In Vitro Shoot Regeneration and Callogenesis of Sechium compositum (Donn. Sm.) C. Jeffrey for Plant Conservation and Secondary Metabolites Product
, Arévalo-Galarza Ma. de Lourdes 1 and Castillo-Juárez Israel 4
Round 1
Reviewer 1 Report (Previous Reviewer 1)
Comments and Suggestions for AuthorsThe papar horticulturae-2950177 by Maria de la Luz et al about the tissue culture of Sechium compositum was carefully read again. Reading the author's responce and comparing the article before and after the revision, the author has made a good revision of the article. I think the manuscript could be accept to publish in Horticulturae.
Comments for author File: 
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Author Response
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Author Response.docx
Reviewer 2 Report (New Reviewer)
Comments and Suggestions for AuthorsBud multiplication and in vitro callogenesis of Sechium compositum (Donn. Sm.) C. Jeffrey from stem and leaf explants
The aim of this MS is develop an in vitro multiplication protocol from bud explants of Sechium compositum (Donn. Sm.) C. Jeffrey for regeneration, propagation and conservation of this wild species to avoid its extinction. As well as to establish a callogenesis protocol from leaves and stems explants in order to have alternative source of secondary metabolites in the future, such as cucurbitacins, phenolic acids and flavonoids of interest.
But some comments should be follow in this MS to be accepted by the journal:
I suggest changing the title to be:
In vitro shoot regeneration and callogenesis of Sechium compositum (Donn. Sm.) C. Jeffrey for plant conservation and secondary metabolites production
Please Paraphrase the abstract to be more clear and precise
The full name of plant growth regulators and its abbreviation as 6-benzylaminopurine (BA), should be written only when you mention them in the first time after that you have to use the abbreviation only without brackets.
Could put the botanical description of this plant, it is tree or herb or what and what is the edible or economical part of this plant species (I think its fruits), refer to this information in the introduction
How old are the plants when you take the explants (buds) for culture establishment and, refer to this in material and methods section
And if you take leaves and stem explant from the in vitro established cultures, refer to this in the experiment of callus induction
Introduction should be divided into 3-4 paragraphs each one speak about one point and the last one is the aim of study
2. Materials and Methods
Line 114-115 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 114 1.2 mg mL-1, it should be in mg L-1, please correct
Line 129: in which country the Sechium 128 P. Br. Germplasm Bank, refer in the text
2.1 plant material line 128- 136 and you have to add if possible the fertilization and irrigation of the tree (the source of explants, bud explants)
Line 175: which explant did you use and how many explants per replicate?
Line 186: Stem and leaf explants were taken, taken from what the in vitro established cultures or from the mother plant, if from cultures how old these cultures? How many explants per repetition
Minor revision
Comments for author File: Comments.pdf
Extensive editing of English language required
Author Response
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Author Response File: Author Response.docx
Reviewer 3 Report (New Reviewer)
Comments and Suggestions for AuthorsIn this manuscript, the authors investigated the effects of phytoregulators on in vitro tissue culture of the wild Schium, namely S. compositum. Such data will be important for the maintenace of such recalcitant wild species. However, there are many mistakes and wrong 　interpretation of the results. So, I cannot recommend to publish this manuscript in the present text.
Major points
(1) Line 284-286: the best treatments ofr number of shoots…..
My comment: Table 3 and Figure 1 indicate that the effects of BA are very variable, and there is no relationship between the concentration of BA and the phenotype obtained. It is quite curious why the best results (shoot height) was obtained on the medium of 1.0 mg/L BA but the worst results was obtained on the medium the 0.8 mg/L. The data showed there is no concrete effects by the BA. In contrast, TDZ showed concentration-dependent effects on the shoot multiplication and rooting. In this respect, S. compositum can response well against the TDZ but not BA.
(2) Figure 2.
My comment: As mentioned above, BA will be unsuitable for induction of shoot multiplication. Therefore, the data of Figure 2 will be meaningless. In addition, this data showed a contradictory data when compared with the Figure 1. In Figure 1, 0.1 mg/L BA did not show any roots and calluses. However, in Figure 2, 0.1 mg/L BA showed root and callus formation which are better than the control MS. Which are the correct?
