Contributions towards the consistent ID of (, )

Brandon Shaw

2019

Dissertation submitted for the degree of Master of Science in Plant and Fungal , Diversity and Conservation awarded by Queen Mary, University of London. https://doi.org/10.34885/71

© The Author. All rights reserved. Contributions towards the consistent ID of powdery mildew (Erysiphales, Ascomycota) species

Brandon Shaw Supervised by Dr Oliver Ellingham and Dr Ester Gaya

Abstract

Powdery mildews (Erysiphales, Ascomycota) are an order of plant pathogenic fungi that infect 10,000 different angiosperm species, including important crop species. Traditional morphological methods of identifying powdery mildews can be difficult and time consuming, as different character states can be hard to distinguish. Molecular techniques using the ITS region fail to provide a species level identification 1 out of every 4 times and often must be used in conjunction with morphology to provide an identification beyond genus. The Mcm7 gene could potentially be used to provide more accurate identifications and build phylogenies incorporating large number of taxa to help understand the systematics of the order. Here, it is found that incorporating Mcm7 and ITS into a phylogeny greatly increases bootstrap support values, suggesting that Mcm7 may be used in the future to understand the systematics of this previously problematic order. ITS is successfully extracted from more and older specimens than Mcm7 and recommendations on using Mcm7 in the lab are made.

Introduction

For the first time in decades, the number of undernourished people in the world is rising and is equal to the number seen in 2010-2011 (FAO, 2019), with the current world population of 7.7 billion expected to reach 9.7 billion in 2050 (United Nations, 2019), now more than ever global food security is an issue at the forefront of our minds. An extra 200,000 billion calories a year by 2050 must be produced (Bebber & Gurr, 2015), one way to improve food security will be to improve crop yield. Currently, pests and plant diseases cause a 20-25% decrease in harvest yield and 10% loss post-harvest resulting in a huge economic loss and posing a serious threat to global food security (Bebber & Gurr, 2015; Martinelli et al., 2015). Pathogenic fungi that have the biggest effect, rapidly spread by human trade and transport with species ranges expanding owing to climate change, one such pathogenic group are the powdery mildews(Bebber & Gurr, 2015; Bebber, Holmes, & Gurr, 2014; Ellingham, et al., 2016).

Powdery mildews are plant pathogenic fungi in the order Erysiphales in the A B Ascomycota with 872 species currently described (Braun & Cook, 2012) characterised by a talcum powder-like patches growing on host leaves, Figure 1. Powdery mildews infect around 10,000 angiosperm species (Braun, 1987) including popular garden plants such as Asteraceae which are infected by the powdery mildew genus Golovinomyces (Takamatsu, et al., 2006) and the genus Figure 1 Talcum powder-like patches of powdery mildew growing which predominantly infects on Ballota sp. (A) and Lupinus sp. (B). Rosaceae(Takamatsu, et al., 2010). More importantly, powdery mildews infect a wide variety of crop plants, Blumeria graminis which infects important grain crops wheat, rye, and barley and has a worldwide distribution (Braun & Cook, 2012; Zhang et al., 2005), Podosphaera xanthii infects crops belonging to the family Cucurbitaceae and has an almost worldwide distribution (Braun & Cook, 2012; Mcgrath, 2001), and necator which is the most common and damaging pathogen of various grapevine cultivars and is found wherever grape is grown (Braun & Cook, 2012; Gadoury et al., 2001).

All 18 genera powdery mildews are obligate biotrophic parasites of angiosperms, 14 genera are epiphytic forming haustoria in the plant epidermal cells, including the most specious genera Erysiphe, and three are endoparasites forming haustoria in plant parenchyma cells (Phyllactinia, Leveillula, and Pleochaeta) (Aghayeva, Abasova, & Takamatsu, 2018). Appendix 1 shows a summary of currently accepted genera within the order Erysiphales. Powdery mildews can be treated in various ways, in the garden, pruning off the infected areas or useing home remedy treatments such as baking soda mixed with soap and water, and diluted milk (Beresford et al., 1996; Bettiol, 1999; Martins et al., 2016) may suffice. On commercial levels however, these treatments are not practical as they may be costly and time consuming. Various fungicides are used, soluble silicone based fungicides are used to help improve host response to infection, increasing the production of antifungal compounds by the plant (Bowen, et al., 1992; Rémus-Borel, Menzies, & Bélanger, 2005). Demethylation inhibitor fungicides are another treatment for powdery mildew however, resistance to these fungicides has been observed, such as in Podosphaera xanthii (Mcgrath, 2001).

Powdery mildews have proven to be a taxonomically challenging group, A B with species level identification using morphological and current molecular techniques being problematic and inaccurate at times, morphological techniques especially requiring an in-depth knowledge of the group and a time using light microscopy to reach a potential ID. In order to develop more targeted fungicides and treatment plans for different Figure 2 Chasmothecia (A) and conidiophore (B) viewed using light powdery mildew species, we must microscopy. first be able to accurately identify them and understand their phylogeny in order to predict the response of more closely related taxa to certain treatments. Morphological characters for identification come from the asexual and sexual life stages (Figure 2), the asexual life stage (anamorph) conidiophore is common and can be found in virtually all powdery mildew samples but provide fewer characters for identification whereas the sexual life stage (teleomorph) chasmothecia are rarer and provide many more characters for identification. Chasmothecia can be rare as they are produced in times of stress such as over wintering, chasmothecia have never been observed in some species such as in the genera , the powdery mildew that infects tomato Oidium neolycopersici has never had chasmothecia observed (Jones, Whipps, & Gurr, 2001). Identification can be further complicated as in the past anamorphs and teleomorphs have been described as different species (Braun, 2012).

The advent of molecular techniques has improved the ease of identifying powdery mildews. The ITS region has traditionally been used in fungal barcoding including the powdery mildews (Schoch et al., 2012). However, the ITS region has is less effective for powdery mildews for barcoding than other fungal groups, highlighted by the fact that it fails to provide a species level identification 1 out of every 4 times (Ellingham, 2017). As a result of the inaccuracy of ITS, few if any large-scale order wide phylogenies with a large number of taxa have been published, instead phylogenies covering the different genera of the powdery mildews are available, or phylogenies across the order but with few taxa (Braun et al., 2006).

Powdery mildews are often thought of as infecting a small number of hosts, typically within the same plant genus or family, however a few species seem to infect a wide variety of hosts from multiple host families or even orders (JCunnington, Lawrie, & Pascoe, 2010). It is suspected that at least some of these could be species complexes and are multiple different species. Current methods are not strong enough to investigate whether this is true, examples of cases like this which require more molecular analysis include Podosphaera tridactyla a suspected species complex infecting the genus Prunus (Takamatsu et al., 2010), Golovinomyces cichoracearum is a well-known species complex, infecting different plant families including Asteraceae and Solanaceae (Cunnington et al., 2010). The inaccuracy of ITS and need for more accurate molecular tools and methods for identifying powdery mildews has been stated multiple times (Attanayake, et al., 2009; Shin, et al., 2019) Mcm7 is a potential gene that could be used to identify powdery mildews more easily, as well as build better phylogenies and barcoding databases and potentially explore species complexes (Ellingham, 2017). Mcm7 is a gene which codes for the mini-chromosome maintenance complex component 7, mini-chromosome maintenance proteins are essential for DNA replication and is therefore well conserved (Kearsey & Labib, 1998) making it a good potential candidate for use in phylogenetic analysis. Mcm7 has been used in other groups within the Ascomycota to investigate phylogenetic relationships (Raja, et al., 2011) including Eurotiomycetes, Lecanoromycetes, , Lichinomycetes, and Sordariomycetes (Schmitt et al., 2009).

This study will make use of the 2215 powdery mildew specimens stored in the RBG Kew fungarium and extractions made during Ellingham, 2017’s citizen science “powdery mildew survey” to test the hypotheses that DNA will be successfully extracted from RBG Kew fungarium using protocols developed by Ivanova et al., (2006) modified for use in the Mycology lab at RBG Kew. Primers used for PCR on fresh powdery mildew samples will be successful for fungarium specimens 10/20/30 years old of both the ITS region and Mcm7.For use in phylogenies it is hypothesised any phylogenies built using Mcm7 will have higher bootstrap support values than those without. It is also hypothesised that DNA barcoding will confirm the stated ID of RBG Kew specimens.

In order to test the hypotheses, the current understanding of the powdery mildew phylogeny must be understood, this will be done by completing a baseline data study collating sequence data of powdery mildew species from online repositories into one large phylogeny. Gap analysis will be carried out on the fungarium collection and samples from the “powdery mildew survey” to identify what specimens are available which have little or no sequence data for both ITS and Mcm7 to determine which specimens and samples will be prioritised for sampling. Phylogenies built using sequence data for Mcm7 from online repositories and data generated in this study will be used to build a phylogenetic tree, this will be compared to a tree built with ITS data from the same specimens and a tree built using both genes concatenated.

Materials and Methods

Databasing and gap analysis

GenBank taxonomy browser was used for the baseline study, which had list of taxa to include in the order wide phylogeny. A database with information of what sequence data is available for each taxa including how many ITS and Mcm7 sequences are available, what other sequences are available, and nomenclature was generated (appendix 2). Following this, sequence data for each taxa was downloaded from various sources sequences generated sequences generated during the “powdery mildew survey” were incorporated first, followed by the UNITE database (https://plutof.ut.ee/) and finally, if sequence data was not available on UNITE it was taken from the GenBank taxonomy browser.

Gap analysis was carried out by searching Herbtrack, the database of RBG Kew’s specimens to identify which specimens were available at Kew which little or no sequence data. Specimens were selected from British and overseas collections from as many genera as possible. Where samples were available from suspected species complexes, multiple were chosen from the biggest variety of host plant families possible. A table of species sampled from the fungarium along with a donation to the “Powdery Mildew Survey” can be seen in appendix 3.

DNA extraction

DNA was extracted from specimens in the RBG Kew fungarium using the enzymatic/ glass filter plate extraction for fungi protocol by Ivanova, et al. (2006). Modified for the Mycology lab at RBG Kew, modifications include the use of a vacuum instead of a centrifuge for washing steps, 250 µl of protein wash buffer used (instead of 180 µl), 300 µl of wash buffer was added to each well which was washed twice (instead of 750 µl once), 50 µl of distilled water was added for the final step, yielding 50 µl for each sample not 60 µl.

A section of infected leaf around 10mm2 was cut from the fungarium specimen, the area selected was the most heavily infected area, where chasmothecia were present as many as possible were included in the sample (up to 30 chasmothecia however, some samples fewer than 10). Samples with less heavily infected areas or no chasmothecia had extra conidia scrapes added to the sample.

Each sample then had 100 µl of lysis buffer and Proteinase K (10:1 solution) added, then covered and stored at 56 oC for 8 to 16 hours. Samples were centrifuged at 500 rcf for 1 minute, 100 µl of Binding mix was added to each sample and mixed by pipetting up and down. Then, 150 µl of the lysate was added to a glass fibre filtration plate and a vacuum was used to bind the DNA to the glass fibre filtration membrane. Then, 250 µl of protein was buffer was added to each sample and vacuumed, 300 µl of wash buffer was added to each well and vacuumed, this step was repeated. The plate was incubated at 56 oC for 30 minutes to evaporate residual ethanol. Finally, 50 µl of 0.1 AE buffer (pre-warmed to 56 oC) was added to each well and was incubated at 56 oC for 5 minutes on top of a 96-well PCR plate, the glass filter plate was then centrifuged at 500 rcf for 5 minutes with the 96-well PCR plate underneath to collect the eluted DNA. Extractions were stores at -20 oC.

PCR amplification

Primers used for the amplification of the ITS region were the powdery mildew specific primers PMITS1 (forward primer, TCGGACTGGCC(T/C)AGGGAGA) and PMITS2 (reverse primer, TCACTCGCCGTTACTGAGGT(Cunnington, et al., 2003). Primer concentration for the PCR reaction was 10 ng µl-1 each reaction contained 0.875 µl of each primer, 12.5 µl of Dream Taq, 6 µl of TBT par, 3.75 µl of distilled water, and 1µl of DNA from the extraction. The thermocycler (Eppendorf vapo.protect) protocol for amplification of ITS was an initial denaturing step at 95 oC for 5 minutes, then 35 cycles of denaturing at 95 oC for 15 seconds, annealing at 50 oC for 20 seconds and, elongation at 72 oC for 60 seconds, and after the last cycle a final elongation stage at 72 oC for 5 minutes.

Primers used for amplification of the Mcm7 gene were Mcm7F2 (forward primer, TGTGATCGRTGYGGDTGTGA) and Mcm7R8 (Reverse primer, TCATYCCRTCRCCCATYTCYTTWG) (Ellingham, 2017). Primer concentration for the PCR reaction was 10 ng µl-1 each reaction contained 0.875 µl of each primer, 12 µl of Dream Taq, 9.75 µl of TBT par, 1 µl of MgCl2, and 1 µl of DNA from the extraction, if this did not work samples were trialled again with 2 µl of MgCl2 in the master mix, reducing the amount of TBT par and Dream Taq by 0.5 µl each. The thermocycler protocol for amplification of Mcm7 was a gradient cycle of an initial denaturing step at 95 oC for 5 minutes, then 10 cycles of denaturing at 95 oC for 30 seconds, annealing at 54 oC for 60 seconds, elongation at 72 oC for 90 seconds, followed by 27 cycles of denaturing at 95 oC for 30 seconds, annealing at 61 oC for 60 seconds, elongation at 72 oC for 90 seconds, and after the last cycle a final elongation stage at 72 oC for 5 minutes. A negative control with no template DNA was included in each PCR carried out during this study.

PCR products were visualised using gel electrophoresis, to make the gel 1.5g of agarose was mixed with 100 ml of TAE x0.5 concentration and microwaved until all the agarose had dissolved, 2.5 µl of ethidium bromide was added. The solution was poured into a gel tray with well combs, once set the gel was moved into a tray containing a buffer of TAE x0.5 and 2 µl of PCR product was added into each well, 2 µl of ladder (ThermoScientific gene ruler #SM1333) was added into the first well of each row instead of DNA, the gel was run at 95 V for 50 minutes and visualised using an analyticjena UVP GelStudio on VisionWorks Acquisition and analysis software version 8.21.18003.9986. PCR products were stored at 4 oC until cleaned for sequencing or at -20 oC for extended periods of time.

