Two New Chalcid Wasps (Hymenoptera: Eulophidae and Megastigmidae) Are Parasitoids of Ophelimus bipolaris (Hymenoptera: Eulophidae) on Eucalyptus in China

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript describes two new chalcid wasps based on morphological features and molecular data. The manuscript is well-written in language, and the SEM photos are good quality. However, few comments were listed as follows,

• In your previous paper published in 2025, you used the COI and 28S genes for species identification. Why was the COI gene not used in this paper?
• For the mesosoma of the female Aprostocetus eucalyptus, the description states "without median line", yet this structure is clearly visible in Fig. 2B. In comparison, the male is described as possessing "a distinct median line", as shown in Fig. 4B. This is highly confusing. Please add corresponding annotations for this feature in the relevant figures.
• The collection data is overly simplistic, lacking key information including altitude and collector details. In addition, this is inconsistent with the relevant information recorded in your Materials and Methods section.
• It is neccessary to make a Chinese key to the species.

For other issues, please refer to the annotated PDF document for detailed comments.

Comments for author File: Comments.pdf

Author Response

Dear Reviewer,

We are greatly thankful for your careful reading, professional comments, and constructive suggestions on our manuscript. These comments are very helpful for us to further improve the quality of the paper. We have revised the manuscript accordingly and responded to each comment point by point as follows.

Comment 1

In your previous paper published in 2025, you used the COI and 28S genes for species identification. Why was the COI gene not used in this paper?

Response:

We appreciate the reviewer’s constructive comment. The 28S rRNA gene is a well‑established and highly reliable molecular marker for species identification in chalcid wasps. It has been widely and successfully used in taxonomic studies of both Aprostocetus and Megastigmus, providing sufficient resolution to accurately distinguish closely related species. In the present study, we therefore selected the 28S rRNA gene as the sole molecular marker, and the molecular results were consistent with morphological evidence.

Comment 2

For the mesosoma of the female Aprostocetus eucalyptus, the description states "without median line", yet this structure is clearly visible in Fig. 2B. In comparison, the male is described as possessing "a distinct median line", as shown in Fig. 4B. This is highly confusing. Please add corresponding annotations for this feature in the relevant figures.

Response:

We sincerely apologize for this inappropriate description and the confusion it caused. We have carefully revised the morphological description of the female mesoscutum to be consistent with the morphological character shown in Figure 2B. All relevant descriptions in the manuscript have been updated accordingly.

Comment 3

The collection data is overly simplistic, lacking key information including altitude and collector details. In addition, this is inconsistent with the relevant information recorded in your Materials and Methods section.

Response:

We apologize for the incomplete collection information and the inconsistency between different sections. We have supplemented the missing key collection data, including altitude, collector, and detailed collection information, in the type material section. Meanwhile, we have carefully checked and revised the Materials and Methods section to ensure full consistency with the supplemented collection data. All relevant information is now complete and standardized.

Comment 4

It is necessary to make a Chinese key to the species.

Response:

We appreciate the reviewer’s kind suggestion. However, according to the general publication policy and scope of this journal, only English taxonomic keys are required and commonly adopted for international audiences. Since the journal primarily targets a global readership, an English dichotomous key to the species has been provided in the manuscript to ensure accessibility for researchers worldwide. For this reason, we respectfully suggest not including a separate Chinese key in the final version. We hope the reviewer can understand this situation.

Comment 5

For other issues, please refer to the annotated PDF document for detailed comments.

Response:

We have carefully checked all the detailed comments and corrections in the annotated PDF, including grammatical modifications, word choice improvements, format adjustments, and expression optimization. All these revisions have been implemented in the revised manuscript.

Once again, we sincerely appreciate your valuable comments and suggestions, which have significantly improved the quality of this manuscript. We hope the revised manuscript now meets the publication standards of the journal.

Sincerely,

The Authors

Reviewer 2 Report

Comments and Suggestions for Authors

Dear authors,

The paper is nicely written and presented, study is sound and with enough  proof for the new species descriptions. I have several comments, that you can find in the pdf file that I attached. Majority are writing suggestions or some minor typing mistakes. However, I must point out two things, that have to be changed or re-written. Please add tables with supporting genetic distances for both phylogenetic analysis, in the manuscript body or as supplementary material. You could also maybe add more sequences of different species of the genus (especially for Aprostocetus, since you described one species last year from the same host), to better support the phylogenetic analysis. Why didn`t you include those sequences?

Furthermore, as you already pointed, two sequences from the GenBank match the two sequences of your newly described species. You need to further explain this in Discussion section. If the 28S is a reliable marker, those sequences are the same species as yours two. Aprostocetus is from India, you can even find insect and plant hosts for that sample on the internet. The other one, Megastigmus is from Australia. If you think these are the same species, based on 99-100% match, then the distribution of the new species is completely different and you also have new data on the biology of the species (in the case of Aprostocetus). If you wish to refrain of assigning these ncbi sequences as the species you describe in this paper, then you need to explain why. In both cases, this has to be discussed in detail in the Discussion section, before your manuscript is accepted.

Comments for author File: Comments.pdf

Author Response

Dear Reviewer,

We greatly appreciate your positive evaluation and constructive suggestions. We have thoroughly revised the manuscript and responded to all your comments as follows.

Comment 1

All writing suggestions, minor typing mistakes, grammatical errors and format inconsistencies in the annotated PDF have been carefully checked and corrected, with all revisions clearly marked in red. We have also added the complete pairwise genetic distance matrices as Supplementary Tables S1 and S2, and cited them in the main text to further support the genetic distinctness of the two new species. In addition, we apologize for the earlier oversight and have now added the 28S rRNA sequence of A. bipolaris (described last year from the same host) to our phylogenetic analysis, updating both the phylogenetic tree and genetic distance table to strengthen the phylogenetic resolution.

Comment 2

Thank you for this valuable and constructive comment. We have added a detailed discussion of the two highly similar NCBI sequences in the revised manuscript, covering their taxonomic implications, biogeographic patterns and host associations. We fully acknowledge that the 99.33% and 100% 28S rRNA sequence similarities strongly suggest potential conspecificity between our new species and the Indian (PV911623) and Australian (MT383732) isolates. However, neither sequence is accompanied by formal morphological descriptions, so definitive confirmation requires direct comparison with authentic voucher specimens from these regions. As requested, we have included the confirmed host association of the Australian isolate (reared from Eucalyptus galls).The validity of our new species remains fully supported by comprehensive integrative taxonomic evidence combining detailed morphological characterization and molecular data. All revisions are clearly marked in red in the manuscript.

Once again, we sincerely appreciate your valuable comments, which have greatly improved the quality of our manuscript.

Sincerely,

The Authors