Effects of Arbuscular Mycorrhizal Fungi on the Physiological Responses and Root Organic Acid Secretion of Tomato (Solanum lycopersicum) Under Cadmium Stress

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

see attached 

Comments for author File: Comments.pdf

Author Response

Dear Editor and reviewers,

Thank you very much for consideration of this manuscript (horticulturae-3894197) and peer review with helpful suggestions. We have revised the manuscript according to your suggestions with red background that you can immediately recognize where the changes have been made. Furthermore, the MS has used the English Editing by MDPI, with the aim of correcting grammatical errors and further enhancing reading fluency.

Now, I will answer reviewers’ comments one by one.

Response to the comments of Reviewer #1:

1. thank you for submitting your article to Horticulturae. Your experiment was very interesting and certainly of great interest. However I found an important flaw. The statistical analyses that you performed is not proper for the purpose of the research. You claim to evaluate the combined effect of cd stress and AMF on physiological and biochemical responses thus I was expecting at least a TWO WAY ANOVA in order to evaluate the combined effect. The effects that you observe are just in relation to the cd, the mycorrhiza or by the interaction of both factors? Also the mycorrhizal frequency and intensity cannot be analyzed with an anova, is a percentage that will not follow a normal distribution, thus you have to do another analyses that consider a binomial distribution (from 0 to 1). I also missed the arbuscules content. You use trouvelout method which retrieves this trait, thus I would add it please. You did a lot of experimental work in terms of measuring phisiologicaI and biochemical parameters, that is awesome, but the analysis of it is not appropriate, please re do it. Please correct your statistics in order to consider a new evaluation of your hard work. Do not discourage please and simply correct it.

Response: Thank you for pointing out this problem in our manuscript. We have corrected this problem according to your suggestions. Thanks very much.

 

2. Avoid abbreviations in the title.

Response: Thank you for your suggestion. We have already corrected this issue according to your suggestions.

 

3. Please add the composition of the soil used, especially its phosphorus content.

Response: Thank you for your suggestion. We have added the composition of the soil used.

 

4. the volume employed(a Cd stress).

Response: We have added it (3 times, 100 mL/time) in 2.1. Plants, growth conditions and experimental design.

 

5. Why did you choose these concentrations?

Response: From previous studies (Xingyu, Z.; Mei, Qing.; Haobo, X.; Jinbao, Tao.; Fangman, L.; Pingfei, Ge.; Yang, Y.; Wenqian, Wang.; Yongen, Lu.; Donald, Grierson.; Zhibiao, Y.; Yuyang, Zhang. Co-Selection of Low Cadmium Accumulation and High Yield During Tomato Improvement. Adv. Sci. 2025, e05138. DOI 10.1002/advs.202505138.), we know that this concentration of cadmium stress has caused significant damage to tomato. So, we chose this concentration (50 μmol/L Cd).

 

6. If possible, it would be interesting to include the lycopene content.

Response: Thank you for your suggestion. As our tomato materials have been exhausted, we are unable to conduct the test for lycopene content. Your suggestion is extremely good. We will give priority to this indicator in our future research.

 

7. Please revise this reference. You should rely on studies that developed the method rather than on those that merely cited the work.

Response: Thank you for your suggestion. We have added more detailed methods for the examination of certain indicators, such as hydrogen peroxide (H₂O₂), superoxide anion (O₂^.-), succinic acid, malic acid, etc.

 

8. Please include the dry matter of stems and roots.

Response: Thank you for your suggestion. As our tomato materials have been exhausted, we are unable to conduct the test for dry matter of stems and roots. We will avoid the occurrence of this problem in our future research. Thanks again.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Peer Review Report

Manuscript ID: horticulturae-3894197
Title: Effects of AMF on the physiological responses and root organic acid secretion of tomato (Solanum lycopersicum) under cadmium stress
Authors: Dejian Zhang et al.

Recommendation: Major Revisions Required

1. Overall Assessment

This manuscript presents a comprehensive and well-designed study investigating the role of Arbuscular Mycorrhizal Fungi (AMF, Diversispora versiformis) in alleviating cadmium (Cd) stress in tomato plants. The research is timely, addresses a significant environmental and agricultural problem, and explores a wide range of physiological and biochemical parameters. The experimental design is robust, the data is generally well-presented, and the results support the conclusion that AMF inoculation enhances Cd tolerance in tomatoes through multiple mechanisms.

The manuscript is suitable for publication in Horticulturae after major revisions to address issues primarily related to language clarity, grammatical errors, data presentation consistency, and a more critical discussion that integrates the findings with the broader literature.

2. Major Corrections Required

1. Language and Grammar:

   • The manuscript requires thorough proofreading by a native English speaker or professional editing service. There are numerous grammatical errors, awkward phrasings, and typos that hinder readability and scientific precision.

   • Examples:

     • P1, L50: "...alleviating the toxic induced by ROS" -> "...alleviating the toxicity induced by ROS"

     • P2, L47: "Study has shown" -> "Studies have shown"

     • P4, Sec 2.1: "after sterilized with" -> "after being sterilized with"; "autoclaved sands" -> "autoclaved sand"; "was included" -> "contained".