PCR clean up and sequencing

Cleaning PCR products for sequencing required three procedures to prepare samples for sanger sequencing first, the enzymatic purification of PCR products, second a cycle sequencing reaction, and third an ethanol precipitation.The enzymatic purification of PCR products mixed 2 µl of ExoFAP diluted to a 1:10 solution and 5 µl of PCR product which was incubated in a thermocycler at 37 oC for 15 minutes and then 85 oC for 15 minutes.

The cycle sequencing reaction which further amplifies DNA products and removes excess dNTPs, ddNTPs, salts, dyes, and enzymes. Two reactions are set up for this stage, one using the forward primer and one using the reverse primer. Each reaction contains 0.5 µl of Applied Biosystems Dye terminator, 1.5 µl Sequencing Buffer 5X, 1 µl of the primer (diluted to 1/10 the concentration used for the original PCR), 2 µl of PCR product from the enzymatic purification step, and 5 µl of distilled water. The thermocycler protocol for the cycle sequencing reaction was 2 minutes at 90 oC followed by 60 cycles of 90 oC for 10 seconds, 50 oC for 10 seconds, then 60 oC for 2 minutes.

The ethanol precipitation used the products from the cycle sequencing reaction, the products were spun briefly and 1.25 µl EDTA (125 mM) was added to each sample and spun down. Then, 15 µl of 100% ethanol was added to each sample and the samples were vortexed before being left at room temperature for 15 minutes. The samples were centrifuged at maximum speed at 4 oC for 45 minutes to freeze a DNA pellet at the bottom of the well. Once the centrifuge had finished the plate was removed (not touching the bottom of the plate so the pellet is not disturbed) and the lid removed and replaced with a paper towel, the plate was spun upside down at 200 g briefly. Then, 15 µl of fresh cold 70% ethanol was added to each well, the lid was placed back on the samples and centrifuged at maximum speed at 4 oC for 15 minutes. The lid was replaced with a paper towel and the plate briefly spun upside down again. The plate was dried at 65 oC for 2 minutes and 30 µl of distilled water was added to each well, warmed at 65 oC for 1 minute and vortexed to dissolve the pellet. The samples were sequenced using Sanger sequencing. Sequences were searched on BLAST.

Phylogeny

Geneious (v 8.1.9) was used to contig forward and reverse sequence data using De Novo assemble, samples which did not automatically contig using this method were manually done where possible by searching for the sequence “GATCATTA” to find the start of the ITS region or “TGTGACT” to find the start of the Mcm7 region, the sequence before this point was trimmed and low quality/ excess bases were trimmed from both ends and De Novo assembly run again to see if sequences would contig. AliView version 1.26 was used to align sequence data using MAFFT for both ITS and Mcm7 .

CIPRES (https://www.phylo.org) was used to run RAxML using the tool “RAxML-HPC2 on XSEDE” to build phylogenies, the model for bootstrapping phase was GTRGAMMA and the number of bootstrap iterations was 1000. FigTree v1.4.3 was used to view phylogenies. The large ITS phylogeny was rooted with Byssoascus striatosporus as in Braun et al., 2006 this was used as this tree contains the basal genera Parauncinula and Caespitotheca. For the smaller phylogenies Blumeria graminis was used as an outgroup and to root the trees. As Hillis & Bull, 1993 suggests that a bootstrap support value of >70% is equal to a >95% probability that the clade is true, therefore this study will use a threshold of 70% as significant.

Results

Voucher information on the taxa downloaded from public databases and sampled from the RBG Kew fungarium collection for a total of 629 taxa, is included in Appendix 2

In total, 209 specimens were sampled during the study, 178 from the RBG Kew fungarium and 31 from the donation to the “Powdery Mildew Survey” (appendix 3). Of the specimens sampled, 148 successfully amplified ITS and 61 successfully amplified Mcm7, all specimens which had Mcm7 amplified also had ITS amplified (appendix 3). Of the samples processed for sequencing 39 generated a useable sequence for ITS and 23 for Mcm7, sequences were then searched on BLAST, a list of the sequences obtained and their accession numbers on GenBank can be seen in appendix 6.

Phylogenies

A tree built using all the available online sequence data of ITS containing 1034 taxa was constructed, with Byssoascus striatosporus as an outgroup and can be seen in appendix 5, this tree has very few significant bootstrap support values, especially in early divergences. Comparing the smaller trees built with either ITS (Figure 4) or Mcm7 (Figure 3) alone shows that both genes still produce trees with few significant bootstrap support values, especially in earlier divergence, ITS has significant bootstrap support values at the start of the Podosphaera genus where Mcm7 does not. Mcm7 has a significant divergence at the start of the genus Neoerysiphe where ITS does not the Mcm7 tree places this genus sister to the genus Erysiphe whereas the ITS tree places it sister to the genus Golovinomyces in the tribe Golovinomyceteae. ITS also has more significant divergence in the genus Golovinomyces which also has significant support (80%) for placing the genus Arthrocladiella sister to the genus Golovinomyces whereas the support in the Mcm7 tree is not significant (31%), support in the genus Erysiphe is low in both trees. The ITS tree places the one representative of the genus Phyllactinia within the genus Neoerysiphe with incredibly low support value (2%) the Mcm7 tree places it sister to the genus Erysiphe with a low, non- significant support value (45%).

When both genes are concatenated and used to build a tree bootstrap support values increase and the support for early divergence becomes significant (Figure 5). In this tree the genus Podosphaera has improved support values than both trees built with single genes. The positioning of genera Neoerysiphe and Golovinomyces is still not significant, Neoerysiphe is placed sister to Erysiphe with low support value (36%) and Golovinomyces sister to Neoerysiphe and Erysiphe with low support value (57%) however, support values within each genera being greatly improved, with most values being significant within apart from where multiple representatives of the same species have been included which is to be expected. Arthrocladiella is placed sister to Golovinomyces with a significant support value (90%). Support values for the genus Erysiphe in this tree are mostly significant, apart from where multiple representatives of the same species are present. Phyllactinia is placed diverging early being sister to tribes Erysiphese and Golovinomyceteae with a significant support value (79%) forming a tribe on its own Phyllactinieae.

Barcoding

All sequences generated were searched on the BLAST database for any matches, appendix 6 shows the results. Blast searches of the ITS sequences generated typically confirmed the ID to genus level and occasionally to species level for specimens such as Erysiphe aquilegiae and Podosphaera xanthii. The blast search of the ITS sequence Golovinomyces cucurbitacearum matched with Podosphaera xanthii and G. cucurbitacearum was placed within the genus Podosphaera in the ITS tree (Figure 4) with significant support values.

A few Mcm7 sequences matched with Mcm7 sequences from other fungal groups Dermatocarpon, Claviceps, and Solenopsora. R odosphaera morsuvae ex Ribes sanguineum odosphaera clandestina ex Crataegus monogyna R odosphaera sp ex Taraxacum officinale odosphaera plantaginis ex lantago lanceolata odosphaera sp ex Taraxacum officinale Tribe odosphaera sp ex Taraxacum officinale odosphaera macrospora ex Tellima grandiflora R odosphaera clandestina ex Malus pumila Cystotheceae odosphaera leucotricha ex Malus domestica R odosphaera dipsacearum ex Dipsacus sp R odosphaera epilobi ex Epilobium hirsutum odosphaera euphorbiaehelioscopiae ex Euphorbia peplus R Golovinomyces cynoglossi ex Myosotis laxa Golovinomyces cynoglossi ex Myosotis arvensis R Golovinomyces sp ex Monarda didyma R Golovinomyces biocellaris ex Salvia officinalis R Golovinomyces magnicellulatus ex Epilobium hirsutum R Golovinomyces magnicellulatus ex hlox paniculata Golovinomyces magnicellulatus ex hlox paniculata Golovinomyces cichoracearum ex ilosella aurantiaca Golovinomyces cynoglossi ex Myosotis arvensis Golovinomyces cynoglossi ex Myosotis sp Golovinomyces cynoglossi ex Myosotis arvensis R Golovinomyces sordidus ex lantago lanceolata R odosphaera amelanchieris ex Amelanchier lamarc ii Golovinomyces sordidus ex lantago lanceolata Golovinomyces sordidus ex lantago lanceolata R Golovinomyces sordidus ex lantago lanceolata Golovinomyces cichoracearum R Golovinomyces sordidus ex lantago major Golovinomyces sordidus ex lantago major Erysiphe polygoni Golovinomyces sordidus ex plantago major Golovinomyces sordidus ex lantago maritima Golovinomyces sordidus ex lantago major Golovinomyces orontii ex Cucurbita peop Golovinomyces sp ex erbascum thapsus R Golovinomyces cynoglossi ex Symphytum xuplandicum Tribe Golovinomyces cynoglossi ex ulmonaria sp Golovinomyces cynoglossi ex Silene dioica Golovinomyces cynoglossi ex ulmonaria sp Golocinomyceteae Golovinomyces sonchicola ex Sonchus oleraceus R Golovinomyces cichoracearum ex Sonchus oleraceus Golovinomyces cichoracearum ex Sonchus oleraceus R Golovinomyces cichoracearum ex Sonchus oleraceus R Golovinomyces depressus ex Arctium minus R Golovinomyces depressus ex Arctium minus R Golovinomyces depressus ex Arctium minus Arthrocladiella mougeotii ex Lycium barbarum b eoerysiphe galii ex Galium aparine eoerysiphe galii ex Galium aparine eoerysiphe galii ex Galium aparine eoerysiphe galeopsidis ex allota nigra eoerysiphe galeopsidis ex Stachys byzantina eoerysiphe galeopsidis ex Acanthus mollis

eoerysiphe galeopsidis ex Lamium sp eoerysiphe galeopsidis ex Lamium sp eoerysiphe galeopsidis ex Stachys arvensis

R eoerysiphe galeopsidis ex Acanthus spinosus eoerysiphe galeopsidis ex Lamium amplexicaule eoerysiphe galeopsidis ex Lamium purpureum eoerysiphe galeopsidis ex Lamium album Rb eoerysiphe galeopsidis ex Lamium purpureum R eoerysiphe galeopsidis ex Lamium purpureum eoerysiphe galeopsidis ex Stachys byzantina b eoerysiphe galeopsidis ex Lamium sp R eoerysiphe galeopsidis ex Stachys sylvatica eoerysiphe galeopsidis ex Lamium sp R eoerysiphe galeopsidis ex Acanthus mollis R eoerysiphe nevoi ex Lapsana communis eoerysiphe nevoi ex Taraxacum officinale eoerysiphe nevoi ex Sonchus arvensis R eoerysiphe nevoi ex Sonchus arvensis eoerysiphe nevoi ex Sonchus arvensis eoerysiphe geranii ex Geranium sp R eoerysiphe geranii ex Geranium phaeum Rb eoerysiphe geranii ex Geranium sp R eoerysiphe geranii ex Geranium sp R eoerysiphe geranii ex Geranium pratense Tribe eoerysiphe joerstadii hyllactinia betulae A C Erysiphe ulmi Phyllactineae R Erysiphe prunastri ex runus spinosa R Erysiphe necator ex itis vinifera R Erysiphe hedwigii ex iburum opulus R Erysiphe sp ex Lonicera japonica Erysiphe sp ex Lonicera periclymenum Erysiphe sp ex Lonicera sp Erysiphe sp ex Lonicera periclymenum Erysiphe aquilegiae ex Caltha palustris Erysiphe aquilegiae ex Ranunculus acris Erysiphe aquilegiae var. ranunculi

R Erysiphe aquilegiae ex Aquilegia sp Erysiphe aquilegiae ex Aquilegia sp Erysiphe aquilegiae ex Ranunculus repens R Erysiphe aquilegiae ex Ranunculus repens Erysiphe aquilegiae ex Aquilegia sp Erysiphe tortilis R Erysiphe aquilegiae ex Ranunculus repens Erysiphe aquilegiae var. ranunculi

R Erysiphe circeae ex Circaea lutetiana R Erysiphe aquilegiae ex Aquilegia sp R Erysiphe catalpae ex Catalpa bignonioides Erysiphe aquilegiae var. ranunculi A C Erysiphe ornata var. europeae A C Rb Erysiphe aquilegiae ex Aquilegia sp R Erysiphe aquilegiae ex Aquilegia sp Erysiphe aquilegiae ex Aquilegia sp R Erysiphe aquilegiae ex Aquilegia vulgaris Erysiphe aquilegiae ex Aquilegia vulgaris R Erysiphe aquilegiae ex Aquilegia vulgaris R Erysiphe aquilegiae ex Delphinium sp Erysiphe aquilegiae var. ranunculi Erysiphe elevata ex Catalpa bignonioides Erysiphe syringae japonicae Erysiphe platani ex latanus x hispanica Erysiphe divaricata A C R Erysiphe cruciferarum ex rassica sp Erysiphe cruciferarum ex rassica sp R Erysiphe cruciferarum astragali R Erysiphe trifoliorum ex Trifolium pratense Erysiphe trifoliorum ex Trifolium pratense R Erysiphe trifoliorum ex Trifolium pratense Tribe Erysiphese R Erysiphe trifoliorum ex Lathyrus pratensis

Erysiphe trifoliorum ex Lathyrus pratensis R Erysiphe trifoliorum ex ypericum sp Erysiphe trifoliorum ex Trifolium pratense Erysiphe trifoliorum ex Trifolium dubicum Erysiphe baptisiae Erysiphe thesii Microsphaera palczews ii Erysiphe guarionii A C Erysiphe berberidis ex erberis sp Erysiphe berberidis ex erberis thunbergii Erysiphe berberidis ex Mahonia moseri R Erysiphe berberidis ex erberis sp R Erysiphe buhrii ex Silene dioica Erysiphe heraclei ex Anthriscus sylvestris Erysiphe heraclei ex eracleum sphondylium Erysiphe heraclei ex eracleum sphondylium Erysiphe heraclei ex eracleum sphondylium

R Erysiphe heraclei ex eracleum sphondylium R Erysiphe heraclei ex eracleum sphondylium R Erysiphe heraclei ex eracleum sphondylium Erysiphe heraclei ex eracleum sphondylium R Erysiphe heraclei ex Anthriscus sylvestris Erysiphe heraclei ex Anthriscus sylvestris Erysiphe heraclei ex Anthriscus sylvestris ex uercus robur Erysiphe alphitoides ex uercus robur R Erysiphe alphitoides ex isteria sinensis

R Erysiphe alphitoides ex uercus robur b Erysiphe alphitoides ex isteria sinensis Erysiphe alphitoides ex uercus robur R Erysiphe alphitoides ex uercus robur b Erysiphe alphitoides ex uercus robur R Erysiphe alphitoides ex uercus robur Erysiphe alphitoides ex uercus robur Erysiphe alphitoides ex isteria sinensis R Erysiphe alphitoides ex uercus robur Erysiphe a ebiae

R Erysiphe euonymicola ex Euonymus japonicus R Erysiphe euonymicola ex Euonymus japonicus Erysiphe friesii Erysiphe euonymicola ex Euonymus japonicus Erysiphe euonymicola ex Euonymus japonicus Erysiphe euonymi japonica Erysiphe euonymicola ex Euonymus japonicus R Erysiphe euonymicola ex Euonymus japonicus Microsphaera japonica Erysiphe alphitoides ex uercus robur lumeria graminis ex oa trivalis Tribe Blumerieae .

