Tapping into the Potential of Underutilized Niger (Guizotia abyssinica (L. f.) Cass.) Through Breeding and Biotechnological Tools
, Niranjan Ravindra Thakur 3, Sunil Shriram Gomashe 1, Munagapati Padmavathy 4 and Stanislaus Antony Ceasar 5,*
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsIn this review ms, authors summarize the potentials of few molecular breeding approaches to improve Niger yield. I have a few concerns that authors need to address, such as:
1. Figure 2 needs an explanation of which factors worked in 2015-2018 to fluctuate the yield and eventually reduced in 2018?
2. In line 90, need a citation to support "Due to its self-incomparable nature, it is difficult to develop an inbred line"
3. In line 131-161, the text is mostly descriptive, irrelevent to support the title. Repeated words as well. Please make it short and insightful to support the title and objectives.
4. In line 187-192, unnecessary content. please rewrite this section.
5. in paragraph 6.2 and 6.3, again I suggest to rewrite with insightful, short, and relevant findings.
6. A figure in 7.2 and 7.3 will add value to describe these sections.
7. In Future strategies, author needs to be concise and to the point. I recommend to rewrite and focus on Niger breeding and biotechnological progress.
Comments on the Quality of English Language
English edits needed
Author Response
Reviewer 1:
In this review ms, authors summarize the potentials of few molecular breeding approaches to improve Niger yield. I have a few concerns that authors need to address, such as:
- Figure 2 needs an explanation of which factors worked in 2015-2018 to fluctuate the yield and eventually reduced in 2018?
Author response: Thank you for your insight. I appreciate your comments on the data. The reason for declining the production has been addressed in the manuscript and supported with a reference.
- In line 90, need a citation to support "Due to its self-incomparable nature, it is difficult to develop an inbred line"
Author response: Thank you for the critical check. A new reference has been added.
- In line 131-161, the text is mostly descriptive, irrelevent to support the title. Repeated words as well. Please make it short and insightful to support the title and objectives.
Author response: Thank you for your valuable feedback on this paragraph. We appreciate your comments. We have revised the section to make it more concise and relevant to the manuscript.
- In line 187-192, unnecessary content. please rewrite this section.
Author response: Thank you for your valuable feedback on this paragraph. We appreciate your comments on this section. We have revised this section to make it concise and more relevant to the manuscript.
- in paragraph 6.2 and 6.3, again I suggest to rewrite with insightful, short, and relevant findings.
Author response: Thanks for your remarks and valuable insight. We concise the 6.2 and 6.3 paragraph with more insights as per your comments.
- A figure in 7.2 and 7.3 will add value to describe these sections.
Author response: Thanks for your valuable suggestion to improve the contents. We have already created figure 7 to describe these sections. However, we didn’t cite the figure in the original submission. Figure 7 is cited in 7.2 and 7.3 in the revised manuscript.
- In Future strategies, author needs to be concise and to the point. I recommend to rewrite and focus on Niger breeding and biotechnological progress.
Author response: Thank you for the critical check. We have revised the “future strategies section” and made it more concise as per the suggestion.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsPlease check the attachment.
Comments for author File: Comments.pdf
Author Response
Reviewer 2:
The represents a comprehensive review for the potential of underutilized Niger through breeding and biotechnological tools. It provides important information for future research endeavors at enhancing application in Niger. The highlight of this manuscript is firstly presenting the urgent need to genetically improve the minor crop of Niger from biotechnological interventions to conventional breeding. However, there are a number of problems with the text that should be addressed.
Major Revisions
- Page 2, Figure 1. ‘Precision phenotyping’ includes ‘Field trial’, so the below Field trial’ and its picture should be deleted.
Author response: Thank you for your critical review of this manuscript. I appreciate your attention to detail regarding Figure 1.
