Dual Role of Acid Rain and Pyricularia oryzae on Growth, Photosynthesis and Chloroplast Ultrastructure in Rice Seedlings
, Qi Wang 1, Huimin Xiang 1,3,4,5,6, Yiliang Liu 1, Hui Wei 1,3,4,5,6 and Zhong Qin 1,3,4,5,6Round 1
Reviewer 1 Report
The author comments appear in word doc file as attached
Comments for author File: 
Comments.pdf
Author Response
Dear Editor,
We are pleased to submit our revised manuscript (agronomy-1527247) entitled “Combined effects of acid rain and Pyricularia oryzae on growth, photosynthesis and chloroplast ultrastructure in rice seedlings”, which we would like you to consider for publication in agronomy.
We appreciate very much your comments and kind corrections on the previous version, and those of your reviewers. We have addressed all of these very carefully. Revisions have been highlighted using the "Track Changes" function in Microsoft Word. Below, it’s our response to the reviewers’ comment.
Please do not hesitate to contact us should you have any other questions.
RESPONSE TO REVIEWER 1 COMMENTS:
The article concerns with the elucidation of growth, photosynthetic characteristics, and chloroplast ultrastructure of rice seedlings decreased under treatments of Pyricularia oryzae , c o Rice blast and Acid rain (pH 2.0, 4.0), and their combination. The experiment seems to be well pursued and brings about the insight and a verification of different effects of rice pathogen and low pH rain on growth and photosynthetic efficiency of rice plant. The authors may go through the comments listed below and answer for each of the queries: 

Point 1: There are very good papers on acid rain and rice blast which talk about interference with PSII. Has interaction been studied before?
Response 1: Thank you very much for your confirmation, the effect of acid rain and rice blast interaction on antioxidant enzymes in rice has indeed been studied before.
Point 2: No mention of variety / genotype on which experiment was conducted. Name the genotype/ stock.
Response 2: Rice variety, Mai Xiang zhan (PI 2006009) belongs to Indica, selected by Rice Research Institute of Guangdong Academy of Agricultural Sciences.
Point 3: Line 98/99: The major query would be that if Resistant and susc. Can be treated as two factors or levels of the same factor. The factorial ANOVA must have been followed which include Acid rain and P. oryzae as two factors.
Response 3: Thank you for pointing out this problem. The P. oryzae infection included two treatments, inoculated and not inoculated with P. oryzae. We supplemented the ANOVA data in the figures (Fig. 1-3).
Point 4: Line 102: whether the expt was done in open field or controlled conditions. If it was done in open controlled chamber, what was the purpose of recording annual ppt.
Response 4: The purpose of our recording rainfall is to measure the amount of acid rainfall for potted plants based on the annual acid rainfall in Guangdong Province to ensure that the amount of acid rain is close to the real situation.
Point 5: Line 149: same as point no. 2
Response 5: We rewrite the sentences to make it clear.
Point 6: ANOVA is missing
Response 6: Thank you for pointing out this problem. We supplemented the ANOVA data in the figures (Fig. 1-3).
Point 7: Line 162 may be modified/ deleted.
Response 7: Thanks for the suggestion, line 162 has been removed.
Point 8: Why was biomass recorded? Why not Harvest index as is more important incase of rice?
Response 8: This is because the index directly related to the photosynthetic characteristics of plants is biomass. Second, considering that the main onset of rice blast is concentrated in the tillering stage of rice, the growth cycle of rice in this experiment was not cultivated until the harvesting stage.
Point 9: How can the effect of soil N and acid rain N be separated out.
Response 9: First, acid rain is sprayed on the rice from top to bottom. On the one hand, acid rain directly contacts the surface of the rice, and on the other hand, the nitrogen ions in acid rain enter the soil. In addition, there are other ions in the acid rain besides nitrogen ions, such as hydrogen ions, which cause soil acidification and eventually affect plant growth. To distinguish the effect of soil nitrogen on the experiment, no exogenous fertilizer was applied throughout the experiment.
Thank you for handling our manuscript and I look forward to hearing from you soon!