(3) Table 4 and Figure 4
My comment: In my opinion, these two data are identical. So most of data should be deleted to avoid repetitation.
Minor points
(1) The authors use “callogenesis” and also “callus formation”. Are there any differences in these two words? I recommend the unified use (I prefer, “callus formation”).
(2) Line 29, twice more roots (80%) and a 14 reduction in the bud…
My comment: I cannot understand this sentence.
(3) Line 34, 4.89
My comment: Readers cannot understand this number. The authors should keep in mind that readers can understand outlines of the results from abstract.
(3) Line 82, studies by [17]; it is unusual citation. “by” should be deleted.
(4) Line 98, [27], plant
(5) Line 102, explant-based callogenesis protocols have not been studied.
My comment: which plant species have not been studied?
(6) Line 275, treatment 2 should be treatment 7?
(7) Table 3, the authors should mention the culture period. (How long cultured?)
(8) Line 336, C) hoot should be C) Shoot
(9) Table 4, Treatments 8 to 14 were obtained by culturing leaf explant, not “sheet”
Comments for author File: Comments.pdf
There are some minor irregular presentation in the text. A moderate revise or editing may be required.
Author Response
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Author Response File: Author Response.pdf
Round 2
Reviewer 3 Report (New Reviewer)
Comments and Suggestions for AuthorsThe authors revised and the manuscript is somewhat improved. However, several description should be still improved. In addition, several sentences are still obsucure for readers. Several my previous queries still remained to be corrected. So I cannot still recommend publication of this mannuscript in the present style.
Line 26-30
In this part, the results of 0.1 mg BA treatment were describe. However, these sentences include the results of two independent experiments. The reader will confuse the corresponding results were seeked in Figure 1 and Table 1. So the authors should clarly mention that these sentences originated from two independent experiments.
In addition, the numbers (such as 9.22 buds or 80% roots) are unclear. These values may indicate the number of buds per explant and the proportion of explants showing rooting.
Line 32
S. compositum should be italic.
Introduction part
There are several subheading in the Introduction part. It is unusual for most journals.
Line 99
S. compositum should be italic.
Line 154
10 L-1 is unclear. 10 g per L of soil?
Line 163-165
However, in the germplasm bank, our plant was established via seed since 2005 (321-05).
I do not understand the relationship between “our plant in the germplasm bank” and the experiment present in this paper. In addition, what is 321-05?
Line 302
The authors do not correct “treatment 2” even though in the response letter they mentioned it had been corrected!
Table3. Line 319
I cannot find the T£ in the Table 3.
page 14, line 57
“formed roots roots and”, remove one “roots”
page 17, line 124
I cannot agree the following sentences, “BA shows a dose-dependent trend” . In addition, line 128, “values” appeared two times, but I cannot understand the “value”. Value is which value? My opinion, the relative low level of both BA and TDZ can efficiently support the shoot formation and their growth. BA did not show any trends of dose dependency in Table 3.
Discussion part,
In several sentences, the authors referred the citation in an unusual manner.
page 18, line 162, certain authors
page 18, line 174, authors like [36]
page 18, line 185, authors like [22] etc.
These references should be corrected. Authors should be removed and first author’s name should be cited.
Comments on the Quality of English LanguageI recommend English editing.
Author Response
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Author Response File: Author Response.pdf
Round 3
Reviewer 3 Report (New Reviewer)
Comments and Suggestions for AuthorsOnly minor comments are provided. I recommend publication after correction of these points.
line 33, did you measure root volume? and where is the corresponding data present? In addition, I cannot find the “14%” in the Result section. The author must introduce 14% in the result section if they want to describe it in the abstract.
line 153, if the authors amended 10 kg of vermicompost per plant every 6 months, change “10 kg-1” to “10 kg”
Table 3, I cannot find “T” in the Table 3. Insert it into the left end of field row.
Comments for author File: Comments.pdf
Minor mistakes still exist. Please address them.
