PMID:30497711 Molecular and biological characteristics of laboratory metconazole-resistant mutants in Fusarium graminearum

[jingluodatacurator]

Curated by @jingluodatacurator

https://canto.phi-base.org/curs/4709bfac76d4d9e6

[jingluodatacurator]

Notes:

Uniprot IDs: I1RBR4, I1RJR2, I1S2M5 (https://www.uniprot.org/uniprotkb?query=Fusarium+Graminearum+AND+CYP51) Strains: BM6, BM50, 2021 (section 2.1) The main expt. for curation: section 2.2. Chemical taming of metconazole-resistant F. graminearum mutants, section 2.5. Pathogenicity, section 3.1. In vitro selection of metconazole-resistant mutants.

[jingluodatacurator]

Hi @CuzickA, initial curation has been completed and pending review, thanks.

[CuzickA]

UniProt checking

The results text indicates that although there are three CYP51 paralogues in Fg, sequence alterations in the metconazole-resistant mutants were only found within CYP51A. Current annotations (see below) have been made to CYP51A genotypes.

Current pathogen phenotype annotations

Therefore checking UniProt for Fg CYP51A looks correct....Yes, it does https://www.uniprot.org/uniprotkb/I1RJR2/entry

I may delete the Fg CYP51B and CYP51C UniProt entries later if not needed for annotation

UPDATE I have now deleted the Fg CYP51B and CYP51C UniProt entries (15_01_2024)

[CuzickA]

Checking pathogen genotypes - These look good.

Expt: F. graminearum metconazole-resistant mutants were generated by multiple rounds of selection on metconazole.

Three wild type strains 2021, BM6 and BM50

Three mutants BM6-3, BM6-4 and BM6-6 showed a single amino acid substitution G443S (GGT→AGT)

The mutant 2021-3 showed a single amino acid substitution D243N (GAT→AAT)

The mutant BM50-7 showed two amino acid substitutions, E103Q (GAA→CAA) and V157L (GTC→CTC), simultaneously.

Expt on expression levels In this study, the expression levels of FgCYP51A, FgCYP51B and FgCYP51C in the metconazole-resistant mutants were compared with their parental strains. The results showed FgCYP51A expression of all the metconazole-resistant mutants significantly upregulated as compared with their corresponding parental strains (Fig. 5A).

(Note text on overexpression of CYP51B and C in some mutant lines but no coding or promoter sequence alteration. Decided not to include in curation for now. 'In addition to FgCYP51A, the mutant BM50-7 showed a significant increase in FgCYP51B expression level (Fig. 5B), whereas the mutant 2021-3 showed a significant increase in both FgCYP51B and FgCYP51C expression levels (Fig. 5C) as compared with their corresponding parental strains, respectively. Overexpression of CYP51 genes is caused by changes in the promoter region of the CYP51 genes by the insertion of tandem repeats or transposable elements in some fungi (Carter et al., 2014; Cools et al., 2012; Price et al., 2015). To furtherly confirm changes of the promotor region of CYP51 genes, the promotor region of FgCYP51A, FgCYP51B and FgCYP51C was amplified and sequenced and no change was observed in these mutants (data not shown).'

[CuzickA]

Checking conditions

changed to

Hi @jingluodatacurator, we need to add the chemical in the conditions and just one time point 3dpi. I think the additional time point you added of 5dpi was how the initial mutants were created.

[CuzickA]

This Pathogen Phenotype section now looks good AC next steps 1) Add new PHIPO and PHI-ECO terms _DONE 15_012024 2) Try to add AE alteration in archetype _DONE but question for Nichola to follow up 15_012024 3) Add new Fg strains 'BM50' and 'BM6' _DONE 15_012024 4) Check PHI phenotype annotations _waiting for JL to make edits 15_012024

[CuzickA]

Checking PHI phenotypes

Current annotations

[CuzickA]

Hi @jingluodatacurator, good job at adding in these PHI phenotype annotations :-). There are a few changes to make please. 1) The Wheat strain should be 'cv. Huaimai 33' instead of 'cv. Chinese Spring' 2) When there are expts like this on different tissue types 'coleoptile' and 'wheat ear' each needs a separate annotation. So one whole annotation for the 'coleoptile' and one whole annotation for the 'wheat ear'. Each can have the corresponding condition, 10dpi and 15 dpi respectively. Another note about tissue type 'ear' which is a bit funny ... the definition of this ontology term from BTO is 'The organ of hearing.' So for wheat ear it would be better to use 'inflorescence'. 3) Please could you also add in the wildtype metagenotype controls. Thank you :-)

