Identification and characterization of a novel multi-stress responsive gene in Arabidopsis

Faiza Tawab; Iqbal Munir; Zeeshan Nasim; Mohammad Sayyar Khan; Saleha Tawab; Adnan Nasim; Aqib Iqbal; Mian Afaq Ahmad; Waqar Ali; Raheel Munir; Maria Munir; Noreen Asim

doi:10.1371/journal.pone.0244030

Peer Review History

Original SubmissionAugust 22, 2020
25 Sep 2020 Decision Letter - Keqiang Wu, Editor PONE-D-20-26347 Identification and characterization of a novel multi-stress responsive gene in Arabidopsis PLOS ONE Dear Dr. Munir, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please submit your revised manuscript by Nov 09 2020 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. 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PLOS requires an ORCID iD for the corresponding author in Editorial Manager on papers submitted after December 6th, 2016. Please ensure that you have an ORCID iD and that it is validated in Editorial Manager. To do this, go to ‘Update my Information’ (in the upper left-hand corner of the main menu), and click on the Fetch/Validate link next to the ORCID field. This will take you to the ORCID site and allow you to create a new iD or authenticate a pre-existing iD in Editorial Manager. Please see the following video for instructions on linking an ORCID iD to your Editorial Manager account: https://www.youtube.com/watch?v=_xcclfuvtxQ 4. Please upload a new copy of Figure S1 as the detail is not clear. Please follow the link for more information: https://blogs.plos.org/plos/2019/06/looking-good-tips-for-creating-your-plos-figures-graphics/ [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Partly Reviewer #2: No ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: No ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: The authors carried out re-analysis of three publically available RNA seq datasets and zeroed in on At4g18280 (here named as GRPL1). The manuscript described the physiological analysis of GRPL1 over expression lines under salt, drought and heat stress. The study does add to knowledge on relatively less explored and functionally uncharacterized protein. However there are several points were this manuscript needs improvements. My observation and comments are as follows. • There is too much of redundancy in introduction section. There are many sentences with similar message or meaning, these needs to be removed. The aim of the study needs to be clearly mentioned. • There is one study on identification and characterization of At4g18280 (here named as GRPL1) ‘Overexpression of a Novel Component Induces HAK5 and Enhances Growth in Arabidopsis’. Adams et al. 2013. The study suggested the role of this gene in pot deficiency and salt stress response. This ref is missing in the whole manuscript. • It would be better to mention the plant /crop to which the SRA data sets belong. The reason or criteria for selecting only these three data sets out of many available for analysis should also be mentioned. • Line 159-160. “leaves of three weeks old were excised to analyse the standardized water content”. What is the meaning of standardized water content?? • Line 187-192: As floral dip method is well known method, instead of detailed description just reference is sufficient to mention the transformation method used. Infact the very ref for floral dip is missing. The details of promoter cloning are also absent. • There is repetition of methods in “Plant Material, Growth Conditions and Stress Treatment” and “Measurement of Physiological and Biochemical Indexes under Salt, Drought and Heat Stress”. These can be combined under one heading. • There is discrepancy in the number of days for drought stress imposition (water with holding) in Line no 156 (20 days), 231 (32 days) and 358 (28 days) . • Result section: • The authors described GO analysis of DEGs and next section started analyzing expression of AT4G18280 (GRPL1) which is too abrupt and requires explanation as to how the authors zeroed in on AT4G18280 (GRPL1). Moreover the first section “Global Gene …..Abiotic Stresses” details can be made shorter. • Again there is redundancy in few lines of section 1 and section 2 of results. • Line no. 277 PEG induces osmotic stress not drought stress. Moreover there is no distinction in the Table (Fig.1a) which is drought and PEG imposed stress. • Line 295: First, the overexpression was analyzed at the… • Fig. 2C should have scale bar for comparison of size. For Fig. 2D data from how many plants was recorded (replicates)? In general no of plants /replicates should be mentioned in all experiments • Instead of “Over-expressed Transgenic Lines”, transgenic lines overexpressing GRPL1 or overexpression lines may be a better term. • In Fig 3 B and C, over expression line 1-4 did not perform better than Wild type. so this should be mentioned instead of generalizing the observation. After how many days the survival % under salt stress was recorded? Should be mentioned in text. • Line 350-353: without understanding of the biological function of GRPL1 , these are too presumptive statements. • Lines 397 -406 these lines should be shifted to section 1 “Global Gene …..Abiotic Stresses” of results. • Lines 410 -459: Discussion of DEGs is not required as these results might have been discussed by the authors of SRA study. Here authors should focus on how many