Figure 3 Tree built from Mcm7 sequences, samples with accessions starting x xx(x) are accessions from the “powdery mildew survey”, samples with accessions starting with AOC come from a donation to the survey and other accession numbers refer to the RBG Kew fungarium accession number. Numbers at the nodes are bootstrap support values. odosphaera myrtillina Sphaerotheca polemonii odosphaera clandestina ex Malus pumila Tribe odosphaera leucotricha ex Malus domestica odosphaera clandestina ex Crataegus monogyna odosphaera dipsacearum sp ex Dipsacus sp Cystotheceae odosphaera euphorbiae helioscopiae ex Euphorbia peplus Golovinomyces cucurbitacearum odosphaera balsaminae odosphaera xanthii odosphaera sp ex Taraxacum officinale ibrodium balsaminae A C Golovinomyces verbasci ex erbascum thapsus Golovinomyces sordidus ex lantago major eoerysiphe galeopsidis ex Lamium sp eoerysiphe galeopsidis ex Stachys byzantina eoerysiphe galeopsidis ex Stachys sylvatica eoerysiphe galeopsidis ex Lamium sp eoerysiphe galeopsidis ex Stachys byzantina eoerysiphe galeopsidis ex Lamium purpureum eoerysiphe galeopsidis ex Lamium sp eoerysiphe galeopsidis ex allota nigra eoerysiphe galeopsidis ex Lamium amplexicaule eoerysiphe galeopsidis ex Stachys arvensis eoerysiphe galeopsidis ex Acanthus spinosus eoerysiphe galeopsidis ex Acanthus mollis eoerysiphe geranii ex Geranium phaeum eoerysiphe geranii ex Geranium pratense hyllactinia betulae A C G Tribe eoerysiphe galii ex Galium aparine eoerysiphe galii ex Galium aparine Golocinomyceteae Golovinomyces cichoracearum eoerysiphe nevoi ex Taraxacum officinale Golovinomyces cichoracearum eoerysiphe nevoi ex Sonchus arvensis eoerysiphe nevoi ex Sonchus arvensis Arthrocladiella mougeotii ex Lycium barbarum Golovinomyces depressus ex Arctium minus Golovinomyces depressus ex Arctium minus Golovinomyces depressus ex Arctium minus Erysiphe echinopis Golovinomyces cichoracearum Golovinomyces biocellaris ex Salvia officinlais Golovinomyces ambrosiae Golovinomyces circumfusus Golovinomyces circumfusus A C Golovinomyces cichoracearum D Golovinomyces cichoracearum Erysiphe verbenicola Golovinomyces orontii ex Cucurbita pepo Golovinomyces cynoglossi ex Silene dioica Golovinomyces veroci A C Golovinomyces sonchicola ex Sonchus oleracues Golovinomyces sonchicola ex Sonchus oleraceus Golovinomyces sonchicola ex Sonchus oleraceus Golovinomyces cynoglossi ex Myosotis sp Golovinomyces cynoglossi ex Myosotis arvensis Golovinomyces magnicellulatus ex hlox paniculata Golovinomyces magnicellulatus ex Epilobium hirsutum Golovinomyces magnicellulatus ex hlox paniculata Golovinomyces cichoracearum ex ilosella aurantiaca Golovinomyces cynoglossi ex Myosotis laxa Golovinomyces cynoglossi ex Myosotis arvensis Golovinomyces macrosporus Erysiphe ulmi A Erysiphe prunastri ex runus spinosus Erysiphe mayorii E Erysiphe necator ex itis vinifera Erysiphe deutziae Microsphaera ornata A Microsphaera penicillata Erysiphe syringae D Erysiphe platani ex latanus x hispanica Erysiphe cichoracearum Erysiphe elevata ex Catalpa bignonioides Microsphaera rayssiae Erysiphe baptisiae C Erysiphe sp ex Trifolium pratense Erysiphe trifoliorum ex Trifolium pratense Erysiphe trifoliorum ex Trifolium pratense Erysiphe trifoliorum ex Lathyrus odoratus Erysiphe trifoliorum ex ypericum sp Erysiphe sp ex Lathyrus pratensis Erysiphe trifoliorum ex Trifolium dubium Erysiphe thesii Erysiphe berberidis ex Mahonia moseri Erysiphe berberidis ex erberis thunbergii Erysiphe berberidis ex erberis sp Erysiphe cruciferarum ex rassica sp Erysiphe cruciferarum ex rassica sp Erysiphe cruciferarum ex Alliaria petiolata Erysiphe heraclei ex eracleum sphondylium Tribe Erysiphese

Erysiphe heraclei ex eracleum sphondylium Erysiphe heraclei ex eracleum sphondylium Erysiphe heraclei ex Anthriscus sylvestris Erysiphe heraclei ex Anthriscus sylvestris Erysiphe buhrii ex Silene dioica Erysiphe heraclei ex eracleum sphondylium Erysiphe heraclei ex eracleum sphondylium Erysiphe heraclei ex eracleum sphondylium Erysiphe heraclei ex Anthriscus sylvestris Erysiphe alphitoides ex uercus robur Erysiphe alphitoides ex uercus robur Erysiphe euonymicola ex Euonymus japonicus Erysiphe friesii Erysiphe euonymi japonici G Erysiphe euonymicola ex Euonymus japonicus Erysiphe alphitoides ex uercus robur Erysiphe alphitoides ex uercus robur Erysiphe a ebiae

Erysiphe alphitoides ex uercus robur Erysiphe alphitoides ex isteria sinensis Erysiphe alphitoides ex uercus robur Erysiphe alphitoides ex uercus robur Erysiphe alphitoides ex uercus robur Erysiphe euonymicola ex Euonymus japonicus Erysiphe euonymicola ex Euonymus japonicus Erysiphe euonymicola ex Euonymus japonicus Erysiphe sp ex Lonicera periclymenum

Erysiphe sp ex Lonicera sp Erysiphe sp ex Lonicera japonica Erysiphe sp ex Lonicera peroclymenum Microsphaera lonicerae C Erysiphe aquilegiae ex Aquilegia sp Erysiphe aquilegiae ex Aquilegia vulgaris Erysiphe aquilegiae ex Aquilegia sp Erysiphe aquilegiae var. ranunculi A C Erysiphe aquilegiae var. ranunculi Erysiphe aquilegiae Delphinium sp Erysiphe aquilegiae ex Ranunculus acris Erysiphe aquilegiae var. ranunculi A C Erysiphe tortilis A seudoidium passiflorae A C Erysiphe aquilegiae var. ranunculi Erysiphe catalpae ex Catalpa bignonioides Erysiphe aquilegiae ex Aquilegia sp Erysiphe aquilegiae ex Ranunculus repens Erysiphe aquilegiae ex Ranunculus repens Erysiphe aquilegiae ex Aquilegia sp Erysiphe aquilegiae ex Caltha palustris lumeria graminis ex oa trivialis Tribe Blumerieae

. igure Tree built from ITS sequences, samples with accessions starting x xx(x) are accessions from the “powdery mildew survey”, samples with accessions starting with AOC come from a donation to the survey and other accession numbers refer to the RBG Kew fungarium accession number. Numbers at the nodes are bootstrap support values. Tribe Cystotheceae Tribe Phyllactinieae

Tribe Erysiphese

Tribe Golovinomyceteae

Figure 5 Tree built from concatenated ITS and Mcm7 sequences, samples with accessions starting x_xx(x) are accessions from the “powdery mildew survey”, samples with accessions starting with AOC come from a donation to the survey and other accession numbers refer to the RBG Kew fungarium accession number. Numbers at the nodes are bootstrap support values. Tribe Blumerieae

Discussion

Phylogenies

A tree built containing sequence data for all available taxa was constructed and can be seen in appendix 5. This tree has incredibly low non-significant support values, especially in earlier divergences. This tree highlights the need for new and improved methods in order to fully understand the systematics of the order Erysiphales on a large scale. Despite this tree having low support values in early divergence, some of the later divergences have stronger support values and place genera in line with current thinking. For example, Leveillula, Pleochaeta, and Phyllactinia form their own monophyletic clade, this is believed to be the case as these are the only endoparasitic genera and have grouped in phylogenies built in the past (Aghayeva et al., 2018; Braun et al., 2006). However, the tree falls short in the placement of basal genera, in the past Parauncinula and Caespitotheca have been considered basal genera that diverged before Blumeria (Braun et al., 2006), however this tree has not supported this placing them together and sister to the genus Phyllactinia.

The trees built generally follow current thinking of the Erysiphales phylogeny, inconsistencies include the placement of the genus Neoerysiphe, which typically forms a distinct clade placed sister to the genus Golovinomyces, forming the monophyletic tribe Golovinomyceteae which is seen in Braun et al., 2006 this is seen in the tree built with ITS but with low support values, in the Mcm7 tree and the concatenated tree the tribe Golovinomyceteae is not monophyletic. The placement of the genera Neoerysiphe and Golovinomyces has low support in all trees built, this could point to more data needing to be generated or other genes may be needed in order to conclusively state where these genera fall. Other genes that have been tested in the past for their potential use in studing powdery mildews include Tsr1, Actin, and β-tubulin (Ellingham, 2017).

Sequence data for one species of Phyllactinia was collected during this study, Phyllactinia betulae, this is the first sequence of Mcm7 for this genus and it formed a monophyletic clade on its own in the concatenated phylogeny (Figure 5) which corresponds to the tribe Phyllactiniaea in the Braun et al., 2006 phylogeny. Collecting more data for this genus along with the other endophytic genera Leveillula and Pleochaeta may help to support this clade and improve support values within an integrated phylogeny of ITS and Mcm7. The concatenated tree also had improved support values for the genus Podospahera, Arthrocladellia, and the most speciose genus Erysiphe which had poor support values in the single gene trees. Overall it is clear that using a tree which integrates ITS and Mcm7 builds the best tree and significantly increases most support values, this suggests that using a concatenated tree would be able to resolve suspected species complexes if the correct sequence data is available.

The position of less speciose genera on the phylogenetic tree of Erysiphales is still questionable, such as Brasiliomyces (Braun & Cook, 2012), and basal genera Parauncinula and Caespitotheca. Gathering Mcm7 sequences for these genera may improve our understanding of their position given that using ITS and Mcm7 together built a relatively strong phylogeny.

Barcoding

ITS did not provide many species level IDs typically only to genus level, this is often the case meaning that morphological analysis is typically needed alongside molecular analysis to ID a species. Interestingly, Golovinomyces cucurbitacearum a species which previously had no sequence data available matched with the species Podosphaera xanthii, both species infect plants in the Cucurbitaceae family and P. xanthii is a suspected species complex. The ITS tree in Figure 4 also places G. cucurbitacearum in the genus Podosphaera, this means it is likely that G. cucurbitacearum belongs to the genus Podosphaera and not Golovinomyces. As P. xanthii is a suspected species complex it is possible that G. cucurbitacearum and P. xanthii are the same species (and therefore G. cucurbitacearum is a synonym of P. xanthii) or that G. cucurbitacearum is a species within the complex of P. xanthii.More sequence data for both species will allow investigation into which is the case.

The searches of Mcm7 sequences did not match any other sequences, which is not surprising as Mcm7 is not widely used across the powdery mildews, the few sequences that did match, matched with Mcm7 sequences from other fungal groups, including Dermatocarpon, Claviceps, and Solenopsora.

A relatively low number of sequences were obtained by the study compared to how many successfully amplified, this may be because of contamination during PCR clean-up protocols, or suboptimal clean-up protocols, possibly too many cycles in the cycle sequencing steps, or it could potentially be because of issues from amplifying various sequences from other organisms on the leaf surface.

Assessing adequacy of Mcm7

In terms of the practicality of using Mcm7 in the lab, optimal protocols still need work. Whereas ITS is commonly used in many labs, Mcm7 is not and as a result the protocols used for working with Mcm7 during this study have changed drastically. Initially, the chelex method (Turan, et al., 2015.; Walsh, et al., 1991) was trialled for extracting DNA from samples, however this was only successful for ITS so the glass filter plate method was adopted. The PCR protocol also went through multiple iterations before reaching the protocol that was used in this study. Initially, less TBT par was used and there was no MgCl2 in the master mix. Increasing the amount of TBT-PAR has been shown to enhance PCR products in the past as it contains a series of compounds which help to “mop up” contaminants, reducing inhibitors (Samarakoon, Wang, & Alford, 2013). Adding MgCl2 to the master mix improves the product as MgCl2 is a cofactor to the DNApolymerase enzyme (Schabmueller et al., 2000). Mcm7 had fewer samples which successfully amplified compared to ITS (61 for Mcm7 compared to 148 for ITS). It was also noted that ITS was successfully amplified from samples which were much older than Mcm7. The oldest sample from which ITS was successfully extracted was from 1893, whereas for Mcm7 the oldest sample was from 1935. In general, samples from across the 19th century (especially the latter half) were relatively successful in extracting ITS compared to Mcm7 which success rates seem to drop rapidly when samples over 30 years old.

It is likely that ITS had a higher amplification success and worked on older specimens more successfully because Mcm7 is a single copy gene which appears in the genome only once, whereas ITS is a region repeated multiple times in the genome (Ellingham, 2017). This means that as the specimens age and the DNA breaks down, as ITS appears multiple times it more likely to survive for longer than Mcm7, which only requires one break to no longer be extractible.

Therefore, it is recommended that for use of Mcm7 in future studies that when using fresh material, the gene should be sampled as soon as possible and, when using collections in studies that the youngest specimens be selected for sampling for the highest likelihood of success.

It also appears that certain genera had lower success in extracting Mcm7 than others, namely Podosphaera had a much lower success rate than other genera. This highlights the need for genera specific primer design in the future, Podosphaera sequences generated in this study may be useful for this. It can expected that if Mcm7 becomes more widely used across various labs that the protocols will be refined to their optimal state in the same way ITS has, this includes optimal PCR protocols and primers. The first Phyllactinia sequence for Mcm7 generated for this study may now be used to generate genus specific primers for Mcm7.