     • P5, Sec 2.2: "were chose" -> "were chosen"; "ranspiration rate" -> "transpiration rate".

     • Throughout: "observably" is used frequently; "significantly" is the standard scientific term.

   • Action: A comprehensive language edit is essential.

2. Data Presentation and Statistics:

   • Table 1 and 2 Confusion: Table 1 (on P6) contains root Cd content, but its title is about plant growth. Table 2 (on P8) contains the root colonization rate, but its title is about root architecture. The data should be reorganized logically. Root Cd content belongs in a section with the colonization data (e.g., a new Table 2). Root colonization rate should be in its own table or with the Cd content data. The current structure is confusing.

   • Figure Citations: Figures 1, 2, and 3 are cited in the results, but the manuscript text refers to "Table 1" also showing root architecture. This is incorrect and refers to Table 2. All citations of tables and figures must be double-checked for accuracy.

   • SD vs. SE: The tables state "means ± SD" (Standard Deviation). For biological replicates with n=5, Standard Error (SE) is often more appropriate to show the precision of the mean estimate. The authors should confirm which is presented and be consistent. The number of replicates (n) should also be clearly stated in the figure/table captions or methods.

   • Statistical Notation: The notation in Table 4 for chlorophyll fluorescence parameters (\(\varphi\)PSII, Fv/Fm') is inconsistent with the text description on P8, L279 (qPSII, Fo'/Fm'). All symbols must be consistent throughout the manuscript.

3. Discussion Section:

   • The discussion is largely a restatement of the results with supporting literature. It needs to be strengthened by:

     • Synthesizing Mechanisms: Better integrate the findings into a cohesive model. How do the improved hormone levels (IAA/ABA) link to the changes in organic acid secretion and antioxidant defense? How does the fungal symbiosis directly or indirectly orchestrate these changes?

     • Critical Analysis: Discuss any unexpected or conflicting results. For example, why did AMF further increase antioxidant enzyme activities under Cd stress (Table 7) compared to non-AMF Cd plants? Is this a sign of priming?

     • Specify the AMF Species: The discussion cites many studies using different AMF species (e.g., R. intraradices). It should be explicitly noted that effects can be species- and host-specific, and that this study specifically demonstrates the efficacy of D. versiformis for tomatoes.

3. Minor Corrections

1. Abstract:

   • The final sentence is cut off: "Under Cd stress, AMF could increase tomato root antioxidant capacity". This needs to be completed.

   • The abstract could be slightly condensed to focus more on the key findings and less on the list of parameters measured.

2. Introduction:

   • The introduction is comprehensive but slightly long. Consider tightening the focus on the specific knowledge gap regarding AMF, tomato, and Cd stress, which is well-stated towards the end.

3. Materials and Methods:

   • Sec 2.1: Specify the concentration (e.g., 75% ethanol) and duration of seed sterilization.

   • Sec 2.1: "the photon flux density set as 644–886 µmol/m²/s" – This is a very high light intensity for seedlings (close to full sunlight). Please verify this value.

   • Sec 2.2: Clarify the source and catalog numbers for all ELISA kits used for hormone analysis.

4. Results:

   • P7, L248: The text says "Figure 2" but the caption says "Root colonization of D. versiformis...". The figure label is (A), but the caption mentions (A) and (B). The figure and its caption must be aligned.

   • When reporting percentages (increase/decrease), it is often helpful to also state the actual p-value or a phrase like "(p < 0.05)" for each claim to reinforce statistical significance.

5. References:

   • The reference list is extensive and mostly appropriate. However, check for consistency in formatting (e.g., journal abbreviations, use of "et al.", bold vs. italic volume numbers). Some DOIs are not hyperlinked correctly in the provided text.

4. Final Decision and Summary

Decision: Major Revisions Required

This study provides valuable insights into the mechanisms of AMF-induced Cd stress tolerance in tomatoes. The scope of the work is impressive, and the findings are of significant interest to the readership of Horticulturae. However, the current manuscript requires major revisions to meet the journal's standards for publication.

The most critical issues to address are the language and grammar, the organization and accuracy of tables and data presentation, and the depth of the discussion. Once these issues are thoroughly addressed, this manuscript will be a strong candidate for acceptance.

I recommend a full re-review of the revised manuscript to ensure all points have been adequately addressed.

Author Response

Dear Editor and reviewers,

Thank you very much for consideration of this manuscript (horticulturae-3894197) and peer review with helpful suggestions. We have revised the manuscript according to your suggestions with red background that you can immediately recognize where the changes have been made. Furthermore, the MS has used the English Editing by MDPI, with the aim of correcting grammatical errors and further enhancing reading fluency.

Now, I will answer reviewers’ comments one by one.

