Effect of Nitrification Inhibitors on Photosynthesis and Nitrogen Metabolism in ‘Sweet Sapphire’ (V. vinifera L.) Grape Seedlings

Round 1

Reviewer 1 Report

Dear Authors,

Please elaborate on Methods and Materials in detail.

Have you validated gene expression using qRT-PCR. It would be good if you can choose 5-6 important genes for this study and validate the transcriptome data.

Comments for author File: Comments.pdf

Author Response

Introduction:

Row 33&34: Avoid starting sentence with “It” too often. You can combine sentences with clause.

Thank you for your suggestion.We have revised the sentence in the manuscript.

What do you suggest by resistant to storage and transport. Do you mean that this variety is resistant to damage caused during storage and transport or post-harvest damage? If yes, then say that clearly.

Thank you for your suggestion. we hold the opinion that resistant to storage and transport mean that this variety is resistant to damage caused during storage and transport or post-harvest damage. And we made corresponding changes in the manuscript.

For example: In the recent years, ‘Sweet sapphire’ is the most popular grape variety due to its excellent quality for storage and transportation, naturally seedless, and comparatively more resistant to disease as compared to other Eurasian species.

Thank you for your suggestion. We have revised rows 31-34 in the manuscript according to your suggestion.

What is high fertilizer and nitrogen? It should be, Sweet sapphire requires more nitrogen or frequent fertilization for growth and development.

Thank you for your suggestion. That's what we're trying to say Sweet sapphire requires more nitrogen or frequent fertilization for growth and development. We have made corresponding corrections in the manuscript.

Row 35# it is fertilizerin American English.

Thank you for pointing out this error and we are sorry for our mistake.The error in row 35 was corrected in the manuscript.

Row 36: Add require before for.

Thank you for pointing out this error and we are sorry for our mistake.The error in row 36 was corrected in the manuscript.

Row 38: what do you mean by relates? Be scientific and use simple words to indicate what you want to mention. You can say “Nitrogen is important for photosynthesis and therefore, it plays significant role in the crop or fruit yield. 

Thank you for your suggestion. We have amended row 38 the manuscript into “Nitrogen is important for photosynthesis and therefore, it plays significant role in the crop or fruit yield”.

Materials and Methods:

How often DMPP was applied during the experiment?

In this experiment, DMPP was applied to the soil once mixed with urea at the first watering, only once.

Explain methods in briefly although you have used services for RNA extraction, transcriptome sequencing.

Thank you for your suggestion. We have perfected the methods of RNA extraction and transcriptome sequencing in rows 144-163 of the manuscript. We apologize for this mistake in the interest of language brevity.

What DPI was used for scanning?

Thank you for your question.We used 300 dpi for scanning.

Explain briefly how root vigor was tested. How much TTC concentrationwas used and how vigor is determined?

Thank you for your suggestion. We have given a brief explanation of the methods used to determine root activity in rows 111-117 of the manuscript. We apologize for this misunderstanding in the interest of language brevity.

For photosynthesis pigment analysis, give details of ratio of extraction, instruments used for the measurements, Write the instrument manufacturer details in the parenthesis.

Thank you for your suggestion. For photosynthesis pigment analysis, we have given details of ratio of extraction in rows 119-124 of the manuscript. We also indicating the instruments and instrument manufacturer details in row 131 and 135. We apologize for this mistake in the interest of language brevity.

Results:

Row 149# “T2 treatment had significantly greater effects than T1 and T3”. Significant greater effect in terms of what? Please rewrite or rephrase to make sentence meaningful.

Thank you for your suggestion. We have made changes to row 181 in the manuscript to change the sentence to T2 treatment had significantly greater effects than T1 and T3 in total biomass, shoot biomass and root biomass. We think that can make sentence meaningful.

Fig 1: Legend. T3 box is smaller. Check the treatment order in the legend.

Thank you for your suggestion. We have checked all the pictures in the article, repaint the picture and consummate the legends. We apologize for this carelessness.

Row 217-221# Why “-“ was used to indicate total number of genes. Is it a range?

Thank you for your suggestion. We have three replicates per treatment, and "-" represents the range of three replicate values. For instance, "22034-22260 genes were identified in the roots" indicate that we identified 22260, 22053 and 22034 genes in the roots.

How many genes were identified in CK?

Thank you for pointing out this carelessness and we are sorry for our mistake.The error in rows 254-258 was corrected in the manuscript.

Did you validate any DEGs by RT-qPCR?

Thank you for your suggestion. We are very sorry for this problem. We reviewed the relevant literature and found that doing RT-qPCR was the next step of research work,We will accomplish the relevant experiments in this part next.

Row 257# Grapevine roots

Thank you for your suggestion. We have replaced grape roots with grapevine roots in rows 295 of the manuscript.

Row 331#: Cite the reference for statement “ NH₄⁺-N can highly stimulate later root formation by inhibiting cell elongation rather than cell division.

Thank you for pointing out this error and we are sorry for our mistake. We have added the statement of citing references in row 371 of the manuscript.

Reviewer 2 Report

The manuscript is well prepared and constructed and a few corrections are needed before further processing.

1. Please include a methodological section on Gene Ontology and KEGG analysis approaches. 

2. Please increase the font size of given figures. At normal 100% page view, some of the figures' axis and color codes are hard to read. 

3. The manuscript has supplementary file submitted, but no mention within the text. Please mention these data in the text. 

4. The discussion on LHC, Psa, Psb, Pet, Nar and Nrt genes and their roles should be more. Please add some more discussion in this section. 

Author Response

1. Please include a methodological section on Gene Ontology and KEGG analysis approaches. 

Thank you for your suggestion. We have perfected the methods of Gene Ontology and KEGG analysis in rows 144-163 of the manuscript. We apologize for this misunderstanding in the interest of language brevity.

2. Please increase the font size of given figures. At normal 100% page view, some of the figures' axis and color codes are hard to read. 

Thank you for your suggestion. We have checked all the pictures in the article, repaint the picture and consummate the legends(Fig. 1-3). We apologize for this carelessness.

3. The manuscript has supplementary file submitted, but no mention within the text. Please mention these data in the text. 

Thank you for your suggestion. We have mentioned supplementary file data in the text in the manuscript. We apologize for this mistake in the interest of language brevity.

4. The discussion on LHC, Psa, Psb, Pet, Nar and Nrt genes and their roles should be more. Please add some more discussion in this section. 

Thank you for your suggestion. We have added to the discussion of the LHC, Psa genes in rows 424-431 of the manuscript, and we are sorry that the discussion of the LHC and Psa genes is not rich enough. But for Psb, Pet, Nar and Nrt genes, we believe that the principles and the relevant literature have been clearly stated.