Conclusions

It has been shown that when used alone Mcm7 and ITS produce weak phylogenies, however when both are concatenated the resulting phylogeny is significantly better. More sequence data is required to fully understand whether Mcm7 can be used to fully understand the Erysiphales phylogeny, especially for underrepresented genera such as Brasiliomyces. More sequence data for the endoparasitic clade containing the genera Leveillula, Pleochaeta, and Phyllactinia may also help strengthen the concatenated phylogeny. The position of Neoerysiphe and Golovinomyces is still questionable and more sequence data or different genes may be needed to resolve these. More sequence data for this concatenated tree is likely to resolve suspected species complexes.

Building a large-scale phylogeny of the Erysiphales may allow comprehensive phylogenetic study to resolve problematic taxa such as suspected species complexes and genera. Powdery mildews pose an important threat to future food security and understanding their taxonomy may allow us to understand better how to treat infections or reduce impact.

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Appendix

Abbreviations

CIPRES – Cyberinfrastructure for Phylogenetic Research ddNTP – Difeoxynucleotides triphosphates DNA – Deoxyribonucleic acid dNTP – nucleoside triphosphate EDTA - Ethylenediaminetetraacetic acid ID - Identification ITS – Internal transcribed spacer Mcm7 – minichromosome maintenance complex component 7 NCBI – National Center for Biotechnology Information PCR – Polymerase chain reaction RBG Kew – Royal Botanic Gardens Kew TAE – Tris-acetate-EDTA UNITE - https://unite.ut.ee/cite.php

Tribe Genus Described Synonyms Erysipheae Erysiphe R. Hedw. ex DC., in Lamarck & de "Alphitomorpha Wallr., Verh. Ges. nat. Freunde Berlin 1(1): 31 (1819) Candolle, Fl. franç., Edn 3 (Paris) Arthrocladia Golovin, Trudy botanicheskogo Instituta im. V.L. Komarova 2: 272 (1805) AN SSSR, Series II, Spor. Rast. 10: 309 (1956) Arthrocladiella Vassilkov, otaniches iĭ Zhurnal : ( ) ulbomicrosphaera A. . ang, Acta Mycol. Sin. 6(2): 74 (1987) Bulbouncinula R.Y. Zheng & G.Q. Chen, Acta microbiol. sin. 19(4): 376 (1979) Calocladia Lév., Annls Sci. Nat., Bot., sér. 3 15: 154 (1851) Erysiphe sect. Trichocladia de Bary, Abh. senckenb. naturforsch. Ges. 7: 411 (1870) Erysiphella Peck, Ann. Rep. N.Y. St. Mus. nat. Hist. 28: 63 (1876) [1875] Erysiphopsis Halst., Bull. Torrey bot. Club 26: 594 (1899) Furcouncinula Z.X. Chen, in Chen, Gao, Hu & Luo, Acta Mycol. Sin. 1(1): 10 (1982) Linkomyces Golovin, Compl. Fl. Champ. Supér. Maroc 5: 127 (1958) Medusosphaera Golovin & Gamalizk., Bot. Mater. Otd. Sporov. Rast. Bot. Inst. Komarova Akad. Nauk S.S.S.R. 15: 91 (1962) Microsphaera Lév., Annls Sci. Nat., Bot., sér. 3 15: 381 (1851) Orthochaeta Sawada, Rep. Govt Res. Inst. Dep. Agric., Formosa 85: 22 (1943) Pseudoidium Y.S. Paul & J.N. Kapoor, Indian Phytopath. 38(4): 762 (1986) [1985] Salmonomyces Chidd., Sydowia 13(1-6): 55 (1959) Setoerysiphe Y. Nomura, Trans. Mycol. Soc. Japan 25(2): 163 (1984) Tigria Trevis., Spighe Paglie 1: 22 (1853) Trichocladia (de Bary) Neger, Flora, Regensburg 88: 350 (1901) Uncinula Lév., Annls Sci. Nat., Bot., sér. 3 15: 151 (1851) Uncinuliella R.Y. Zheng & G.Q. Chen, Acta microbiol. sin. 19(3): 283 (1979)" Golovinomyceteae Golovinomyces (U. Braun) V.P. Heluta, Biol. Zh. Erysiphe sect. Golovinomyces U. Braun, Feddes Repert. 88(9-10): 659 Armenii 41: 357 (1988) (1978) [1977] Golovinomyceteae Arthrocladiella assil ov, otaniches iĭ Zhurnal Index fungorum has Arthrocladiella listed as syn for Erysiphe 45: 1368 (1960) Golovinomyceteae Neoerysiphe U. Braun, Schlechtendalia 3: 50 (1999) Phyllactinieae Phyllactinia Lév., Annls Sci. Nat., Bot., sér. 3 15: 144 (1851) Phyllactinieae Leveillula G. Arnaud, Annls Épiphyt. 7: 92 (1921) Phyllactinieae Pleochaeta Sacc. & Speg., in Saccardo, "Queirozia Viégas & Cardoso, Revista Soc. Brazil agron. 7: 5 (1944) Michelia 2(no. 7): 373 (1881) Uncinulopsis Sawada, Trans. Formosa Nat. Hist. Soc. 6: 33 (1916)" Cystotheceae Sawadaea Miyabe, in Sawada, Spec. Bull. Agric. Exp. Station Formosa 9: 49 (1914) Cystotheceae Cystotheca Berk. & M.A. Curtis, Proc. Amer. Lanomyces Gäum., Ann. Jard. Bot. Buitenzorg 32: 46 (1922) Acad. Arts & Sci. 4: 130 (1860) Cystotheceae Podosphaera Kunze, in Kunze & Schmidt, "Albigo Ehrh. ex Reum, Oekon. Botan.: 68 (1833) Desetangsia Nieuwl., Mykologische Hefte (Leipzig) 2: Am. Midl. Nat. 4: 385 (1916) Euoidium Y.S. Paul & J.N. Kapoor, Indian 111 (1823) Phytopath. 38(4): 761 (1986) [1985] Kokkalera Ponnappa, Sydowia 23(1- 6): 4 (1970) [1969] Leucothallia Trevis., Spighe Paglie 1: 23 (1853) Sphaerotheca Lév., Annls Sci. Nat., Bot., sér. 3 15: 138 (1851) " Blumerieae Blumeria Golovin ex Speer, Sydowia 27(1- Blumeria Golovin, Sborník Rab. Inst. Prikl. Zool. Fitopat. 5: 124 (1958) 6): 2 (1975) [1973-1974] Oidium Link, in Willdenow, Sp. pl., Edn 4 "Acrosporium Nees, Syst. Pilze (Würzburg): 53 (1816) [1816-17] 6(1): 121 (1824) Amphiblistrum Corda, Icon. fung. (Prague) 1: 11 (1837) Graciloidium (R.T.A. Cook, A.J. Inman & C. Billings) R.T.A. Cook & U. Braun, Taxonomic Manual of the Erysiphales (Powdery Mildews): 349 (2012) Octagoidium (R.T.A. Cook, A.J. Inman & C. Billings) R.T.A. Cook & U. Braun, Taxonomic Manual of the Erysiphales (Powdery Mildews): 172 (2012) Oideum Ehrenb., Sylv. mycol. berol. (Berlin): 10 (1818) Oidium Link, Mag. Gesell. naturf. Freunde, Berlin 3(1-2): 18 (1809) Oidium Sacc., Michelia 2(no. 6): 15 (1880) Oidium subgen. Graciloidium R.T.A. Cook, A.J. Inman & C. Billings, Mycol. Res. 101(8): 998 (1997) Oidium subgen. Octagoidium R.T.A. Cook, A.J. Inman & C. Billings, Mycol. Res. 101(8): 998 (1997) Oospora Wallr., Fl. crypt. Germ. (Norimbergae) 2: 182 (1833) Toruloidea Sumst., Mycologia 5(2): 53 (1913)" Setoidium (R.T.A. Cook, A.J. Inman & C. Oidium subgen. Setoidium R.T.A. Cook, A.J. Inman & C. Billings, Mycol. Billings) R.T.A. Cook & U. Braun, Res. 101(8): 998 (1997) Taxonomic Manual of the Erysiphales (Powdery Mildews): 92 (2012) Striatoidium (R.T.A. Cook, A.J. Inman & C. Oidium subgen. Striatoidium R.T.A. Cook, A.J. Inman & C. Billings, Mycol. Billings) R.T.A. Cook & U. Braun, Res. 101(8): 998 (1997) Taxonomic Manual of the Erysiphales (Powdery Mildews): 339 (2012) Fibroidium (R.T.A. Cook, A.J. Inman & C. Oidium subgen. Fibroidium R.T.A. Cook, A.J. Inman & C. Billings, Mycol. Billings) R.T.A. Cook & U. Braun, Res. 101(8): 998 (1997) Taxonomic Manual of the Erysiphales (Powdery Mildews): 99 (2012)

Microidium (To-anun & S. Takam.) To-anun & Oidium subgen. Microidium To-anun & S. Takam., Mycoscience 46(1): 7 S. Takam., Taxonomic Manual of (2005) the Erysiphales (Powdery Mildews): 624 (2012) Brasiliomyces Viégas, Bragantia 4(1-6): 17 "Californiomyces U. Braun, Nova Hedwigia 34(3 & 4): 688 (1981) (1944) Salmonia S. Blumer & E. Müll., Phytopath. Z. 50: 382 (1964)" Caespitotheca S. Takam. & U. Braun, Mycol. Res. 109(8): 907 (2005) Parauncinula S. Takam. & U. Braun, Uncinulella Hara, (1936) Mycoscience 46(1): 14 (2005)

Appendix 1 – Summary of Currently accepted powdery mildew genera with synonymy

Genus Species Number of # in ITS Mcm7 Actin B- Tsr1 Other Synonymy/ other notes seqs Fungarium tubulin Neoerysiphe galeopsidis 107(70) 93 50+ 10+ 10+ 4 2 Erysiphe galeopsidis 109(70) 8 50+ 10+ 10+ 4 2 Neoerysiphe galeopsidis (pages for both on NCBI) Erysiphe ranunculi 113(76 6 50+ 10+ 10+ 10+ 1 syn E.aquilegiae UNITE) Erysiphe aquilegiae 113(76 3 50+ 10+ 10+ 10+ 1 UNITE) Microsphaera alphitoides 270 (206 1 50+ 20 7 6 11 10 syn E.alphitoides UNITE) Erysiphe alphitoides 270 (206 163 50+ 10 7 6 11 10 UNITE) Erysiphe heraclei 105(60) 27 50+ 10 6 8 9 Erysiphe umbelliferarum f. 105(60) 1 50+ 10 6 8 9 syn E.heraclei selini Erysiphe orontii 455(146) 52 50+ 1 5 OEC126 precursor Golovinomyces orontii 455(146) 8 50+ 1 5 Oidium begoniae 455(146) 1 50+ 1 5 syn G. orontii Erysiphe necator 2626(88) 1 50+ 1 1 1 1 14-alpha demethylase resistant allele Leveillula taurica 166(97) 126 50+ Podosphaera fusca 145(74) 63 50+ 2 cytochrome b Erysiphe cichoracearum 13581(115) 45 50+ whole genome shotgun syn G. cichoracearum (thousands) Sphaerotheca fusca 145(74) 17 50+ 2 cytochrome b syn P.fusca Golovinomyces cichoracearum 13581(115) 13 50+ whole genome shotgun (thousands) Podosphaera pannosa 114(93) 10 50+ 3 3 Sphaerotheca pannosa 114(93) 9 50+ 3 3 syn P.pannosa Sphaerotheca verbenae 145(74) 8 50+ 2 cytochrome b syn P.fusca

Podosphaera xanthii 382(306) 4 50+ 3 2 GTPase, elongation factor 1, syn P. fusca (all data taked from cytochrome b P. xanthii because of high no. entries) Oidium obductum 166(97) 4 50+ syn L. taurica Erysiphe cichoracearum var. 13581(115) 2 50+ whole genome shotgun syn G. cichoracearum latispora (thousands) Leveillula solanacearum 166(97) 2 50+ syn L.taurica Leveillula taurica f. pegani 166(97) 1 50+ Oidium neolycopersici 3984(95) 0 50+ 1 whole genome shotgun, chitin syn (similar synthase Ps.neolycopersici OE2013PM21 to) final id not O. neolycopersici Sphaerotheca cucurbitae 145(74) 0 50+ 2 cytochrome b syn P.fusca (+4 ITS from Sp. cucurbitae) Erysiphe quercicola 346(192) 0 50+ none listedas final id Oidium heveae 276(64) 0 50+ whole genome shotgun, cutinase gene, HO-73 MAP kinase Erysiphe cruciferarum 59(38) 22 40+ 3 1 1 4 OE2013PM18 not ided at E.cruciferarum Erysiphe biocellata 46(47) 8 40+ 2 2 syn Golovinomyces biocellatus Golovinomyces biocellatus 46(47) 0 40+ 2 2 Erysiphe pisi 87(50) 119 40+ 1 2 1 Adenylate cyclase gene,cAMP-dependent proten kinase, other protein kinases, various other genes,whole genome shotgun Sawadaea bicornis 65(45) 15 40+ 7 5 3 Podosphaera tridactyla 51(34) 11 40+ 1 2 2 Podosphaera aphanis 56(47) 6 40+ 2 5 Uncinula aceris 65(45) 6 40+ 7 5 3 syn S.bicornis Uncinula bicornis 65(45) 5 40+ 7 5 3 syn S.bicornis