Response to the comments of Reviewer #2:

Overall Assessment

This manuscript presents a comprehensive and well-designed study investigating the role of Arbuscular Mycorrhizal Fungi (AMF, Diversispora versiformis) in alleviating cadmium (Cd) stress in tomato plants. The research is timely, addresses a significant environmental and agricultural problem, and explores a wide range of physiological and biochemical parameters. The experimental design is robust, the data is generally well-presented, and the results support the conclusion that AMF inoculation enhances Cd tolerance in tomatoes through multiple mechanisms.

The manuscript is suitable for publication in Horticulturae after major revisions to address issues primarily related to language clarity, grammatical errors, data presentation consistency, and a more critical discussion that integrates the findings with the broader literature.

 

1. Language and Grammar:

The manuscript requires thorough proofreading by a native English speaker or professional editing service. There are numerous grammatical errors, awkward phrasings, and typos that hinder readability and scientific precision.

Examples:

P1, L50: "...alleviating the toxic induced by ROS" -> "...alleviating the toxicity induced by ROS"

P2, L47: "Study has shown" -> "Studies have shown"

P4, Sec 2.1: "after sterilized with" -> "after being sterilized with"; "autoclaved sands" -> "autoclaved sand"; "was included" -> "contained".

P5, Sec 2.2: "were chose" -> "were chosen"; "ranspiration rate" -> "transpiration rate".

Throughout: "observably" is used frequently; "significantly" is the standard scientific term.

Action: A comprehensive language edit is essential.

Response: Thanks very much. All the errors you mentioned have been corrected. Your suggestion is of great value to this MS. So, we has used the English Editing by MDPI, with the aim of correcting grammatical errors and further enhancing reading fluency.

 

2. Data Presentation and Statistics:

Table 1 and 2 Confusion: Table 1 (on P6) contains root Cd content, but its title is about plant growth. Table 2 (on P8) contains the root colonization rate, but its title is about root architecture. The data should be reorganized logically. Root Cd content belongs in a section with the colonization data (e.g., a new Table 2). Root colonization rate should be in its own table or with the Cd content data. The current structure is confusing.

Figure Citations: Figures 1, 2, and 3 are cited in the results, but the manuscript text refers to "Table 1" also showing root architecture. This is incorrect and refers to Table 2. All citations of tables and figures must be double-checked for accuracy.

SD vs. SE: The tables state "means ± SD" (Standard Deviation). For biological replicates with n=5, Standard Error (SE) is often more appropriate to show the precision of the mean estimate. The authors should confirm which is presented and be consistent. The number of replicates (n) should also be clearly stated in the figure/table captions or methods.

Statistical Notation: The notation in Table 4 for chlorophyll fluorescence parameters (\(\varphi\)PSII, Fv/Fm') is inconsistent with the text description on P8, L279 (qPSII, Fo'/Fm'). All symbols must be consistent throughout the manuscript..

Response: Thanks very much. All the errors you mentioned have been corrected. For example, we have reorganized Table 2 and placed Root colonization rate and Cd content together. All citations of tables and figures have double-checked for accuracy. The number of replicates (n=5) have be clearly stated in the all tables notes. Once again, I would like to express my gratitude for your correction.

 

3. Discussion Section:

The discussion is largely a restatement of the results with supporting literature. It needs to be strengthened by:

Synthesizing Mechanisms: Better integrate the findings into a cohesive model. How do the improved hormone levels (IAA/ABA) link to the changes in organic acid secretion and antioxidant defense? How does the fungal symbiosis directly or indirectly orchestrate these changes?

Critical Analysis: Discuss any unexpected or conflicting results. For example, why did AMF further increase antioxidant enzyme activities under Cd stress (Table 7) compared to non-AMF Cd plants? Is this a sign of priming?

Specify the AMF Species: The discussion cites many studies using different AMF species (e.g., R. intraradices). It should be explicitly noted that effects can be species- and host-specific, and that this study specifically demonstrates the efficacy of D. versiformis for tomatoes.

Response: Thank you for your comments. We made some additions and revisions to the Discussion. Furthermore, Our study showed that Cd stress triggered an outbreak of ROS in tomato roots, causing damage to the cell membranes and the outflow of cytoplasm, and subsequently leading to membrane lipid peroxidation. However AMF significantly increased antioxidant enzymes activities (POD, SOD, CAT, etc.) to reduce ROS level (such as hydrogen peroxide, superoxide anion radicals, etc.), thereby alleviating the damage caused by Cd stress to tomato seedlings. This is one of the mechanisms of AMF in alleviating cadmium stress in tomatoes as discussed in this study. And you are right that our Discussion cites many studies using different AMF species. There is no doubt that the interaction effect is definitely influenced by the species of plants and the species of AMF. Therefore, during our Discussion, we clearly stated that D. versiformis significantly enhanced the cadmium stress resistance of tomatoes.

 

4. Minor Corrections

4.1 Abstract:

The final sentence is cut off: "Under Cd stress, AMF could increase tomato root antioxidant capacity". This needs to be completed.

The abstract could be slightly condensed to focus more on the key findings and less on the list of parameters measured.

4.2 Introduction:

The introduction is comprehensive but slightly long. Consider tightening the focus on the specific knowledge gap regarding AMF, tomato, and Cd stress, which is well-stated towards the end.

