Identification of Auchenorrhyncha Nymphs Using DNA Barcoding and Phylogenetic Analysis of the Most Common Genera Collected in Olive Fields
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe manuscript provides valuable field and genetic data on Auchenorrhyncha communities; however, several key areas require improvement to fully realize its potential, as highlighted in the original comments.
Comments for author File: 
Comments.pdf
Author Response
Comment 1. No mention of how sites were chosen (random? representative?) so please clarify site selection rationale and environmental variation.
Thanks a lot for this comment. Please check if is it more clear now.
Comment 2. The author(s) say PCR products were visualized, but size (~680 bp) should be stated.
It is mentioned in the previous paragraph. Although I added here too
Comment 3 Needs clarification-were all species equally represented?
We didn’t add anything here, since the number of adults of each species is presented in table 1 in the results, and the number of nymphs collected were in table 2. Some collected in high numbers and some only few specimens collected.
Comment 4 Do the author(s) mean "unidentified"? Psammotettix nymphs could not be resolved to species due to overlapping barcodes.
We rephrased
Comment 5 Psammotettix nymphs could not be resolved to species due to overlapping barcodes.
We rephrased the sentence
Comment 6. The Discussion section was found to repeat many of the results without synthesizing their broader implications. It is recommended to focus more on the study’s limitations, ecological interpretations, and potential future applications.
Thank you for the comment, you are totally right. Please check the discussion, since we added more comments, highlighted in red.
Comment 7. Haplotype data is only briefly mentioned in relation to Neophilaenus. This section could be strengthened by expanding the discussion to include insights into gene flow or potential local adaptation. Consider adding a paragraph on population structure and possible biogeographic barriers within Lesvos Island, which may influence the genetic differentiation of the studied populations.
We added more comments on this topic in the text.
Comment 8 The interpretation of congeneric vs. intraspecific divergence thresholds lacks references or sufficient justification. Please cite commonly accepted threshold values from relevant studies (e.g., COI divergence in Hemiptera or similar insect taxa) to support the delineation of species boundaries.
You are correct. We included thresholds from a study about Hemiptera.
Reviewer 2 Report
Comments and Suggestions for AuthorsThis article is highly relevant as it helps to identify immature individuals (and some females) of a taxonomic group that plays a significant role in plant pathogen transmission.
However, some aspects need further discussion before publication:
- Firstly, the authors collected four nymphs of Philaenus signatus and 66 of P. spumarius (see Table 1) yet did not conduct any molecular analysis to distinguish between the two species. This is despite the authors themselves pointing out that these are the main vectors of Xylella fastidiosa in Greece and the Mediterranean. I would like to know the reason for this omission. Secondly, how did the authors distinguish the nymphs of the two species when they did not collect any adults of P. signatus (see Table 1)? Furthermore, why is this information not included in Figure 1 or as supplementary material when the main objective of the article is to distinguish the immature stages of Cicadomorpha and Fulgoromorpha?
- The authors state that Jassargus (Pontojargus) kurdicus was recorded for the first time in Greece and that they were unable to find matches in the databases. However, they correctly included the accession number when they sequenced the specimens. Having collected eight nymphs which they state were identified morphologically, why did they not also include a specimen photo in Figure 1 or as supplementary material?
Some minor corrections are also needed:
- A section of the Methods and Materials (L103–109 and 112–115) is plagiarised from article #26, which is cited, but this section will need to be rewritten so that it is not copied verbatim.
- The English needs improvement, and contractions should be avoided.
- Fig. 2 is labelled 1 in the caption.
- The totals for adults and nymphs in Table 2 are in the wrong place.
- The authors refer to '53 species from the Malaise traps and 38 additional species identified morphologically from sweep nets' (lines 142–145). However, there are actually 58 species (and 13 genera).
- L239–242: This is a 'copy-paste' of part of the journal's guidelines and should be removed.
Author Response
Comment-1: Firstly, the authors collected four nymphs of Philaenus signatus and 66 of P. spumarius (see Table 1) yet did not conduct any molecular analysis to distinguish between the two species. This is despite the authors themselves pointing out that these are the main vectors of Xylella fastidiosa in Greece and the Mediterranean. I would like to know the reason for this omission. Secondly, how did the authors distinguish the nymphs of the two species when they did not collect any adults of P. signatus (see Table 1)? Furthermore, why is this information not included in Figure 1 or as supplementary material when the main objective of the article is to distinguish the immature stages of Cicadomorpha and Fulgoromorpha?
Response-1: We understand your confusion, but the misconception is that 4 P. signatus and 66 P.spumarius on table 1 are Adults (please check also the caption of table 1). All these, are identified adults. We hadn’t collected any nymphs of Aphrophorids since they live in spittles on the plants so we couldn’t collect them with sweep net or malaise.
Comment-2: The authors state that Jassargus (Pontojargus) kurdicus was recorded for the first time in Greece and that they were unable to find matches in the databases. However, they correctly included the accession number when they sequenced the specimens. Having collected eight nymphs which they state were identified morphologically, why did they not also include a specimen photo in Figure 1 or as supplementary material?
Response 2: Also is the same misconception as the first one. 8 adults were collected and were identified after the observation of male genitalia. Otherwise, it would be included in the Figure with the rest nymphs. Nevertheless, I added photo (Fig 3) of adult and aedeagus since it’s a rare species, not existing in Europe.
Comment-3: A section of the Methods and Materials (L103–109 and 112–115) is plagiarised from article #26, which is cited, but this section will need to be rewritten so that it is not copied verbatim.
Thank you for pointing this out. We made some changes to avoid plagiarism
Comment-4: The English needs improvement, and contractions should be avoided.
The manuscript was revised by a native English speaker.
Comment-5: - Fig. 2 is labelled 1 in the caption. Thank you a lot. We corrected
Comment-6: - The totals for adults and nymphs in Table 2 are in the wrong place. Thank you a lot again. We corrected.
Comment-7: - The authors refer to '53 species from the Malaise traps and 38 additional species identified morphologically from sweep nets' (lines 142–145). However, there are actually 58 species (and 13 genera).
Response 7: That’s a good point. The total number of collected species is 58 since some common species were collected with both methods. By that, we mean that e.g. P. spumarius was collected both with malaise and with sweep net. So the total number of species collected with malaise were 58 and 38 species with sweep net.
- L239–242: This is a 'copy-paste' of part of the journal's guidelines and should be removed.
You are totally right! We missed it. Thank you again.
Round 2
Reviewer 2 Report
Comments and Suggestions for AuthorsI have no further comments on the manuscript. As far as I am concerned, it can be published once the new corrections have been made.