Author Response
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Author Response File: Author Response.docx
This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe paper by de la Luz et al studied the tissue culture characteristics of Sechium compositum under two stages experiment. The study aims to develop an in vitro multiplication protocol from bud explants, as well as a callogenesis protocol for Sechium compositum. The employed method is overall adapted and the data set sound.
Therefore I believe the dataset is underexploited since the study and its results are very little put into Horticulturae and ecological context, and it would be possible to dig much deeper into processes and thus to make the study interesting for a much larger audience.
Specific comments
Abstract:
1) The third sentence, What important knowledge lack about S. compositum? Do you mean there is a lack of awareness of the multifaceted importance of this plant, but people have destroyed the habitat of this plant? Or people overuse this plant? So this plant is at risk of reduced genetic diversity.
2)Replace " multiplication of seedlings" to "seedlings propagation" in the fifth sentence.
3) The keywords should include “tissue culture”.
Result:
4) In the page eighth, I think treatments 5, 6, 8, and 10 had a remarkable different callus and root formation than the control according to the figure 1.
Discussion
5)In the page 19, the first sentence of the first paragraph of discussion, you said previous work showed similar responses to the BA, TDZ, etc. You should cite at least some of them.
6)"Authors such as..." replace to the fisrt name of the first authors.
7)You should compare the culture conditions with other Sechium species in Discussion.
Comments for author File: Comments.pdf
Author Response
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Author Response File: Author Response.docx
Reviewer 2 Report
Comments and Suggestions for AuthorsThis manuscript tested the effect of different plant growth regulators such as BA, TDZ, and 2, 4-D, for the in vitro multiplication and callogenesis of Sechium compositum (Cucurbitaceae) and gave a conclusion " MS supplementing with 0.1 mg L-1 of BA will optimize the multiplication and regeneration of seedlings and 2.0 mg L-1 of 2,4-D was the best callus-inducing growth regulator". It has been known that BA and 2, 4-D were the growth regulator par excellence for the optimization of the induction, multiplication, or regeneration. The novelty of research work is average, but it is of great meaningful. The English language writing of this manuscript is fine and no issues were detected. Here are my suggestions as follows:
1. The marks A and B in the Figure and Figure 4 legend are inconsistent, please check.
2. The references are not in uniform format and need to be carefully checked. Please check all the reference.
The manuscript needs a minor revision before considering for accept to publish.
Author Response
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Author Response File: Author Response.docx
Reviewer 3 Report
Comments and Suggestions for Authors1. The Introduction needs to be simplified. And the authors devote a lot of space to describing the importance of the conservation of plant genetic resources before introducing Sechium compositum. It is not necessary to include 10 paragraphs in Introduction. Please simplify Introduction and write a total of 3-4 paragraphs.
2. In 2. Materials and Methods and 2.2.1. The literature cited by the author [42] is from 1962, and there is no latest study on concentration?
3. In 4 Discussion. The author's citation of references is not very standardized, such as in 4.1, line 2-3, ‘Authors such as [36]’, and others. Such descriptions are not standardized, and it is recommended to change.
4. In 5 Conclusions. The authors give the single BA concentration with the best proliferative effect (0.1 mg L-1). But in the abstract, the authors give four optimal concentrations (0.1, 0.2, 0.4, and 1.0), which is unknown for this difference and suggests correction.
5. Some of the references in the bibliography are a bit old and would like to be replaced.
Comments for author File: Comments.pdf
Author Response
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Author Response File: Author Response.docx
Reviewer 4 Report
Comments and Suggestions for AuthorsThe study shows results of shoot and callus induction in Sechium compositum. The introduction is adequate and informative. Methods needs serious revision. Results and discussion needs to be deeply revised. Please see comments in pdf version.
Comments for author File: Comments.pdf
English needs revision, improper wording is used making the revision difficult.
Author Response
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Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsI didn't receive a standardized point-to-point response to assess whether the article was revised or not as the previous suggestion.
Reviewer 4 Report
Comments and Suggestions for AuthorsI have revieved previous version of this manuscript and I see some progress, however paper still needs improvements. The results and especially discussion section needs solid revision. All detailed comments are given in pdf file.
Comments for author File: Comments.pdf
The English needs revision.