4.3 Materials and Methods:

Sec 2.1: Specify the concentration (e.g., 75% ethanol) and duration of seed sterilization.

Sec 2.1: "the photon flux density set as 644–886 µmol/m²/s" – This is a very high light intensity for seedlings (close to full sunlight). Please verify this value.

Sec 2.2: Clarify the source and catalog numbers for all ELISA kits used for hormone analysis.

4.4 Results:

P7, L248: The text says "Figure 2" but the caption says "Root colonization of D. versiformis...". The figure label is (A), but the caption mentions (A) and (B). The figure and its caption must be aligned.

When reporting percentages (increase/decrease), it is often helpful to also state the actual p-value or a phrase like "(p < 0.05)" for each claim to reinforce statistical significance.

4.5 References:

The reference list is extensive and mostly appropriate. However, check for consistency in formatting (e.g., journal abbreviations, use of "et al.", bold vs. italic volume numbers). Some DOIs are not hyperlinked correctly in the provided text.

Response: Thanks very much. All the errors you mentioned have been corrected. Thanks again.

 

5. Final Decision and Summary

Decision: Major Revisions Required

This study provides valuable insights into the mechanisms of AMF-induced Cd stress tolerance in tomatoes. The scope of the work is impressive, and the findings are of significant interest to the readership of Horticulturae. However, the current manuscript requires major revisions to meet the journal's standards for publication.

The most critical issues to address are the language and grammar, the organization and accuracy of tables and data presentation, and the depth of the discussion. Once these issues are thoroughly addressed, this manuscript will be a strong candidate for acceptance.

I recommend a full re-review of the revised manuscript to ensure all points have been adequately addressed.

Response: Thank you very much for your positive feedback on this MS and for providing many excellent suggestions. We have made revisions to the MS based on your suggestions, and at the same time, we has used the English Editing by MDPI, with the aim of correcting grammatical errors and further enhancing reading fluency.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

Comments to the Author:

Title: Effects of AMF on the physiological responses and root organic acid secretion of tomato (Solanum lycopersicum) under cadmium stress

 

Overview and general recommendation:

The manuscript deals with an important topic related to the effects of AMF on the physiological responses and root organic acid secretion of tomato (Solanum lycopersicum) under cadmium stress. The manuscript technically sounds well and shows high novelty. However, it needs major linguistic adjustments; therefore, I invite the authors to pass their manuscript to a native English speaker for editing and revision. In this regard, the needed adjustments are highlighted in “Minor comments” section. Also, the list of reference shall be up-to-date; references older than 2020 (last five years of publication) shall be replaced by newer ones.

Although it outlines properly the subject under study as well as its aims and main findings, the Abstract section needs to be concise and summarized. Better keywords shall be provided. The Introduction section was clear, well formulated, and outlined the main subject under study, along with the aims of the undergone research. The Materials and Methods section is generally clear, well written, and encloses most of the information related to the adopted methodology, and statistical analysis. However, in paragraph 2.2., authors shall describe briefly the main methods adopted. Regarding the Results section, the scientific analysis of the findings was well performed. Percentages of improvements/variation were highlighted. Although authors compared their findings with previously published ones in literature, the Discussion section showed in some places a repetition of the findings declaration; this point should be seriously taken into consideration when making corrections. An appropriate Conclusions section was added in which authors summarized the findings of their study and suggested further related research being based on the raised assumptions.

My comments and queries for authors are detailed below in “Major comments” and “Minor comments” sections.

 

Major comments:

1.  The manuscript needs major linguistic adjustments; accordingly, I invite the authors to pass their manuscript to a native English speaker for editing and revision. Most needed adjustments are highlighted in “Minor comments” section.
2. The list of reference shall be up-to-date; references older than 2020 (last five years of publication) shall be replaced by newer ones.
3. Abstract: Although it outlines properly the subject under study as well as its aims and main findings, the Abstract section needs to be concise and summarized.
4. Keywords: Better keywords shall be provided.
5. Materials and Methods, 2.2. Determination and methods: Kindly describe briefly the main methods adopted in paragraph 2.2.
6. Discussion: The Discussion section showed in some places a repetition of the findings declaration; this point should be seriously taken into consideration when making corrections.

 

Minor comments:

1. Introduction: Page 2, line 78: Reference 5 is a little bit old; accordingly, kindly replace it by a more recent one (last five years of publication).
2. Introduction: Page 2, lines 80–85: “Gibberellins… [14]”: The references used for these statements are old; accordingly, kindly replace them by recent ones (last five years of publication).
3. Introduction: Page 2, lines 87–90: “Physiologically… [13]”: The sentence is a little bit cumbersome; accordingly, kindly reformulate in order to make it clearer and more aiming.
4. Introduction: Page 3, line 94: Kindly adjust as follow: “symbiotic fungi”.
5. Introduction: Page 3, line 95: Kindly adjust as follow: “are capable”.
6. Introduction: Page 3, lines 104–106: “For instance… [21]”: The reference used for this statement is old; accordingly, kindly replace it by a more recent one (last five years of publication).
7. Introduction: Page 3, lines 108–109: “It is… [22]”: Same recommendation as in the previous comment.
8. Introduction: Page 3, line 135: Kindly adjust as follow: “focused”.
9. Materials and Methods, 2.1. Plants, growth conditions and experimental design: Page 4, line 147: Kindly adjust as follow: “and then was used”.
10. Materials and Methods, 2.1. Plants, growth conditions and experimental design: Page 4, line 148: Kindly adjust as follow: “Tomato seeds”.
11. Materials and Methods, 2.1. Plants, growth conditions and experimental design: Page 4, lines 149–150: Kindly adjust as follow: “after being sterilized”.
12. Materials and Methods, 2.1. Plants, growth conditions and experimental design: Page 4, lines 151–152: Kindly adjust as follow: “3-leaf tomato seedlings”.
13. Materials and Methods, 2.1. Plants, growth conditions and experimental design: Page 4, line 153: Kindly remove “was” before “included”.
14. Materials and Methods, 2.1. Plants, growth conditions and experimental design: Page 4, line 156: Kindly adjust as follow: “non-inoculated”.
15. Materials and Methods, 2.1. Plants, growth conditions and experimental design: Page 4, line 161: Kindly replace “was began” by “was initiated”.
16. Materials and Methods, 2.1. Plants, growth conditions and experimental design: Page 4, line 163: Kindly replace “viz” by “i.e.”
17. Materials and Methods, 2.2. Determination and methods, Page 4, line 176: Kindly adjust as follow: “were chosen”.
18. Materials and Methods, 2.2. Determination and methods, Page 5, line 195: Kindly adjust as follow: “which were in accordance”.
19. Materials and Methods, 2.3. Statistical analysis, Page 5, lines 203–204: “We employed… (8.1v)”: Kindly avoid the first voice form of the sentence and adopt the impersonal form instead.
20. Materials and Methods, 2.3. Statistical analysis, Page 5, line 205: Kindly adjust as follow: “was analyzed”.
21. Results, 3.1. Effects of AMF on the agronomic traits of tomatoes under Cd stress conditions: Page 6, line 225: Kindly adjust as follow: “growth performance of Solanum lycopersicum”.
22. Results, 3.2. The development of AMF in tomato root systems and the changes in Cd content: Page 6, line 233: Kindly replace “have” by “had”.
23. Results, 3.2. The development of AMF in tomato root systems and the changes in Cd content: Page 6, line 235: Kindly write the Latin name in Italic form.
24. Results, 3.2. The development of AMF in tomato root systems and the changes in Cd content: Page 6, line 240: Kindly adjust as follow: “being significantly higher”.
25. Results, 3.3. Effect of AMF on the root architecture of tomato under Cd conditions: Page 7, line 257: Kindly adjust as follow: “no significant effect in terms of root volume”.
26. Results, 3.3. Effect of AMF on the root architecture of tomato under Cd conditions: Page 7, line 259: Kindly remove “increasing it”.
27. Results, 3.4. Effect of of AMF on chlorophyll contents and chlorophyll fluorescence parameters of tomato leaves under Cd conditions: Page 9, line 297: Kindly add “respectively” after “9.09%”.
28. Results, 3.5. Effects of AMF on photosynthesis of tomato leaves under Cd conditions: Page 9, line 315: Kindly add “respectively” after “15.59%”.
29. Results, 3.6. Effect of AMF on the ROS level in tomato roots under Cd conditions: Page 10, lines 326 and 330: Kindly add “respectively” after “32.77%” and “77.07”.
30. Results, 3.7. Effects of AMF on the antioxidant enzyme activities in tomato roots under Cd conditions: Page 10, lines 341 and 345: Kindly add “respectively” after “30.04%” and “1.32 times”.
31. Results, 3.8. Effects of AMF on the content of osmotic regulatory substances in tomato roots under Cd conditions: Page 11, lines 360 and 366: Kindly add “respectively” after “49.27%” and “1.11 times”.
32. Results, 3.10. Effects of AMF on the content of succinic acid and malic acid in tomato roots under Cd conditions: Page 12, line 399 and 403: Kindly add “respectively: after “30.59%” and “2.63%”.
33. Discussion: Page 13, line 413: Kindly adjust as follow: “refers”.
34. Discussion: Page 13, lines 424-427: “The study… [35]”: The reference used for this statement is old; accordingly, kindly replace it by a more recent one (last five years of publication).
