Deciphering the Role of Stay-Green Trait to Mitigate Terminal Heat Stress in Bread Wheat

Round 1

Reviewer 1 Report

Interesting study providing evidence that the stay green trait in wheat can help mitigate increasing temperatures (terminal heat stress). The research shows good rigour with two years of field experiments in Islamabad, Pakistan and a glasshouse experiment in South Dakota, USA.

The main strengths are the strong international collaboration and complementary research between the National Agricultural Research Centre in Islambad for field experiments and South Dakota State University with its excellent molecular biology and glasshouse facilities. A minor weakness is that only a few genotypes were screened in the greenhouse experiment (but this could be due to the high costs of the molecular characterisation). The aims need to be stated more clearly at the end of the Introduction (not just the method).

Below are some minor specific suggestions:

Pg 2, Ln 54 - Replace "reduce" with "reduced"

Pg 2, Ln 83 - Replace "Current" with "The current"

Pg 3, Ln 101-109 - Need to clearly state aims at the end of the Introduction.

Pg 3, Ln 116 - Replace "Plantation" with "Planting"

Pg 16, Ln 408 - Replace "Current" with "The current"

Pg 16, Ln 422 - Replace "Greenhouse" with "The Greenhouse"

Pg 16, Ln 430 - Replace "Despite of" with "Despite"

Pg 17, Ln 486 - Replace "Present" with "The present"

Author Response

Response to Reviewer 1 Comments

General Response

The authors thank the reviewer for the generous comments and in-depth review of the manuscript. The manuscript has been revised considering the significant shortcomings highlighted by the reviewer. Our responses are given in point by point manner below. We hope that the reviewer will find the revised manuscript fulfilling the rigorous criteria of publication in “Agronomy”.

Point by Point Response

Point 1: Pg 2, Ln 54 - Replace "reduce" with "reduced"

Response 1: Replaced “reduce” with “reduced” (Pg 2, Ln 60).

 

Point 2: Pg 2, Ln 83 - Replace "Current" with "The current"

Response 2: Replaced “Current” with “The current” (Pg 2, Ln 90).

 

Point 3: Pg 3, Ln 101-109 - Need to clearly state aims at the end of the Introduction.

Response 3: The aims of the study have been clearly stated at the end of the Introduction (Pg 3, Ln 118-121).

 

Point 4: Pg 3, Ln 116 - Replace "Plantation" with "Planting"

Response 4: Replaced “Plantation” with “Planting” (Pg 3, Ln 130).

 

Point 5: Pg 16, Ln 408 - Replace "Current" with "The current"

Response 5: Replaced “Current” with “The current” (Pg 16, Ln 518).

 

Point 6: Pg 16, Ln 422 - Replace "Greenhouse" with "The Greenhouse"

Response 6: Replaced “Greenhouse” with “The greenhouse” (Pg 16, Ln 532).

 

Point 7: Pg 16, Ln 430 - Replace "Despite of" with "Despite"

Response 7: Replaced “Despite of” with “Despite” (Pg 16, Ln 540).

 

Point 8: Pg 17, Ln 486 - Replace "Present" with "The present"

Response 8: Replaced “Present” with “The present” (Pg 17, Ln 599-600).

Author Response File: Author Response.docx

Reviewer 2 Report

Overall, the manuscript is novel, and fits within the journal guidelines. However, in my opinion, moderate English changes are required. Also, there are several minor issues that the authors should address before publication.

Minor concerns:

Line 2 and in the whole manuscript: please change "stay green" to "stay-green".

Line 18: Please change "Present study was aimed to" to "The present study aimed to reveal".

Line 21: Please mention the stay-green types.

Line 22: Please change "Results" to "The results".

Line 24: Please change "In greenhouse" to "In the greenhouse".

Line 28: Please define Cao, Cab, SGR, and RCCR.

Lines 83 and 84: Please change "Current study is focused on" to "The current study focused on".

Line 85 and in the whole manuscript: Please use "," before "the conjunction and" in a list of three or more items.

Lines 86-98: Please remove unnecessary relative pronouns ("that") in this paragraph.

Line 101: This sentence needs a reference.

Lines 101- 109: This paragraph is not clear.

Lines 111 and 236: Please change "Phenotypic characterization" to "Phenotyping"

Line 113: This sentence is not clear.

Line 118: Please change "field experiment" to "The field experiment".

Line 121: Please change "The traits evaluated were" to "The evaluated traits were".

Line 143: Please add "The" before greenhouse experiment.

Lines 143-153: This paragraph is not clear.

Lines 154 and 160: Please change "was" to "were".