Microsphaera diffusa 46(72) 3 40+ syn E.diffusa Erysiphe glycines var. 46(72) 2 40+ syn E.diffusa lespedezae Erysiphe diffusa 46(72) 0 40+ Podosphaera clandestina 47(39) 13 35+ 1 2 1 3 Golovinomyces monardae 40(29) 0 35+ Erysiphe trifoliorum 48(30) 2 30+ 5 3 3 3 Podosphaera leucotricha 38 16 30+ 2 3 Podosphaera macrospora 35(32) 3 30+ 1 1 1 Sphaerotheca alpina f. macrospora 35(32) 3 30+ 1 1 1 syn P.macrospora Phyllactinia fraxini 34 35 30+ 1 1 Golovinomyces ambrosiae 37(40) 8 30+ 1 chitin synthase live culture on UNITE Erysiphe macleayae 32(23) 2 30+ Erysiphe cynoglossi 40(20) 8 25+ 9 4 Sphaerotheca erigerontis- 42(26) 3 25+ 3 3 5 syn P. e-c OE2016PMCS67 the canadensis only final id as P.erigerontis- canadensis Podosphaera erigerontis- 42(26) 0 25+ 3 3 5 OE2016PMCS67 the only final canadensis id as P.erigerontis-canadensis Oidium farinosum 38 1 25+ 2 3 cytochrome b syn P.leucotricha Microsphaera platani 30(28) 7 25+ 1 1 1 1 syn E.platani +6 ITS from M.platani Erysiphe platani 30(28) 5 25+ 1 1 1 1 Microsphaera platani (Oidium 30(28) 1 25+ 1 1 1 1 syn E.platani +6 ITS from stage) M.platani Erysiphe polygoni 36(26) 37 25+ 3 3 cytochrome b Sphaerotheca fuliginea 30(8) 16 25+ syn P. fuliginea Sphaerotheca spiraeae 26(23) 5 25+ syn P.spiraeae Oidium erysiphoides 30(8) 2 25+ syn P.fuliginea Podosphaera spiraeae 26(23) 1 25+ Erysiphe lespedezae 27(15) 0 25+

Golovinomyces spadiceus 29(21) 0 25+ Podosphaera fuliginea 30(8) 0 25+ Golovinomyces asperifolii 21 0 21+ Golovinomyces cynoglossi 40(20) 21 20+ 9 4 Erysiphe elevata 28(3) 9 20+ 1 1 2 Microsphaera elevata 28(3) 2 20+ 1 1 2 syn E.elevata Erysiphe trifolii 28(27) 69 20+ Phosphoribosyltransferase gene, tRNA-dihydrouridine, CDP-diacylglycerol-3- phosphate 3- phosphatidyltransferase gene Sawadaea tulasnei 30(21) 21 20+ 1 Microsphaera trifolii 28(27) 10 20+ Phosphoribosyltransferase syn E.trifolii gene, tRNA-dihydrouridine, CDP-diacylglycerol-3- phosphate 3- phosphatidyltransferase gene Erysiphe adunca 27 7 20+ 1 1 Erysiphe pulchra 3096(14) 2 20+ MicrosatelliteEP066,64,60,etc, whole genome shotgun (many-1000) Erysiphe martii 28(27) 1 20+ Phosphoribosyltransferase syn E.trifolii gene, tRNA-dihydrouridine, CDP-diacylglycerol-3- phosphate 3- phosphatidyltransferase gene Erysiphe hiratae 25(17) 0 20+ Erysiphe japonica 23(44) 0 20+ Erysiphe longiappendiculata 21(5) 0 20+ Microidium phyllanthi 21(25) 0 20+ Erysiphe sordida 35(13) 7 15+ 9 8 1 1 syn G.sordidus Golovinomyces sordidus 35(13) 4 15+ 9 8 1 1

Erysiphe magnicellulata 19(14) 9 15+ 3 syn G. magnicellulatus Erysiphe lonicerae 26(23) 5 15+ 3 1 1 2 Not on Loci database but Mcm7 done Golovinomyces magnicellulatus 19(14) 4 15+ 3 Arthrocladiella mougeotii 23 8 15+ 1 2 1 0 syn E.mougeotii Blumeria graminis 65826 5 15+ 1 5+ 5+ 5+ whole genome shotgun Microsphaera mougeotii 23 3 15+ 1 2 1 0 syn E.mougeotii (data taken from A.mougeotii) Phyllactinia guttata 20(17) 38 15+ Erysiphe artemisiae 20(17) 17 15+ syn G.artemisiae Erysiphe convolvuli 30(19) 6 15+ 2 3 nicotnate phosphoribosyltransferase gene, tRNA-dihydrouridine synthase b gene Erysiphe glycines 16(12) 6 15+ syn E. diffusa Erysiphe betae 18(17) 4 15+ Final id not E.betae Phyllactinia corylea 20(17) 4 15+ syn Ph. guttata Golovinomyces artemisiae 20(17) 3 15+ Phyllactinia betulae 20(17) 2 15+ syn Ph. guttata +4 ITS from Ph. betulae Erysiphe arcuata 19(12 0 15+ 1 1 1 Final IDof 2013PM13 not UNITE) E.arcuta Golovinomyces asterum 19(15) 0 15+ Phyllactinia berberidis 20(17) 0 15+ syn Ph. guttata +2 ITS from Ph. berberidis Cystotheca kusanoi 31 0 15+ Erysiphe australiana 16(12) 0 15+ Erysiphe sedi 20(13) 0 15+ Erysiphe uncinuloides 19(6) 0 15+ Euoidium longipes 24(18) 0 15+ 1 1 OE2014PM110CS ided as O.longipes, syn of Eu.longipes

Phyllactinia enkianthi 16(8) 0 15+ Pseudoidium neolycopersici 16(75) 0 15+ Sawadaea polyfida 17(11) 0 15+ Neoerysiphe geranii 17(11) 3 10+ 4 Golovinomyces sonchicola 21(10) 1 10+ 4 2 1 No final id as G.sonchicola Microsphaera berberidis 21(12) 12 10+ 3 2 3 1 syn E.berberidis Erysiphe depressa 21(11) 6 10+ 3 3 1 syn G.depressus Golovinomyces depressus 21(11) 5 10+ 3 3 1 Erysiphe berberidis 21(12) 4 10+ 3 2 3 1 Erysiphe buhrii 17(13) 4 10+ 1 1 1 1 Uncinula prunastri 14(8) 3 10+ 1 syn E.prunastri Erysiphe hyperici 13(10) 3 10+ 1 1 Not on Loci database but Mcm7 done Microsphaera hypericacearum 13(10) 3 10+ 1 1 synE.hyperici Not on Loci database but Mcm7 done Podosphaera plantaginis 13(10) 1 10+ 1 1 Mcm7 done but not on loci database Erysiphe prunastri 14(8) 0 10+ 1 Microsphaera palczewskii 12(9) 13 10+ syn E.palczewskii Erysiphe syringae 17(10) 7 10+ 1 MAT1-1-1, MAT1-2-1, Cytochrome, alpha-demethylase Podosphaera fugax 24(13) 6 10+ 4 3 4 Phyllactinia mali 11(8) 3 10+ Erysiphe pseudogracilis 13(6) 2 10+ Erysiphe syringae-japonicae 12(10) 1 10+ 1 Golovinomyces bolayi 11 0 10+ Erysiphe azerbaijanica 3(12) 0 10+ Erysiphe longifilamentosa 11(5) 0 10+ Erysiphe monoperidiata 13(6) 0 10+ Erysiphe palczewskii 12(9) 0 10+ Parauncinula polyspora 11 0 10+

Phyllactinia antarctica 11(6) 0 10+ Phyllactinia juglandis 11(8) 0 10+ Pleochaeta shiraiana 11(7) 0 10+ Podosphaera prunicola 12(11) 0 10+ Golovinomyces neosalviae 14 0 14 Pseudoidium javanicum 14(5) 0 14 Phyllactinia fraxinicola 11(8) 0 11 Neoerysiphe galii 13(9) 3 10 2 1 Erysiphe galii 13(9) 2 10 2 1 syn N.galii Erysiphe hypophylla 10(18) 16 10 None have final id as E.hypophylla Microsphaera hypophylla 10(18) 0 10 syn E.hypophylla +1 ITS from M.hypophylla (None have final id as E.hypophylla) Erysiphe blasti 10(11) 0 10 Erysiphe paeoniae 10(9) 0 10 Phyllactinia broussonetiae- 10(6) 0 10 kaempferi Podosphaera epilobii 12(8) 2 9 1 1 1 Podosphaera balsaminae 9(7) 15 9 Erysiphe magnifica 9(7) 5 9 Uncinula septata 9(7) 1 9 syn Pa.septata Oidium lycopersici 25(8) 0 9 syn Eu. lycopersici OE2013PM21 final id not O.lycopersici Erysiphe castaneigena 9(8) 0 9 Erysiphe corylopsidis 9(5) 0 9 Erysiphe ligustri 16(6) 0 9 3 Chitin synthase, cytochrome P450 Erysiphe mori 9(14) 0 9

Microsphaera sinensis 9(8) 0 9 syn E.castaneigena +1 ITS from M.sinesis Parauncinula septata 9(7) 0 9 Phyllactinia actinidiae 9(8) 0 9 Erysiphe hedwigii 11(9) 4 8 1 1 1 Microsphaera hedwigii 11(9) 4 8 1 1 1 syn E.hedwigii (+1 ITS from M.hedwigii) Erysiphe friesii 8(5) 3 8 Erysiphe viciae-unijugae 8(6) 3 8 Erysiphe limonii 8(6) 1 8 Oidium mangiferae 8 0 8 Pseudoidium pedaliacearum 8 0 8 Erysiphe baliensis 5(8) 0 8 Erysiphe berberidicola 8(6) 0 8 Erysiphe corylacearum 8(5) 0 8 Erysiphe magnoliae 8(6) 0 8 Erysiphe magnoliicola 8(9) 0 8 Erysiphe salmonii 8(5) 0 8 Erysiphe takamatsui 8(3) 0 8 Microidium phyllanthi- 18(8) 0 8 reticulati Neoerysiphe hiratae 8(7) 0 8 Neoerysiphe kerribeeensis 8 0 8 Phyllactinia alni 8(4) 0 8 Podosphaera astericola 8(5) 0 8 Podosphaera macularis 13(7) 35 7 MAT 1-1-1, SLA2, MAT 1-1- 3 Sphaerotheca humuli 13(7) 10 7 MAT 1-1-1, SLA2, MAT 1-1- syn P. macularis (+1 ITS 3 fromSp. humuli) Golovinomyces fischeri 11(6) 6 7 3 1 (OE2013PM33 final id not G.fischeri)

Microsphaera symphoricarpi 7(7) 5 7 syn E.symphoricarpi Sphaerotheca macularis 13(7) 3 7 MAT 1-1-1, SLA2, MAT 1-1- syn P.macularis 3 Erysiphe fischeri 11(6) 2 7 3 1 syn G.fischeri (OE2013PM33 final id not G.fischeri) Podosphaera amelanchieris 7(5) 2 7 Golovinomyces montagnei 8(5) 1 7 1 Erysiphe symphoricarpi 7(7) 1 7 Phyllactinia populi 7(5) 1 7 Cystotheca lanestris 7 0 7 (10 on NCBI) Erysiphe aucubae 7(5) 0 7 Erysiphe eschscholziae 7(4) 0 7 Erysiphe gracilis 7(34) 0 7 Erysiphe kenjiana 7 0 7 Phyllactinia kakicola 7(3) 0 7 Pseudoidium hortensiae 6(7) 0 7 Sawadaea nankinensis 7 0 7 Sphaerotheca mors-uvae 10(6) 5 6 1 1 2 synP.mors-uvae Golovinomyces verbasci 9(5) 3 6 1 1 1 Erysiphe verbasci 9(5) 2 6 1 1 1 syn G.verbasci Podosphaera mors-uvae 10(6) 1 6 1 1 2 Uncinula flexuosa 6(6) 21 6 syn E.flexuosa Erysiphe howeana 6(1) 10 6 Erysiphe magellanica 6 9 6 Erysiphe flexuosa 6(6) 6 6 Microsphaera ornata 6 4 6 syn E.ornata Microsphaera ornata var. europaea 6 2 6 syn E.ornata Golovinomyces chrysanthemi 6 1 6 Golovinomyces macrocarpus 6(4) 1 6 Leveillula duriaei 6(4) 1 6 Phyllactinia magnoliae 6(3) 1 6

Uncinula salicis var. miyabei 6(5) 1 6 syn E.miyabei Erysiphe epigena 6(15) 0 6 Erysiphe hypogena 6(5) 0 6 Erysiphe longissima 6(3) 0 6 Erysiphe miyabei 6(5) 0 6 Erysiphe ornata 6 0 6 Golovinomyces riedlianus 6(4) 0 6 Leveillula lactucae-serriolae 6(1) 0 6 Parauncinula uncinata 6 0 6 Phyllactinia dalbergiae 6(4) 0 6 Phyllactinia eupteleae 6(4) 0 6 Phyllactinia obclavata 6(2) 0 6 Phyllactinia roboris 6(3) 0 6 Phyllactinia salmonii 6(3) 0 6 Setoidium castanopsidis 6(3) 0 6 syn C. castanopsidis (no record on here) Erysiphe circaeae 6(5) 4 5 1 Erysiphe euonymi 5(2) 11 5 Erysiphe akebiae 11 (5 2 5 3 3 UNITE) Erysiphe carpinicola 5(4) 2 5 Erysiphe deutziae 5(4) 2 5 Erysiphe vanbruntiana 5 2 5 Phyllactinia ailanthi 5(4) 2 5 Leveillula cylindrospora 5(4) 1 5 Uncinula fraxini 5(3) 1 5 syn E.fraxinicola Uncinula verniciferae 5(4) 1 5 syn E.verniciferae Golovinomyces asperifoliorum 5 0 5 Erysiphe acantholimonis 5 0 5 Erysiphe asiatica 5(3 0 5 UNITE)

Erysiphe caricae-papayae 5 0 5 Erysiphe carpini-laxiflorae 5(4) 0 5 Erysiphe fraxinicola 5(3) 0 5 Erysiphe havrylenkoana 5(2) 0 5 Erysiphe izuensis 5(4) 0 5 Erysiphe kissiana 5(2) 0 5 Erysiphe pseudolonicerae 6(5) 0 5 Erysiphe simulans 3(5) 0 5 Erysiphe verniciferae 5(4) 0 5 Erysiphe wallrothii 5(4) 0 5 Neoerysiphe cumminsiana 5 0 5 Phyllactinia broussonetiae- 5(2) 0 5 papyriferae Phyllactinia cassiae-fistulae 5(2) 0 5 Phyllactinia gmelinae 5(2) 0 5 Phyllactinia pyri-serotinae 5(4) 0 5 Podosphaera minor 5(1) 0 5 Microsphaera ludens 10(4) 2 4 2 1 2 1 syn E.ludens Not on Loci database but Mcm7 done Erysiphe ludens 10(4) 0 4 2 1 2 1 Not on Loci database but Mcm7 done Podosphaera euphorbiae- 6(3) 0 4 1 1 helioscopiae Erysiphe urticae 4 13 4 Erysiphe valerianae 5(3) 3 4 1 syn G. valerianae Erysiphe astragali 4 2 4 Erysiphe kusanoi 4(33) 1 4 Golovinomyces valerianae 5(3) 1 4 1 Microsphaera japonica 4(5) 1 4 syn E.cornicola Phyllactinia toonae 4(2) 1 4 Erysiphe trina 4 0 4