35. Discussion: Page 13, line 434: Kindly adjust as follow: “when AMF reduced”.
36. Discussion: Page 13, line 436: Kindly adjust the sentence as follow: “… Zhang et al. [12] who found that AMF…”
37. Discussion: Page 13, line 439: Kindly adjust as follow: “indicated that”.
38. Discussion: Page 13, line 442: Kindly adjust as follow: “which weakens”.
39. Discussion: Page 13, line 454: Kindly adjust as follow: “and found that”.
40. Discussion: Page 14, lines 475-477: “In PSII… [49]”: The reference used for this statement is a little bit old; accordingly, kindly replace it by a more recent one (last five years of publication).
41. Discussion: Page 14, line 478: Kindly write the Latin name in Italic form.
42. Discussion: Page 14, line 480: Kindly adjust as follow: “indicating that”.
43. Discussion: Page 14, line 491: Kindly adjust as follow: “increased”.
44. Discussion: Page 14, line 492: Kindly adjust as follow: “indicated that”.
45. Discussion: Page 14, line 504: Kindly write the Latin name in Italic form.
46. Discussion: Page 14, line 505: Kindly adjust the sentence as follow: “… agreement with this study where Cd acts…”.
47. Discussion: Page 15, line 513: Kindly adjust as follow: “those of this study.”
48. Discussion: Page 15, line 532: Kindly adjust as follow: “triggered”.
49. Discussion: Page 15, line 533: KIndly adjust as follow: “indicated”.
50. Discussion: Page 15, lines 535-539: “This study… seedlings”: The sentence is badly written in standard English; accordingly, kindly reformulate it.
51. Discussion: Page 15, line 541: Kindly adjust as follow: “by enhancing”.
52. Discussion: Page 15, line 544: Kindly adjust as follow: “induces”.
53. Discussion: Page 15, lines 548-551: “AMF… [58]”: The reference used for this statement is old; accordingly, kindly replace it by a more recent one (last five years of publication).
54. Discussion: Page 15, line 552: Kindly adjust as follow: “holds”.
55. Discussion: Page 15, lines 551-553: “At early… [39,59]”: Reference 59 is very old; accordingly, kindly replace it by a more recent one (last five years of publication).
56. Discussion: Page 15, line 556: Kindly adjust as follow: “this study”.
57. Discussion: Page 15, lines 556-559: “Regarding… [59]”: The reference used for this statement is very old; accordingly, kindly replace it by a more recent one (last five years of publication).
58. Discussion: Page 15, line 561: Kindly adjust the sentence as follow: “The latter not only maintain…”
59. Discussion: Page 16, line 568: Kindly adjust the sentence as follow: “…of this study where Cd stress…”
60. Discussion: Page 16, line 568: Reference 61 is lacking in text !! Also. The one provided in the list of references is old; accordingly, kindly replace it by a more recent one (last five years of publication).
61. Discussion: Page 16, lines 570-571: Kindly adjust the sentence as follow: “Such an increase is an adaptive response…”
62. Discussion: Page 16, lines 573-576: “This study… seedlings”: The sentence is a little bit cumbersome; accordingly, kindly reformulate in order to make it clearer and more aiming.
63. Discussion: Page 16, lines 585-586: “Strigolactone… [62]”: The reference used for this statement is old; accordingly, kindly replace it by a more recent one (last five years of publication).
64. Discussion: Page 16, line 588: Kindly adjust as follow: “treated”.
65. Discussion: Page 16, line 591: Kindly adjust as follow: “The increased effect”.
66. Discussion: Page 16, line 592: Kindly adjust as follow: “has strengthened”.
67. Discussion: Page 16, lines 598-600: “When plants… [64]”: The reference used for this statement is very old; accordingly, kindly replace it by the following recent and reliable one: “doi:10.1016/j.heliyon.2024.e27138”.
68. Discussion: Page 16, line 607: Kindly adjust as follow: “may have inhibited”.
69. Discussion: Pages 16-17, lines 612-615: “Low… [66]”: The reference used for this statement is very old; accordingly, kindly replace it by a more recent one (last five years of publication).
70. Discussion: Page 17, lines 622-624: “This might… [68]”: The reference used for this statement is relatively old; accordingly, kindly replace it by the following recent and reliable one: “doi:10.1007/s42729-025-02265-7”.
71. Discussion: Page 17, lines 624-628: “The low-molecular-weight… value”: These statements lack reliable sources (references); accordingly, kindly provide them.
72. Discussion: Page 17, line 637: Kindly adjust as follow: “indicated”.
73. Discussion: Page 17, line 643: Kindly adjust as follow: “are as follows.”
74. Discussion: Page 17, lines 643-646: “First… host plants”: The sentence is a little bit cumbersome; accordingly, kindly reformulate in order to make it clearer and more aiming.
75. Discussion: Page 17, line 648: Kindly adjust as follow: “indicated”.
76. Conclusions: Page 17, line 656: Kindly adjust as follow: “could disrupt”.
77. Conclusions: Page 17, lines  658-661: “Furthermore… etc”: The sentence is badly written in standard English; accordingly, kindly reformulate it.
78. Conclusions: Page 17, line 661: Kindly adjust as follow: “demonstrated”.

