Effect of hydrogen peroxide concentration on lettuce seed germination

1. What do you like most about this manuscript? 

I really enjoyed reading about your study and learned new information. I didn’t know that hydrogen peroxide (at low concentrations) could help seed germination. This was interesting to learn, and it has some real-world applications I wasn’t aware of!

I applaud your efforts on this project. You clearly put thought and time into your experimental design and this preprint. What I liked most about the manuscript was the experimental design. I believe the biggest strengths of this study are:

1. A control group: distilled water with no hydrogen peroxide

2. Replicates: each treatment had 20 seeds 

3. Controlled variables: temperature, light, and water 

4. Descriptions of calculations: average radicle length, uncertainty, germination percentage, and seed vigor index

I also liked how you collected and analyzed the data. The measurements used were simple yet effective because they helped answer the research question. Including photos in Figures 2 and 3 also offered strong visual evidence of how varying hydrogen peroxide concentration affected seed germination.

2. How effectively did the background information support the research question? What additional information would be helpful for the reader? 

Based on the length of this manuscript, the number of references included was enough to demonstrate that you explored this topic. Further, most - if not all - of these sources seem to relate to the main topic of the study. In the Introduction, I thought your explanations about seed germination and priming with hydrogen peroxide were helpful to understand your research question. The flow from a process (germination) to factors that influence it (varying hydrogen peroxide concentrations) made a strong case for your experimental design. However, I wanted to know more about why this study was needed - that is, what is the big picture of this research topic? For example, is it important to study the effects of varying hydrogen peroxide concentrations on lettuce seed germination to improve or support healthy agricultural crops? Or, why did you choose lettuce instead of a different plant? Explaining your motivation for this study (why it matters) would strengthen the connection between the background information and research question.

3. What are the strengths of the hypothesis? How could it be improved?

The hypothesis was easy to find in the Introduction. It was also appropriately written in that it is both testable and falsifiable, two qualities of good scientific hypotheses. However, I was confused on whether germination percentage, seed vigor index, and radicle length are part of or separate from “germination efficiency.” You could address this comment by either defining germination efficiency or listing each variable (and its measurement) separately. 

4. What are the strengths of the experimental design? Can you think of accessible experiments or analysis that would help the authors strengthen their study? 

As mentioned earlier, I believe a strength of this manuscript is the experimental design. Although I was curious why, after soaking the seeds in distilled water or hydrogen peroxide solutions, the seeds were kept in a dark cupboard rather than growing them under light. Also, I wondered why the lettuce seeds were sourced from different vendors rather than using the same type of seed throughout all treatments and the control. Could different seed varieties have influenced the results?

While not necessary, you could use some statistical analyses to support your claims. I encourage you to visit our website for student-friendly resources on statistics.

5. What are the successes and the areas for improvement in the Discussion? Consider factors such as how clearly results are summarized, how well the results support the conclusions, how key takeaways are communicated, how limitations or next steps are discussed, etc.

While I liked the information presented in the Introduction, I found some of the content in the Discussion to be less helpful for learning more about this particular study and its results, and some paragraphs were difficult to follow along. For example, although the paragraph about genes and gene expression was interesting, it didn’t add much to my understanding about this specific study. To improve clarity, I suggest you reduce how much new information is presented in the Discussion because sometimes too much information distracts from the main ideas. So, if you revise your manuscript, I suggest you consider what ideas are essential to understand the study and its results versus what ideas are related but not necessarily relevant. Nonetheless, I appreciated your efforts to explain your results relative to other studies and liked your inferences about why you may have obtained these results. For example, I liked the explanation you offered for “why seeds primed with 0.75% and 1% H₂O₂ performed worse than the control in this study.” 

6. What changes can the authors make to improve how they are verbally or visually communicating their scientific knowledge, results, and reasoning? Could someone who has never heard of this topic understand the main message of the paper? 

Based on eiRxiv’s guidelines, you included the necessary information in each section, and the manuscript mostly flows together. However, there are two ways your scientific communication could be improved. First, while the terms used are relevant to the topic, some paragraphs felt jargony or too technical. Defining terms or simplifying language would make this preprint more accessible while still demonstrating rigor and quality. Second, in the Results section, I recommend you include actual data instead of just mentioning the figures and tables. This will help support your claims by “showing” rather than telling. For example, you could list some of the average radicle lengths from Table 1. Aside from this and other suggestions addressed above, I thought the writing was good! I also liked the figures and tables and found them to be informative - they helped highlight patterns or trends in the data.