Leaf–Air Temperature Difference as a Nondestructive Indicator of Waterlogging Tolerance in Cassava Genotypes

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Dear authors,

I congratulate you on the article "Leaf-Air Temperature Difference as a Non-Destructive Indicator of Waterlogging Tolerance in Cassava Genotypes", which addresses a relevant and innovative topic for the field of plant physiology and  sustainability. Following the review , the article has significant potential, but a number of improvements are needed to achieve the journal's standards.

These adjustments will increase the scientific impact of the paper and ensure compliance with editorial requirements. I encourage you to integrate these recommendations and I am convinced that the article will make a valuable contribution to the field.

Good thoughts,

 

Abstract

Clarification of the hypothesis in line 21 is necessary in order to explicitly mention  ΔT and the link with physiological performance, avoiding the ambiguity of the term 'cooler canopy'. Add key data to lines 24–27 is important to include numerical values (ΔT, correlation coefficients) and statistical significance, which will support the claims and increase the scientific impact

1.Introduction

Line 38-42: The global introduction is relevant, but too general and long; I recommend condensing and connecting more clearly with the specific problem of cassava under embalming, in order to avoid logical fragmentation between the global food context and the physiological study.

Lines 74-85 and 105-112: Text contains long sentences and repetitions ("temperature raises may be due to stomatal closure" occurs twice); I recommend reformulating it into shorter sentences, standardizing the technical terms (SPAD, Fv/Fm, ΔT ) and correcting syntax errors for readability.

Line 123-125: Wording "In over-research, major crops and cereals..." is unclear and unidiomatic; I recommend replacing it with "In extensively studied major crops and cereals..." and clarifying the sentence to highlight the contrast with cassava, avoiding the phrase "poorly unknown" which is redundant and incorrect - suggest "remain poorly understood in cassava, particularly regarding anaerobic responses under waterlogging conditions ."

2.Materials and Methods

Line 145-163: The description of the origin of the cultivars is very long and contains speculative information ("the exact circumstances... are unknown")  which reduces the clarity of the section. I recommend condensing the essential information (name, source, relevant characteristics) and moving the historical details to a note or appendix to keep the focus on the materials used.

Line 174-179: The experimental design is presented correctly, but critical details for reproducibility are missing, such as the total number of plants per treatment, the mode of randomization,  and the light conditions in the greenhouse. I recommend that you fill in this information and clarify if "three replicas" refer to the level of the pot or individual plant.

Line 186-187: The VPD calculation is mentioned, but the method or formula used is not explained; I recommend including the formula for VPD and specifying whether the values were daily or instantaneous averages, for transparency and reproducibility.

Line 192-193: It is mentioned that the measurements were made on "one plant/replica (n = 1)", which raises problems of representativeness. Clarify whether this choice was statistically justified and suggest including more leaves per replica for robustness.

Line 195-196: The mention 'block temperature 28°C' can introduce artifacts into the ΔT. Explain how this setting affects the leaf-to-air difference and whether corrections have been made to the actual ambient temperature.

Line 228-232: Statistical analysis is briefly described; I recommend specifying the presupposition tests ( normality, homoscedasticity) and justifying the use of unifactorial ANOVA in the context of repeated data, possibly suggesting mixed models for accuracy.

3.Results

Line 239-241: The presentation of temperature and humidity is correct, but the daily variation ranges and standard deviations are missing in the text (not just in the figure).

I recommend including these details for transparency and avoiding reliance solely on charts.

Line 262-264: The statistical significance is mentioned (p < 0.001 and p < 0.01), but the exact values of the F-statistic or the degree of freedom is not specified.

I recommend including these details in the text or table for transparency and statistical validity.

Line 268-272: Temperature increase percentages and ΔT are shown, however it is unclear whether these are mean ± SD or crude values; I recommend adding confidence intervals and indicating whether the differences are significant between cultivation,  not just between treatments.

Line 336-343: The percentage reduction values for Fv/Fm are very small (0%–7.6%), but it is not clear whether these differences are biologically relevant or only statistically significant. Introduce a paragraph on discussing biological relevance and including confidence intervals to support interpretation.

Line 404-411: The description of the correlations is general and does not include the exact values of the r-coefficients or the significance levels for each relationship; I recommend the numerical presentation of the most important correlations (e.g. r = −0.85 between ΔT and Pn) and the clear indication of the significance  thresholds for transparency.

