. Ruda
8. Tomato
9. Venus Flytrap

We have some examples of their responses below.

After December, and with the end of classes, we continued with two of the students, Danae and Derek, as interns during January-March 2023. I worked mainly with Derek in the beginning of our manuscript writing, working on the introduction of the paper and doing an analysis of the literature together (where we tried playing a little with ChatGPT-4 to ask some questions about the plant physiology literature, but it didn’t help us much due to how technical and paywalled the scientific literature is). Étienne worked primarily with Danae on the final data collection we needed to complete the research.

By the end of March, our data collection with the high school students was complete, and we continued the process of writing the manuscript, reviewing the literature, improving the Python software to make the analysis more automatic, and preparing the figures. Also, during July 2023, we had the opportunity to collaborate with a science foundation in Belgrade, Serbia, and two undergraduates, one from Serbia and one from the US, collected additional data for the paper. In total we had 398 recordings from 16 plant species and 5 countries represented in this study: Chile, Germany, Serbia, South Korea, and the United States.

The Final Pass: Polishing and Publication

The process of finishing the paper was long, but predictable. We started writing from the blank page on January 11, working a few days a week on the paper, and on September 27, 2023, we sent the paper to the journal Plant Signaling and Behavior, where many of the well-known members of the new generation of scientists who study botanical physiology publish (learning in plants, communication between plants, etc.). Fingers crossed, for everyone involved, it was our first attempt to publish in plant physiology (we the puppies in the lions’ den). 

On October 24, 2023, on a beautiful day that I will not forget, we received an email from the editor, František Baluška, a professor of plant physiology at the University of Bonn, Germany, and well known in the community. With my heart rate shooting through the roof, I read the line: “We consider your work appropriate for publication if you address the comments of the reviewers.” The text sounds very dry and anodyne, but I was ecstatic with the news, as well as the entire team. We had 6 reviewers with many questions and doubts that we needed to clarify, and it would be a lot of work to respond to the comments, but “the door was open.”

Étienne and I congratulated the entire team on their success, and, in the months that followed, he and I worked carefully on the revisions. We needed to clarify our methodology, analyze ~20 more papers, and add more context to the manuscript. Our response letter was 16 pages, and written in the highly formal, scholarly, and polite manner even by scientific manuscript standards. On January 14, 2024, we sent the revised manuscript. Fingers crossed again.

On January 22, we received another email from the editor: “We are happy to inform you that your work has been accepted for publication. You should hear soon from our production team about final preparations for publication and receive the proofs soon.”

I wrote the entire team on WhatsApp immediately. Fireworks exploded in our minds. Another moment of happiness. Normally in science publication there are two rounds of revisions, but in this case, we were done with just one round! Completely accepted. We did it!

During the weeks of February and March, we worked with the magazine’s production team to verify and polish the proofs (the form of the manuscript that will be published), and on March 12, we sent the final revisions. On March 18, our work was released and available to everyone around the world to read. I have been publishing papers since 2002, but, dear readers, this paper is the paper I am most professionally proud of. We had fulfilled the core mission of Backyard Brains, that high school students themselves can contribute to the scientific literature, not just learn about science, but also add new knowledge to the world.

I’ve discussed the whole process with my scientific colleagues in Chile, and one of them, Daniela Flores, a biology graduate student from U-Chile, said something illuminating: “It would be great that this happens more and more frequently, but the key is to find a school willing to take the risk of doing original scientific research, and also the scientists who want to work with the school long term.” A paragraph in the discussion of our paper is a result of this exchange, and is essential to put this work in context: a call to action to the scientific community:

“It is a non-trivial part of this work that high school students did the majority of the data collection, showing the advantages that plant electrophysiology has for introducing young students to the scientific process of data collection, data analysis, and manuscript writing, something most aspiring scientists are only exposed to in late undergraduate education at the earliest, if lucky. The participation of professional scientists on a consistent, long-term basis with young researchers is critical for such initiatives. Young students can record data and discuss results, but analyzing the literature and the technical writing is the biggest challenge, which requires active oversight and collaboration by scientist colleagues.”

Again, what we restate is that it is not at all impossible for high school students to have an accepted scientific paper even before going to university, but the active participation of the scientific community is needed. Our hope is that this paper serves as a model for other scientists and high schools that aim to add new knowledge to the world. Together we can make ideas and curiosity become something real and continue to bring scientific culture to humanity, regardless of whether you are starting your studies at a high school or if you are at the peak of your career at an emblematic science institution.

Acknowledgments

We must forever thank the faculty and students of Colegio (High School) Alberto Blest Gana School for working with us (Chile), and the Center for the Promotion of Science and the Nordeus Foundation (Serbia).

Post-data I: What now? We are continuing with the research, working with an undergraduate student in biochemistry from the University of Santiago, Carla Contreras, and we are analyzing the speed of these electrical signals. Stay tuned dear readers….