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A new approach to species delimitation in Septoria - CBS - KNAW

A new approach to species delimitation in Septoria - CBS - KNAW

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Verkley et al.much more profound difference is seen between cultures of thetwo <strong>species</strong>, with colonies of S. galeopsidis on OA be<strong>in</strong>g opaqueand dark olivaceous-black even at the marg<strong>in</strong>, while colonies ofS. lamiicola are more translucent yellowish <strong>to</strong> ochreous, becom<strong>in</strong>gdarker only due <strong>to</strong> the formation of pycnidia. Priest (2006) po<strong>in</strong>ted<strong>to</strong>wards differences <strong>in</strong> conidial width and conidiogenesis betweenS. lamiicola and S. galeopsidis, but hav<strong>in</strong>g compared both <strong>species</strong>morphologically <strong>in</strong> planta and <strong>in</strong> vitro, we conclude that these<strong>species</strong> cannot be dist<strong>in</strong>guished us<strong>in</strong>g these criteria. These two<strong>species</strong> are, however, readily dist<strong>in</strong>guished by DNA sequencedata, and the multilocus phylogeny provides evidence for a closerelationship with S. matricariae (<strong>CBS</strong> 109000, <strong>CBS</strong> 109001), whileother Sep<strong>to</strong>ria occurr<strong>in</strong>g on the same plant family as S. lamiicola(Lamiaceae) are all much more distant (Fig. 2). The Austrianand Dutch collections of S. lamiicola on L. album are sufficientlyhomogenous <strong>to</strong> consider them conspecific.Sep<strong>to</strong>ria leucanthemi Sacc. & Speg., <strong>in</strong> Saccardo, Michelia1: 191. 1878. Fig. 25.≡ Rhabdospora leucanthemi (Sacc. & Speg.) Petr., Sydowia 11: 351.1957.For addditional synonyms see Punithal<strong>in</strong>gam (1967b).Description <strong>in</strong> planta: Symp<strong>to</strong>ms leaf spots hologenous orepigenous, scattered, circular <strong>to</strong> irregular, pale <strong>to</strong> dull brownthroughout or with whitish central area, <strong>in</strong>def<strong>in</strong>ite with concentriczones or delimited by a slightly darker marg<strong>in</strong>. Conidiomatapycnidial, predom<strong>in</strong>antly epiphyllous, numerous scattered <strong>in</strong> eachleaf spot, subglobose <strong>to</strong> globose, brown <strong>to</strong> black, semi-immersed,130–220(–240) µm diam; ostiolum central, circular, 35–100 µmwide, surround<strong>in</strong>g cells dark brown and with more thickenedwalls; conidiomatal wall about 8–12.5 µm thick, composed of ahomogenous tissue of hyal<strong>in</strong>e, angular cells, 2.5–5 µm diam withrelatively th<strong>in</strong>, hyal<strong>in</strong>e walls, surrounded by a layer of pale <strong>to</strong> darkbrown angular <strong>to</strong> more irregular cells, 3–6.5 µm diam with slightlythickened walls. Conidiogenous cells hyal<strong>in</strong>e, discrete, rarely<strong>in</strong>tegrated <strong>in</strong> 1–2-septate conidiophores, cyl<strong>in</strong>drical <strong>to</strong> doliiform,or ampulliform, holoblastic, proliferat<strong>in</strong>g percurrently with <strong>in</strong>dist<strong>in</strong>ctannellations, or sympodially, 6–18 × 4–6.5(–7.5) µm. Conidia filiform<strong>to</strong> filiform-cyl<strong>in</strong>drical, straight, curved, sometimes slightly flexuous,attenuated gradually <strong>to</strong> a narrowly rounded <strong>to</strong> po<strong>in</strong>ted apex, widestnear the base, where attenuat<strong>in</strong>g abruptly or more gradually <strong>in</strong><strong>to</strong> anarrowly truncate base, (5–)6–13-septate (later secondary septaare developed <strong>in</strong> some cells), not constricted around the septa,hyal<strong>in</strong>e, contents with several m<strong>in</strong>ute oil-droplets and granularmaterial <strong>in</strong> each cell <strong>in</strong> the liv<strong>in</strong>g state, with m<strong>in</strong>ute oil-droplets andgranular contents <strong>in</strong> the rehydrated state, (67–)80–100(–125) ×2.5–3.0(–3.5) µm (rehydrated). Sexual morph unknown.Description <strong>in</strong> vitro: Colonies on OA 6–8(–11) mm diam <strong>in</strong> 2 wk(11–14(–17) mm <strong>in</strong> 3 wk), with an even, glabrous, colourlessmarg<strong>in</strong>; colonies spread<strong>in</strong>g, the surface plane, immersed myceliumpale buff, later more or less rosy-buff; <strong>in</strong> the centre complexes ofpycnidial conidiomata with pale brown or olivaceous walls releasemasses of pale whitish <strong>to</strong> buff conidial slime; reverse concolorous,but honey <strong>in</strong> the centre. Colonies on CMA 9–11(–13) mm diam <strong>in</strong>2 wk (15–18 mm <strong>in</strong> 3 wk), as on OA, but with some white diffuseand high aerial hyphae <strong>in</strong> the centre, and later more elevated <strong>in</strong> thecentre; reverse <strong>in</strong> the centre hazel <strong>to</strong> fawn after 3 wk; conidiomatamuch more numerous and larger than on OA, develop<strong>in</strong>g <strong>in</strong>concentric or random patterns as discrete, large