Regulation on the Induction of Protocorm-like Bodies and Callus in Dendrobium officinale
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe submitted paper aims to improve the in vitro propagation of Dendrobium officinale through callus proliferation and PLB induction. The paper contains insights and some interesting findings. However, improvements are necessary to reach the publishable version.
1- Introduction: The clarity of lines 86-88 should be improved
2- Materials and methods: The first time that MS medium is cited (line 138), the authors should use the full citation (Murashige and Skoog), and MS should be put into brackets. Was the complete MS formulation (macro- and micronutrients and vitamins) used? The agar brand should be cited. Please use the term potato extract. and banana homogenate. The light intensity should be expressed as photosynthetic photon flux density (PPFD), and the kind of lamps used should be specified. I suggest recasting the Materials and Methods section by reducing the repetitions (e.g. the authors could have one paragraph to describe the explant preparation, including the explanation on how the explants were taken from the in vitro seedlings; then the basal medium used for all the experiments should be specified and in different sub-paragraphs, the authors could explain the specific additions for the different trials; the same could be done for the culture conditions and in the last paragraph were statistical analyses are explained, the number of explants for each experiments and repetitions could be specified).
3- Results- I found this section redundant; I suggest condensing it and avoiding repeating concepts already stated in the text, particularly in the Materials and methods section.
4- Figures- I suggest splitting Figure 1A from Figure 1 B and C. In addition, the legends of all figures should specify the conditions of each experiment.
5- Discussion- The authors should avoid casting a too speculative discussion by emphasizing the obtained results.
Author Response
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Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsI commend the authors for their dedication to the experiment and the preparation of the manuscript. Below are some observations:
Line 28: “… 2000 lux …” — I suggest that the authors replace light intensity from “lux” to “Photosynthetic Photon Flux Density (PPFD, in µmol/m²/s)”;
Line 34: add keywords that are not already in the title;
Line 67: the term “in vitro” is not italicized — please check its formatting throughout the manuscript;
Lines 136–137: add the duration of the disinfestation process;
Line 138: include the name and citation for the MS medium (Murashige & Skoog, 1962); also add this reference to the reference list;
Lines 147–148: specify the standardized plant size used in the experiment;
Line 164: “…dehydrated in a graded ethanol series…” — please specify this step, including the ethanol concentrations and the time spent at each concentration.
Author Response
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Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsThe manuscript submitted for review concerns the optimization of micropropagation of an important species Dendrobium officinale with ornamental significance, as well as medicinal and edible value. The work is well written; however, I have several comments:
In the Materials and Methods section, please clarify:
- the type of agar and its manufacturer;
- details regarding the form of potato and banana, so that the procedure can be reproducible;
- information on lighting: number and type of light sources, manufacturer, parameters, method of measurement (device used, distance of plants from the light source, and at what distance the measurement was taken). Ideally, the PPFD value should be provided.
In the Results:
1. in section 4.1, it could be expanded with examples of the effect of explant position on the plant in other species, not necessarily from Orchidaceae;
2. in section 4.2, it could be expanded with information on whether other auxins were used for the induction of PLBs and callus in orchids, and what the effects were.
Author Response
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Author Response File: Author Response.pdf
Reviewer 4 Report
Comments and Suggestions for AuthorsDear Authors,
I read the manuscript carefully and the study is relevant and also the experimental setup is clear; however, several points related to statistical analysis, experimental design, and interpretation need clarification.
The main issue concerns the statistical treatment of the data. Although three replicates are mentioned (Sections 2.2–2.6), it is not specified whether the values in Figures 1 and 3–5 represent mean ± SD or SE. In addition, one-way ANOVA is indicated (Section 2.7), but no post hoc test is specified, despite multiple comparisons being shown (different letters above bars). Please specify the post hoc test used and define in the figure legends what the error bars and significance letters represent.
Regarding the effect of 2,4-D (Section 3.2), the conclusion that higher concentrations promote PLB induction is based on a limited range (1.0–3.0 mg/L). While a trend is observed, this statement should be interpreted more cautiously and not generalized. Similarly, temperature, light intensity, and photoperiod (Sections 3.3–3.4) are evaluated independently, but their interactions are not considered; therefore, statements identifying “optimal” conditions (e.g., 25/22 °C and 2000 lux) are not fully supported and this limitation should be stated.
Finally, the use of a single evaluation time point (40 days; Section 2.2) may overlook important differences in induction dynamics, given that developmental changes are already described at earlier stages (7–20 days; Section 3.1). In addition, some statements in the Discussion and Conclusions (e.g., “systematically reveals”, “key regulatory factors”) are stronger than supported by the experimental scope and should be moderated.
Author Response
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Author Response File: Author Response.pdf
