Germination and the Biochemical Response of Pumpkin Seeds to Different Concentrations of Humic Acid under Cadmium Stress
Round 1
Reviewer 1 Report
Series of concentrations of humic acid are tested to alleviate cadmium stress on germination of pumpkin seeds, and analyzed reason why it was buffered through biochemical responses. Enzymes's activity are good indicator to evaluate cadmium biological toxicity on germination, tolerable mechanism of pumpkin seeds to different cadmium stress are explained well in the manuscript. In addition, lipid and protein stimulation are powerful to substantiate the mechanism.
Something was forgotten before ‘acid’ in the last sentence at Abstract.
Author Response
Hallo,
Thank you both reviewers for their valuable comments. We did incorporate all suggested changes in the new version of the manuscript.
We rewrite the Abstract.
We add chemicals specification in the section M&M.
And also we did change the graphs and add missing into both Tables.
Also we would like to answer reviewer comment on bio stimulation property of humic acid.
We know that humic acid has got this property but we did not perform any greenhouse or field experiments with pumpkin so that why we are not discussing this aspect here.
With greetings,
Beata Dedicova
Author Response File: 
Author Response.docx
Reviewer 2 Report
The MS by Aghbolaghi et al. reports the effect of humic acid on germination of pumpkin seeds under Cd stress. Overall, the description is not clearly stated, and the objectives, and discussion are not well rationalized.
The authors cite as their rationale for their study the previous studies in which pumpkins absorb a small amount of nitrate or heavy metals. However, they do not explain the botanical and agronomical significance of the absorption of small amounts of nitrate acid and heavy metals. It is also not clear why they chose to focus on cadmium among heavy metals, and what significance such high concentrations of 100-200 mg/L have. No description what type of Cd salt was used. It stated that humic acid is effective in alleviating heavy metal stress, but there is no description of the type of humic acid used, nor is there any explanation of the significance of using that concentration.
The analyses and presentation of the data is arbitrary. Since the present experiment was a factorial experiment combining three levels of Cd concentration and five levels of humic acid concentration, ANOVA for these factors should be conducted first. There is no scientific significance for the curve fittings in Figure 1-3. In the methods section, it is stated that seeds were sampled daily for biochemical analysis, but Table 1 does not show the daily measurements. There are numerous other inadequacies.
Author Response
Hallo,
Thank you both reviewers for their valuable comments. We did incorporate all suggested changes in the new version of the manuscript.
We rewrite the Abstract.
We add chemicals specification in the section M&M.
And also we did change the graphs and add missing into both Tables.
Also we would like to answer reviewer comment on bio stimulation property of humic acid.
We know that humic acid has got this property but we did not perform any greenhouse or field experiments with pumpkin so that why we are not discussing this aspect here.
With greetings,
Beata Dedicova
Author Response File: Author Response.docx
Reviewer 3 Report
In this study, authors investigated the effects of the application of humic acid on the germination and physiological response in pumpkin seeds under cadmium stress. The paper was properly drafted, however, the research is lack of innovation. The authors should clarify why this research is important, why the antioxidative enzymes was determined in this study. Especially in the background part, the relationship between the all the physiological parameters and cadmium stress should be elucidated. Besides, the abstract was written poorly. Abstract is not an mini-review, it should include the main findings and conclusions.
Author Response
Hallo,
Thank you both reviewers for their valuable comments. We did incorporate all suggested changes in the new version of the manuscript.
We rewrite the Abstract.
We add chemicals specification in the section M&M.
And also we did change the graphs and add missing into both Tables.
Also we would like to answer reviewer comment on bio stimulation property of humic acid.
We know that humic acid has got this property but we did not perform any greenhouse or field experiments with pumpkin so that why we are not discussing this aspect here.
With greetings,
Beata Dedicova
Author Response File: Author Response.docx
Round 2
Reviewer 2 Report
Although the abstract of the MS was revised, the authors did not response to my comments at all. The main text, figures, and tables are almost the same as before. So the MS is not acceptable for publication.
Author Response
Hallo,
Thank you both reviewers for their valuable comments. We did incorporate all suggested changes in the new version of the manuscript.
- Abstract has been changed.
- Just to clarified why we used Fisher's LSD method. This method is used in ANOVA to create confidence intervals for all pairwise differences between factor level means while controlling the individual error rate to a significance level you specify. Fisher's LSD method then uses the individual error rate and number of comparisons to calculate the simultaneous confidence level for all confidence intervals. This simultaneous confidence level is the probability that all confidence intervals contain the true difference. It is important to consider the family error rate when making multiple comparisons because your chances of committing a type I error for a series of comparisons is greater than the error rate for any one comparison alone.
Reviewer 3 Report
The abstract is too long, please make it brief and concise.
Author Response
Hallo,
Thank you both reviewers for their valuable comments. We did incorporate all suggested changes in the new version of the manuscript.
Abstract has been changed.
Author Response File: Author Response.docx