To clarify, both high-throughput genotyping and precision phenotyping are included in the "selection, characterization, and screening" section. Additionally, at the same time the figure illustrates that neglected and underutilized species can be directly incorporated into breeding and field trials, as indicated by the multiple colorful arrows.
- Page 2, ‘Production volume of Niger seeds across India’ should expand to global such as Kenya, Uganda, Sudan, Malawi etc., otherwise the information is only useful for India.
Author response: Thank you for your insight. We have added references about the Ethiopian region, including information on Niger production and varieties cultivated in the Ethiopian region based on CSA. The newly added content is shown below.
“2. 1 Production volume of Niger seeds in Ethiopia
Niger seed production has a long history in Ethiopia and remains a significant crop, accounting for 50–60% of the country's oil crop output. According to Central Statistical Agency (CSA) of Ethiopia, over the last 18 years, its production has doubled, rising from 1.04 million quintals to approximately 3 million quintals. This improvement is likely due to the adoption of improved varieties and enhanced agronomic practices such as fertilizer application, timely planting, site selection, and effective field management [11]. This suggests potential to boost production and productivity by developing suitable varieties and improving agronomic practices though breeding and biotechnological tools.”
- Page 3, Line 78-84, please further analyze the reason for the production decrease of Niger during 2013 to 2023.
Author response: Thank you for your insight. I appreciate your comments. The reason for declining production has been addressed in the revised manuscript. The newly added content is shown below.
“From the year 2013, the production of Niger is decreasing, but in 2017, the production was slightly increasing and again gets decline. Niger has a high oil content of around 40%, with a valuable fatty acid composition. However, its high unsaturated fatty acid content leads to poor keeping quality, which limits its wider use. Factors such as a lack of government interest in tribal cuisine, limited genetic variation, susceptibility to diseases, lack of diverse varieties, shattering, self-incompatibility, less or low responsiveness to management inputs [8], and little research attention have mainly contributed to a decline in production and have mainly remained as an underutilized oilseed crop [9]. There is a need to increase the production of Niger.”
- Page 3, Figure 2, the data should be updated to 2023.
Author response: Thanks for your valuable comments and the data has been updated up to 2024 which also covers 2023.
- Page 4, Figure 3 lacks scientific significance and thus it can be removed.
Author response: Thank you for your valuable remarks. The figure illustrates the sibling procedure used to prevent accessions from crossing with each other. This germplasm has been conserved by Indian Government’s Indian Council of Agricultural Research-National Bureau of Plant Genetic Resources (ICAR-NBPGR) Campus, Akola, India, which is why it is categorized under the heading "Conservation of Niger Germplasm." So, we would like to retain this figure; kindly approve the same.
- Page 7, Line 252-282, it is not necessary to descript the details of Agrobacterium- mediated genetic transformation of Niger. Please concise to Murthy et al. method, acceptor material of UNS-4, the difference from other crops in genetic transformation, and Figure 5.
Author response: Thank you for your valuable comments. We agree with your suggestions. We focused on Murthy et al’s method on UNS-4 variety and made concise in the revision as per your suggestions. We have added more insights on the future direction of genetic engineering in Niger. We have also compared and discussed the protocol optimization for Niger transformation in future direction. The newly added content is shown below.
“Only one report is available so far on the Agrobacterium-mediated genetic transformation of Niger. Murthy et al. [41] reported the protocol for Agrobacterium-mediated genetic transformation of Niger using hypocotyl and cotyledon explants of UNS-4 variety. Seven-day-old seedlings were used as explants for transformation. The explants were cultured on MS medium with varying concentrations of 6-benzylaminopurine (BA) for shoot regeneration. Agrobacterium tumefaciens strain EHA101, with the binary vector pIG121Hm, was used for transformation. The T-DNA region contained nptII and hpt genes as selectable markers, and GUS as a reporter gene, driven by the CaMV 35S promoter [41]. Co-cultivation occurred for 3 days at 22°C. Following co-cultivation, explants were transferred to selection and regeneration media. After successful regeneration, the plants were acclimatized in a greenhouse [41]. Agrobacterium-mediated transformation of Niger using seedling explants is depicted in Figure 5.
