AFLP-Based Genetic Structure of Lithuanian Populations of Small Balsam (Impatiens parviflora DC.) in Relation to Habitat Characteristics
, Kristė Stravinskaitė 1, Edvina Krokaitė 1,*, Rasa Janulionienė 1, Tomas Rekašius 2,3, Algimantas Paulauskas 1, Vitas Marozas 4 and Eugenija Kupčinskienė 1
Round 1
Reviewer 1 Report
Overall, the good efforts were made by the authors. My comments on the manuscript for compliance for the acceptance of the manuscript.
1. AFLP, can be good markers system to understand genetic diversity the plant species with no sequence data. However, underestimate the genetic diversity due the its inability to resolve the recessive gene diversity, possibly be the reason of the low genetic diversity in the present case.
2. Any justification of the high gene flow in the geographically separated population due to the natural barrios and various anthropogenic activities.
3. What was the outcome of the previous studies (Lithuanian populations of I. parviflora have been already examined by randomly amplified polymorphic DNA (RAPD) [30] and inter-simple sequence repeats 82 (ISSR) [33]). Besides, the coverage do we get the additional information to understand and stargaze the invasiveness. Because, due to availability of cost effective, gel free methods are preferred to avoid the multiple efforts on the similar projects.
4. Any justification of 17 genetic populations in the between three different analysis (Bayesian structure analysis, Coordinate, AMOVA analysis )
5. Overall the efforts are required to maintain the clearly of the data interpretation and presentation.
Author Response
Dear reviewer,
We would like to thank you very much for your professional and positive review of our article.
Our answers are as follows:
English of the article has been revised.
Introduction has been updated providing wider information about research of invasive woody species in Lithuania (rows 62-64) and about our main outcomes from the previous study (rows 93-101).
Result and discussion chapters underwent major revision in many places.
- In the discussion, rows 534-562, 572-575 were added explaining various possible reasons of low variability of populations at AFLP loci.
- Assumptions about the most distinct population in PCoA plot were provided in the rows 547-552.
- Our AFLP loci assessment was done by capillary electrophoresis as it is mentioned in the methods (devices and labelled primers): rows 180-184 and Table 1.
Our main outcomes from the previous study were included in the introduction (rows 93-101).
In case comparisons were absent in the initial paper, present study results were compared to the former results in the discussions adding following rows: 537-539, 626, 640-643, 645-648, 709-710, 745-747.
- Assumptions concerning the number of Bayessian gene clusters, rows 609-612.
- Data presentation and clarification has been done moving excessive for the discussion results (in many places) to the result chapter also adding new assumptions and comparisons in respect to the former research (our and the other authors). In addition, Table 4 was created, moving results from the plain text into the table concerning statistical evaluations of three biotopes according to EIV values and coverage.
Please see the attachment and please, have a look to the renewed paper.
The insights you provide are very useful for improving the current article and will be very valuable getting ideas for our future work and its presentation.
Thanking once more for your time spent reading our article and wishing best.
On behalf of the article authors
Eugenija Kupčinskienė
Author Response File: 
Author Response.docx
Reviewer 2 Report
-Lineas 538-540. It is that there is no robust reason/hypothesis why this has to happen. The possible relationship between both variables is not well supported in the manuscript.
- There are several paragraphs in the text, especially in the discussion section, that should be included in the results section or should be eliminated.
- The values obtained in the PCoA are low, so their interpretation must be done with caution, as they indicate that there is no clear pattern or trend.
- Please indicate in the method section what is the reference for the names and authors of the species?
- Some more specific comments appear in the manuscript.
Please see attached file.
Comments for author File: 
Comments.pdf
Author Response
Dear reviewer,
We would like to thank you very much for your comprehensive, concise and sincere positive review of our article.
Our answers are as follows:
English of the article has been revised
1. Former rows 538-540. The sites with the most similar assemblages of herbaceous plant species (JUp, Pre, and KAS) (Figure 5) did not coincide with the sites of the closest genetically populations of I. parviflora (Pal, Zag, Sve, and KMa, or DRa, KVa and Nid) (Figure 2).
Present paper: rows 121-130, 663-669, and 756-758 have been added and revised talking about the bases for comparisons of molecular and phytocenological data.
2. Removal of some paragraphs from the discussion chapter to result chapter was extremely useful and it has done for all sentences defined by you. In the present paper underlined by you rows have been corrected:
Rows 57-66 species were arranged alphabetically.
Introduction has been updated providing wider information about research of invasive woody species in Lithuania (rows 62-64) and about our main outcomes from the previous study (rows 92-101).
3. In the discussion rows 534-562 were added explaining various possible reasons of low variability of populations at AFLP loci.
4. Species names were checked according to the World Flora Online (it has been added to the methods, in the row 197) and as a consequence part of the names has been changed in the Figure 5.
5. Result and discussion chapters underwent major revision in many places as it is stated in the 2 answer. Data clarification has been done moving excessive for the discussion results (in many places) to the result chapter also adding new assumptions and comparisons in respect to the former research (our and the other authors) adding following rows: 537-539, 626, 640-643, 645-648, 709-710, 745-747. In addition, Table 4 was created, moving results from the plain text into the table concerning statistical evaluations of three biotopes according to EIV values and coverage.
Please see the attachment.
Finalising our answer we should state, that in addition to your big input into present paper, your insights provided will be very important carrying out our future research.
On behalf of the authors of the paper
Eugenija Kupčinskienė
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
Based on the inputs provided by authors, I do not have further questions. Revision may be accepted for the publication.
Reviewer 2 Report
I consider that the manuscript has been corrected and can be accepted for publication
