Effects of Salinity-Stress on Seed Germination and Growth Physiology of Quinclorac-Resistant Echinochloa crus-galli (L.) Beauv
Round 1
Reviewer 1 Report
Overall a well presented study with results that can be used as a platform for future weed management in rice fields. It’s a descriptive study which several parameters measured. However, there are minor changes that needs to be done before this can be finalized for publication.
E. crus-galli should be checked throughout the manuscript for italics font. Same applies to Oryza sativa
Figures are not clear at 100% zoom. Is there a way to increase the font size, perhaps one below the other instead of side-by-side? (Especially Figs 1-5)
Authors might want to include year of the publications mentioned as in-text references?
Introduction
Line 48-52- check sentence formations and punctuations.
Materials and Methods
Line 77-All biotype Y E. crus-galli plants died at 21 days after the treatment of……….
Line 81- the seeds were propagated………………
Line 98- Wu et al., (year??)
Line 106- what was used to seal the plates? Parafilm?
Line 126- weighed?
Line 131- Bradford (year), 28 °Cline 146- Oryza sativa (in italics)
Line 157- I am assuming this is where results start, however, authors missed to separate the sections and title them.
Line 185- Figures should be standalone entities and thus abbreviations should be included in the figure legends too. This makes the figures readable and can be easy to understand.
Line 191- NaCl
The higher concentrations………… this sentence should be rewritten
line 231- while for the treats????
Line 258- Karimi et al (year?)
Line 285- the plants grown under………
This sentence needs to be rewritten
Line 334- This research showed….
Here authors need to explicitly mention the main findings of the study and check for grammar too
Line 336- the mechanism is said to be considered as concluded. I think it’s a very strong point to make, perhaps authors can change this sentence with future scope of validation required for this study?
Overall, conclusion needs to be rewritten completely
Author Response
May 10, 2022
Dear Reviewer:
Thank you very much for your reviewers’ comments concerning our manuscript entitled ‘Effects of salinity-stress on seed germination and growth physiology of quinclorac-resistant Echinochloa crus-galli (L.) Beauv’ (Manuscript ID: agronomy-1702685). Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied your comments carefully and have made corrections which we hope meet with approval. We checked throughout the manuscript for italics font of E crus-galli and Oryza sativa, and corrected incorrect writing (Line 145-146, 431, 435). We also modified the figures (Figs 1-5) for increasing the font size. Besides, we have carefully revised the manuscript to make sure it is properly formatted according to suggestion of the editor and the viewers.
Revised portions are marked in blue in the paper. The main corrections in the paper and the responds to the reviewer’s comments are as flowing:
Introduction
Line 48-52- check sentence formations and punctuations.
Answer: Thanks for your comments. We revised this sentence as “Nowadays, although the rapid development of herbicide resistance in E. crus-galli was developing [10-12], but a better adaptability of the weed to the extreme environment is reported [13-14], and the adverse influence on seed germination of E. crus-galli was observed [15]”.(Line 48-51)
Materials and Methods
Line 77-All biotype Y E. crus-galli plants died at 21 days after the treatment of……….
Line 81- the seeds were propagated………………
Answer: Thanks for your comments, I stand corrected (Line 75-76, 79).
Line 98- Wu et al., (year??)
Answer: 2019 (Line 96).
Line 106- what was used to seal the plates? Parafilm?
Answer: Yes. We revised this sentence as “was properly sealed by parafilm.”(Line 104).
Line 126- weighed?
Answer: Yes. Thank you for pointing out this mistake in writing. The correct expression is “The fresh leaves were removed and weighed up”(Line 124).
Line 131- Bradford (year), 28 °C line 146- Oryza sativa (in italics)
Answer: Thanks for your comments, I stand corrected (Line 129, 143-144).
Line 157- I am assuming this is where results start, however, authors missed to separate the sections and title them.
Answer: Thanks for your comments. In this part, we added title “3. Results” (Line 158).
Line 185- Figures should be standalone entities and thus abbreviations should be included in the figure legends too. This makes the figures readable and can be easy to understand.