4.Discussion:

Line 418-424: The discussion about morphological effects is clear, but the integration of statistical values (p, confidence intervals) and comparison with other studies on cassava are missing.

I recommend that you add these details and avoid excessive repetition of the findings already presented in the Results section.

Line 471-485: Discussion of leaf temperature increase and ΔT is relevant, but should include numerical correlations (r, p) to support claims and avoid generalizations ("more evident at Tokunoshima-white") without statistical test; I recommend integrating the data from Figure 6 and clarifying the physiological mechanism with updated references.

Line 493-499: The explanation for the decrease in Pn, E and gs is correct, but the integration of physiological mechanisms (e.g. effects of hypoxia on water and CO₂ transport) is missing and it is not mentioned whether these decreases are directly correlated with ΔT; I recommend explicitly linking with the data in Figure 6 and including correlation coefficients to support the claims.

Line 518-523: Numerical correlations are presented, which is a strong point, but the text does not explain the biological difference between correlations under WW and WL. Interpret these differences (why relationships are stronger under WL) and discuss the implications for the phenotyping of embalming tolerance.

Line 533-545: The conclusion on ΔT as an indicator is formulated too general and repetitive, with many external references; I recommend summarizing it in a clear sentence, avoiding redundancy and emphasizing limitations  (low number of genotypes, controlled conditions), in order to keep the balance between enthusiasm and scientific rigor.

5. Conclusions

Line 576-579: The statement "ΔT offers strong potential as a practical parameter for rapid screening" is too categorical considering that the study included only two cultivations and controlled conditions; I recommend rephrasing in a more cautious tone, such as "ΔT shows promise as an indicator, but requires validation across diverse genotypes and environments".

Lines 582-583: Phrase "This is so unique in the sense that it handles a special under researched root crop..." it is non-academic and redundant; I recommend eliminating or rephrasing it in a concise scientific style, avoiding subjective and unjustified expressions.

References

1. Journal of Crop Sciences 669–670 (Ref. 31): Japan Journal of Crop Sciences 2020. 89: p. 277-287." The journal is not italicized, and pages are written with "-" instead of "–". Correct with: Japan J. Crop Sci. 2020, 89, 277–287.
2. There are errors in the name of the authors and their order. Authors are listed incompletely or with punctuation errors. Example: Line 607–609 (Ref. 1): "More, S., J, , et al." Here we have double commas and missing initials. Correct with: More, S.; J.; et al.
3. Some titles contain errors or unnecessary repetition. Line 669–670 (Ref. 31): "prtevailing a temperate zone..." Problem: "prtevailing" → "prevailing".
4. Presence of unsupported platforms and lack of mandatory data (publisher, ISBN, DOI). Example is: Line 619–620 (Ref. 6): "ReseaerchGate". This is NOT an official MDPI source. I recommend removing "ResearchGate" and keeping only the conference title. Line 680–681 (Ref. 36): Manual without publisher and ISBN.
5. Mixture of languages. Line 617–618 (Ref. 5): "multiscale characterization..." Problem: Missing capital letters at the beginning of the title. Correct: "Multiscale Characterization of Inland Valley Agro-Ecosystems in West Africa."
6. I recommend that you check each bibliographic resource separately, so that in the end it is up to the standards of the journal.

Author Response

I have uploaded Cover letter to the responses

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors!

The manuscript is interesting, has potential for application in agriculture. The experimental design is appropriate to test the hypothesis. The manuscript’s results are reproducible based on the details given in the methods section. The article`s references are mostly old publications (out of the last 5 years). I think they are relevant. The topic is specific, but  could it be possible to change the old references to the newer ones? The reference list does not include an excessive number of self-citations. The conclusions are consistent with the evidence and arguments presented.

The text of the manuscript is not clear, you should revise it.

My comments:

L 58, 64, 70, … You should change the references in the text according to the instruction for authors.

L 145. Add Japan after Faculty of Agriculture.

L 167. You should write Hyponica nutrient solution composition or the manufacturer.

L 193. Why did you use the  youngest leaf for measurement? Add your reasons.

Methods. How did you measure the Leaf Temperature? Add information about the appliance. 