[CuzickA]

AE alteration in archetype

From Nichola's Table S2

From Mair et al 2016

[CuzickA]

AE alteration in archetype

FgCYP51A(aaG443S)[Overexpression] ~'G476S; Cyp51A; ASPEFU'~

FgCYP51A(aaD243N)[Overexpression] ~'264N; Cyp51A; ASPEFU'~

FgCYP51A(aaE103Q, V157L)[Overexpression] ~'E121Q, 175L; Cyp51A; ASPEFU'~

Info here based on Nichola's TableS2. THIS MAY BE INCORRECT. FgCYP51A mapping to Equivalent codon in Zymoseptoria tritici CYP51B.

NEED INFO for FgCYP51A mapping to Equivalent codon in CYP51A Aspergillus fumigatus.

I will try FRAST.

[CuzickA]

Obtain FASTA for FRAST

Search Genbank for FGSG_04092 FgCYP51A

https://www.ncbi.nlm.nih.gov/nuccore/XM_011323246.1

[CuzickA]

FRAST aligned against archetype CYP51A

FgCYP51A(aaG443S)[Overexpression] 'G448S; Cyp51A; ASPEFU'

AE added to session

FgCYP51A(aaD243N)[Overexpression] 'D245N; Cyp51A; ASPEFU'

AE added to session

[CuzickA]

FgCYP51A(aaE103Q, V157L)[Overexpression] 'E105Q, 159?; Cyp51A; ASPEFU' This second alteration is tricky and I will need to ask Nichola. TO ADD AS E105Q, 159L; Cyp51A; ASPEFU' AE not currently added to session

It looks as though FgCYP51A residue 157 corresponds with archetype AfCYP51A residue 159. However the alteration recorded in FgCYP51A V157L indicates that the original AA was 'V' and the mutation 'L', whilst in the archetype AfCYP51A the AA is already 'L'. Seems like there is natural variation between Fg and Af CYP511A at this residue and that the mutation in Fg alters the sequence to align with Af.

Does the suggested AE seem reasonable or is it better to omit this AE for the double mutant?

[CuzickA]

Next steps for this session

1) @jingluodatacurator to make edits to PHI phenotypes based on AC feedback above. 2) AC to check edits made in point 1) 3) AC to check AE alteration in archetype with Nichola.

[jingluodatacurator]

Summary of expt. :

(2.2. Chemical taming of metconazole-resistant F. graminearum mutants & 2.3. Cross-resistance) metconazole-resistant mutants were generated from three wild type strains by chemical taming. Mycelial plugs cut from 2-day-old colony of three wild type strains 2021, BM6 and BM50 were transferred to PDA plates containing 0.05 μg mL−1 metconazole. After incubation for 5 days at 25 °C, a growing sector was transferred to PDA plates containing a twofold increased concentration of metconazole. After ten rounds of selection at increasing concentrations of fungicide (from 0.05 to 25.6 μg mL−1), metconazole-adapted mutants were isolated and transferred to no-fungicide PDA plates. After cultured for 3 days at 25 °C in the dark, a single mycelial tip was transferred to new PDA plates. After twenty generations of continuous subculture, metconazole-resistant mutants were obtained and stored in 4 °C for further use. Mycelial plugs (5 mm in diameter) cut from the colony grown for 2 days on PDA plates at 25 °C were placed on PDA plates. Plates were incubated in the dark at 25 °C for 3 days, and colony diameters were measured. The 50% effective concentration (EC50) was calculated by fitting mycelial growth inhibition rates against the log-transformed fungicide concentrations. Each concentration had three replicates for each strain.

(2.5. Pathogenicity) Pathogenicity was assessed on wheat seedlings in the laboratory and wheat ears in the field respectively as previously described with minor modification (Liu et al., 2013; Yang et al., 2018). For wheat seedlings test, the coleoptiles of 3-day-old seedlings (cv. Huaimai 33) were cut, and the cut surfaces were inoculated with 2 μL of spore suspensions (105 spores/mL). Water-inoculated coleoptiles served as controls. After the seedlings had grown for additional 10 days at RH 95% and 25 °C, the length of the lesion for each coleoptile was determined. The test was repeated twice with 20 replicates with each strain or mutant. For wheat ears test, the wheat ears (cv. Huaimai 33) at the beginning of anthesis were inoculated with 10 μL of spore suspensions (105 spores/mL). Water-inoculated wheat ears served as controls. After inoculation for 15 days, disease symptoms were assessed. Each strain or mutant was represented by 18 replicate wheat ears.