genes they short listed infact a list of genes should have been more appropriate (in results). And then they can discuss validation of their selected gene GRPL1 by qRT-PCR (lines 459-462). • Line 474-476: Authors have discussed propidium iodide staining for recording no of cells however there is no data presented. Similarly Line 528-529 electrolyte leakage observation under salt stress is mentioned however no data is given. • Conclusion can be made more specific and short. Reviewer #2: Reviewer 1 The author identified a gene GRPL1 which was induced by multiple stresses by querying transcriptome results of multiple stresses. The transgenic Arabidopsis was obtained by overexpression of GRPL1. It was proved that GRPL1 can improve the resistance to high salt, drought and heat stress through resistance identification and analysis of various physiological indexes. It is a new idea to identify genes involved in multiple stress resistance processes. However, the quality of the data in this paper is not high, especially the pictures are not standardized. It is difficult to understand that GRPL1 is regulated by ABA2 gene. Therefore, this paper can not be published in the PLOS ONE at present, and we hope to revise it. Suggestions for revision are as follows: 1. Why didn't we use GRPL1 mutants for phenotypic analysis. 2. When analyzing transcriptome data, the reason why GRPL1 gene was chosen is not clear. 3. Paragraph in P303-308 is too short, so it is suggested to combine with other results. 4. The discussion is too long. 5. There are conjectured results in the conclusion part, which are not supported by data, so it is suggested to remove them. 6. Fig2C is lack of scale, the size of pictures is different, and there is no clear picture of salt tolerance. 7. Fig.4A is not clear. 8. The primers of Tables 1 were incomplete and there were no QRT PCR primers. 9. ABA2 gene is alcohol dehydrogenase. It is difficult to understand why ABA2 mutation affects GRPL1 expression, because ABA2 is not a transcription factor. ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. 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Revision 1
13 Nov 2020 Author Response Reviewer 1: 1. There is too much of redundancy in introduction section. There are many sentences with similar message or meaning, these needs to be removed. The aim of the study needs to be clearly mentioned. We thank Reviewer 1 for highlighting the redundancy in introduction section. We revised introduction section of the manuscript and also clearly included the aim of this study. 2. There is one study on identification and characterization of At4g18280 (here named as GRPL1) ‘Overexpression of a Novel Component Induces HAK5 and Enhances Growth in Arabidopsis’. Adams et al. 2013. The study suggested the role of this gene in pot deficiency and salt stress response. This ref is missing in the whole manuscript. We thank Reviewer 1 for his suggestion, we cited the work of Adams et al., 2013 in the revised version. 3. It would be better to mention the plant /crop to which the SRA data sets belong. The reason or criteria for selecting only these three data sets out of many available for analysis should also be mentioned. All the RNA-seq datasets belong to Arabidopsis thaliana, this information was added to the material and methods section (line83). A number of facts influenced the selection of these datasets, for instance, we wanted to have datasets that contain at least two or three biological replicates, the samples used should be of similar developmental stage, ideally the level and duration of the specific abiotic stress should be similar, and also our limited computational resources. We added this information in the methodology section (line 83-85). 4. Line 159-160. “leaves of three weeks old were excised to analyse the standardized water content”. What is the meaning of standardized water content?? We thank Reviewer 1 for pointing out this confusion. By standardized water content we meant “initial water content”. However, to make it clearer, we corrected and modified the water loss analysis methodology (line 152-158). 5. Line 187-192: As floral dip method is well known method, instead of detailed description just reference is sufficient to mention the transformation method used. Infact the very ref for floral dip is missing. The details of promoter cloning are also absent. We agree with Reviewer 1, as the floral dip is a commonly used and well-known procedure, we excluded the detailed description (line 187-191). We added details of promoter length and cloning strategy in the methodology (line 110-113). 6. There is repetition of methods in “Plant Material, Growth Conditions and Stress Treatment” and “Measurement of Physiological and Biochemical Indexes under Salt, Drought and Heat Stress”. These can be combined under one heading. As suggested by the Reviewer 1, we combined both “Plant Material, Growth Conditions and Stress Treatment” and “Measurement of Physiological and Biochemical Indexes under Salt, Drought and Heat Stress” sections under one heading of “Plant Material, Growth Conditions and Stress Response Analyses”. 