Comments on the Quality of English Language

The manuscript needs major linguistic adjustments; therefore, I invite the authors to pass their manuscript to a native English speaker for editing and revision. In this regard, the needed adjustments are highlighted in “Minor comments” section.

Author Response

Dear Editor and reviewers,

Thank you very much for consideration of this manuscript (horticulturae-3894197) and peer review with helpful suggestions. We have revised the manuscript according to your suggestions with red background that you can immediately recognize where the changes have been made. Furthermore, the MS has used the English Editing by MDPI, with the aim of correcting grammatical errors and further enhancing reading fluency.

Now, I will answer reviewers’ comments one by one.

Response to the comments of Reviewer #3:

 

The manuscript deals with an important topic related to the effects of AMF on the physiological responses and root organic acid secretion of tomato (Solanum lycopersicum) under cadmium stress. The manuscript technically sounds well and shows high novelty. However, it needs major linguistic adjustments; therefore, I invite the authors to pass their manuscript to a native English speaker for editing and revision. In this regard, the needed adjustments are highlighted in “Minor comments” section. Also, the list of reference shall be up-to-date; references older than 2020 (last five years of publication) shall be replaced by newer ones.

Although it outlines properly the subject under study as well as its aims and main findings, the Abstract section needs to be concise and summarized. Better keywords shall be provided. The Introduction section was clear, well formulated, and outlined the main subject under study, along with the aims of the undergone research. The Materials and Methods section is generally clear, well written, and encloses most of the information related to the adopted methodology, and statistical analysis. However, in paragraph 2.2., authors shall describe briefly the main methods adopted. Regarding the Results section, the scientific analysis of the findings was well performed. Percentages of improvements/variation were highlighted. Although authors compared their findings with previously published ones in literature, the Discussion section showed in some places a repetition of the findings declaration; this point should be seriously taken into consideration when making corrections. An appropriate Conclusions section was added in which authors summarized the findings of their study and suggested further related research being based on the raised assumptions.

My comments and queries for authors are detailed below in “Major comments” and “Minor comments” sections.

 

1. Major comments:

1.1 The manuscript needs major linguistic adjustments; accordingly, I invite the authors to pass their manuscript to a native English speaker for editing and revision. Most needed adjustments are highlighted in “Minor comments” section.

1.2 The list of reference shall be up-to-date; references older than 2020 (last five years of publication) shall be replaced by newer ones.

1.3 Abstract: Although it outlines properly the subject under study as well as its aims and main findings, the Abstract section needs to be concise and summarized.

1.4 Keywords: Better keywords shall be provided.

1.5 Materials and Methods, 2.2. Determination and methods: Kindly describe briefly the main methods adopted in paragraph 2.2.

1.6 Discussion: The Discussion section showed in some places a repetition of the findings declaration; this point should be seriously taken into consideration when making corrections.

Response: Based on your suggestions, we has used the English Editing by MDPI, with the aim of correcting grammatical errors and further enhancing reading fluency. The Abstract section have been concise and summarized. Better keywords have been provided. We provided detailed descriptions of some experimental methods. Also, we made appropriate modifications to the discussed sections. Thank you very much for your suggestion.

 

2. Minor comments.

Response: Thank you very much for offering so many minor suggestions. We are deeply impressed by your meticulous work. We have already revised all the issues you raised. We would like to express our deepest gratitude for the help you have provided.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

 Accept in present form

Author Response

Dear reviewer,

   Thank you very much for your assistance with this MS.

    Wish you all the best in your work and good health.

Reviewer 2 Report

Comments and Suggestions for Authors

The modifications in the paper appear to be well done and appropriate for publication. The authors have addressed comments and reviewer suggestions by confirming the intended meanings were retained and improving clarity throughout the manuscript. The manuscript includes detailed experimental design, comprehensive data analysis, and thorough discussion supported by relevant citations. The presentation of figures, tables, and results on the effects of arbuscular mycorrhizal fungi (AMF) on tomato plants under cadmium stress is clear and scientifically sound.

Specifically, the authors provided clear evidence that AMF inoculation:

• Promotes growth and physiological performance of tomato seedlings exposed to cadmium stress.

• Reduces cadmium uptake in roots.

• Enhances photosynthesis, chlorophyll content, and antioxidant enzyme activity.

• Modulates reactive oxygen species and osmotic regulatory substances to alleviate cadmium toxicity.

The conclusions align well with the data presented and the manuscript is formatted professionally for the journal. Based on the detailed content and revisions shown in the file, the paper is suitable for publication pending the editor's final review.

Author Response

Dear reviewer,

   Thank you very much for your assistance with this MS.

    Wish you all the best in your work and good health.

Reviewer 3 Report

Comments and Suggestions for Authors

Comments to the Author:

Title: Effects of AMF on the physiological responses and root organic acid secretion of tomato (Solanum lycopersicum) under cadmium stress

 

Overview and general recommendation:

Although authors made significant improvements to their manuscript, still lots of work is needed at this stage.

My comments and queries for authors are detailed below in “Major comments” and “Minor comments” sections.

 

Major comments:

1. The manuscript needs some linguistic adjustments. Most needed adjustments are highlighted in “Minor comments” section.
2. The list of reference shall be up-to-date; references older than 2020 (last five years of publication) shall be replaced by newer ones.
3. Abstract: Although it outlines properly the subject under study as well as its aims and main findings, the Abstract section needs to be concise and summarized.
4. Keywords: Better keywords shall be provided.
5. Discussion: The Discussion section showed in some places a repetition of the findings declaration; this point should be seriously taken into consideration when making corrections.