Answer: Thanks for your comments. In the figure legends (Figure 1), we supplied “ CH, the E. crus-galli biotype collected from Chunhua, Hunan province, China; QR, the E. crus-galli biotype collected from Chunhua, Hunan province, China, which were bred for four generations from 2014 to 2017, and resistant to quinclorac; HS, the E. crus-galli biotype collected from Hanshou, Hunan province, China; SM, the E. crus-galli biotype collected from Shimen, Hunan province, China; HK, the E. crus-galli biotype collected from Hekou, Hunan province, China; DC, the E. crus-galli biotype collected from Dingcheng, Hunan province, China; QS, the E. crus-galli biotype collected from Chunhua, Hunan province, China, which were bred for four generations from 2014 to 2017, and sensitive to quinclorac. The same as below.(Line 193-200)
Besides, we modified the figures (Figs 1-5) for increasing the font size.(Line 185-189)
Line 191- NaCl
The higher concentrations………… this sentence should be rewritten
Answer: Thanks for your comments. The sentence was revised as “The higher concentration of NaCl, the greater inhibitory effect on shoots and roots of E. crus-galli.” (Line 203-204)
line 231- while for the treats????
Answer: The sentence was revised as “While without NaCl-stress”.(Line 245)
Line 258- Karimi et al (year?)
Answer: Karimi et al (2018)(Line 283).
Line 285- the plants grown under………
This sentence needs to be rewritten
Answer: The sentence was revised as “Different plants have different tolerance to salt sress, compared with cowpea, cotton is more tolerant to salt.”(Line 315-316)
Line 334- This research showed….
Here authors need to explicitly mention the main findings of the study and check for grammar too
Line 336- the mechanism is said to be considered as concluded. I think it’s a very strong point to make, perhaps authors can change this sentence with future scope of validation required for this study?
Overall, conclusion needs to be rewritten completely
Answer: Thanks for your comments. We have rewritten the conclusion, this part is modified as “Overall, we have identifed the effects of salt stress on germination, seedling growth and their GST activity of E. crus-galli populations with different quinclorac resistance levels. While under salt stress, the higher level of quinclorac-resistant leading to the greater inhibitory effect on seed germination, shoots and roots length, and fresh weight of E. crus-galli , but the lower GST activity. Besides, through transcriptome sequencing, we found many DEGs between resistance and sensitve E. crus-galli, which regulating the response to salt stress. We will carry out functional verification for key genes in the future, which will lay a foundation in the management of the resistant weeds. ”(Line 352-359)
We appreciate for warm work of editor and reviewers earnestly, and hope that the correction will meet with approval.
Once again, thank you very much for your comments and suggestion, we are looking forward to hearing from you soon.
Sincerely yours,
Lamei Wu
Lamei Wu (corresponding author)
Hunan Agricultural Biotechnology Research Institute
Hunan Academy of Agricultural Sciences
Changsha, Hunan 410125, P. R. CHINA
E-mail: [email protected]
Tel: 86-0731-84696075; Fax: 86-0731-84696075
Author Response File: 
Author Response.doc
Reviewer 2 Report
The manuscript entitled "Effects of salinity stress on seed germination and growth physiology of quinclorac-resistant Echinochloa crus-galli (L.) Beauv" is very interesting. The study is scientifically systematic and logical, and the data set is likely necessary for the general understanding of the salinity effect on resistant or susceptible plants. The originality of the work is not questionable. The title reflects the manuscript's content; the abstract is concise; it presents the purpose, general materials and methods, summarised results, and the conclusion. In the Introduction section authors provide enough background information. However, a better narrative could be achieved to better explain critical statements of the investigated problem. The study design is reasonable. Results should be presented with more clearance, figures are too small, and the letters representing statistical significance are hardly visible. What statistical analyses were used? The letters that represent statistical differences should be explained in the figure caption. The results of statistical tests should be presented in the body of the text (F, p values, etc.). Also, the results of transcriptomics should be better explained in the text. The authors stated in the discussion section that they revealed downregulation of Cytochrome P450, PEPcase, and POD. Where can this be seen in the results? Based on this, the authors then conclude the mechanism of quinclorac resistance? I suggest clarifying this in all three sections.