7. There is discrepancy in the number of days for drought stress imposition (water with holding) in Line no 156 (20 days), 231 (32 days) and 358 (28 days) . We thank Reviewer 1 for pointing out these typos, the correct number of days is 20. We did corrections in line no 231 (line 254 now and 156 (line 150 now). Result section: 8. The authors described GO analysis of DEGs and next section started analyzing expression of AT4G18280 (GRPL1) which is too abrupt and requires explanation as to how the authors zeroed in on AT4G18280 (GRPL1). Moreover the first section “Global Gene …..Abiotic Stresses” details can be made shorter. We thank Reviewer 1 for this valuable recommendation, we modified this section and also added more detailed description of selecting GRPL1 as a candidate gene (line 173-180). 9. Again there is redundancy in few lines of section 1 and section 2 of results. As recommended by Reviewer 1, we combined section 1 and 2 of results 10. Line no. 277 PEG induces osmotic stress not drought stress. Moreover there is no distinction in the Table (Fig.1a) which is drought and PEG imposed stress. Correction was made to line no 277 (182 now) by changing “drought” to “osmotic”. Distinction was made to the table which now shows the drought and PEG imposed stress. 11. Line 295: First, the overexpression was analyzed at the… We modified the sentence as suggested by Reviewer 1 (now line no 198) 12. Fig. 2C should have scale bar for comparison of size. For Fig. 2D data from how many plants was recorded (replicates)? In general no of plants /replicates should be mentioned in all experiments We put the scale bar on each picture. For Fresh weight experiment, we weighed 10 plants for each genotype. We added this information in the figure legend and methodology section. 13. Instead of “Over-expressed Transgenic Lines”, transgenic lines overexpressing GRPL1 or overexpression lines may be a better term. As per the recommendation of Reviewer 1, “Over-expressed Transgenic Lines” was replaced with “overexpression lines” throughout the text. 14. In Fig 3 B and C, over expression line 1-4 did not perform better than Wild type. so this should be mentioned instead of generalizing the observation. After how many days the survival % under salt stress was recorded? Should be mentioned in text. AsReviewer 1recommended, we added the information of line 1-4 not performing better than WT plants (232-233). The survival percentage was recorded after two weeks of exposure to salt stress. We modified the text as well (line no 199). 15. Line 350-353: without understanding of the biological function of GRPL1, these are too presumptive statements. We agree with Reviewer 1 and toned-down our statement as below: “Although the GRPL1-mediated tolerance mechanism is still elusive, there is a possibility that” (line 234-237) 16. Lines 397 -406 these lines should be shifted to section 1 “Global Gene …..Abiotic Stresses” of results. We removed the redundant and out of the context part of discussion, to give more focus to significance of GRPL1 in stress tolerance 17. Lines 410 -459: Discussion of DEGs is not required as these results might have been discussed by the authors of SRA study. Here authors should focus on how many genes they short listed infact a list of genes should have been more appropriate (in results). And then they can discuss validation of their selected gene GRPL1 by qRT-PCR (lines 459-462). We thank Reviewer 1 for this suggestion and modified the discussion part accordingly. 18. Line 474-476: Authors have discussed propidium iodide staining for recording no of cells however there is no data presented. Similarly Line 528-529 electrolyte leakage observation under salt stress is mentioned however no data is given. We thank Reviewer 1 for pointing out this mistake. We removed the description related to propidium iodide and electrolyte leakage under salt stress. 19. Conclusion can be made more specific and short. As suggested by Reviewer 1, we modified and shortened the discussion part (413-418) Reviewer #2: The author identified a gene GRPL1 which was induced by multiple stresses by querying transcriptome results of multiple stresses. The transgenic Arabidopsis was obtained by overexpression of GRPL1. It was proved that GRPL1 can improve the resistance to high salt, drought and heat stress through resistance identification and analysis of various physiological indexes. It is a new idea to identify genes involved in multiple stress resistance processes. However, the quality of the data in this paper is not high, especially the pictures are not standardized. It is difficult to understand that GRPL1 is regulated by ABA2 gene. Therefore, this paper can not be published in the PLOS ONE at present, and we hope to revise it. Suggestions for revision are as follows: 1. Why didn't we use GRPL1 mutants for phenotypic analysis. We did check two mutant lines (SALK_112442 and SALK_15781) for GRPL1, but unfortunately both mutant lines only showed a slight knock-down (~5-10%) of GRPL1. These lines have T-DNA insertion in 5′ and 3′ UTRs, respectively. Unfortunately, there are no available lines that has T-DNA insertion in the coding region. Therefore, we excluded them from further experiments. 2. When analyzing transcriptome data, the reason why GRPL1 gene was chosen is not clear. We thank Reviewer 2 for highlighting this point. We modified the section of results focusing on shortlisting of GRPL1 (line 173-180) 3. Paragraph in P303-308 is too short, so it is suggested to combine with other results. We thank Reviewer 2 for this valuable suggestion. We combined this section of results with the previous heading. 