Evaluation of Factors Affecting Direct Organogenesis in a Somatic Tissue Culture of Sinningia speciosa (Lodd.) Hiern
Round 1
Reviewer 1 Report
The paper is clearly and concisely written, and the topic is interesting and current. Some minor revisions are needed as follow bellow:
Line 9-11: In this study, the influence of genotype, concentration of thidiazuron (TDZ), and explant position on the culture medium in organogenesis in a somatic tissue culture of two gloxinia cultivars … the verb is missing in the sentence
Line 69-91: … g L–1, m–2 s–1 …. Use the superscript … should be applied through the text
Line 96: adventitious shoots/total number of explants) x 100%) … one more bracket is missing after 100%
Line 81: adventitious shoot number were recorded ... please specify how big (long) shoots were (approximately) in order to be counted
Line 94: Please specify at this place what do you consider as a treatment! Is it C x T x P combination
Line 95, Figure 1 a): Percentage of explant regeneration vs. shoot formation frequency … consider to uniform the terms (this is the same, isn’t it?)
Line 179-185: I suggest forming the section Conclusions
Author Response
Please see the attachment.
Author Response File: 
Author Response.docx
Reviewer 2 Report
The studies presented in the manuscript entitled “Evaluation of factors affecting direct organogenesis in somatic tissue culture of Sinningia speciosa” refers to the establishment of effective method of micropropagation via organogenesis of two gloxinia cultivars ommiting callus phase. The authors obtained shoot regeneration in the presence of thidiazuron (TDZ) directly on the cultured in vitro mature young leaves as explants. The genotype, TDZ concentration, position of explant on the medium etc. significantly affected the shoot regeneration frequency. It was found that the supplementation of the medium with 3.5µM TDZ increased the effectiveness of organogenesis and that 4.0µM TDZ resulted in the higher number of shoots per explant. Obtained shoots were able to regenerate roots on the medium enriched with 0.1 mg L-1 NAA and finally develop into the plantlets similar to the donor plants. In my opinion the presented results are interesting, and the manuscript text is well organized and easy to read. The obtained results are clearly presented, properly analyzed and interpreted. Nevertheless, some additional information should be include in the text before publishing:
- Introduction
Page 2, lines 59-60
Explain why the development of a callus-free reproduction method is so important to the breeding of gloxinia? Somaclonal variability is more likely to occur in callus cultures, which promotes diversity and is desirable in breeding. Is the goal in yours study to reduce this variability by omitting callus phase in the culture?
Add citations which demonstrated the attempts to obtain gloxinia cultures without callus phase you are writing about, please.
- Materials and Methods
- Plant material
Page 2, line 79
I suggest add the photo to show how looked like explants from adaxial and abaxial sides.
2.2 Root Formation and Plantlet Acclimatization
Page 2, lines 86-92
Add more detailed information, please how many shoots were obtained from each variant, and how many of them were rooted and acclimatized finally in greenhouse.
- Results and Discussion
3.1. Adventitious Shoot Formation from Leaf Explants
Page 5, line 128
Correct the description of the interaction from “A x T x P” to “C x T x P”
3.2 Root Formation and Plantlets Acclimatization
Page 2, lines 175-178
I suggest to include into manuscript the data concerning rooting and acclimatization, if possible and to develop a little the discussion in this point. To what extent were the other protocols less effective as compared to the presented one for this species? It would be also valuable to add the photos both the rooted and the successfully obtained regenerants.
A few detailed corrections have been included in the manuscript text.
Comments for author File: 
Comments.pdf
Author Response
Please see the attachment.
Author Response File: Author Response.docx
