DNA Barcoding Data Reveal Important Overlooked Diversity of Cortinarius sensu lato (Agaricales, Basidiomycota) in the Romanian Carpathians

Round 1

Reviewer 1 Report

This is an interesting piece of work. The authors focused on the genus Cortinarius s.l. in the Romanian Carpathian area employing molecular phylogenetic and morphological methods. Based on their 234 collections, they sequenced the nrDNA ITS region. These sequences were then compared to the data available in public databases. Their phylogenetic analyses and morphological observations identified 109 species representing 40 sections and 3 clades of the genus. Among the species, 43 were documented from Romania, while 66 species are reported as new to the country. It is so nice that almost all the collections can be identified to species based on ITS sequences only. Such fungal species identification can’t be realized in many other parts of the world, or even in North America, where the Cortinarius was studied for a long time. The reviewer would recommend to accept the manuscript after a minor revision. The followings should be improved.

1.     Lines 54-55: change “two taxonomic school have” to “two taxonomic schools have”

2.     Lines 97-98: “it is useful in differentiating species and for analyzing the relationships among them” The reviewer believes that it (ITS sequence) is useful in differentiating species, but not for analyzing the relationships among the species. Many phylogenetic relationships can’t be resolved by the ITS sequence without any doubt. Please reword the sentence.

3.     Lines 229-231: Different authors may have different concept about the sections. It would be informative if relative literature can be cited here.

4.     Table 3: It is strongly suggested to round the measurements of the basidiospores. For example, round “9.42” to 9.5. The values calculated by soft, such as ”12.43–13.11 × 7.97–8.65 µm”, look very nice, but not reflect the reality. Generally speaking, under a light microscope resolution, the smallest value that we can measure with confidence is 0.5 µm. 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

A few comments to help the Authors rewrite the paper:

 Line: 124 (Table 1) - no information after whom the species names of fungi are given; according to Index Fungorum or Mycobank Database?

Line: 133-137 (Figure 1) - no caption for the last two photographs (I and J); the authors of these photographs were also not given.

Line: 138 - Table 1 - what do the numbers 1, 2 given next to the names of fungi mean: C. delibutus 1, C. delibutus 2, C. trivialis 1, C. trivialis 1, ...? This information is missing under the table and in the Material and Methods chapter.

Line: 153 - Table 2 - ???. Compare lines 254 (Table 2) and 263 (Table 3). Check the order in which tables are cited throughout the article.

Line: 254 - Table 2 - proposes to include in the supplement along with the list of cited articles.

Line: 268-272 – I don't understand what the authors of the article meant. Why did they give the mushroom species that could not be identified "a provisional name", if they still require further taxonomic and phylogenetic research. Why are they not included in the article as Cortinarius sp. 1, Cortinarius sp. 2, ….. which would be correct.

On what basis do the authors believe that these species are new to science, since they themselves write that they require further taxonomic and phylogenetic research?

What about Cortinarius magicus aff. ? (compare table 3) – does this species also require further taxonomic and phylogenetic studies? No comments from the Authors.

Fungal species that require further taxonomic and phylogenetic analysis should be omitted from this article. If the authors want to include them in this article, they should complete the taxonomic and phylogenetic analyzes and provide the correct taxonomic names of the fungi.

 

Line: 273 - Discussion

The discussion is quite laconic, it is more of a summary than a discussion. There are no conclusions that result from the conducted research.

There is no comparison, for example, to the number of species of the genus Cortinarius known from countries neighboring Romania, or from other European countries, e.g. Norway, Finland or Austria. Are there any rare, endangered or red lists of fungi in Romania or other European countries among the identified species of mushrooms?

Some wordings/conclusions are incomprehensible, e.g. “One of the major importance of bringing DNA barcoding into the study of biodiversity is the detection of cryptic or semi-cryptic species.”

“During our work, two phlegmacioid (Cortinarius aff. glaucopus, C. aff. sublilacinopes), and one myxacioid (C. aff. vibratilis) species were recognized as potentially new (semi-cryptic) species to science, but further taxonomic and phylogenetic investigations and morphological comparisons with materials from other countries are necessary to resolve their true identity.”  - a statement not supported by reliable analyses, the authors contradict themselves - compare the comments above.

 

“We also hypothesize that other groups of macrofungi will be shown to be more species rich when accurate investigations combining molecular techniques (e.g., DNA barcoding) together with morphological identification methods will be applied in the future.” – this has been known for a long time!

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

I have no comments anymore.