Figures. The column shading and legend are poorly distinguishable from each other. Perhaps it's worth increasing the font size of the legend and change the hatching of the columns. The data are interpreted appropriately and consistently throughout the manuscript. The statistical analysis is sufficient.

You should decipher abbreviations DAT (L 221), TrtD (L 286).

L 281. In this section there was an idea that the cultivars did not differ among treatments. However the sentence :”These results revealed that the effects of WL treatment on the Pn, E, gs, Ci, and PWUE  values of two cassava cultivars at the early growth stage were cultivar-specific…” (L 323) has the different meaning. You should check and correct the meaning of the sentences in this section.

L 478-490 and 514-516. The sentence “This result was consistent with Pan, Jiang [47] who reported that, leaf temperature was higher in the sensitive genotypes than that in the tolerant genotypes after waterlogging.” was repeated twice.

Comments on the Quality of English Language

Dear Authors!

You should check the text of the manuscript for typos and small mistakes. For example L 194 - the word dat.

L 60. You should change “…become saturated,….” to “…become saturated with water, ….”.

L 105. Rephrase the sentence “…Barón [24] increases in leaf temperatures… ” to “…Barón [24], the increase in leaf temperatures…”.

L 123. You should change “In over-research, major crops and cereals…” to “In over-research of major crops and cereals…”

Author Response

I have Cover letter to the responses

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The submitted manuscript addresses the current and important issue of assessing flooding tolerance in cassava using the leaf-to-air temperature difference (ΔT) as a non-destructive physiological indicator. The work's substantive assumptions are sound and well-grounded in the literature, and the interpretation of the relationships between reduced transpiration, stomatal closure, and increased leaf temperature is consistent with current knowledge. The conclusions regarding the usefulness of ΔT as a screening parameter for flooding tolerance are logical and supported by correlational analyses and physiological data, although it should be emphasized that the genotype range is very narrow and does not fully justify far-reaching generalizations.
However, the work suffers from numerous editorial and linguistic shortcomings. There are grammatical errors, typos, stylistic inconsistencies, and repetitive wording, particularly in the introductory and discussion sections. Some sentences are imprecise or overly worded, making the text difficult to understand. There are also minor terminological inconsistencies and inconsistencies in the notation of units and symbols (e.g., CO₂, ΔT, VPD). The statistical methods are generally described correctly, but the number of biological replicates in the context of correlational analyses requires further clarification. In summary, the work has a solid scientific foundation but requires careful linguistic and editorial revisions, as well as moderation in formulating any conclusions of a practical nature.

1. Lines 57–59 – the sentence is broken and incoherent: “hailstorms, salinity, and heat stress (More, V Ravi [1].” – missing parentheses and logical closure.
2. Lines 92–109 – the fragment repeatedly repeats the same idea about an increase in leaf temperature due to stomatal closure; there are syntactic errors and semantic chaos (e.g., lines 105–109 contain a practically illogical sentence).
3. Lines 123–125 – the phrase “these relationship changes remain poorly unknown” is stylistically and logically incorrect; it should be “poorly understood.”
4. Lines 149–160 – the description of the origin of the varieties is inconsistent, linguistically sloppy, and based on assumptions (“were assumed to be”), which requires simplification and a clear separation of facts from hypotheses.
5. Lines 193–199 – numerous errors in the notation of units and symbols (e.g., "LI- 6,400XT," "µ mol−1," inconsistent spacing and superscripts/subscripts).
6. Lines 231–232 – editorial error: “Turkey’s HSD test” instead of Tukey’s HSD test.
7. Lines 581–583 – colloquial, non-academic style in the conclusion (“This is so unique in the sense…”), unacceptable in a scientific journal.
8. Lines 21–23 and 29–31 – overly far-reaching conclusions about “better tolerance” based on examining only two local, closely related genotypes.
9. Lines 98–99 – logical inconsistency: the statement that “reduced stomatal conductance contributes to lower leaf temperatures” is physiologically incorrect (it should be the other way around).
10. Lines 120–122 – the suggestion of a positive relationship between higher ΔT and yield is inappropriate for flood-stress conditions and requires stronger context or a more restrictive interpretation.
11. Lines 323–325 – the authors write about "cultivar-specific effects," despite the lack of significant differences between cultivars in many analyses (contradictory to the results).
12. Lines 571–576 – the application conclusions (breeding screening) are disproportionate to the scale of the experiment and should be clearly marked as preliminary.