[jingluodatacurator]

Uniprot IDs: author provided 'based on the sequence of the CYP51 genes deposited in KEGG F. graminearum genome database (CYP51A, FGSG_04092; CYP51B, FGSG_01000; CYP51C, FGSG_11024), however FGSG_04092 could not locate any Uniprot IDs. Therefore '( Fusarium ( graminearum cyp51A ) )' was suggested and searched in the Uniprot search box to find ID I1RJR2 for CYP51A.

Strains: wild strains BM6, BM50, 2021 were used for chemical taming mutation expt.

[jingluodatacurator]

Genotype creation:

BM6-3, BM6-4, BM6-6, BM50-7, 2021-3 have been curated for metconazole. Should they also be curated for other 5 fungicides? @CuzickA as mentioned in 3.2 Cross-resistance, 'compared to the parental strains BM50-7 and 2021, the mutants BM50-7 and 2021-3 was much less sensitive to the six tested azole fungicides with resistance factors (RF) ranging from 3.17 to 6.42 and from 6.95 to 36.46, respectively (Table 2). This indicated that the mutants BM50-7 and 2021-3 had a positive cross resistance between metconazole and other five tested DMI fungicides. The mutants BM6-3, BM6-4 and BM6-6 from the parental strain BM6 also exerted a positive cross resistance between metconazole and epoxiconazole, difenoconazole or tebuconazole, but no cross resistance between metconazole and ipconazole or prochloraz (Table 2).'

[jingluodatacurator]

PHI phenotype creation:

Wild type genotypes have been curated for both BM50 and 2021 strains. Wild type metagenotype controls have been curated for both BM50 and 2021 strains, each been curated for wheat seedlings test and wheat ears test. PHI genotypes have been curated for both BM50 and 2021 strains, each been curated for wheat seedlings test and wheat ears test.

[jingluodatacurator]

Curation completed pending review by @CuzickA thanks.

https://canto.phi-base.org/curs/4709bfac76d4d9e6

[CuzickA]

Current PHI annotations

[CuzickA]

Hi @jingluodatacurator, these PHI annotations are looking pretty good. Just a few more changes to make please.

Feedback 1) Please change the infects tissue 'ear' as noted above

Another note about tissue type 'ear' which is a bit funny ... the definition of this ontology term from BTO is 'The organ of hearing.' So for wheat ear it would be better to use 'inflorescence'.

2) In the cases where there is infective ability 'reduced virulence' the PHIPO terms should be 'decreased extent of pathogen-associated host lesions' instead of 'presence of pathogen-associated host lesions'. This corresponds to the authors text 'the mutants BM50-7 and 2021-3 produced significantly decreased diseased lesions on both wheat coleoptiles and wheat ears (Fig. 4).' which could be added into the comment section of the annotation please. 3) I think the PHI phenotypes could also be curated for 'BM6'. The text to add to the comment would be 'Compared with the corresponding parental strains, the mutants BM6-3, BM6-4 and BM6-6 produced similar diseased lesions,'. In this both the wild type and mutant annotations would be 'presence of pathogen-associated host lesions'.

[CuzickA]

Genotype creation:

BM6-3, BM6-4, BM6-6, BM50-7, 2021-3 have been curated for metconazole. Should they also be curated for other 5 fungicides? @CuzickA as mentioned in 3.2 Cross-resistance, 'compared to the parental strains BM50-7 and 2021, the mutants BM50-7 and 2021-3 was much less sensitive to the six tested azole fungicides with resistance factors (RF) ranging from 3.17 to 6.42 and from 6.95 to 36.46, respectively (Table 2). This indicated that the mutants BM50-7 and 2021-3 had a positive cross resistance between metconazole and other five tested DMI fungicides. The mutants BM6-3, BM6-4 and BM6-6 from the parental strain BM6 also exerted a positive cross resistance between metconazole and epoxiconazole, difenoconazole or tebuconazole, but no cross resistance between metconazole and ipconazole or prochloraz (Table 2).'

