Well-Known, Misidentified, or Unnamed? A DNA Barcode-Based Reassessment of the Lepidoptera Fauna of Cyprus Supported by Morphology

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

1. The article contains Unidentified Species, so it is best to list relevant morphological plates to facilitate the classification of related taxa for scholars to judge when reading.
2. Please carefully verify the accuracy of some publicly available genetic information. Currently, many publicly available genetic information are subject to human error due to weak taxonomic research by the providers, especially in some Families where errors may occur. Therefore, it needs to be treated with great caution.

Author Response

Thank you very much for taking the time to review this manuscript thoroughly. Please find the detailed responses below and the corresponding revisions/corrections highlighted/in track changes in the re-submitted files.

1. The article contains Unidentified Species, so it is best to list relevant morphological plates to facilitate the classification of related taxa for scholars to judge when reading.

Response 1: This is fundamentally a very important point; however, it would go far beyond the scope of our work on a checklist of Linnaean species to add morphological plates to unidentified species. The more than 100 species or clusters that cannot yet be reliably identified are intended to be analyzed and determined step by step in comprehensive revisionary studies. Moreover, the majority of the species that are currently not yet identifiable are also visually documented via BOLD. In addition, further sequencing is planned for the species that have not yet been genetically studied. We have, however, included a brief note on our further plans in the discussion.

1. Please carefully verify the accuracy of some publicly available genetic information. Currently, many publicly available genetic information are subject to human error due to weak taxonomic research by the providers, especially in some Families where errors may occur. Therefore, it needs to be treated with great caution.

Response 2: We also thank you for this point, which is relevant to many studies and often underestimated. We fully agree, and this is a key reason why we fundamentally rely primarily on our own verifiable data. Accordingly, the vast majority of sequences actually originate from studies conducted by the directly involved authors or from reliable and verified sources.

 

Reviewer 2 Report

Comments and Suggestions for Authors

Very important contribution to the Cypriot, and Mediterranean fauna. I cannot suggest any essential changes in the manuscript. However, the Authors can consider my short notes and suggestions made in "bubbles" (see: revised mscr.).

 

Comments for author File: Comments.pdf

Author Response

Thank you very much for taking the time to review this manuscript thoroughly. Please find the corresponding revisions/corrections based on your notes highlighted/in track changes in the re-submitted files.

Very important contribution to the Cypriot, and Mediterranean fauna. I cannot suggest any essential changes in the manuscript. However, the Authors can consider my short notes and suggestions made in "bubbles" (see: revised mscr.).

 

Reviewer 3 Report

Comments and Suggestions for Authors

The manuscript under review represents an example of a well-conducted and beautifully written work devoted to the lepidopteran fauna of Cyprus. The authors set out to verify and expand the list of lepidopteran species in Cyprus, relying on years of research and the use of DNA barcoding methodology. Although the researchers were able to obtain DNA barcodes for only about half of the island's species, the study results are impressive in the number of new discoveries for the island's fauna. More importantly, the use of DNA barcodes allowed the creation of a verified species list versus a list based on anecdotic evidence, as species identifications by previous authors are difficult to confirm or refute.

Nevertheless, it should be noted that this is the first step in creating a barcode library for Cyprus, as approximately half of the species remain unstudied, and a significant number remain unidentified because similar or identical barcodes are not still available in the BOLD database.

I have no fundamental comments about the manuscript, and I believe that after minor revision, the work can and should be published.

Minor comments

The work is not purely molecular. It includes not only DNA barcoding, but also a variety of morphological and morphology-based identifications. In principle, this is rather a positive aspect that was left out of the title.

Table S1 likely has some encoding issues. I don't understand what #N/A in column D means. Perhaps I or the authors should somehow change or fix the cell format in the Excel spreadsheet?