Overall, the paper highlights the importance of finding better management of the resistant weed, but the authors should critically question their findings: how will an increase in salinity reflect crop yield simultaneously? The referencing is accurate, adequate, and balanced with older and more recent literature.
Comments for author File: 
Comments.pdf
Author Response
May 10, 2022
Dear Reviewer:
Thank you very much for your reviewers’ comments concerning our manuscript entitled ‘Effects of salinity-stress on seed germination and growth physiology of quinclorac-resistant Echinochloa crus-galli (L.) Beauv’ (Manuscript ID: agronomy-1702685). Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied your comments carefully and have made corrections which we hope meet with approval. We have carefully revised the manuscript to make sure it is properly formatted according to suggestion of the editor and the viewers.
Revised portions are marked in blue in the paper. The main corrections in the paper and the responds to the reviewer’s comments are as flowing:
Results should be presented with more clearance, figures are too small, and the letters representing statistical significance are hardly visible. What statistical analyses were used? The letters that represent statistical differences should be explained in the figure caption. The results of statistical tests should be presented in the body of the text (F, p values, etc.). Also, the results of transcriptomics should be better explained in the text. The authors stated in the discussion section that they revealed down-regulation of Cytochrome P450, PEPcase, and POD. Where can this be seen in the results? Based on this, the authors then conclude the mechanism of quinclorac resistance? I suggest clarifying this in all three sections.
Answer: Thanks for your comments. We modified the figures (Figs 1-5) for increasing the font size, and enlarged the letters which representing statistical significance. Data were submitted to analysis of variance, the averages of the germination rate, the shoot length, the root length and the fresh weight were were compared by the Duncan's multiple-range test at 5% signifcance, all data were calculated using SPSS 10.0. The figures were drawn using Sigma Plot 10.0 (Line 154-157). In the figure caption, we explained the letters that represent statistical differences as “Lower case letters is expressed as least square means ± standard error (P<0.05).”(Line 200). In the part of 3.6., we explained the results of transcriptomics as “Following NaCl treatment in the QR and QS biotypes of E. crus-galli, 563 DEGs in the QR samples relative to the QS samples were annotated into KEGG pathways. Pathway enrichment analysis found the most highly enriched pathways included the “linoleate 9S-lipoxygenase” (K15718), “solute carrier family” (K14709、K14445), “arogenate/prephenate dehydratase” (K05359), “monodehydroascorbate reductase” (K08232), “Cytochrome P450” (K05280), “peroxidase”(K00430), cellulose synthase A (K10999) and “glutathione S-transferase” (K00799)_pathways. 4 herbicide-related species such as “Cytochrome P450”, “glutathione S-transferase” were found to be down-accumulated. 7 DEGs associated with “defense response” were found response to salt stress, ,which involved in the regulatory of stress processes” (Line 271-280).
Overall, the paper highlights the importance of finding better management of the resistant weed, but the authors should critically question their findings: how will an increase in salinity reflect crop yield simultaneously? The referencing is accurate, adequate, and balanced with older and more recent literature.
Answer: Thanks for your comments. We revised the effect of salt stress on yield as “Is salinity not benefit for crops ? While compared with no irrigation, saline water application can increase crop yields but may lead to some adverse effects on soil properties and plant growth. Water salinity less than 3.2 dS m−1 stimulated root growth in the soil layers 10–40 cm at the heading stage without obvious yield reduction or salt accumulation, and the interaction effects of irrigation amount and water salinity had no statistically significant (p > 0.05) effects on root parameters and yield [30]”(Line 291-296).
We appreciate for warm work of reviewer earnestly, and hope that the correction will meet with approval.
Once again, thank you very much for your comments and suggestion, we are looking forward to hearing from you soon.
Sincerely yours,
Lamei Wu
Lamei Wu (corresponding author)
Hunan Agricultural Biotechnology Research Institute
Hunan Academy of Agricultural Sciences
Changsha, Hunan 410125, P. R. CHINA
E-mail: [email protected]
Tel: 86-0731-84696075; Fax: 86-0731-84696075
Author Response File: Author Response.doc