Hi @jingluodatacurator, yes you could go ahead and curate these. As a general rule we are trying to capture the main information from the title and abstract of the article.

I think it would also be possible to curate 'pathogen phenotypes' without fungicide

Fig 3 information

Text from the abstract relating to these expts 'Biological fitness of the mutants conferring D243N mutation or E103Q&V157 L mutations significantly decreased in comparison to the corresponding parental strains, suggesting a fitness penalty. The mutants conferring G443S mutation had no change in biological fitness as compared with the parental strain, indicating that the G443S mutation may emerge in field resistant populations of F. graminearum in the future.'

[CuzickA]

Summary of next steps for @jingluodatacurator

1) Make PHI phenotype edits as suggested above. 2) Add pathogen phenotypes with additional fungicides. 3) Add pathogen phenotypes for growth and sporulation without fungicide in Fig 3. (This will make the interpretation of the PHI phenotype annotations more useful to data users. Eg reduced in vitro fungal growth is likely to lead to reduced in planta growth and less disease lesions)

AC next steps 1) check new annotations from points 1-3 above 2) check AE archetype with Nichola.

[jingluodatacurator]

Hi @CuzickA, steps 1-3 above have been completed as instructed, thanks.

[CuzickA]

Hi @jingluodatacurator, feedback on '2. Add pathogen phenotypes with additional fungicides.'

The comment needs to refer to the annotation please. It looks like it has been copied from the previous annotation. eg This comment describes 'metaconazole resistance' but the annotation is for 'resistance to epoxyconazole'. A better sentence to copy from the publication for the comment would be 'The mutants BM6-3, BM6-4 and BM6-6 from the parental strain BM6 also exerted a positive cross resistance between metconazole and epoxiconazole, difenoconazole or tebuconazole, but no cross resistance between metconazole and ipconazole or prochloraz (Table 2).'

Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

[CuzickA]

Continued feedback on for @jingluodatacurator on '2. Add pathogen phenotypes with additional fungicides.'

The actual pathogen phenotype annotations look fine :-).

You could also curate Ipconazole and Prochloraz

[CuzickA]

Hi @jingluodatacurator, I don't see any new annotations for '3. Add pathogen phenotypes for growth and sporulation without fungicide in Fig 3. (This will make the interpretation of the PHI phenotype annotations more useful to data users. Eg reduced in vitro fungal growth is likely to lead to reduced in planta growth and less disease lesions)'

Please add these.

[CuzickA]

Checking PHI phenotypes

The above annotations look good. @jingluodatacurator, please add into the comment section the following text from the publication 'Compared with the corresponding parental strains, the mutants BM6-3, BM6-4 and BM6-6 produced similar diseased lesions, but the mutants BM50-7 and 2021-3 produced significantly decreased diseased lesions on both wheat coleoptiles and wheat ears (Fig. 4).'

[CuzickA]

You are getting the hang of these metagenotype annotations now, well done @jingluodatacurator.

Just a small thing to add here please

The mutant can have AE infective ability 'unaffected pathogenicity' as the phenotype is the same as the wild type control.

[CuzickA]

Summary of next steps for @jingluodatacurator

1) Please address feedback comments on steps 1-3 above.

AC next steps

1) check JL edits to feedback 2) check AE archetype with Nichola. 3) make disease annotations Now DONE below

[CuzickA]

I have added 'disease annotations'

Hi @jingluodatacurator, you may be interested in these. Here is the help documentation on this type of annotation https://canto.phi-base.org/docs/disease_annotation

[jingluodatacurator]

Hi @jingluodatacurator, feedback on '2. Add pathogen phenotypes with additional fungicides.'

The comment needs to refer to the annotation please. It looks like it has been copied from the previous annotation. eg This comment describes 'metaconazole resistance' but the annotation is for 'resistance to epoxyconazole'. A better sentence to copy from the publication for the comment would be 'The mutants BM6-3, BM6-4 and BM6-6 from the parental strain BM6 also exerted a positive cross resistance between metconazole and epoxiconazole, difenoconazole or tebuconazole, but no cross resistance between metconazole and ipconazole or prochloraz (Table 2).'

Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

Hi @CuzickA, I have updated the comments for the pathogen phenotypes with additional fungicides thanks.