Podosphaera ferruginea 4(5) 0 4 Erysiphe cornicola 4(5) 0 4 Erysiphe huayinensis 4(2) 0 4 Erysiphe javanica 4(3) 0 4 Erysiphe linderae 4(12) 0 4 Erysiphe liquidambaris 4(6) 0 4 Erysiphe multappendicis 4(3) 0 4 Golovinomyces salviae 4(2) 0 4 Phyllactinia alangii 4(2) 0 4 Phyllactinia linderae 4(2) 0 4 Phyllactinia paliuri 4(3) 0 4 Phyllactinia poinsettiae 4(3) 0 4 Pleochaeta indica 4 0 4 Podosphaera caricicola 4(3) 0 4 Podosphaera elsholtziae 4(2) 0 4 No record on IF Podosphaera longiseta 4(1) 0 4 Pseudoidium nyctaginacearum 4(2) 0 4 Erysiphe catalpae 5(3) 4 3 1 1 More ided based on morpho alone Podosphaera dipsacacearum 6(3) 3 3 1 2 Mcm7 done but not on loci database Leveillula lanuginosa 3 10 3 Erysiphe azaleae 4(2) 8 3 1 Podosphaera erodii 3(2) 6 3 Erysiphe baeumleri 3(1) 4 3 Erysiphe ulmariae 3 4 3 Sphaerotheca euphorbiae-hirtae 3(2) 4 3 synP.euphorbiae-hirtae Erysiphe lythri 3 3 3 Erysiphe divaricata 3 1 3 Erysiphe densa 3(1) 1 3 Erysiphe mayorii 3 1 3

Erysiphe russellii 3(1) 1 3 Golovinomyces circumfusus 3(1) 1 3 Leveillula lactucarum 3 1 3 Microsphaera juglandis 3 1 3 syn E.juglandis Oidium ericinum 4(2) 1 3 1 syn E. azaleae Podosphaera phtheirospermi 3(1) 1 3 Podosphaera aphanis var. aphanis 3(4) 1 3 Fibroidium diosteae 3 0 3 Erysiphe acaenae 3 0 3 Erysiphe carpini-cordatae 3(2) 0 3 Erysiphe fallax 3(2) 0 3 Erysiphe juglandis 3 0 3 Erysiphe liriodendri 3(2) 0 3 Erysiphe monascogera 3(4) 0 3 Erysiphe oehrensii 3(2) 0 3 Erysiphe oleosa 3(1) 0 3 Erysiphe patagoniaca 3 0 3 Erysiphe pileae 3(1) 0 3 Erysiphe sidae 3(5) 0 3 Erysiphe thaxteri 3(2) 0 3 Erysiphe viburni-plicati 3 0 3 Euoidium mutisiae 3 0 3 Euoidium reginae 3 0 3 Golovinomyces adenophorae 3(2) 0 3 Leveillula saxaouli 3(2) 0 3 Phyllactinia lagerstroemiae 3(5) 0 3 Pleochaeta polychaeta 3(2) 0 3 Podosphaera euphorbiae-hirtae 3(2) 0 3 Podosphaera negeri 3 0 3 Podosphaera perseae- 3(2) 0 3 No record on IF americanae

Pseudoidium hardenbergiae 3(2) 0 3 Microsphaera sparsa 2(3) 7 2 syn E. viburni +1 ITS from M.sparsa OE2016PMCS53 final id is E.hedwigii Microsphaera viburni 2(3) 3 2 syn E.viburni OE2016PMCS53 final id is E.hedwigii Microsphaera alni 2(1) 3 2 syn Ph.alnicola Erysiphe bivonae syn 2 1 2 syn E.clandestina UNITE Leveillula clavata 2(3) 1 2 Neoerysiphe joerstadii 2(1) 1 2 Phyllactinia leveilluloides 2(1) 1 2 Phyllactinia orbicularis 2(1) 1 2 Uncinula sengokui 2(1) 1 2 syn E.sengokui Uncinula actinidiae 2 1 2 syn E.actinidiae Erysiphe epimedii 2 0 2 Golovinomyces euphorbiicola 2 0 2 Podosphaera cayratiae 2 0 2 Erysiphe viburni 2(3) 0 2 OE2016PMCS53 final id is E.hedwigii Brasiliomyces malachrae 2 0 2 Cystotheca tjibodensis 2 0 2 Cystotheca wrightii 2 0 2 (5 on NCBI) Erysiphe abbreviata 2 0 2 Erysiphe abeliicola 2 0 2 Erysiphe actinidiae 2 0 2 (3 on NCBI) Erysiphe aphananthes 2 0 2 Erysiphe berchemiae 2(1) 0 2 Erysiphe betulina 2(1) 0 2 Erysiphe bremeri 2(1) 0 2 Erysiphe clandestina 2 0 2

Erysiphe fernandoae 2(3) 0 2 Erysiphe frickii 2 0 2 Erysiphe ljubarskii 2 0 2 Erysiphe malvae 2 0 2 Erysiphe michikoae 2(3) 0 2 Erysiphe miranda 2 0 2 Erysiphe nothofagi 2 0 2 Erysiphe paracarpinicola 2(1) 0 2 Erysiphe peruviana 2 0 2 Erysiphe schizandrae 2(1) 0 2 Erysiphe sengokui 2(1) 0 2 Erysiphe sinomenii 2(1) 0 2 Erysiphe staphyleae 2 0 2 Erysiphe tectonae 2 0 2 Erysiphe weigelae 2 0 2 Erysiphe zelkowae 2(1) 0 2 Fibroidium maculatae (2)1 0 2 No record on IF Golovinomyces arabidis 2(1) 0 2 Golovinomyces calceolariae 2 0 2 Golovinomyces leuceriae 2 0 2 Golovinomyces ocimi 2 0 2 Golovinomyces sparsus 0(2) 0 2 Leveillula chrozophorae 2 0 2 Leveillula elaeagni 2 0 2 Leveillula rubiae 2(1) 0 2 Leveillula simonianii 2(1) 0 2 Phyllactinia actinidiae- 2 0 2 latifoliae Phyllactinia alnicola 2(1) 0 2 Phyllactinia angulata 2(1) 0 2 Phyllactinia bougainvilleae 2(1) 0 2

Phyllactinia carpinicola 2(1) 0 2 Phyllactinia corni 2 0 2 Phyllactinia hemipteleae 2 0 2 Phyllactinia mimosae 2(1) 0 2 Phyllactinia mori-macrourae 2(1) 0 2 Phyllactinia moricola 2(13) 0 2 Phyllactinia parietariae 2(1) 0 2 Phyllactinia philadelphi 2 0 2 Phyllactinia pterostyracis 2(1) 0 2 Phyllactinia sapii 2(1) 0 2 Phyllactinia takamatsui 2(3) 0 2 Phyllactinia terminaliae 2(1) 0 2 Podosphaera collomiae 2(1) 0 2 Podosphaera girardiniae 2(1) 0 2 Podosphaera gunnerae 2 0 2 Podosphaera hibiscicola 2(1) 0 2 Podosphaera intermedia 2(1) 0 2 Podosphaera paracurvispora 2(1) 0 2 No record on IF Podosphaera photiniae 2(1) 0 2 Sawadaea koelreuteriae 2 0 2 Sawadaea negundinis 2 0 2 Sphaerotheca euphorbiae 1 26 1 syn P.euphorbiae Erysiphe tortilis 2(1) 17 1 1 OE2014PM83CS ided as E.tortillis but no seq data Microsphaera penicillata 1 8 1 syn E.penicillata Erysiphe knautiae 1 5 1 Erysiphe lycopsidis 2 4 1 1 Erysiphe vanbruntiana var. 1 4 1 sambuci-racemosae Microsphaera vaccinii 1 2 1 syn E.penicillata

Erysiphe hyoscyami 1 1 1 syn G. hyoscyami, E. hyoscyami in herbtrack Brasiliomyces trinus 1 1 1 Erysiphe rayssiae 1 1 1 Microsphaera magnusii 1 1 1 syn E.magnusii Microsphaera menispermi 1 1 1 syn E.menispermi Microsphaera rayssiae 1 1 1 syn E.rayssiae Microsphaera hommae 1 1 1 syn E.corylacearum Oidium caricae 1 1 1 syn Ps.caricae Phyllactinia pyri 1(3) 1 1 Erysiphe circinata 1 0 1 (Takamatsuella circinata?-IF) Erysiphe coriariigena 1 0 1 Erysiphe discariae 1 0 1 Erysiphe ovidiae 1 0 1 Erysiphe rhamnicola 1 0 1 Erysiphe robiniae 1 0 1 Erysiphe rosae 1 0 1 Euoidium fuegianum 1 0 1 Golovinomyces hyoscyami 1 0 1 Pleochaeta turbinata 1 0 1 Podosphaera verbenae 0 1 No record on IF Pseudoidium ipomoeae 1 0 1 No record on IF Sawadaea aesculi 1 0 1 Striatoidium aloysiae 1 0 1 syn N.aloysiae (no entry) Striatoidium baccharidis 1 0 1 syn N.bacchardis (no entry) Striatoidium maquii 1 0 1 syn N.maquii (no entry) Bulbomicroidium bauhiniicola 1 0 1 (no index fungorum) Caespitotheca forestalis 1 0 1 (no index fungorum)(3 seq on NCBI) Erysiphe balbisiae 1 0 1 Erysiphe begoniicola 1 0 1

Erysiphe caricae 1(6) 0 1 Erysiphe carpophila 1 0 1 Erysiphe chloranthi 1 0 1 Erysiphe clethrae 1 0 1 Erysiphe coriariae 1 0 1 Erysiphe diervillae 1(2) 0 1 Erysiphe erlangshanensis 1 0 1 Erysiphe euphorbiae 1 0 1 Erysiphe fimbriata 1 0 1 Erysiphe guarinonii 1(1) 0 1 Erysiphe helwingiae 1 0 1 Erysiphe hommae 1 0 1 syn E.corylacearum??? Erysiphe hydrangeae 1 0 1 Erysiphe magnusii 1 0 1 Erysiphe menispermi 1 0 1 Erysiphe myoschili 1 0 1 Erysiphe myzodendri 1 0 1 Erysiphe nishidana 1 0 1 Erysiphe nomurae 1 0 1 Erysiphe orixae 1 0 1 Erysiphe panacis 1 0 1 Erysiphe penicillata 1 0 1 Erysiphe phyllanthi 1 0 1 Erysiphe ribicola 1 0 1 Erysiphe robiniicola 0(1) 0 1 Erysiphe rodgersiae 1 0 1 Erysiphe schizophragmatis 1 0 1 Erysiphe sesbaniae 1 0 1 Erysiphe togashiana 1 0 1 Erysiphe wadae 1(3) 0 1

Fibroidium balsaminae 1 0 1 Golovinomyces echinopis 1 0 1 Golovinomyces inulae 1 0 1 Golovinomyces verbenae 0(1) 0 1 Leveillula buddlejae 1 0 1 Leveillula contractirostris 1 0 1 Leveillula jaczewskii 1 0 1 Leveillula lorantii 1 0 1 No record on IF Leveillula picridis 1 0 1 Leveillula verbasci 1 0 1 Oidium cinnamomi 1 0 1 syn Ps. cinnamomi Oidium citri 1 0 1 Oidium ixodiae 1 0 1 syn Eu.ixodiae Phyllactinia adesmiae 1 0 1 Phyllactinia ampulliformis 1 0 1 Phyllactinia babayanii 1 0 1 Phyllactinia caricicola 1 0 1 Phyllactinia carpini 1 0 1 Phyllactinia chubutiana 1(2) 0 1 Phyllactinia durantae 1 0 1 Pleochaeta prosopidis 1 0 1 Podosphaera caricae-papayae 0(1) 0 1 Podosphaera carpesiicola 1 0 1 Podosphaera cercidiphylli 1 0 1 Podosphaera curvispora 1(2) 0 1 Podosphaera diclipterae 1 0 1 Podosphaera lini 1(2) 0 1 Podosphaera phaseoli 1 0 1 Podosphaera pseudofusca 1 0 1 Podosphaera salatai 1 0 1 Podosphaera senecionis 3(1) 0 1 1 1

Podosphaera sparsa 1 0 1 Podosphaera stephanandrae 1 0 1 Pseudoidium boroniae 1 0 1 Erysiphe euonymicola 29(16) 0 6 1 5 Erysiphe intermedia 6(4) 2 4 1 1 Microsphaera trifolii var. 6(4) 1 4 1 1 syn E.intermedia intermedia Podosphaera filipendulae 8(5) 0 2 3 2 1 OE2014PM33CS final id not P. filipendulae Microsphaera grossulariae 28 Microsphaera euphorbiae 21 syn E.euphorbiicola Uncinula adunca 21 syn E. adunca Erysiphe aquilegiae var. 20 syn E. aquilegiae ranunculi Podosphaera myrtillina 17 Podosphaera euphorbiae 14 Sphaerotheca aphanis 13 syn P. aphanis Golovinomyces cucurbitacearum 11 Microsphaera azaleae 10 syn E. azaleae Sphaerotheca balsaminae 9 syn P. balsaminae Sphaerotheca epilobii 8 syn P. epilobii Microsphaera friesii 8 syn E. friesii Erysiphe euonymi-japonici 7 listed on NCBI but no sequences Erysiphe acalyphae 7 Microsphaera ravenelii 7 syn E.ravenelii Erysiphe sparsa 6 G.sparsa Uncinula parvula 6 syn E.parvula Erysiphe capreae 5 Podosphaera biuncinata 5

Erysiphe aquilegiae var. 5 aquilegiae Golovinomyces asterum var. 5 syn G. asterum solidaginis Golovinomyces magnicellulata 5 No record - magnicellulatus? Microsphaera pulchra 5 syn E. pulchra Oidium hortensiae 7(6) 5 syn Ps. hortensiae Erysiphe verbenicola 4 Leveillula leguminosarum 4 Microsphaera euonymi 4 syn E. euonymi Sphaerotheca fugax 4 syn P. fugax Uncinula necator 4 E. necator Sphaerotheca plantaginis 4 syn P. plantaginis Erysiphe ulmi 3 4 Erysiphe graminis 3 Blumeria graminis Oidium lauracearum 3 syn Ps.lauracearum Sphaerotheca polemonii 3 syn P. polemonii Erysiphe adunca var. regularis 3 syn E. adunca Microsphaera lonicerae 3 syn E. lonicerae Erysiphe pisi var. cruchetiana 3 syn E. pisi Microsphaera vanbruntiana 3 syn E. vanbruntiana Microsphaera vanbruntiana var. 3 syn E. vanbruntiana sambuci-racemosae Oidium carpini 3 OE2013PM3 final id not O. carpini Erysiphe actinostemmatis 2 Erysiphe baptisiae 2 Erysiphe galegae 2 Erysiphe pseudoregularis 2 Erysiphe thesii 2 Specimens on UNITE (no sequence)