Comments on the Quality of English Language

the main review included this information

Author Response

i have uploaded Cover letter to the responses

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Dear Authors,

I would like to congratulate you on the completion of the article “Leaf-Air Temperature Difference as a Non-Destructive Indicator of Waterlogging Tolerance in Cassava Genotypes”, which addresses a relevant and innovative topic in the field of plant physiology and sustainability.

I would also like to commend you for the professional and open manner in which you responded to the recommendations and comments received. Your constructive approach and willingness to improve the manuscript reflect a genuine commitment to research quality.

I wish you much success in your ongoing scientific work and in further developing this promising topic. I am confident that your efforts will contribute to strengthening the value of the article and its impact within the academic community .

Warm regards!

Author Response

On behalf of Authors, i would like to thanks and appreciate your constructive recommendations and comments which we believed have improved the quality of our manuscript.

We wish you success in safeguarding, controlling and improving qualities of scientific work.

Best regards

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors!

My suggestions are mainly design corrections:

1) L 50. You should change “…as described by [4]…” to “…as described by Andriesse et al. (1994)], comprise the upper reaches… [4].”.

L 67. Change the sentence to: “The work of Setter, T. et al. (2009) indicated that …”.

L 87. Change the sentence to: “ Blum A. J. et al (1982) assessed canopy…[18]”.

L 104. Change the sentence to: “According to Pineda M.M. et al. (2020) [24], the increase in leaf temperatures  

may be due to stomatal closure.”.

L 188. Change the sentence: “…calculated according to Jones, H.G. (1999)[31],…”.

L 441-445. Change the sentences to “Similarly, Ledent J. (2002) reported that prolonged drought reduced leaf number and size while enhancing the retention of already expanded leaves [34] , and Suárez L. and V. Mederos (2011) noted that leaf production and longevity are varietal traits strongly influenced by environmental conditions [35]. In cassava, leaf loss is considered a strategy to conserve water [36, 37]. Furthermore,  Aina O.O. et al. (2007) demonstrated…[38].”

L 463-473. Change the sentences to “According to Long S. et al. (1994) , the Fv/Fm ratio is a key parameter for detecting damage to photosystem II (PSII) and identifying the occurrence of photoinhibition [40]. …The stability of PSII at 0, 4, and 8 DAT suggests that oxidative stress was limited during early waterlogging, which may be attributed to effective ROS scavenging and the absence of chlorophyll degradation, as supported by SPAD data (Fig. 4b),  Nishizawa A.Y. et al. (2008) reported that ROS scavengers reduce oxidative damage under abiotic stress [43] while   Сao  et al. (2022) found that …[44]”.

Similar corrections should be made on L 488, 492, 495, 497, 499, 503, 508, 522, 537, 541.

2) You should write drought (L 137), light (L 172), leaf (L 177), treatment (L 207), soil (L 250), light intensity (L 251), bars (L 276, 329, 354, 398), treatment duration (L 282), waterlogging (L 425) with small letters.

L 75. Change the sentence to “…reduction in net photosynthetic rate (Pn) under waterlogging.”.

3) L 241. You should correct m³/m to m³/m³.

4) You used large P letter in Abstract and in the text (L 404, etc), but also you wrote small p (L 259, etc) in the text. You should write the same parameter uniformly.

5) You should check the references and their design. For example, instead of Chen, et al., Transcriptomic profiling suggests candidate molecular responses to waterlogging in cassava. PLoS One, 2022. 17(1): p. e0261086I found Cao, M., Zheng, L., Li, J., Mao, Y., Zhang, R., Niu, X., ... & Chen, Y. (2022). Transcriptomic profiling suggests candidate molecular responses to waterlogging in cassava. PloS One, 17(1), e0261086.

Comments on the Quality of English Language

I didn`t find any language mistakes.

Author Response

We have uploaded cover letter of point by point responses

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

Accepted 

Author Response

On behalf of Authors, i would like to thank and appreciate you for the constructive recommendations, suggestions and comments which have improve the quality of our manuscript further. We wish you success in filtering and doing improvement process that helps maintain the integrity and credibility of academic publishing. 

Best regards