Establishment and Optimization of Micropropagation System for Southern Highbush Blueberry

Round 1

Reviewer 1 Report

Overall, the manuscript present considerable results but some points need to be improved.

·         In the abstract and keywords: please explain some abbreviation (eg, IBA, WPM).

·         The introduction is well written, but the objectives of the study must be clearer and in relation with the results obtained.

·         Figure 1 in Material and methods you should explain this figure and interpretate and mention in the text, it is better to start the paragraph with text then the figure

·         L126: “running water” you mean tap water?

·         L162: please remove the full name and keep just abbreviations.

·         Statistical analysis: please explain more the statistical analysis did you use ANOVA? Did you test the normality of your data? how much replication you have for each experimental?

·         Figure 2: your essay here is missing the unprocessed control (it is very important to compare your results)

·         Please in the caption of figures 3, 4 and 7 explain the abbreviation

·         Figure 5: teterment?

·         In the conclusion you shouldn’t repeat the results you should conclude from your results please improve the conclusion

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Reviewer 2 Report

Dear Authors,

You can find the evaluations of your work in the text below or in the attached document.

Kind regards

Abstract:

- In the last sentence of this section, the contributions of this study to the literature can be briefly mentioned.

Line 17: ….the in vitro culture, a modified woody….

Line 18: …in woody plant medium with (NH4)2SO4 facilitated the proliferation of blueberry shoots…

Introduction:

- Latin words such as in vitro and ex vitro should be written in italics in the text.

Line 30: …..is associated with a reduced risk …

Line 35: …. market has generated an - increased demand….

Line 39: … allows for the rapid production…

Line 56: … Quinoa seeds soaked in a 20% sodium hypochlorite solution…

Line 77: …. a survival rate of 89.7% and a shoot formation rate …

Line 80: … Furthermore, the addition of indole-3-butyric acid (IBA)…

Materials and Methods:

- Why did you decide to use “4% NaOCl” in surface sterilization? Did you get it from a reference where this concentration works better for surface sterilization of southern highbush blueberry 'ZY09'? If you used a reference, you should cite it.

Line 110: …. (Step 109 4) Ex vitro rooting and acclimatization….

Results:

Line 204: The lowest contamination rate of 0.00% was observed…

Line 206: ….the highest differentiation rate of 93.33% was achieved…

Line 278: …fresh root weight, and a dry root weight…

Discussion:

- The data available is large, this section should be expanded using more recent references.

Line 346: ….is considered suitable for the ex vitro rooting of blueberries…

Conclusion:

OK

Tables:

- Since the tables (Table 1, 2, 3, and 4) describe the method, it would be better if they were given as a supplenetary file instead of in the text.

Figures:

- The position of Figure 1 is in the wrong place in the text. Figure 1 should be given after subtitle “2.6.2. Screening of growth regulators for root induction”.

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Reviewer 3 Report

The MS entitled “Establishment and optimization of micropropagation system for southern highbush blueberry” with authors Yuting Wang, Xiaoyun Zhang, Zhehao Jiang, Xiaolong Yang, Xiaojuan Liu, Xi Ou, Wei Su and Riyuan Chen presents interesting data but needs improvement to be published.

Vaccinium sp. consists of approximately 450 species and has a long history in Europe and America of use as food and medicine. They are considered one of the richest sources of antioxidants. Traditional approaches to growing blueberries are time- and labor-intensive. Plant tissue culture technologies open up new possibilities for germplasm maintenance. Using in vitro techniques, it is possible to obtain a large number of homogeneous plants in a short period of time.

Vaccinium corymbosum L. is one of the main fruit crops domesticated in the twentieth century. Selection of cultivars for research is based on scientific and commercial value. Many studies have investigated the in vitro cultivation of blueberries, some of them recommending different variants of modified WPM medium. Appropriate selection of cytokinins determines successful micropropagation.

The article follows the order required by the journal: Introduction, Materials and Methods, Results, Discussion, Conclusions. Abstract is informative; the Introduction is good and to the point to the purpose; the purpose of the study should be more clearly stated. Over 57% of the cited literature is from the last 10 years. However, the quality of the manuscript will improve if the authors use clearer and more precise sentences, as they are difficult to understand. It is essential to improve Material and Methods, and data presentation.

I have some comments on the text:

L13 - Throughout the manuscript ex vitro and in vitro should be in Italics.

L106 – In my opinion, the sequence of individual points should be the following: Plant material; Decontamination of explants; Media composition (to describe all variant nutrient media in appropriate order, removing repetitions as in the lines: L147, L157, L168, L191 etc); Conditions for in vitro cultures and plant adaptation ….etc.

L142 – Please, write the Мanufacturer of the nutrient media.

L177 – Why do the authors use such high humidity and how is it achieved, Please, describe.

L210 – The text is not clear on how the Induction rate is calculated.

L223 – I would recommend the authors of try to combine the individual graphics into one or 2 such.

L239 – I would recommend the authors of try to combine the individual graphics into such of one.

Why the authors report the Fresh and Dry weight indicator - the meaning of these indicators is not explained in the discussion.

L283 – In the legend below the figure, Shoot parameters are described, but in the graphic – Root parameters.

L315 – The sentence is not clear.

L326 – Please, revise this passage, avoiding repetitions.

L466 – Please write in correct form.

 L363 - The authors conclude that they have developed a protocol. When a protocol is published it is correct to describe it step by step in the Results. Please, indicate how many plants are produced from one explant?

The authors should carefully and critically review publications on the subject and highlight more clearly their novel contribution.

Author Response

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Author Response File: Author Response.docx

Reviewer 4 Report

The study of the influence of the conditions of sterilization of the initial extrapalants, the composition of nutrient media and their hormonal composition is important for the development of a protocol for in vitro micropropagation of plants. In the presented work, this was done for blueberries, as a producer of active secondary metabolites and a widely used food product.

The work was carried out using methods for assessing the morphological and physiological state of plants. The data is presented correctly and clearly for readers.

Small remarks:

- line 22 of the text - the abbreviated name of the hormone is used;

- figure 6 - it is desirable to indicate the substrates used (similar to other figures);

- line 336 - data on the differentiation of the kidneys are not presented in the manuscript (see the results section), therefore, the conclusion about the effect of thidiazuron on this process needs to be corrected.

Author Response

请参阅附件。

Author Response File: Author Response.docx

Round 2

Reviewer 3 Report

Thanks to the authors for considering my comments.