Integrative Taxonomy and Synonymization of Aculus mosoniensis (Acari: Eriophyidae), a Potential Biological Control Agent for Tree of Heaven (Ailanthus altissima)

Round 1

Reviewer 1 Report

Comments

Line 63   Not all Eriophyoidea show high specificity to host plants.

I suggest to insert the word "Many" before "Eriophyid mites .... are highly specialized.

Line 222  ichnocarpi  -  fide Amrine & Stasny, 1994 Catalog the spelling is  ichnocarpae. Please check.

also lines 320,337 and table 2

Otherwise an excellent research, well written/presented. All figures are necessary.

Author Response

Line 63 Not all Eriophyoidea show high specificity to host plants. I suggest to insert the word "Many" before "Eriophyid mites ....are highly specialized.

The suggestion was partially accepted and “the most of them are” was added on line 63.

Line 222 ichnocarpi - fide Amrine & Stasny, 1994 Catalog thespelling is ichnocarpae. Please check. Also lines 320,337 and table 2

Thank you. Aculus ichnocarpi has been replaced by Aculus ichnocarpae in the text (line 222), in the figure 5 and in the corresponding caption ( line 333), table 2 and corresponding caption (line 337). To be honest, the epithet should be changed in ichnocarpi (genitive case, masculine gender), but this is not the context in which this change could be done.

Reviewer 2 Report

The MS is focused on the synonymy and seasonal dimorphism of two eriophyd mite species. The author show the synonymy of 2 previously described species based on material from Europe and Asia and provide new data on the studied mites obtained with the aid of conventional light microscopy, scanning electrom microscopy and sequencing of 2 marker genes (cox1 and ITS1). The MS is well-written, although its relatively simple content could be explained in a shorter form. I think that this MS could be accepted almost as it is. Some additional remarks are below.

63 Eriophyid mites (Prostigmata: Eriophyoidea) are highly specialized...

In the light of recent molecular multigene phylogenetics and phylogenomics, it seems incorrect ot state that eriophyoids belong to Prostigmata.

271 Figure 1. Drawings of the female internal genitalia are inadequate, spermathecae should be drawn - they are missed now.

285. Figure 2B. Is it leg I or leg II?

340. In the ITS1 alignment generated from all analysed specimens, due to the presence of indels sequences varied in length from 440 bp (Colombes in France) to 449 bp...

Could you clarify whether these indels made the alignment ambiguous or not? In other words, did you have problems aligning the iTS1 sequences because of uncertainty caused by these indels or not?

347. Figure 6.

I suggest using an additional more distant out-group, because this would help you to see a good support for and test the monophyly of the investigated species (like in Figure 5). You could also provide a combined tree based on 2 gene sequences, which in this case would possibly be more informative. It is not a problem that you have not sequences for both genes for all samples, because modern soft treat such cases as missing data and this usually does not affect the total evidence analysis.

Fig5 & 6: be-headed arrows looks a bit wierd and confusing, because their arrowheads indicate somthing uncertain; may be you could use brackets, simple lines or color coding.

Probably some images showing the damage symptoms (if any) or healthy host plant would make this MS more atractive for readers. It is also not clear from the text, why the selected mites could be used as biocontrol agents, because their impact on the host does not mention in the text. I think it could be briefly mentioned in the Introduction what kind of effect they produce on the host that make them appropriate for controlling the plant.

Author Response

63 Eriophyid mites (Prostigmata: Eriophyoidea) are highly specialized... In the light of recent molecular multigene phylogenetics andphylogenomics, it seems incorrect ot state that eriophyoidsbelong to Prostigmata.

It is right. According to Bolton et al., 2017 based on cladistic analysis of 103 morphological traits, and Klimov et all., 2018 and Klimov et all., 2022 results of molecular phylogenetic analyses (RNA and protein genes) Eriophyoidea are included in the Sarcoptiformes clade closed to Endeostigmata and Nematacicoidea. We fixed it.

271 Figure 1. Drawings of the female internal genitalia are inadequate, spermathecae should be drawn - they are missed now.

Fixed.

285. Figure 2B. Is it leg I or leg II?

It is clearly reported in the Caption that it is Leg I. The drawings is the classical scheme used in this case in which the differences between leg I and leg II are stereoptiped.

Line 340. In the ITS1 alignment generated from all analysed specimens, due to the presence of indels sequences varied in length from 440 bp (Colombes in France) to 449 bp...

Could you clarify whether these indels made the alignment ambiguous or not? In other words, did you have problems aligning the iTS1 sequences because of uncertainty caused by these indels or not?

This is a good point. In fact these indels did not make the alignment ambiguous. Nonetheless they are not consistently present or identical when we repeated alignments. Our approach was not to take into account these indels based on the assumption that the phylogenetic reconstruction would be more accurate if it does not rely on the entire sequence.

Line 347. Figure 6.

I suggest using an additional more distant out-group, because this would help you to see a good support for and test the monophyly of the investigated species (like in Figure 5). You could also provide a combined tree based on 2 gene sequences, which in this case would possibly be more informative. It is not a problem that you have not sequences for both genes for all samples, because modern soft treat such cases as missing data and this usually does not affect the total evidence analysis.

Given the limited amount of phylogenetic signal as reflected in the short internal branches, and potential confounding non-phylogenetic signals in the clones (although we have applied a conservative approach by keeping only clones represented at least twice to avoid inclusion of spurious mutation raised from PCR and cloning error which can produce a false spectrum of genomic diversity), given the relatively long phylogenetic distances between the nearest outgroups ( in the Aculus or Aculops) and the ingroup, we agree with the reviewer that the choice of the proxy outgroup is critical, which was not the case for the CO1. The choice of Aculus cercidis remains the best as with more distant outgroups, internal branches of the group are collapsing. We could have also proposed an unrooted phylogenetic reconstruction to alleviate this issue. Moreover, as we also estimated the pairwise distances between congeners, it seems to us that it makes more sense to keep Aculus cercidis for the phylogenetic reconstruction than another taxa outside the genus. Regarding the suggestion of concantenating the two regions, we fully agree that this is an approach more commonly used than before as suggested by Wiens in 2006 (Journal of Biomedical Informatics 39 (2006) 34–42). We are providing a phylogenetic tree with concatenated sequences which to us is not very convincing as the ingroup is totally lacking of support. The observed pattern could be attributed to different factors. Lumping genes or sequences with contrasting evolutionary rates such as ITS1 and CO1, could induce a risk of distorting phylogenetic signal. We have lumped sequences of which many  are missing including the important paratype . As suggested by Wiens (2006), highly incomplete taxa can be accurately placed in phylogenies, as long as many characters have been sampled overall, which is not our case , as we have a total of 1111 characters of which 658 are missing in  case of CO1 or 453 in case of ITS1. An  alternative could be to add this tree in a supplementary material if the reviewer insists on it.

Fig5 & 6: be-headed arrows looks a bit wierd and confusing, because their arrowheads indicate somthing uncertain; may be you could use brackets, simple lines or color coding.

We agreed with the reviewer and put the haplotype id at each tip in the tree