Flowering Time and Physiological Reaction of Dendrobium nobile Lindl in Response to TDZ Application

Round 1

Reviewer 1 Report

The reviewed article (ID - horticulturae-2009157) presents the results of research on the influence of Thidiazuron (TDC) on flowering time and selected morphological and biochemical parameters of Dendrobium nobile plants. The paper discusses aspects of flower production that may be of interest to Horticulturae Journal readers.

Although the structure of the manuscript does not raise any major objections in general, several issues require clarification before the work is ready for publication. Below are my comments in relation to the chapters of the manuscript:

Results: Statement on Lines 142-143 is incorrect - please check fig 1.

Discussion: Too many elements of the literature review and too few references to own research results in a confrontation with the results of other similar studies. Some longer excerpts are completely devoid of reference to their own results. For example, lines 225-257 or 264-277. I suggest editing this section a bit and referring to your own results while discussing.

Material and Methods: The authors used 3 concentrations of TDC in the study, which they describe in detail, however, the information on the control is imprecise. What was the control of how many plants in how many repetitions? Also, the designation of the control at work and in tables should be unambiguous - the authors use two designations (CK or 0mgL-1). I suggest clarifying the description in the methodology and standardizing these markings in the manuscript and tables, and entering zero values, not dashes, in the tables. While discussing the obtained results, I suggest referring them to the control, which will allow readers to better assess the effect of different concentrations of TDC.

There is often no information on how to present the results of specific analyzes/measurements. E.g. section 4.4 - how are the measurement results given (in cm). Similarly, the description of the other methods should be reviewed.

In Section 4.6 the authors write about the conducted interaction analyzes, however, in the whole manuscript there is not even one piece of information about possible interactions. There is no information on how many factor ANOVA was used in the study. Interaction analysis suggests at least 2 factors. However, the authors below Tables 2 and 3 points to a one-way ANOVA. Therefore, it was not possible to investigate the interaction.

Conclusions: The authors included elements of the methodology and should focus only on the main conclusions of the work. See lines 391-392.

In general: The authors introduce many abbreviations that should be explained  when first mentioned in the paper. In the tables, which should be understandable without the text of the manuscript also independently, the meaning of the abbreviations should be explained. The work contains many minor editing errors that require correction. I have made my comments in the pdf attachment of the manuscript and hope, they may be helpful during the proofreading.

Comments for author File: Comments.pdf

Author Response

Grateful for your careful and precise work. The revisions are currently made as requested. The current amendments are shown with track changes.

1.Results: Statement on Lines 142-143 is incorrect - please check fig 1.

We agree with you.There are significant differences among three treatments here.

2.Discussion: Too many elements of the literature review and too few references to own research results in a confrontation with the results of other similar studies. Some longer excerpts are completely devoid of reference to their own results. For example, lines 225-257 or 264-277. I suggest editing this section a bit and referring to your own results while discussing.

Thanks for a good suggestion. Simplify these paragraphs now. Some excerpts (Line 230-232, line 236-237) irrelevant are actually improper in Discussion, so they are deleted now. Some literatures (line 230-232,236-237,238-240,) are moved to Introduction Part. Attributive clause (line 268-270) is deleted because of its irrelevant encumbrance. The paragraph structure（from line 271-277）is logically readjusted and simplified.

3.Material and Methods: The authors used 3 concentrations of TDC in the study, which they describe in detail, however, the information on the control is imprecise. What was the control of how many plants in how many repetitions? Also, the designation of the control at work and in tables should be unambiguous - the authors use two designations (CK or 0mgL-1). I suggest clarifying the description in the methodology and standardizing these markings in the manuscript and tables, and entering zero values, not dashes, in the tables. While discussing the obtained results, I suggest referring them to the control, which will allow readers to better assess the effect of different concentrations of TDC.

Relative details about the control and sampling method are supplemented now. The control is changed to 0mg/L. All standard descriptions and SD are given in the tables and figures.

4.There is often no information on how to present the results of specific analyzes/measurements. E.g. section 4.4 - how are the measurement results given (in cm). Similarly, the description of the other methods should be reviewed.

All required information is finished in sections.

5.In Section 4.6 the authors write about the conducted interaction analyzes, however, in the whole manuscript there is not even one piece of information about possible interactions. There is no information on how many factor ANOVA was used in the study. Interaction analysis suggests at least 2 factors. However, the authors below Tables 2 and 3 points to a one-way ANOVA. Therefore, it was not possible to investigate the interaction.

Thank you for your careful review. We rechecked that part and corrected it now.

6.Conclusions: The authors included elements of the methodology and should focus only on the main conclusions of the work. See lines 391-392.

We agree with you. The mentioned part is deleted.

7.In general: The authors introduce many abbreviations that should be explained when first mentioned in the paper. In the tables, which should be understandable without the text of the manuscript also independently, the meaning of the abbreviations should be explained. The work contains many minor editing errors that require correction. I have made my comments in the pdf attachment of the manuscript and hope, they may be helpful during the proofreading.

