Chrysofolia kunmingensis E.F. Yang, 2023

Lei, Yiming, Yang, Erfu, Dao, Chengjiao, Li, Xi, Karunarathna, Samantha C., Tibpromma, Saowaluck & Yan, Kai, 2023, Endophytic fungi associated with Coriaria nepalensis in an abandoned open-pit phosphate mine in Yunnan, P. R. China, Phytotaxa 616 (1), pp. 27-46 : 31

Chrysofolia kunmingensis E.F. Yang , sp. nov. ( Figure 2 )

Index Fungorum number: IF900234

Etymology: The name reflects the location Kunming from where the holotype was collected.

Holotype: HKAS 129065

Endophytic on fresh leaves of Coriaria nepalensis , sporulated after half month. Sexual morph: Unknown. Asexual morph on PDA: Mycelium 2–5 µm thick, hyaline, septate, branched, thick-welled, conidial masses and conidiomata were observed in PDA culture after one month. Conidiomata 175–236 × 190–260 µm high (x = 205 × 225 µm; n = 20), pycnidial, separate to aggregated, exuding orange or red slimy conidial mass, without neck. Conidiophores reduced to conidiogenous cells. Conidiogenous cell 5–8 × 1.5–3 μm (x = 6.5 × 2.5 µm; n = 10), hyaline, cylindrical to ampulliform, straight to curved, wider at the base, smooth-walled. Conidia 8–14 × 3–6 μm (x = 11 × 5 µm; n = 20), hyaline, one-celled, ellipsoidal to irregular, straight, apex obtuse, base with flattened hilum, guttulate, smooth-walled.

Culture characteristics: Colonies on PDA, reaching around 30 mm diameter after 2 weeks at 20–25 ° C. Obverse: orange to red slimy conidial mass, raised, effuse, circular; fluffy, white mycelium at the centre, lowly dense at the outer parts, with abundant conidiomata. Reverse: orange-red in the centre, dark red at the outer parts, without pigments produced in PDA.

Material examined: China, Yunnan Province, Kunming, Kunyang, an abandoned and Open-Pit Phosphate Mine, on healthy leaves of Coriaria nepalensis , 22 August 2022, C. J. Dao, 2 MS1 L6 , ( HKAS 129065 , holotype), ex-type KUNCC23-13350 , KUNCC23-13351 . GenBank number : KUNCC23-13350 = ITS: OR094460 , LSU: OR094455 , tub2: OR095639 ; KUNCC23-13351 = ITS: OR094461 , LSU: OR094456 , tub2: OR095640 .

Notes: Based on BLASTn results of LSU and ITS, our isolates of Chrysofolia kunmingensis ( KUNCC 23-13350, 23-13351) are 99%–100% similar to C. colombiana ( CBS 139909, CPC 24986, LCM 988.01) and Chrysofolia barringtoniae (SDBR-CMUENBA048). However, C. barringtoniae (SDBR-CMUENBA048) lack tef1-α and tub2 genes in GenBank, while only C. colombiana ( CBS 139909) has tef1-α gene. The BLASTn top similarity of our isolate ( KUNCC 23-13350, 23-13351) was 87% (228/261 bp; 7 gaps) with C. colombiana ( CBS 139909). The multi-gene phylogenetic trees also indicated that our isolate ( KUNCC 23-13350, 23-13351) clustered between C. barringtoniae sister to C. colombiana and formed an independent clade with 94% in ML, 1.0 in BI statistical supports ( Figure 1 ). In morphology, our isolate has different conidia (ellipsoidal to irregular vs. uniform, ellipsoidal) compared to known Chrysofolia species and releases slimy conidial mass in PDA. However, C. colombiana exudes a yellow slimy conidial mass ( Crous et al. 2015); while C. barringtoniae exudes a brown slimy conidial mass ( Suwannarach et al. 2016). Based on BLASTn results, phylogenetic analyses, and morphological differences, we introduce a new endophytic fungus, C. kunmingensis isolated from healthy leaves of Coriaria nepalensis .

Figures

FIGURE 1. Phylogram generated from Maximum Likelihood analysis based on a combined LSU, ITS, tef1-α and tub2 sequence datasets. The 22 strains are included in the combined gene analysis 2460 total character including gaps (LSU: 1–892 bp, ITS: 893–1580 bp, tef1-α: 1581–2065 bp, tub2: 2066–2598 bp). Tree topology of the ML analysis was similar to the BI. The matrix had distinct alignment patterns, with the final ML optimization likelihood value of -11028.098478 (ln). All free model parameters were estimated by RAxML model, with 982 distinct alignment patterns and 24.33% of undetermined characters or gaps. Estimated base frequencies were as follows: A = 0.236385, C =0.275681, G = 0.277948, T = 0.209986, with substitution rates AC = 1.657800, AG = 2.641351, AT = 1.759057, CG = 1.214763, CT = 5.269496, GT = 1.000000. The gamma distribution shape parameter alpha = 0.662280 and the Tree-Length = 1.116951. The final average standard deviation of split frequencies at the end of total MCMC generations was calculated as 0.008871 in BI analysis. The species introduced in this study are indicated in red, and the type strains are indicated in bold. Bootstrap values greater than 60% (ML, Left) and Bayesian posterior probabilities (BI, right) greater than 0.95 are given at the nodes. Hypens (-) represent values less than 60% in ML/0.95 in BI.

FIGURE 2. Chrysofolia kunmingensis (HKAS 129065, holotype). (a,b) Obverse and reverse of colonies on PDA; (c,d) Close up of the fruiting bodies on PDA; (e) Mycelium; (f) Conidiomata; (g–j) Conidiogenous cells bearing conidia which stained by congo red reagent; (k) Conidia stained by congo red reagent.