[CuzickA]

Hi @jingluodatacurator, feedback on '2. Add pathogen phenotypes with additional fungicides.' The comment needs to refer to the annotation please. It looks like it has been copied from the previous annotation. eg This comment describes 'metaconazole resistance' but the annotation is for 'resistance to epoxyconazole'. A better sentence to copy from the publication for the comment would be 'The mutants BM6-3, BM6-4 and BM6-6 from the parental strain BM6 also exerted a positive cross resistance between metconazole and epoxiconazole, difenoconazole or tebuconazole, but no cross resistance between metconazole and ipconazole or prochloraz (Table 2).' Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

Hi @CuzickA, I have updated the comments for the pathogen phenotypes with additional fungicides thanks.

Thanks @jingluodatacurator,

Updated comment looks good for the BM6 strain mutant with other fungicides but does not make sense for the 2021 and BM50 strain mutants with other fungicides (screenshot below).

Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

You will need to look in the paper yourself and find a suitable sentence to add into the comment here please. Otherwise this is making the same mistake as before with just copying the comment from the previous entry. Thanks.

[jingluodatacurator]

Hi @jingluodatacurator, feedback on '2. Add pathogen phenotypes with additional fungicides.' The comment needs to refer to the annotation please. It looks like it has been copied from the previous annotation. eg This comment describes 'metaconazole resistance' but the annotation is for 'resistance to epoxyconazole'. A better sentence to copy from the publication for the comment would be 'The mutants BM6-3, BM6-4 and BM6-6 from the parental strain BM6 also exerted a positive cross resistance between metconazole and epoxiconazole, difenoconazole or tebuconazole, but no cross resistance between metconazole and ipconazole or prochloraz (Table 2).' Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

Hi @CuzickA, I have updated the comments for the pathogen phenotypes with additional fungicides thanks.

Thanks @jingluodatacurator,

Updated comment looks good for the BM6 strain mutant with other fungicides but does not make sense for the 2021 and BM50 strain mutants with other fungicides (screenshot below).

Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

You will need to look in the paper yourself and find a suitable sentence to add into the comment here please. Otherwise this is making the same mistake as before with just copying the comment from the previous entry. Thanks.

Thanks @CuzickA for pointing it out. Changes have been made accordingly and I will try to be more careful adding suitable comments in the future. Related notes have also been added in my personal curation checklist.

[CuzickA]

Hi @jingluodatacurator, please could you look through this ticket and make a note of the outstanding changes that need to be added to the session still. Thanks.

[jingluodatacurator]

Hi @jingluodatacurator, I don't see any new annotations for '3. Add pathogen phenotypes for growth and sporulation without fungicide in Fig 3. (This will make the interpretation of the PHI phenotype annotations more useful to data users. Eg reduced in vitro fungal growth is likely to lead to reduced in planta growth and less disease lesions)'

Please add these.

Hi @CuzickA, can we have a chat to go through it please in our meeting tomorrow, thanks.

[jingluodatacurator]

Continued feedback on for @jingluodatacurator on '2. Add pathogen phenotypes with additional fungicides.'

The actual pathogen phenotype annotations look fine :-).

You could also curate Ipconazole and Prochloraz

Hi @CuzickA, additional curations for Ipconazole and Prochloraz have been added and related notes been added in my personal curation checklist.

[jingluodatacurator]

Hi @jingluodatacurator, I don't see any new annotations for '3. Add pathogen phenotypes for growth and sporulation without fungicide in Fig 3. (This will make the interpretation of the PHI phenotype annotations more useful to data users. Eg reduced in vitro fungal growth is likely to lead to reduced in planta growth and less disease lesions)'

Please add these.

Hi @CuzickA, pathogen phenotypes without fungicide have been curated please check. And related notes been added into my personal curation checklist.

[CuzickA]

Continued feedback on for @jingluodatacurator on '2. Add pathogen phenotypes with additional fungicides.' The actual pathogen phenotype annotations look fine :-). You could also curate Ipconazole and Prochloraz

Hi @CuzickA, additional curations for Ipconazole and Prochloraz have been added and related notes been added in my personal curation checklist.

Hi @jingluodatacurator, these look good thanks.