acervuloid (lateralmost discoid <strong>to</strong> cupulate) stromata with olivaceous sterile tissues,releas<strong>in</strong>g large masses of pale white <strong>to</strong> pale buff conidial slime.Colonies on MEA 7–10 mm diam <strong>in</strong> 2 wk (14–17 mm <strong>in</strong> 3 wk),with an even, colourless, glabrous marg<strong>in</strong>; colonies restricted,irregularly pustulate <strong>to</strong> hemispherical, the bumpy surfaceconsist<strong>in</strong>g of numerous protrud<strong>in</strong>g conidiomatal <strong>in</strong>itials, appear<strong>in</strong>gdark, with sepia, dark brick and c<strong>in</strong>namon t<strong>in</strong>ges, aerial myceliummostly absent, locally dense, pure white and woolly; reverse mostlysepia <strong>to</strong> fawn or v<strong>in</strong>aceous buff. Sporulation only observed afterabout 7 wk. Colonies on CHA 7–13 mm diam <strong>in</strong> 2 wk (15–20 mm<strong>in</strong> 3 wk), with an even, glabrous, pale v<strong>in</strong>aceous buff marg<strong>in</strong>;colonies restricted, irregularly pustulate <strong>to</strong> conical, the surfacebumpy, immersed mycelium honey <strong>to</strong> hazel, covered by dense <strong>to</strong>diffuse, pure white, woolly aerial mycelium; conidiomata sparselydevelop<strong>in</strong>g at the surface after 2 wk, the wall slightly darker thanthe surround<strong>in</strong>g hyphae, releas<strong>in</strong>g pale white conidial slime (evenafter 3 wk); reverse c<strong>in</strong>namon <strong>in</strong> the centre, v<strong>in</strong>aceous buff or paleochreous at the marg<strong>in</strong>.Conidiomata and conidiogenous cells as <strong>in</strong> planta. Conidia as<strong>in</strong> planta, 5–13(–17)-septate, 70–125(–175) × 3–4 µm (OA).Hosts: Various <strong>species</strong> of the genera Chrysanthemum, Tagetes,Achillea, Centaurea and Helianthus (Waddell & Weber 1963,Punithal<strong>in</strong>gam 1967b, c).Material exam<strong>in</strong>ed: Austria, Tirol, Ober Inntal, Samnaun Gruppe, Böderweg onLazidalm, on liv<strong>in</strong>g leaves of Chrysanthemum leucanthemum, 8 Aug. 2000, G.Verkley 1055, <strong>CBS</strong> H-21173, liv<strong>in</strong>g cultures <strong>CBS</strong> 109090, 109091; Same substr.,Tirol, Zanderstal near Spiss, 11 Aug. 2000, G. Verkley 1069, <strong>CBS</strong> H-21170, liv<strong>in</strong>gcultures <strong>CBS</strong> 109083, 109086. Germany, Hamburg, on liv<strong>in</strong>g leaves of Chr.maximum, Sep. 1958, R. Schneider s.n. <strong>CBS</strong> H-18111, liv<strong>in</strong>g culture <strong>CBS</strong> 353.58 =BBA 8504 = IMI 91322. New Zealand, Coromandel distr., Coromandel pen<strong>in</strong>sula,Waikawau, coast along St. Hwy 25, on liv<strong>in</strong>g leaves of Chr. leucanthemum, 22 Jan.2003, G. Verkley 1826, <strong>CBS</strong> H-21247; same substr., North Island, Coromandel,Tairua Forest, along roadside of St. Hway 25, near cross<strong>in</strong>g 25A, 23 Jan. 2003, G.Verkley 1842b, <strong>CBS</strong> H-21243, liv<strong>in</strong>g culture <strong>CBS</strong> 113112.Notes: The six stra<strong>in</strong>s studied here showed m<strong>in</strong>or differences <strong>in</strong>morphological characters and DNA sequences, which show highestsimilarity <strong>to</strong> sequences of <strong>CBS</strong> 128621, an isolate orig<strong>in</strong>at<strong>in</strong>g fromCirsium setidens <strong>in</strong> South Korea, and identified as S. cirsii (Fig. 2).Sep<strong>to</strong>ria leucanthemi is also closely related <strong>to</strong> a number of otherSep<strong>to</strong>ria <strong>species</strong> from Asteraceae, such as S. senecionis and S.putrida (Senecio spp.), S. obesa (Chrysanthemum spp., Artemisia)and S. astericola (Aster sp.). It is confirmed here that Sep<strong>to</strong>riaobesa, which has been regarded as a synonym of S. leucanthemiby Jørstad (1965), should be treated as a separate <strong>species</strong> (seealso the note on S. obesa).Sep<strong>to</strong>ria lycoc<strong>to</strong>ni Speg. ex Sacc., Michelia 2 : 167. 1880.Fig. 26.Description <strong>in</strong> planta: Symp<strong>to</strong>ms leaf spots epigenous, numerous,circular <strong>to</strong> irregular, s<strong>in</strong>gle or confluent, white <strong>to</strong> pale greyish,surrounded by an <strong>in</strong>itially red, then dark brown <strong>to</strong> black and thickenedborder. Conidiomata pycnidial, epiphyllous, <strong>in</strong>conspicuous, up <strong>to</strong> afew <strong>in</strong> each leaf spot, globose <strong>to</strong> subglobose, brown, immersed,90–145(–220) µm diam; ostiolum central, circular, more or lesspapillate, 25–55 µm wide; conidiomatal wall 17–35 µm thick,composed of textura angularis, differentiated layers absent, thecells mostly 3.5–5(–11) µm diam, the outer cells with brown,somewhat thickened walls, the <strong>in</strong>ner cells with th<strong>in</strong>ner, hyali<strong>new</strong>alls. Conidiogenous cells hyal<strong>in</strong>e, cyl<strong>in</strong>drical, or elongatedampulliform with a relatively narrow neck which widens at the <strong>to</strong>p,266

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