Erysiphe echinopis 2 Erysiphe erodii 2 Erysiphe hyphophylla 2 sp error E. hypophylla? Erysiphe polygonii 2 sp error? E.polygoni? Leveillula lichenoides 2 No record on IF Microsphaera extensa 2 syn E.extensa Microsphaera sambucicola 2 Microsphaera alni var. yamadae 2 syn E.yamadae Oidium aureum 2 syn Botryobasidium aureum (not pm?) Oidium candicans 2 Oidium conspersum 2 syn Botryobasidium conspersum Podosphaera parietariae 2 Uncinula australis 2 syn E.australis Uncinula delavayi 2 syn E.delavayi Erysiphe asperifoliorum 2 syn G. asperifoliorum Microsphaera baeumleri 2 syn E. baeumleri Golovinomyces biocellaris 2 sp error? biocellatus? Uncinula circinata 2 Takamatsuella circinata Sphaerotheca dipsacacearum 2 syn P. dipsacacearum Microsphaera divaricata 2 syn E. divaricata Microsphaera magnifica 2 syn E. magnifica Erysiphe pisi var. pisi 2 Microsphaera syringae 2 syn E. syringae Brasiliomyces trina 0 1 B. trinus (orthographic variant?) Erysiphe aggregata 1 Specimens listed on UNITE Erysiphe alangii 1 Erysiphe grossulariae 1 Erysiphe jatrophae 1 Erysiphe pseudacaciae 1

Erysiphe vernalis 1 Erysiphe poonaensis 1 syn G. poonaensis Erysiphe trinae 1 E. trina? sp error? Erysiphe communis f. 1 paeoniae Erysiphe oronthii 1 sp error? G.orontii? Golovinomyces depressum 1 sp error? G.depressus? Leveillula helichrysi 1 Leveillula lanata 1 Leveillula lappae 1 Leveillula scrophulariacearum 1 Microsphaera ellisii 1 syn E.ellisii Microsphaera extensa var. curta 1 syn E.extensa Microsphaera friesii var. friesii 1 syn E.friesii Microsphaera miyabeana 1 syn E.miyabeana Microsphaera nemopanthi 1 syn E.nemopanthi Microsphaera peckii 1 syn E.peckii Microsphaera semitosta 1 syn E.semitosta Microsphaera juglandis-nigrae 1 syn E.juglandis-nigrae Microsphaera ludens var. lathyri 1 syn E. lathyricola Oidium hiratae 1 syn Fibroidium hiratae Oidium ipomoeae 1 syn Ps. ipomoeae Oidium necator 1 sp error? no record on IF Phyllactinia juglandis- 1 mandshuricae Podosphaera myrtillina var. 1 myrtillina Podosphaera schlechtendalii 1 Podosphaera thalictri 1 Sphaerotheca pruinosa 1 syn P. pruinosa Sphaerotheca erigerontis 1

Uncinula clintonii 1 syn E.clintonii Uncinula incrassata 1 syn E.incrassata Uncinula adunca var. 1 syn E. mandshurica mandshurica Microsphaera akebiae 1 syn E. akebiae Golovinomyces asterum var. asterum 1 Microsphaera astragali 1 syn E. astragali Golovinomyces cichoracearum var. 1 cichoracearum Podosphaera clandestina var. 1 syn P. clandestina aucupariae Erysiphe convolvuli var. 1 convolvuli Podosphaera dipsacaceaum 1 sp error? dipsacaceaum Oidium euonymi-japonici 1 syn E. euonymi-japonici Sphaerotheca parietariae 1 syn P. parietariae Microsphaera russellii 1 syn E. russellii Phyllactinia syringae 1 Microsphaera syringae-japonicae 1 syn E. syringae-japonicae Microsphaera tortilis 1 syn E. tortilis Erysiphe trifolii var. 1 syn E. intermedia intermedia Neoerysiphe nevoi 22(18) 1 Erysiphe katumotoi 0 Has NCBI page but no sequence data Erysiphe pseudocarpinicola 0 Has page on NCBI but no sequence data Oidium anacardii 0 syn Ps. anacardii Oidium bixae 0 Phyllactinia marissalii 0

Podosphaera alpina no seq on 0 No final ids as P.alpina UNITE or NCBI Phyllactinia hamamelidis 6(3) 0 Appendix 2 – sequence data currently availiable

Genus Species PCR PCR Accession Host Family Association Country Year ITS Mcm7 Brasiliomyces trinus 168965 Fagaceae Quercus agrifolia USA 1981 Brasiliomyces trina 13561 Fagaceae Quercus agrifolia USA 1946 Brasiliomyces setosus + 263212 Sapindaceae Sapindus oahuensis Hawaii 1984 Brasiliomyces entadae 263213 Fabaceae Entada spicata South Africa 1964 Erysiphe carpinicola 167378 Betulaceae Carpinus betulus France 2010 Erysiphe cichoracearum + + 159957 Asteraceae Cirsium oleraceum Czech Republic 2008 Erysiphe cichoracearum m 45852 Asteraceae Dahlia sp. Portugal 1996 Erysiphe cichoracearum + 49581 Asteraceae Gnaphalium ?luteoalbum Portugal 1996 Erysiphe fischeri 85396 Asteraceae Senecio viscosus Sweden 1988 Erysiphe cichoracearum 85397 Asteraceae Sonchus palustris, Sweden 1984 Phragmites Erysiphe cichoracearum 208317 Asteraceae Inula Asia (Eastern or Indian 1961 Subcontinent) Erysiphe cichoracearum f 208322 Asteraceae Serratula Asia (Eastern) 1902 Erysiphe cichoracearum f 208309 Asteraceae Carthamus Asia (Indian 1984 Subcontinent) Erysiphe cichoracearum f 208315 Asteraceae Guizotia Asia (Indian 1988 Subcontinent) Erysiphe poonaensis + 208314 Asteraceae Goniocaulon Asia (Indian 1954 Subcontinent) Erysiphe cichoracearum 208313 Asteraceae Crepis Asia (Middle East) 1930 Erysiphe cichoracearum 128467 Asteraceae Saussurea sp. China 1960

Erysiphe convolvuli var. 128468 Convolvulace Convolvulus arvensis China 1935 convolvuli a Erysiphe actinostemmat 208327 Cucurbitacea Cucurbitaceae Asia (Eastern) is e Erysiphe acalyphae 128465 Euphorbiacea Acalyphe brachystachya China 1959 e Erysiphe viciae- m 208387 Fabaceae Vicia Asia (Eastern) 1904 unijugae Erysiphe cichoracearum + 208385 Fabaceae Tephrosia Asia (Indian 1978 Subcontinent) Erysiphe cichoracearum 208248 Asteraceae Helianthus Africa 1932 Erysiphe cichoracearum 208250 Asteraceae Vernonia Africa 1963 Erysiphe densa 208247 Asteraceae Compositae Algeria 1819 Erysiphe cichoracearum 208251 Asteraceae Xanthium Egypt 1917 Erysiphe cichoracearum 95154 Asteraceae Erechtites hieracifolia USA 1968 Erysiphe hypophylla 234452 Fagaceae Quercus frainetto Romania 1974 Erysiphe syringae- + + 191155 Oleaceae Ligustrum vulgare Germany 2010 japonicae Erysiphe macleayae + m 233464 Papaveraceae Chelidonium majus Germany 2014 (Papaveraceae) Erysiphe aquilegiae var. + 162122 Ranunculace Aconitum variegatum Austria 2008 ranunculi ae Erysiphe aquilegiae var. + m 117413 Ranunculace Ranunculus aconitifolius Austria 1994 ranunculi ae Erysiphe aquilegiae var. + + 160004 Ranunculace Ranunculus repens Czech Republic 2008 ranunculi ae Erysiphe pseudoregulari + 191154 Salicaceae Salix caprea Germany 2010 s Erysiphe cichoracearum 208300 Solanaceae Solanaceae Peru 1975 Leveillula lanuginosa f 208450 Apiaceae Umbelliferae Europe 1975 Leveillula lanuginosa 208449 Apiaceae Umbelliferae Europe 1975

Leveillula lichenoides 208479 Apiaceae Umbelliferae Africa 1900 Leveillula lanata f 208469 Euphorbiacea Euphorbiaceae North Africa 1976 e Leveillula clavata m 208468 Euphorbiacea Euphorbiaceae Africa 1950 e Leveillula leguminosaru f 208541 Fabaceae Leguminosae Asia 1976 m Leveillula helichrysi 233465 Asteraceae Helichrysum arenarium Germany 2013 (Asteraceae) Leveillula cylindrospora 208397 Chenopodiac Kochia Europe 1956 eae Leveillula duriaei 168956 Lamiaceae Phlomis sp. France 1879 Leveillula scrophulariace 208426 Scrophulariac Scrophulariaceae Europe 1966 arum eae Microsphaera vaccinii + 95166 Ericaceae Rhododendron viscosum USA 1968 Microsphaera euphorbiae 208555 Euphorbiacea Euphorbiaceae America borealis 1909 e Microsphaera juglandis- 169063 Juglandaceae Juglans nigra USA 1908 nigrae Microsphaera menispermi f 169064 Menispermac Menispermum USA 1874 ea canadensis Microsphaera semitosta 169065 Rubiaceae Cephalanthus USA 1879 occidentalis Microsphaera semitosta 263251 Rubiaceae Cephalanthus 1908 occidentalis Microsphaera diffusa 95160 Fabaceae Lespedeza capitata USA 1969 Microsphaera ludens var. f 169049 Fabaceae Latyrus odoratus USA 1909 lathyri Microsphaera ravenelii m 169042 Fabaceae Gleditschia sp. USA 1885 Microsphaera extensa m 95162 Fagaceae Quercus robur USA 1969

Microsphaera nemopanthi + 95163 Aquifoliacea Nemopanthus USA 1968 e mucronatus Microsphaera ellisii 95161 Betulaceae Ostrya virginiana USA 1961 Microsphaera peckii + 169082 Bignoniaceae Campsis radicans USA 1968 Microsphaera japonica + + 196796 Celastraceae Euonymus japonicus China 1991 Microsphaera miyabeana 169077 Styracaceae Styrax japonica Japan 1899 Microsphaera friesii var. + m 53761 Rhamnaceae Rhamnus cathartica Austria 1995 friesii Microsphaera palczewskii + + 125285 Fabaceae Caragana arborescens Finland 2004 Neoerysiphe joerstadii + m 187656 Asteraceae Phagnalon graecum Greece 2012 Phyllactinia leveilluloides 249398 Fagaceae Quercus potosina Mexico 2015 Phyllactinia juglandis- 128480 Juglandaceae Juglans mandshurica China 1978 mandshuricae Phyllactinia magnoliae 196478 Magnoliacea Liriodendron chinensis China 2001 e Phyllactinia toonae m 196801 Meliaceae Toona sinensis China 1992 Phyllactinia pyri 196798 Rosaceae Pyrus pyrifolia China 1992 Phyllactinia populi f 128483 Saliaceae Populus canadensis China 1983 Phyllactinia ailanthi + 196799 Simaroubace Picrasma quassioides China 1992 ae Podosphaera biuncinata 169068 Hamamelidac Hamamelis virginiana USA 1891 eae Podosphaera parietariae 192641 Urticaceae Parietaria pensylvanica Germany 2012 Sphaerotheca fusca + 165240 Asteraceae Calendula officinalis Czech Republic 2008 Sphaerotheca fusca + 160500 Asteraceae Senecio ovatus Czech Republic 2008 Sphaerotheca fusca + 160506 Asteraceae Sonchus oleraceus Czech Republic 2008 Sphaerotheca fusca 31867 Asteraceae Taraxacum officinale Norway 1994 agg. Sphaerotheca balsaminae + 160010 Balsaminacea Impatiens sp. Czech Republic 2008 e

Sphaerotheca fusca + 103641 Orobanchace Melampyrum pratense Italy 2002 ae Sphaerotheca fusca 128508 Asteraceae Lactuca sp. China 1960 Sphaerotheca euphorbiae- 208592 Euphorbiacea Euphorbiaceae Asia 1986 hirtae e Sphaerotheca fusca 95202 Asteraceae Bidens frondosa USA 1965 Sphaerotheca pruinosa 95204 Asteraceae Rhus glabra USA 1977 Uncinula actinidiae 168937 Actinidiaceae Actinidia arguta Japan 1890 Uncinula verniciferae + 128512 Anacardiacea Cotinus coggygria China 1982 e Uncinula salicis var. 168938 Betulaceae Alnus japonica Japan 1894 miyabei Uncinula sengokui + 168939 Celastraceae Celastrus articulatus Japan 1895 Uncinula septata 196797 Fagaceae Quercus glandulifera China 1992 Uncinula fraxini + 168940 Oleaceae Fraxinus longicuspis Japan 1893 Uncinula delavayi 168941 Simaroubace Ailanthus sp. China 1900 ae Uncinula prunastri + 160499 Rosaceae Prunus spinosa Czech Republic 2008 Uncinula incrassata 168946 Fabaceae Pterocarpus mellifera Mozambique [Gazaland] 1906 Uncinula australis 168948 Myrtaceae Eugenia sp. Paraguay 1883 Erysiphe guarionii + + AOC15 Laburnum anagyroides Wales 2016 Erysiphe ornata var. + AOC16 Betula pubescens Wales 2018 europaea Erysiphe divaricata + + AOC17 Frangula alnus var. alnus Wales 2015

Genus Species Acces PCR ITS PCR Mcm7 Host Family Association Count Year sion ry no Erysiphe alangii 20313 + + Apocynaceae Vinca major Wales 1999 8