Thanks for your thoughtful work. The comments　in pdf format helps us to find errors and　revise easily．We finished them as required.

Author Response File: Author Response.pdf

Reviewer 2 Report

In the introduction, the authors do not very convincingly lead to the choice of TDZ as a stimulant. There is a need for more coverage of the use and specifics of other stimulants. In addition, the authors rightly note that TDZ acts through auxins and other hormones. However, the study of the hormonal status of plants in this study was not undertaken. Why? Why do the authors look at the not very clear physiological characteristics indirectly associated with flowering, and not the level of endogenous hormones that directly regulate the transition to flowering?
 In all figures (except the second one) and in the tables there are no values ​​of standard (mean) deviations in the experiment. It is highly doubtful that such small values ​​are indicated by the authors as significantly different. It is necessary to add bars to the charts and SD values ​​to the tables. Why was Fisher's LSD test chosen?
In figure one, in the legend, I see the designations for 4 bars, and on the chart there are three of them in each group! It's puzzling - where are the values ​​for the control variant?
The measurement of chlorophyll and soluble sugars is described very poorly by the authors. This section requires a more detailed description of the principles of the method and the procedures to be followed. What is the repeatability of measurements of all physiological parameters?
Thus, the article raises a number of serious questions that require major improvements. From proving the consistency of the methods used, and explaining the lack of use of the study of hormone levels, to providing correct digital data of the experiment.

Author Response

Grateful for your careful and precise work. The revisions are currently made as requested. The current amendments are shown with track changes.

1.In the introduction, the authors do not very convincingly lead to the choice of TDZ as a stimulant. There is a need for more coverage of the use and specifics of other stimulants. In addition, the authors rightly note that TDZ acts through auxins and other hormones. However, the study of the hormonal status of plants in this study was not undertaken. Why? Why do the authors look at the not very clear physiological characteristics indirectly associated with flowering, and not the level of endogenous hormones that directly regulate the transition to flowering?

Generally, flowering switch in higher plants is profoundly affected by various pathways, including the growth regulator hormone, endogenous carbohydrate levels, and some autonomous pathways. We agree with the reviewer comment on hormone effect on flowering. Usually, the hormone status is traditional way to check the effects of the exogenous hormone. In our research, we aim to endogenous carbohydrate and chlorophyll pathway caused by TDZ as root-irrigation protocol. As to three physiological indexes, current studies provide convincing evidence to confirm their involvement in flowering gene expression. They are directly relevant to the floral induction. More details are shown in the Introduction section.

2.In all figures (except the second one) and in the tables there are no values ​​of standard (mean) deviations in the experiment. It is highly doubtful that such small values ​​are indicated by the authors as significantly different. It is necessary to add bars to the charts and SD values ​​to the tables. Why was Fisher's LSD test chosen?

Thank you. Your suggestion helps us to improve the data preciseness in the paper. Now the standard deviation is supplemented in the tables and figures. Our research is undertaken based on the hypothesis of the previous study, which carbohydrate, chlorophyll level involve in the floral differentiation of Dendrobium. LSD test is optimal for pairwise comparison of differences in means  on　condition　that　the experimental regime involves control group and treatment group.

3.In figure one, in the legend, I see the designations for 4 bars, and on the chart there are three of them in each group! It's puzzling - where are the values ​​for the control variant?

Because there is no floral bud in control group (0mg L^-1). In Figure 1, the value of each floral quantitative index in control group is zero. Only three not four could be shown on the chart.

4.The measurement of chlorophyll and soluble sugars is described very poorly by the authors. This section requires a more detailed description of the principles of the method and the procedures to be followed. What is the repeatability of measurements of all physiological parameters?

Thank you. We have supplemented the relative measurement information on chlorophyll and soluble sugar in Methods section. Measurement process and more details are described in the revision.

Thanks for your thoughtful work. We finished them as required.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

I approve the changes made by the authors to the article. The only remark on the dimension of the data in the text, for example, line 208 - 7.60, and line 216 - 43.6, but next to corrected 14.44. Increase the number of digits after the decimal point throughout the article, it should be the same everywhere, it is necessary to observe the bit depth of the data.

Author Response

Dear Reviewer，

        Grateful for your careful and precise work. The revisions are currently made as requested. The current amendments are shown with track changes(Blue track）.

       1.The only remark on the dimension of the data in the text, for example, line 208 - 7.60, and line 216 - 43.6, but next to corrected 14.44. Increase the number of digits after the decimal point throughout the article, it should be the same everywhere, it is necessary to observe the bit depth of the data.

       Answer：We had been increased the number of digits after the decimal point throughout the article and it added to two decimal places(like line 216-43.6 modified line217-43.64)

      Thank you again for your revision comments to make our manuscript more standardized.