In this study, the transformation frequency of cotyledon was 15%, whereas, the transformation frequency of hypocotyl was only 3% [41]. No other report in currently available on Niger transformation.
Future research should focus on developing high-throughput, efficient, and cost-effective transformation systems. This necessitates optimizing existing protocol, exploring novel gene-editing technologies like CRISPR/Cas9, and investigating alternative explant sources. To improve Niger genetic transformation efficiency, a multi-faceted approach is necessary. Expanding the range of explants tested beyond those used in Murthy et al. [ 41] is crucial. Different tissues may have varying responses to transformation, necessitating the optimization of tissue culture conditions for regeneration. This involves refining media composition, hormone concentrations, and culture parameters to improve shoot and root development from transformed explants. A comparative analysis of Agrobacterium strains is important due to their differing virulence and transformation efficiency. Exploring diverse plasmids with various selectable markers and replication origins will enhance transformation success. Additionally, testing different promoters for transgene expression is crucial for consistency. A systematic investigation of these factors will help identify optimal conditions for efficient and reliable Niger transformation.”
- Please exchange the parts of 6.2. Genetic transformation and 6.3. Molecular marker technology. In general, molecular marker technology is used to get inter breeding materials while genetic transformation to produce breeding materials. Furthermore, genetic transformation is logically more close to genome editing.
Author response: Thank you for your valuable comments. We have updated the sections, with 6.2 now focusing on Molecular Marker Technology and 6.3 on Genetic Transformation.
- Page 9, Line 332-355, please list the researches of MAS in Niger in a table to simplify the descriptions.
Author response: Thanks for the useful suggestion to strengthen the article. While searching the literature, we came to know that only a limited number of studies are available which may not be helpful for generating larger table. However, we have explained all the works in the text.
- Page 12-13, Line 461-505, this part seems not related to Niger. It’s suggested that either combined the methods to Niger, or delete them.
Author response: Thank you for your suggestion on CRISPR tools explained in these lines. Since the CRISPR tools are still far from reaching the Niger crop, readers of this Niger article (after publication) should be introduced to these tools. Also, reviewer-1 has approved keeping these and even asked to make figures for these. So, we would like to keep these sections. However, we have included the roles of base editing and prime editing for Niger improvement in the revised article and also cited figure 7 as per the Reviewer-1’s suggestion.
Minor Essential Revisions
- Too much keywords, please reduce to about five keywords.
Author response: Thank you for your suggestion on the keywords. Keywords are now reduced to five.
- References should be standardized. Latin names and genes should be italics. Some references with DOI, some without. Some title presents in capitalizing the first letter of each word and some in capitalizing the first letter of the first word. Some references list issue No., some without. Please unify them in one format.
Author response: Thank you for the critical check of references. Now the references section has been refined as suggested.
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsThe review manuscript deals with the use of biotechnological and classical tools to underline the potential of underutilized Niger (Guizotia abyssinica (L.f.) Cass.).
In this manuscript authors present different potential tools to improve and therefore help to give the potential of this species.
The idea is really interesting and originalbut lack originality. As examples some references are given with the same topic are already published and not cited in this manuscript.
https://doi.org/10.30574/wjarr.2021.12.2.0579
https://doi.org/10.1186/s43170-022-00121-7
https://doi.org/10.1016/j.biombioe.2012.03.025
Moreover, some illustrations are not specific for the topic, for example
Figues 1and 5 are applicable for any plant species... what is the "plus" for niger?
Figure 6 the interest of this figure in this manuscript???
Therefore, this manuscript, even presenting some part of interest, it needs deep modification and structuration to highlight novelty and originality.
Author Response
Reviewer 3:
The review manuscript deals with the use of biotechnological and classical tools to underline the potential of underutilized Niger (Guizotia abyssinica (L.f.) Cass.).