Erysiphe knautiae 23260 Caprifoliaceae Scabiosa atropurpurea Engla 2016 0 nd Erysiphe symphoricarpi 16649 + Caprifoliaceae Symphoricarpos albus Engla 2008 8 nd Erysiphe vanbruntiana 16688 + Adoxaceae Sambucus racemosus Scotla 2010 var. sambuci- 8 nd racemosae Erysiphe mayorii 28055 + Asteraceae Cirsium vulgare Engla 1994 nd Erysiphe euonymi 22710 + + Celastraceae Euonymus europaeus Chann 2015 3 el Island s Erysiphe euonymi- 22710 + + Celastraceae Euonymus japonicus Chann 2015 japonici 5 el Island s Erysiphe pulchra 16454 + Cornaceae Cornus florida Engla 2009 9 nd Erysiphe tortilis 22729 + + Cornaceae Cornus sanguinea Engla 2016 8 nd Erysiphe azaleae 22730 + Ericaceae Azalea (Rhododendron) Engla 2016 7 cv. nd Erysiphe baptisiae 19421 + + Fabaceae Baptisia 'Purple Smoke' Engla 2014 4 nd Erysiphe galegae 12084 m Fabaceae Galega officinalis Engla 2003 3 nd Erysiphe martii 16194 + Fabaceae Melilotus sp. Engla 1985 4 nd Erysiphe pseudacaciae 18462 + Fabaceae Robinia Britis 1991 h Isles

Erysiphe grossulariae 19893 + Grossulariaceae Ribes sanguineum Engla 2014 4 nd Erysiphe deutziae 14002 + Hydrangeaceae Deutzia scabra Engla 2006 4 'candidissima' nd Erysiphe akebiae 23502 + + Lardizabalaceae Akebia quinata Engla 2017 4 nd Erysiphe lythri 25145 Lythraceae Lythrum salicaria Engla 2018 1 nd Erysiphe magnifica 19359 + Magnoliaceae Magnolia sp. Engla 2014 0 nd Erysiphe syringae 22705 + + Oleaceae Chann 2015 0 el Island s Erysiphe howeana 22708 + + Onagraceae Oenothera stricta Chann 2015 0 el Island s Erysiphe limonii 7147 + Plumbaginaceae Limonium vulgare Engla 1985 nd Erysiphe polygonii 23629 m Polygonaceae Polygonum aviculare Engla 1993 9 nd Erysiphe aquilegiae var. 23260 + + Ranunculaceae Aquilegia cv. Engla 2016 aquilegiae 9 nd Erysiphe aquilegiae var. 11774 + + Ranunculaceae Clematis argentilucida Engla 2003 ranunculi 8 nd Erysiphe aquilegiae var. 16763 + + Ranunculaceae Ranunculus acris Engla 2010 ranunculi 2 nd Erysiphe aquilegiae var. 17300 + + Ranunculaceae Delphinium 'Kennington Engla 2011 ranunculi 2 Classic' nd Erysiphe aquilegiae var. 19919 + + Ranunculaceae Consolida ajacis Engla 2015 ranunculi 1 nd Erysiphe aquilegiae var. 19954 + + Ranunculaceae Thalictrum flavum Engla 2015 ranunculi 2 nd Erysiphe friesii 25759 + + Rhamnaceae Rhamnus alaternus Engla 2018 3 nd Erysiphe capreae 20196 m Saliaceae Salix x multinervis Scotla 2014 4 nd Erysiphe thesii 20669 + + Santalaceae Thesium humifusum Engla 2016 0 nd Erysiphe ulmi 20065 + + Ulmaceae Ulmus procera Engla 2015 3 nd Erysiphe urticae 25757 Urticaceae Urtica dioica Engla 2018 0 nd Erysiphe verbenicola 22731 + + Verbenaceae Verbena bonariensis Engla 2016 6 nd Golovinomyc artemisiae 24940 + Asteraceae Artemisia vulgaris Scotla 2016 es 0 nd Golovinomyc ambrosiae (cf.) 23281 + Asteraceae Helianthus cv. Lemon Engla 2016 es 2 Queen nd Golovinomyc cichoracearum 19247 + + Asteraceae Lapsana communis Engla 2014 es 8 nd Golovinomyc cichoracearum 19953 + + Asteraceae Centaurea macrocephala Engla 2015 es 1 nd Erysiphe cichoracearum 6969 Asteraceae Eupatorium cannabinum Engla 1985 nd Golovinomyc cichoracearum 23492 Asteraceae Arctium lappa Engla 1947 es 9 nd Golovinomyc cichoracearum 22703 + m Asteraceae Carduus tenuiflorus Chann 2015 es 8 el Island s Golovinomyc cichoracearum 22704 m m Asteraceae Hypochaeris radicata Chann 2015 es 2 el Island s Golovinomyc cichoracearum 22711 + + Asteraceae Lactuca serriola Chann 2015 es 9 el Island s Golovinomyc cichoracearum 22713 + + Asteraceae Picris hieracoides Chann 2015 es 6 el Island s Erysiphe cichoracearum 10697 m Asteraceae Sonchus oleraceus Engla 1995 1 nd Erysiphe cichoracearum 14103 + Asteraceae Tragopogon pratensis Engla 2006 2 nd Erysiphe cichoracearum 11774 Asteraceae Helianthus epiphyllus Engla 2003 var. latispora 2 'Gold & Silver' nd Erysiphe echinopis 15869 + + Asteraceae Echinops ritro 'Taplow Engla 2008 4 Blue' nd Golovinomyc circumfusus 20460 + + Asteraceae Eupatorium cannabinum Chann 2015 es (cf.) 9 el Island s Golovinomyc fischeri 23535 + Asteraceae Jacobaea vulgaris Chann 2015 es 6 el Island s Golovinomyc macrosporus 22702 + Asteraceae Glebionis segetum (= Chann 2015 es (cf.) 5 Chrysanthemum el segetum) Island s Golovinomyc cucurbitacearu 23260 + + Cucurbitaceae Lagenaria siceraria Engla 2016 es m 3 nd Golovinomyc cucurbitacearu 23260 + m Cucurbitaceae Cucurbita maxima cv. Engla 2016 es m 6 Amphora nd Golovinomyc cichoracearum 23492 Ranunculaceae Clematis jackmanii Engla 1947 es 6 nd Leveillula lappae 22706 + + Asteraceae Carduus tenuiflorus Chann 2015 0 el Island s Microsphaera ornata 14191 + + Betulaceae Betula pubescens Engla 2006 4 nd Microsphaera penicillata 19501 + m Betulaceae Alnus glutinosa Chann 2014 7 el Island s Microsphaera lonicerae 64681 + + Caprifoliaceae Lonicera periclymenum Engla 1999 nd Microsphaera astragali 78620 + + Fabaceae Astragalus glyciphyllos Engla 2000 nd Microsphaera rayssiae 64365 + m Fabaceae Spartium junceum Engla 1999 nd Phyllactinia orbicularis 19091 m Fagaceae Fagus sylvatica sp. Engla 1999 4 nd Phyllactinia corylea 5154 m + Oleaceae Fraxinus excelsior Engla 1935 nd Phyllactinia mali 12536 Rosaceae Crataegus monogyna Engla 2004 4 nd Podosphaera xanthii 16650 m Asteraceae Taraxacuym officinale Engla 2008 0 agg. nd Podosphaera xanthii (cf.) 19953 + Asteraceae Calendula officinalis Engla 2015 6 nd Podosphaera balsaminae 20370 + m Balsaminaceae Impatiens capensis Engla 2015 6 nd

Sphaerotheca macularis 19150 + Cannabaceae Engla 2005 4 nd Podosphaera phtheirospermi 23329 + Orobanchaceae Melampyrum pratense Scotla 2016 3 nd Sphaerotheca fusca 13320 Solanaceae Physalis alkekengii Engla 2005 4 nd Sphaerotheca polemonii 16454 + Polemoniaceae Polemonium caeruleum Engla 2009 4 nd Podosphaera myrtillina 19268 + Ericaceae Vaccinium myrtillus Scotla 2013 6 nd Sphaerotheca euphorbiae 13974 + Euphorbiaceae Euphorbia peplus Engla 2005 0 nd Sphaerotheca humuli 5156 m Asteraceae Eupatorium cannabinium Engla 1945 nd Sphaerotheca fusca 11600 + m Asteraceae Matricaria matricarioides Engla 2003 1 nd Sphaerotheca fusca 14191 + Asteraceae Pulicaria dysenterica Engla 2006 9 nd Podosphaera aphanis var. 20289 Rosaceae Alchemilla alpina Scotla 2014 aphanis 2 nd Podosphaera amelanchieris 19406 + Rosaceae Amelanchier lamarckii Scotla 2014 8 nd Podosphaera macularis 20772 Cannabaceae Cannabaceae Britis 1982 8 h Isles Sphaeratheca macluris 53295 + Cannabaceae Humulus lupus Engla 1997 nd Podosphaera macularis 20772 + Cannabaceae Cannabaceae Britis 1986 5 h Isles Phyllactinia marissalii AOC1 + Acer pseudoplatanus 2013 Phyllactinia marissalii AOC2 + m Acer platanoides 2011 Phyllactinia marissalii AOC3 + Acer campestre 2018 Phyllactinia betulae AOC4 + + Betula pubescens 2016

Phyllactinia betulae AOC5 m + Betula kamtschatica 2014 Phyllactinia betulae AOC6 + + Betula celtiberica 2018 Fibroidium balsaminae AOC7 + Impatiens capensis 2016 Pseudoidium passiflorae AOC8 + + Passiflora caerulea 2012 Podosphaera aucupariae AOC9 + + Sorbus aucuparia 2017 Podosphaera myrtillina var. AOC1 m + Vaccinium myrtillus 2018 myrtillina 0 Golovinomyc veroci?? AOC1 + Vereaceum chaixii ?? 2011 es 1 Golovinomyc circumfusus AOC1 + + Eupatorium cannabinum 2015 es 2 Erysiphe aquilegiae var. AOC1 + Aconitum napellus 2013 ranunculi 3 Erysiphe aquilegiae var. AOC1 + Clematis x jackmanii 2013 ranunculi 4

Erysiphe ulmariae 62366 Erysiphe ulmariae 7372 Podosphaera biuncinata N/A Microsphaera diffusa 95159 m Microsphaera diffusa 12645 m 0 Erysiphe divaricata AOC1 + 8 Eryspihe penicillata AOC1 + 9 Eryspihe penicillata AOC2 + 0 Erysiphe aquilegiae var AOC2 + + aquilegiae 1

Erysiphe aquilegiae var AOC2 + aquilegiae 2 Erysiphe aquilegiae var AOC2 + ranunculi 3 Erysiphe hypophylla AOC2 + 4 Erysiphe aquilegiae var AOC2 + + ranunculi 5 Erysiphe aquilegiae var AOC2 + AOC26 ranunculi 6 Erysiphe baeumleri AOC2 + AOC27 7 Erysiphe aquilegiae var AOC2 + AOC28 ranunculi 8 Erysiphe ornata var. A3 + AOC29 europeae Erysiphe ornata var. B3 + + AOC30 europeae Erysiphe betae C3 + + AOC31

Appendix 3 – Specimens sampled for DNA extraction from the RBG Kew fungarium, accession numbers starting with AOC indicate samples obtained from a donation to the owdery Mildew Survey. A “+” in a CR column indicates a positive result, an “m” is a wea band, and an “f” is a faint band (that was then repeated with more MgCl2 to attempt to gain a stronger band.

Appendix 5 – Phylogeny of all ITS data available online to view please visit https://drive.google.com/file/d/1xNmeCSZSYcc0TE5Kn8mQTg7bLWmDASHg/view Species Accession ITS MCM7 BLAST result Erysiphe AOC13 + Erysiphe aquilegiae var. aquilegiae ranunculi Erysiphe 158694 + Golovinomyces echinopis sp. Erysiphe akebiae 235024 + + Erysiphe sp. Erysiphe 199191 + + Erysiphe sp. aquilegiae var. ranunculi Erysiphe 1995442 + + Erysiphe sp. aquilegiae var. ranunculi Erysiphe AOC14 + Erysiphe sp. aquilegiae var. ranunculi Erysiphe baptisiae 194214 + + Erysiphe sp. Erysiphe 106971 + Erysiphe sp. cichoracearum Erysiphe deutziae 140024 + Erysiphe sp. Erysiphe 227105 + + Erysiphe sp. euonymi-japonica Erysiphe fresii 257593 + + Erysiphe sp. Erysipe mayorii 28055 + Erysiphe sp. Erysiphe syringae 227050 + + Erysiphe sp. Erysiphe thesii 206690 + + Erysiphe sp. Erysiphe tortilis 227298 + + Mcm7 of Claviceps sp., ITS Erysiphe sp. Erysiphe ulmi 200653 + + Erysiphe sp. Erysiphe 227316 + Golovinomyces verbenicola sp. Fibrodium AOC7 + Phyllactinia sp. balsaminae Golovinomyces 232812 + Golovinomyces ambrosiae sp. Golovinomyces 192478 + Neoerysiphe sp. cichoracearum Golovinomyces 227136 + Golovinomyces cichoracearum sp. Golovinomyces 204609 + Golovinomyces circumfusus sp. Golovinomyces AOC12 + Golovinomyces circumfusus sp. Golocinomyces 232606 + Podosphaera cucurbiutacearu xanthii m Golovinomyces 227025 + Golovinomyces macrosporus sp.

Golovinomyces 227038 + Golovinomyces cichoracearum sp. Golovinomyces 227042 + Neoerysiphe sp. cichoracearum Golovinomyces 227119 + Golovinomyces cichoracearum sp. Golovinomyces AOC11 + Golovinomyces veroci sp. Microsphaera 195017 + Erysiphe sp. penicillata Microsphaera 64365 + Erysiphe sp. rayssiae Microsphaera 64681 + Erysiphe sp. lonicerae Microsphaera 141941 + Erysiphe sp. ornata Phyllactinia AOC6 + + Mcm7 of betulae Solenopsora sp., ITS Phyllactinia sp. Podosphaera 192686 + Podosphaera sp. myrtillina Podosphaera 166500 + Podosphaera xanthii xanthii Podosphaera 203706 + Podosphaera sp. balsaminae Pseudoidium AOC8 + Erysiphe/ passiflorae Pseudoidium sp. Sphaerotheca 164544 + Podosphaera sp. polemonii Erysiphe 160004 + aquilegiae var. ranunculi Erysiphe AOC25 + Mcm7 of aquilegiae var. Claviceps sp. ranunculi Erysiphe AOC17 + Mcm7 of divaricata Dermatocarpon sp. Erysiphe guarionii AOC15 + Erysiphe ornata AOC30 + var. europeae Erysiphe polygoni 14_85 + Erysiphe 191155 + syringae- japonicae Golovinomyces 14_104 + cichoracearum Microsphaera 78620 + astragali

Microsphaera 196796 + japonica Micrspphaera 125285 + palczewskii Neoerysiphe 187656 + joerstadii

Appendix 6