 

Minor comments:

1. Introduction: Page 2, line 79: Reference 5 is a little bit old; accordingly, kindly replace it by a more recent one (last five years of publication).
2. Introduction: Page 2, lines 81–86: “Gibberellins… [14]”: The references used for these statements are old; accordingly, kindly replace them by recent ones (last five years of publication).Moreover, kindly write properly all references in the list of references following the journal’s guidelines.
3. Introduction: Page 3, lines 103–106: “For instance… [21]”: The reference used for this statement is old; accordingly, kindly replace it by a more recent one (last five years of publication).
4. Introduction: Page 3, lines 108–109: “It is… [22]”: Same recommendation as in the previous comment.
5. Discussion: Page 13, lines 453-456: “These findings… [35]”: The reference used for this statement is old; accordingly, kindly replace it by a more recent one (last five years of publication).
6. Discussion: Page 13, line 470: Kindly adjust as follow: “indicated that”.
7. Discussion: Page 14, lines 506-508: “In PSII… [49]”: The reference used for this statement is a little bit old; accordingly, kindly replace it by a more recent one (last five years of publication).
8. Discussion: Page 15, lines 580-582: “AMF… [58]”: The reference used for this statement is old; accordingly, kindly replace it by a more recent one (last five years of publication).
9. Discussion: Page 15, lines 582-585: “In the early… [39,59]”: Reference 59 is very old; accordingly, kindly replace it by a more recent one (last five years of publication).
10. Discussion: Page 15, lines 587-590: “Regarding… [59]”: The reference used for this statement is very old; accordingly, kindly replace it by a more recent one (last five years of publication).
11. Discussion: Page 16, line 599: Reference 61 is lacking in text !! Also. The one provided in the list of references is old; accordingly, kindly replace it by a more recent one (last five years of publication).
12. Discussion: Page 16, lines 614-615: “Strigolactone… [62]”: The reference used for this statement is old; accordingly, kindly replace it by a more recent one (last five years of publication).
13. Discussion: Page 16, lines 627-629: “When plants… [64]”: The reference used for this statement is very old; accordingly, kindly replace it by the following recent and reliable one: “doi:10.1016/j.heliyon.2024.e27138”.
14. Discussion: Page 17, lines 640-644: “Low… [66]”: The reference used for this statement is very old; accordingly, kindly replace it by a more recent one (last five years of publication).
15. Discussion: Page 17, lines 655-656: “Among them… [68]”: The reference used for this statement is relatively old (and was removed from the reference list); accordingly, kindly replace it by the following recent and reliable one: “doi:10.1007/s42729-025-02265-7”.

Comments on the Quality of English Language

The manuscript needs some linguistic adjustments. Most needed adjustments are highlighted in “Minor comments” section in the attached report.

Author Response

Dear Editor and reviewer,

Thank you very much for consideration of this manuscript (horticulturae-3894197) and peer review with helpful suggestions again. We have revised the manuscript according to your suggestions with red background that you can immediately recognize where the changes have been made.

Now, I will answer reviewer’ comments one by one.

Response to the comments of Reviewer #3:

1. The manuscript needs some linguistic adjustments. Most needed adjustments are highlighted in “Minor comments” section.

Response: Thank you very much for offering so many minor suggestions. We have carefully revised the entire text according to your suggestions.

 

2. The list of reference shall be up-to-date; references older than 2020 (last five years of publication) shall be replaced by newer ones.

Response: Thank you. All references older than 2020 (last five years of publication) have been replaced by newer ones. However, the fifth (2018) and thirtieth (2019) references must be cited in this MS. I hope you can understand.

 

3. Abstract: Although it outlines properly the subject under study as well as its aims and main findings, the Abstract section needs to be concise and summarized.

Response: The Abstract section have been concise and summarized again.

 

4. Keywords: Better keywords shall be provided.

Response: Better keywords have been provided again.

 

5. Discussion: The Discussion section showed in some places a repetition of the findings declaration; this point should be seriously taken into consideration when making corrections.

Response: We made appropriate modifications to the Discussed sections.

 

6. Minor comments.

Response: Thank you very much for offering so many minor suggestions again. We have already revised all the issues you raised. And, you mentioned that “Discussion: Page 16, lines 627-629: “When plants… [64]”: The reference used for this statement is very old; accordingly, kindly replace it by the following recent and reliable one: “doi:10.1016/j.heliyon.2024.e27138”.”. However, I cannot found this reference (doi:10.1016/j.heliyon.2024.e27138). So, I replace it by another reference, which published in 2024 (Ahmad Ansari, M. K.; Iqbal, M.; Ahmad, M.; Munir, M.; Gaffar, S.A.; Chaachouay, N. Heavy metal stress and cellular antioxidant systems of plants: a review. Agric. Rev. 2024, 45, 400-409. DOI 10.18805/ag.RF-321.). 

Thank you very much for your assistance with this MS.

Wish you all the best in your work and good health.

 

Author Response File: Author Response.pdf