4. The discussion is too long. As recommended by the Reviewer 2, we modified the discussion part to make it more concise. 5. There are conjectured results in the conclusion part, which are not supported by data, so it is suggested to remove them. We modified the conclusion part as recommended. 6. Fig2C is lack of scale, the size of pictures is different, and there is no clear picture of salt tolerance. We thank Reviewer 2 for pointing this out, we added a scale bar on each picture of Figure 2C. We also updated the picture of salt tolerance (Fig. 4D) 7. Fig.4A is not clear. We understand the concern of Reviewer 2 but the objective of showing this picture was to show the difference in color. As from the figure one can see that the transgenic lines were still alive (greenish) compared to the wild-type plants (brownish). 8. The primers of Tables 1 were incomplete and there were no QRT PCR primers. We thank Reviewer 2 for pointing out this mistake, we updated the list of primers in supplementary Table 1. 9. ABA2 gene is alcohol dehydrogenase. It is difficult to understand why ABA2 mutation affects GRPL1 expression, because ABA2 is not a transcription factor. ABA2 gene is an alcohol dehydrogenase and mutation in this gene results in blockage of xanthoxin to ABA aldehyde, a key step in ABA-biosynthesis resulting in low ABA production. ABA positively regulates the induction of drought-inducible genes. The signaling mechanism is well-known. Briefly, ABA inhibits the PP2C-dependent negative regulation of the SnRK2 protein kinase (Umezawa et al., 2010, PCP), as a result the SnRK2 can phosphorylate its downstream target transcription factors (for instance ABFs) which in turn induces the target genes. The fact that the ABA-deficient mutant is unable to induce the expression of GRPL1 in response to drought stress suggests that ABA is required for the induction of GRPL1 and that it acts downstream of ABA. We added this information in Discussion section (line 402-409). Attachments Attachment Submitted filename: Response to reviewer comments.docx https://doi.org/10.1371/journal.pone.0244030.r002
23 Nov 2020 Decision Letter - Keqiang Wu, Editor PONE-D-20-26347R1 Identification and characterization of a novel multi-stress responsive gene in Arabidopsis PLOS ONE Dear Dr. Munir, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please submit your revised manuscript by Jan 07 2021 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, Keqiang Wu, Ph.D Academic Editor PLOS ONE [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: All comments have been addressed Reviewer #2: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: Line 58 full form of GRPs should be mentioned (when it is mentioned first time) Ilalics for gene names Line 65-67 may be deleted Method of calculation for qRT-PCR should be included in the expression analysis section Mention vector background for overexpression construct, Generation at which the transgenic lines were analysed Line 151 the word dessication should be replaced by dehydration or drought FIG 2C the legend mentions scale bar as 1 cm however the picture does not appear to be in same scale, hence should be otherwise removed Reviewer #2: (No Response) ****** 7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0244030.r003
Revision 2
1 Dec 2020 Author Response Response to Reviewers Reviewer #1 Line 58 full form of GRPs should be mentioned (when it is mentioned first time) We thank Reviewer 1 for highlighting this; the full form of GRPs is now mentioned in line 58. Ilalics for gene names All gene names have been italicized. Line 65-67 may be deleted Lines 65-67 have been removed. Method of calculation for qRT-PCR should be included in the expression analysis section. The qPCR quantification is mentioned in lines 101-111. Mention vector background for overexpression construct The vector background is mentioned in line 129. Generation at which the transgenic lines were analysed It is mentioned in line 136. Line 151 the word dessication should be replaced by dehydration or drought The dessication word is replaced by dehydration in the manuscript. FIG 2C the legend mentions scale bar as 1 cm however the picture does not appear to be in same scale, hence should be otherwise removed Although the scale bar represents 1cm, their lengths are different in both images, i.e. the WT’s scale bar of 1cm has an actual size of 0.489cm whereas the one for transgenic plants is 0.351cm. Attachments Attachment Submitted filename: Response to Reviewer R1.docx https://doi.org/10.1371/journal.pone.0244030.r004
2 Dec 2020 Decision Letter - Keqiang Wu, Editor Identification and characterization of a novel multi-stress responsive gene in Arabidopsis PONE-D-20-26347R2 Dear Dr. Munir, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Keqiang Wu, Ph.D Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0244030.r005
Formally Accepted
9 Dec 2020 Acceptance Letter - Keqiang Wu, Editor PONE-D-20-26347R2 Identification and characterization of a novel multi-stress responsive gene in Arabidopsis Dear Dr. Munir: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Professor Keqiang Wu Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0244030.r006

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