In this manuscript authors present different potential tools to improve and therefore help to give the potential of this species.
The idea is really interesting and original but lack originality. As examples some references are given with the same topic are already published and not cited in this manuscript.
https://doi.org/10.30574/wjarr.2021.12.2.0579
https://doi.org/10.1186/s43170-022-00121-7
DOI:10.20431/2454-6224.0603002
DOI:10.20431/2454-6224.0702003
https://doi.org/10.1016/j.biombioe.2012.03.025
Author response: Thank you for positive feedback and useful suggestions to improve the manuscript. The above references have been added in the revised manuscript.
Moreover, some illustrations are not specific for the topic, for example
Figures 1 and 5 are applicable for any plant species... what is the "plus" for niger?
Author response: Thank you for your insightful feedback on Figures 1 and 5. I appreciate your concern regarding their applicability to any plant species. We have specified the name “Niger” in legends to make it applicable for the focus crop.
To further clarify, while these figures outline general breeding strategies, they specifically highlight the unique aspects of Niger. For instance, Figure 1 emphasizes the integration of high-throughput genotyping and precision phenotyping tailored for Niger’s genetic resources, including neglected and underutilized species. This approach is crucial for developing resilient cultivars that address local agricultural challenges.
Figure 5 illustrates the Agrobacterium-mediated transformation process specifically adapted for Niger, showcasing steps that are essential for successful genetic transformation in this species. The figure also indicates how neglected and underutilized species can be directly utilized in breeding efforts, as represented by the colorful arrows.
We hope this clarification addresses your concerns regarding the "plus" for Niger in these illustrations.
Figure 6 the interest of this figure in this manuscript???
Therefore, this manuscript, even presenting some part of interest, it needs deep modification and structuration to highlight novelty and originality.
Author response: Thank you for your insightful feedback on Figure 6. As explained, these figures may help readers of Niger article to understand the basic principles and apply them in the future since tools like CRISPR/Cas are too far from reaching Niger crops. Also, reviewer-1 has approved keeping these and even asked to make more figures for CRISPR tools like base and prime editing. So, we would like to keep these sections. However, we have included the roles of base editing and prime editing for Niger improvement in the revised article as per Reviewer-1’s suggestion.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsFigure 3 legend is not self-descriptive. Please do it so.
Text in all figures eg fig 1, 2 should be same front size and make it readable.
Others are looking fine. Thank you for your edits.
Comments on the Quality of English LanguageEnglish needs to improve throughout the ms
Author Response
Reviewer 1:
Comments and Suggestions for Authors
Figure 3 legend is not self-descriptive. Please do it so.
RESPONSE: Thanks for the suggestion. We have expanded the description for figure 3 in the revised article to make it self-descriptive.
Text in all figures eg fig 1, 2 should be same front size and make it readable.
RESPONSE: Thanks for the suggestion to improve the figures. We have revised the font sizes and kept the same font size in these figures. We hope these are now readable. Since Figure 1 has many small icons and images, we cannot increase the font size beyond this.
Others are looking fine. Thank you for your edits.
RESPONSE: Many thanks for the positive feedback and recommending our article for publication. We thank you for your time to check the article carefully and offer many suggestions to improve it.
Comments on the Quality of English Language: English needs to improve throughout the ms
RESPONSE: We have tightly proof-read the article once again and fixed any English errors.
Reviewer 3 Report
Comments and Suggestions for AuthorsDear Authors,
Thank you for considering carefully all remarks.
The concerns have been addressed according the recommendations. The manuscript is now acceptable
Author Response
Reviewer 3:
Comments and Suggestions for Authors
Thank you for considering carefully all remarks.
The concerns have been addressed according the recommendations. The manuscript is now acceptable
RESPONSE: Many thanks for the positive feedback and recommending our article for publication. We thank you for your time to check the article carefully and offer many suggestions to improve it.