Thank you for adding in the new term suggestions. In the future, when you request a new PHIPO term, please could you write out the whole term name in the box to avoid ambiguity. Current

so instead of just adding 'Ipconazole', please add 'resistance to Ipconazole'. Thank you :-)

[CuzickA]

Hi @jingluodatacurator, I don't see any new annotations for '3. Add pathogen phenotypes for growth and sporulation without fungicide in Fig 3. (This will make the interpretation of the PHI phenotype annotations more useful to data users. Eg reduced in vitro fungal growth is likely to lead to reduced in planta growth and less disease lesions)' Please add these.

Hi @CuzickA, pathogen phenotypes without fungicide have been curated please check. And related notes been added into my personal curation checklist.

Hi @jingluodatacurator, thanks for adding the hyphal growth phenotypes. Please could you also have a go at curating the 'sporulation' data from Fig 3B. We did this for another paper recently. These annotations will be recorded as the numbers of asexual spores.

[jingluodatacurator]

Hi @jingluodatacurator, I don't see any new annotations for '3. Add pathogen phenotypes for growth and sporulation without fungicide in Fig 3. (This will make the interpretation of the PHI phenotype annotations more useful to data users. Eg reduced in vitro fungal growth is likely to lead to reduced in planta growth and less disease lesions)' Please add these.

Hi @CuzickA, pathogen phenotypes without fungicide have been curated please check. And related notes been added into my personal curation checklist.

Hi @jingluodatacurator, thanks for adding the hyphal growth phenotypes. Please could you also have a go at curating the 'sporulation' data from Fig 3B. We did this for another paper recently. These annotations will be recorded as the numbers of asexual spores.

Hi @CuzickA, sporulation data from Figure 2&3 has been curated, please check thanks.

[CuzickA]

Hi @jingluodatacurator, I don't see any new annotations for '3. Add pathogen phenotypes for growth and sporulation without fungicide in Fig 3. (This will make the interpretation of the PHI phenotype annotations more useful to data users. Eg reduced in vitro fungal growth is likely to lead to reduced in planta growth and less disease lesions)' Please add these.

Hi @CuzickA, pathogen phenotypes without fungicide have been curated please check. And related notes been added into my personal curation checklist.

Hi @jingluodatacurator, thanks for adding the hyphal growth phenotypes. Please could you also have a go at curating the 'sporulation' data from Fig 3B. We did this for another paper recently. These annotations will be recorded as the numbers of asexual spores.

Hi @CuzickA, sporulation data from Figure 2&3 has been curated, please check thanks.

Hi @jingluodatacurator, thanks for having a go at this but it is not quite right. Please could you have another go? 1) Only Fig 3B should be added to the 'figure column' as the Fig 2 data has the chemical added. 2) The PHIPO terms you have used relate to spore germination. As I mentioned in comment above these should be PHIPO terms relating to the numbers of asexual spores.

[CuzickA]

I have now added the new PHIPO and PHI-ECO terms.

Next steps for this session 1) @jingluodatacurator to make changes to sporulation annotations based on comment above. 2) AC to check whole session through again once 1) has been done, as there have been many changes. 3) AC to check AE alteration in archetype with Nichola.

[jingluodatacurator]

Hi @jingluodatacurator, I don't see any new annotations for '3. Add pathogen phenotypes for growth and sporulation without fungicide in Fig 3. (This will make the interpretation of the PHI phenotype annotations more useful to data users. Eg reduced in vitro fungal growth is likely to lead to reduced in planta growth and less disease lesions)' Please add these.

Hi @CuzickA, pathogen phenotypes without fungicide have been curated please check. And related notes been added into my personal curation checklist.

Hi @jingluodatacurator, thanks for adding the hyphal growth phenotypes. Please could you also have a go at curating the 'sporulation' data from Fig 3B. We did this for another paper recently. These annotations will be recorded as the numbers of asexual spores.

Hi @CuzickA, sporulation data from Figure 2&3 has been curated, please check thanks.

Hi @jingluodatacurator, thanks for having a go at this but it is not quite right. Please could you have another go?

1. Only Fig 3B should be added to the 'figure column' as the Fig 2 data has the chemical added.
2. The PHIPO terms you have used relate to spore germination. As I mentioned in comment above these should be PHIPO terms relating to the numbers of asexual spores.

Hi @CuzickA, changes have been made according to points 1&2 above thanks.