Lines 163 and 329: Please change "Molecular characterization" to "Genotyping".

Lines 177-183: This paragraph is not clear.

Line 239; Please change " A scale" to "This rating scale".

In the whole manuscript: Using both American and British English forms is confusing. For example "Among" and "Amongst".

Line 422; Please add "The" before greenhouse experiment.

 

 

 

 

 

 

 

 

 

 

Author Response

Response to Reviewer 2 Comments

General Response

We express immense gratitude for such an in-depth review of the manuscript. The manuscript has been edited for all the minor issues as suggested. The responses are given in point by point manner below. Moreover, the manuscript has been proof read prior to resubmission to avoid any flaw. We hope the revised manuscript fulfils the rigorous criteria of publication in Agronomy.

Point by Point Response

Point 1: Line 2 and in the whole manuscript: please change "stay green" to "stay-green".

Response 1: Changed "stay green" to "stay-green" in the whole manuscript.

 

Point 2: Line 18: Please change "Present study was aimed to" to "The present study aimed to reveal".

Response 2: Changed "Present study was aimed to" to "The present study aimed to reveal" (Line 18).

 

Point 3: Line 21: Please mention the stay-green types.

Response 3: Stay green types have been mentioned for both field and greenhouse experiments (Line 19-23).

 

Point 4: Line 22: Please change "Results" to "The results".

Response 4: Changed "Results" to "The results" (Line 23).

 

Point 5: Line 24: Please change "In greenhouse" to "In the greenhouse".

Response 5: Changed "In greenhouse" to "In the greenhouse" (Line 26).

 

Point 6: Line 28: Please define Cao, Cab, SGR, and RCCR.

Response 6: Cao, Cab, SGR, and RCCR genes have been defined (Full form) (Line 30-31).

 

Point 7: Lines 83 and 84: Please change "Current study is focused on" to "The current study focused on".

Response 7: Changed "Current study is focused on" to "The current study focused on" (Line 90).

 

Point 8: Line 85 and in the whole manuscript: Please use "," before "the conjunction and" in a list of three or more items.

Response 8: Added “,” before “the conjunction and” in the list of three or more items in the whole manuscript.

 

Point 9: Lines 86-98: Please remove unnecessary relative pronouns ("that") in this paragraph.

Response 9: Removed unnecessary relative pronouns (“that” and “which”) (Line 93-114).

 

Point 10: Line 101: This sentence needs a reference.

Response 10: Reference added for  “Stay-green trait seems to improve grain yield under high temperature stress by retaining chlorophyll content, improving photosynthetic capacity, and extending grain filling period” (Line 115-117).

 

Point 11: Lines 101- 109: This paragraph is not clear.

Response 11: The paragraph has been changed to make aims of the study clear (Line 115-123).

 

Point 12: Lines 111 and 236: Please change "Phenotypic characterization" to "Phenotyping"

Response 12: Changed "Phenotypic characterization" to "Phenotyping" (Line 125 and 314).

 

Point 13: Line 113: This sentence is not clear.

Response 13: The sentence has been revised (Line 127-130).

 

Point 14: Line 118: Please change "field experiment" to "The field experiment".

Response 14: Changed "field experiment" to "The field experiment" (Line 133).

 

Point 15: Line 121: Please change "The traits evaluated were" to "The evaluated traits were".

Response 15: Changed "The traits evaluated were" to "The evaluated traits were" (Line 136).

 

Point 16: Line 143: Please add "The" before greenhouse experiment.

Response 16: Added "The" before greenhouse experiment (Line 175).

 

Point 17: Lines 143-153: This paragraph is not clear.

Response 17: The paragraph has been revised as suggested (Line 175-187).

 

Point 18: Lines 154 and 160: Please change "was" to "were".

Response 18: Changed "was" to "were" as suggested (Line 188 and 196).

 

Point 19: Lines 163 and 329: Please change "Molecular characterization" to "Genotyping".

Response 19: Changed "Molecular characterization" to "Genotyping" (Line 197 and 424).

 

Point 20: Lines 177-183: This paragraph is not clear.

Response 20: The paragraph has been revised (Line 211-219).

 

Point 21: Line 239; Please change " A scale" to "This rating scale".

Response 21: Changed " A scale" to "This rating scale" (Line 317).

 

Point 22: In the whole manuscript: Using both American and British English forms is confusing. For example "Among" and "Amongst".

Response 22: The whole manuscript has been thoroughly revised. “Amongst” replaced by “among” (Line 326, 329, 474, 477, 607).

 

Point 23: Line 422; Please add "The" before greenhouse experiment.