I couldn't find a reference in the manuscript to the important work:

John E., Wiemers M., Makris C., Russell P. 2013. The Pontia daplidice (Linnaeus, 1758) / Pontia edusa (Fabricius, 1777) complex (Lepidoptera: Pieridae): confirmation of the presence of Pontia daplidice in Cyprus, and of Cleome iberica DC. as a new host plant for this species in the Levant. Entomologist's Gazette 64(2):69-78.

This work, based on DNA barcoding, provides first evidence for presence of Pontia daplidice in Cyprus.

dx.doi.org/10.5883/DS-LEPI-210 CYPR. I was unable to open this reference.

A significant number of species remain unidentified. It's unlikely that they all represent new species. What are the plans for their analysis? Will they be distributed among groups and specialists for identification and possible description?

In comparison with Crete, it might be worth noting that the mountains on Crete are higher and reach the (sub)alpine zone, which supports high-altitude fauna of Western Asian origin and additional endemic species. In Cyprus, the mountains barely reach these heights, which is unfortunate, and the fauna is somewhat impoverished. This may also influence the level of endemism.

Author Response

Thank you very much for taking the time to review this manuscript thoroughly. Please find the detailed responses below and the corresponding revisions/corrections highlighted/in track changes in the re-submitted files.

The manuscript under review represents an example of a well-conducted and beautifully written work devoted to the lepidopteran fauna of Cyprus. The authors set out to verify and expand the list of lepidopteran species in Cyprus, relying on years of research and the use of DNA barcoding methodology. Although the researchers were able to obtain DNA barcodes for only about half of the island's species, the study results are impressive in the number of new discoveries for the island's fauna. More importantly, the use of DNA barcodes allowed the creation of a verified species list versus a list based on anecdotic evidence, as species identifications by previous authors are difficult to confirm or refute.

Nevertheless, it should be noted that this is the first step in creating a barcode library for Cyprus, as approximately half of the species remain unstudied, and a significant number remain unidentified because similar or identical barcodes are not still available in the BOLD database.

I have no fundamental comments about the manuscript, and I believe that after minor revision, the work can and should be published.

Minor comments

The work is not purely molecular. It includes not only DNA barcoding, but also a variety of morphological and morphology-based identifications. In principle, this is rather a positive aspect that was left out of the title.

Response 1: Many thanks for pointing to this problem. We have accordingly expanded the title

Table S1 likely has some encoding issues. I don't understand what #N/A in column D means. Perhaps I or the authors should somehow change or fix the cell format in the Excel spreadsheet?

Response 2: We have deleted #NV which in fact was unnecessary

I couldn't find a reference in the manuscript to the important work:

John E., Wiemers M., Makris C., Russell P. 2013. The Pontia daplidice (Linnaeus, 1758) / Pontia edusa (Fabricius, 1777) complex (Lepidoptera: Pieridae): confirmation of the presence of Pontia daplidice in Cyprus, and of Cleome iberica DC. as a new host plant for this species in the Levant. Entomologist's Gazette 64(2):69-78.

This work, based on DNA barcoding, provides first evidence for presence of Pontia daplidice in Cyprus.

Response 3: We have added this work to the manuscript

dx.doi.org/10.5883/DS-LEPI-210 CYPR. I was unable to open this reference.

Response 4: The relevant link is now active

A significant number of species remain unidentified. It's unlikely that they all represent new species. What are the plans for their analysis? Will they be distributed among groups and specialists for identification and possible description?

Response 5: These problems can only be resolved through further comprehensive studies, partly within the framework of generic revisions. Naturally, this work will be carried out in cooperation among specialists from different fields. In this regard, we have added an additional remark to the manuscript.

In comparison with Crete, it might be worth noting that the mountains on Crete are higher and reach the (sub)alpine zone, which supports high-altitude fauna of Western Asian origin and additional endemic species. In Cyprus, the mountains barely reach these heights, which is unfortunate, and the fauna is somewhat impoverished. This may also influence the level of endemism.

Response 6: In fact, only a small fraction of Crete’s endemic species is known from the (poorly studied) alpine zone, and none of the known species occurs exclusively in this altitudinal region. However, we have added a supplementary remark to the manuscript.