[CuzickA]

Checking PHI phenotypes

The above annotations look good. @jingluodatacurator, please add into the comment section the following text from the publication 'Compared with the corresponding parental strains, the mutants BM6-3, BM6-4 and BM6-6 produced similar diseased lesions, but the mutants BM50-7 and 2021-3 produced significantly decreased diseased lesions on both wheat coleoptiles and wheat ears (Fig. 4).'

This comment had not been added. I have now done this.

[CuzickA]

Hi @jingluodatacurator, feedback on '2. Add pathogen phenotypes with additional fungicides.' The comment needs to refer to the annotation please. It looks like it has been copied from the previous annotation. eg This comment describes 'metaconazole resistance' but the annotation is for 'resistance to epoxyconazole'. A better sentence to copy from the publication for the comment would be 'The mutants BM6-3, BM6-4 and BM6-6 from the parental strain BM6 also exerted a positive cross resistance between metconazole and epoxiconazole, difenoconazole or tebuconazole, but no cross resistance between metconazole and ipconazole or prochloraz (Table 2).' Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

Hi @CuzickA, I have updated the comments for the pathogen phenotypes with additional fungicides thanks.

Thanks @jingluodatacurator, Updated comment looks good for the BM6 strain mutant with other fungicides but does not make sense for the 2021 and BM50 strain mutants with other fungicides (screenshot below).

Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

You will need to look in the paper yourself and find a suitable sentence to add into the comment here please. Otherwise this is making the same mistake as before with just copying the comment from the previous entry. Thanks.

Thanks @CuzickA for pointing it out. Changes have been made accordingly and I will try to be more careful adding suitable comments in the future. Related notes have also been added in my personal curation checklist.

Hi @jingluodatacurator, I was checking through the comments and some of them were still incorrect. I have now made the changes. It is important to think about what each annotation is reporting on from the article and which statement in the article supports it.

[CuzickA]

I have checked through this session again and all now looks fine.

Next steps 1) AC to check AE alteration in archetype with Nichola.

Then session will be ready to approve.

[jingluodatacurator]

Checking PHI phenotypes The above annotations look good. @jingluodatacurator, please add into the comment section the following text from the publication 'Compared with the corresponding parental strains, the mutants BM6-3, BM6-4 and BM6-6 produced similar diseased lesions, but the mutants BM50-7 and 2021-3 produced significantly decreased diseased lesions on both wheat coleoptiles and wheat ears (Fig. 4).'

This comment had not been added. I have now done this.

Thanks @CuzickA, I thought I have added them but looks like I missed some out.

[jingluodatacurator]

Hi @jingluodatacurator, feedback on '2. Add pathogen phenotypes with additional fungicides.' The comment needs to refer to the annotation please. It looks like it has been copied from the previous annotation. eg This comment describes 'metaconazole resistance' but the annotation is for 'resistance to epoxyconazole'. A better sentence to copy from the publication for the comment would be 'The mutants BM6-3, BM6-4 and BM6-6 from the parental strain BM6 also exerted a positive cross resistance between metconazole and epoxiconazole, difenoconazole or tebuconazole, but no cross resistance between metconazole and ipconazole or prochloraz (Table 2).' Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

Hi @CuzickA, I have updated the comments for the pathogen phenotypes with additional fungicides thanks.

Thanks @jingluodatacurator, Updated comment looks good for the BM6 strain mutant with other fungicides but does not make sense for the 2021 and BM50 strain mutants with other fungicides (screenshot below).

Please update the comments for the 'pathogen phenotypes with additional fungicides' using the corresponding text from the publication.

You will need to look in the paper yourself and find a suitable sentence to add into the comment here please. Otherwise this is making the same mistake as before with just copying the comment from the previous entry. Thanks.

Thanks @CuzickA for pointing it out. Changes have been made accordingly and I will try to be more careful adding suitable comments in the future. Related notes have also been added in my personal curation checklist.

Hi @jingluodatacurator, I was checking through the comments and some of them were still incorrect. I have now made the changes. It is important to think about what each annotation is reporting on from the article and which statement in the article supports it.

Thanks @CuzickA for checking through and correcting, I might have not completely understood the text in some cases therefore made mistakes. I will double check on each comment in the future and raise questions if not completely understood.

[CuzickA]

Nichola has checked the AE.

I have added FgCYP51A(aaE103Q, V157L)[Overexpression] 'E105Q, 159L; Cyp51A; ASPEFU'