Response 23: Added "The" before greenhouse experiment (Line 532).

Author Response File: Author Response.docx

Reviewer 3 Report

Sadia et al. presented a study from physiological and molecular perspective on understanding the connection between stay green trait and heat stress. The manuscript is mostly well written, but there are some small flaws authors should raise attention to improve. I have some minor suggestions for the improvement of the manuscript as below.

1. L20, should be greenhouse not glasshouse.
2. L251, it is better to use “among” not “amongst”.
3. Table 1 should be moved to supplementary table.
4. Please indicating variables on y-axis.
5. Figure 6, please label y-axis name. Also even though you have labeled letter in different time points, it is confused to tell the difference between treatment for one genotype and between genotype, which comparison those letters should stand for?
6. Table 3 is not necessary in main text and should be moved to supplementary table.
7. Figure 9, it is not clear which genotype did you pick to represent its relative groups?

Author Response

Response to Reviewer 3 Comments

General Response

The authors thank the reviewer for the generous comments and significant suggestions on the manuscript. The manuscript has been edited according to the suggestions by the worthy reviewer. Our responses are given in point by point manner below. We hope that the revised manuscript is now suitable for publication.

Point by Point Response

Point 1: L20, should be greenhouse not glasshouse.

Response 1: Replaced “glasshouse” with “greenhouse” (L22).

 

Point 2: L251, it is better to use “among” not “amongst”.

Response 2: Replaced “amongst” with “among” as suggested (Line 326, 329, 474, 477, 607).

 

Point 3: Table 1 should be moved to supplementary table.

Response 3: Table 1 removed from the manuscript main text and has been moved to supplementary materials (Table S3).

 

Point 4: Please indicating variables on y-axis.

Response 4: The variables (abbreviation) with units are mentioned on y-axis and are depicted in detail (full form) in figures captions.

 

Point 5: Figure 6, please label y-axis name. Also even though you have labeled letter in different time points, it is confused to tell the difference between treatment for one genotype and between genotype, which comparison those letters should stand for?

Response 5: Figure 6a represents chlorophyll content and y-axis is labeled. Figure 6b represents photosynthetic efficiency of photosystem II and abbreviation “ØII” has been replaced with “Photosynthetic efficiency”. The lowercase letters represent interaction effect between genotypes and treatments. The analysis of variance (ANOVA) was performed separately for each time point i.e., 0 days after anthesis (0DAA), 7 days after anthesis (7DAA), and 14 days after anthesis (14DAA). The details of ANOVA results are given in the supplementary materials (Table S5). For convenience, ANOVA results of chlorophyll content (SPAD Value) at 0DAA, 7DAA, and 14 DAA have been added below;

CHL_0DAA

Category	LS means	Lower bound (95%)	Upper bound (95%)	Groups
FSG*S	45.1403	41.2066	49.0740	A
FSG*C	45.7625	41.8288	49.6962	A
NSG*S	46.0056	42.0719	49.9392	A
NSG*C	46.0156	42.0819	49.9492	A
NFSG*S	47.5898	43.6561	51.5235	A
NFSG*C	47.5898	43.6561	51.5235	A

CHL_7DAA

Category	LS means	Lower bound (95%)	Upper bound (95%)	Groups
NSG*S	37.4417	33.6071	41.2762	A
FSG*C	47.8571	44.0226	51.6917		B
FSG*S	48.7500	44.9154	52.5846		B
NFSG*S	50.2500	46.4154	54.0846		B
NFSG*C	50.8859	47.0513	54.7204		B
NSG*C	51.3389	47.5043	55.1735		B

CHL_14DAA

Category	LS means	Lower bound (95%)	Upper bound (95%)	Groups
NSG*S	8.4250	3.9074	12.9426	A
NFSG*S	46.9333	42.4157	51.4509		B
FSG*S	47.1933	42.6757	51.7109		B
FSG*C	47.7462	43.2286	52.2637		B
NSG*C	50.1333	45.6157	54.6509		B
NFSG*C	50.7792	46.2616	55.2968		B

 

Point 6: Table 3 is not necessary in main text and should be moved to supplementary table.

Response 6: Table 3 removed from the main text and moved to supplementary materials (Table S6).

 

Point 7: Figure 9, it is not clear which genotype did you pick to represent its relative groups?

Response 7: The genotypes subjected to molecular characterization (Nepal-38, SG-30 and Sonalika) were those selected for greenhouse experiment. The genotypes are mentioned in the greenhouse experiment (L175-177 and 179-180). However, added the genotypes representing each type in figure 9 caption (L496-497).

Author Response File: Author Response.docx