Yours sincerely,
Hong-Ru Li
South China Agricultural University
No. 483 Wushan Road, Tianhe District, Guangzhou
510642, China
Reviewer 2 Report
1.Introduction
Assumption/hypothesis of the experiment should be given at the end of introduction at L.No.80.
2Materials and methods
Need to mention factors and their levels at L.No. 148. Then factorial ANOVA needs to be performed.
- Results
Refer to LNo. 162 ?
Rewrite LNo. 167-168 to make it clear.
Rewrite L.No. 167 and 168 to make it clear
Rewrite L.No. 182-183 to make it clear
Rewrite L.No. 216 & 217 to make it clear
L.No. 261 to 264 to make it clear
- Conclusion
Rewrite L.No. 389-390 to make it clear
Author Response
RESPONSE TO REVIEWER 2 COMMENTS:
Point 1: Assumption/hypothesis of the experiment should be given at the end of introduction at L.No.80.
Response 1: Thank you very much for your suggestions!
Point 2: Need to mention factors and their levels at L.No. 148. Then factorial ANOVA needs to be performed.
Response 2: Thank you for pointing out this problem. We supplemented the ANOVA data in the figures (Fig. 1-3).
Point 3: Rewrite LNo. 167-168 to make it clear.
Rewrite L.No. 182-183 to make it clear
Rewrite L.No. 216 & 217 to make it clear
L.No. 261 to 264 to make it clear
Response 3: Thank you for pointing out this problem. We rewrite the above sentences to make it clear (Lines 167-168, 182-183, 216 & 217 and 261- 264).
Point 4: Rewrite L.No. 389-390 to make it clear.
Response 4: Thank you for pointing out this problem. We rewrite L.No. 389-390 to make it clear.
Thank you for handling our manuscript and I look forward to hearing from you soon!
Yours sincerely,
Hong-Ru Li
South China Agricultural University
No. 483 Wushan Road, Tianhe District, Guangzhou
510642, China
Reviewer 3 Report
Combined effects of acid rain and Pyricularia oryzae on growth, photosynthesis and chloroplast ultrastructure in rice seedlings
- Title could be changed to be more attracted like (dual role of acid rain and Pyricularia oryzae on the growth parameters, photosynthesis, and chloroplast ultrastructure in rice seedlings)
- Line 99: please clarify the susceptible and unsusceptible cultivars name, Unsusceptible means resistant????
- Line 162-163: where the title of the table
Data in the table should be revised especially the leaf are with treatment pH 2.0 + P. oryzae
- Line 182: The Pn, Ci, Tr and Gs in rice seedlings treated with pH 2.0 showed significance decrease compared to control.
- Lines 182-191: Please rewrite the paragraph.
- Lines 196-209: Please rewrite the paragraph.
The research is interesting, novel but the manuscript needs enhancement especially in the results part.
Comments for author File: Comments.pdf
Author Response
RESPONSE TO REVIEWER 3 COMMENTS:
Point 1: Title could be changed to be more attracted like (dual role of acid rain and Pyricularia oryzae on the growth parameters, photosynthesis, and chloroplast ultrastructure in rice seedlings). 

Response 1: Thank you very much for your suggestions! The title has been revised.
Point 2: Line 99: please clarify the susceptible and unsusceptible cultivars name, Unsusceptible means resistant????
Response 2: The P. oryzae infection included two treatments, inoculated and not inoculated with P. oryzae.
Point 3: Line 162-163: where the title of the table (Data in the table should be revised especially the leaf area with treatment pH 2.0 + P. oryzae).
Response 3: Thank you for pointing out this problem. Added the title and corrected the name of pathogen in this article including the abbreviations in the figure and table.
Point 4: Lines 182-191: Please rewrite the paragraph.
Response 4: Thank you for pointing out this problem. We corrected those mistakes point by point (Lines 182-191).
Point 5: Lines 196-209: Please rewrite the paragraph.
Response 5: Thank you for pointing out this problem. We corrected those mistakes point by point (Lines 196-209).
Thank you for handling our manuscript and I look forward to hearing from you soon!
Yours sincerely,
Hong-Ru Li
South China Agricultural University
No. 483 Wushan Road, Tianhe District, Guangzhou
510642, China
