Morphological and molecular analyses reveal three new species and one new record of endophytic fungi from Rhododendron decorum in northwestern Yunnan, China
Keywords:
Endophytic fungi, Morphology, Multigene, New species, Phylogeny, Rhododendron, TaxonomyAbstract
Northwestern Yunnan is one of the most biodiversity-rich regions in the world. Rhododendron decorum is a dominant evergreen shrub or small tree in the subalpine coniferous forests and rhododendron shrublands of this region, playing an important role in maintaining ecosystem functions and serving as a traditional medicinal and edible resource for local communities. During an investigation of the endophytic fungal diversity associated with R. decorum in northwestern Yunnan, sixteen fungal strains were isolated and identified using the tissue isolation method, of which fourteen isolates belonged to Sordariomycetes and two to Leotiomycetes. Based on morphological characteristics and multigene phylogenetic analyses, three new species are introduced in this study, viz., Pestalotiopsis decori, P. jianchuanensis, and Pezicula gaoligongensis. In addition, Colletotrichum godetiae is reported for the first time as an endophyte of R. decorum. Although Trichothecium roseum has been previously recorded from this host, it is redescribed and illustrated in detail in the present study. These findings contribute to the knowledge of the taxonomic diversity and host associations of endophytic fungi associated with Rhododendron decorum in northwestern Yunnan.
References
Bao DF, Zhang JY, Lu YZ, Tian XG, Hyde K, Liu N, Xiao Y, Luo Z, Karunarathna S, Samarakoon M et al (2025) Endophytic fungi associated with medicinal ferns in Guizhou Province, China I: Morpho-molecular characterization of culturable endophytic fungi associated with Dicranopteris spp. Mycosphere 16(1):2259–2455.
https://doi.org/10.5943/mycosphere/16/1/13
Bhat DJ, McKenzie EHC, Bahkali AH, Hyde KD (2012) A multi-locus backbone tree for Pestalotiopsis, with a polyphasic characterization of 14 new species. Fungal Diversity 56(1):95–129.
https://doi.org/10.1007/s13225-012-0198-1
Capella-Gutiérrez S, Silla-Martínez JM, Gabaldón T (2009) trimAl: A tool for automated alignment trimming in large-scale phylogenetic analyses. Bioinformatics 25(15):1972–1973.
https://doi.org/10.1093/bioinformatics/btp348
Carbone I, Kohn LM (1999) A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia 91:553–556.
https://doi.org/10.1080/00275514.1999.12061051
Chethana KWT, Manawasinghe IS, Hurdeal VG, Bhunjun CS, Appadoo MA, Gentekaki E, Raspé O, Promputtha I, Hyde KD (2021) What are fungal species and how to delineate them? Fungal Diversity 109:1–25.
https://doi.org/10.1007/s13225-021-00483-9
Crous PW, Groenewald JZ, Risède JM, Simoneau P, Hyde KD (2004) Calonectria species and their Cylindrocladium anamorphs, species with sphaeropedunculate vesicles. Studies in Mycology 50:415–430.
https://doi.org/10.3114/sim.55.1.213
Damm U, Cannon PF, Woudenberg JH, Crous PW (2012) The Colletotrichum acutatum species complex. Studies in Mycology 73(1):37–113.
https://doi.org/10.3114/sim0010
De Silva DD, Crous PW, Ades PK, Hyde KD, Taylor PWJ (2017) Life styles of Colletotrichum species and implications for plant biosecurity. Fungal Biology Reviews 31:155–168. https://doi.org/10.1016/j.fbr.2017.05.001
De Silva NI, Brooks S, Lumyong S, Hyde KD (2019) Use of endophytes as biocontrol agents. Fungal Biology Reviews 33:133–148.
https://doi.org/10.1016/j.fbr.2018.10.001
Deepak H, Virk V (2022) Optimization of surface sterilization method for the isolation of endophytic fungi associated with Curcuma longa L. and their antibacterial activity. Journal of Advanced Biotechnology and Experimental Therapeutics 5(2):334-346.
https://doi.org/10.5455/jabet.2022.d119
Fang MY, Fang RC, He MY, Hu LZ et al (2005) Rhododendron. Flora China 14:260–455.
Fontana DC, de Paula S, Torres AG, de Souza VHM, Pascholati SF, Schmidt D, Dourado Neto D (2021) Endophytic Fungi: Biological Control and Induced Resistance to Phytopathogens and Abiotic Stresses. Pathogens 10(5):570.
https://doi.org/10.3390/pathogens10050570
Gao FK, Dai CC, Liu XZ (2010) Mechanisms of fungal endophytes in plant protection against pathogens. African Journal of Microbiology Research 4(13):1346–1351.
Georgian E, Emshwiller E (2016) Rhododendron Uses and Distribution of this Knowledge within Ethnic Groups in Northwest Yunnan Province, China. Open J Soc Sci 4:138-150.
https://doi.org/10.4236/jss.2016.41018
Glez-Peña D, Gómez-Blanco D, Reboiro-Jato M, Fdez-Riverola F, Posada D (2010) ALTER: program-oriented conversion of DNA and protein alignments. Nucleic Acids Research 38:W14-W18. https://doi.org/10.1093/nar/gkq321
Gu R, Bao DF, Shen HW, Su XJ, Li YX, Luo ZL (2022) Endophytic Pestalotiopsis species associated with Rhododendron in Cangshan Mountain, Yunnan Province, China. Frontiers in Microbiology 13:1016782.
https://doi.org/10.3389/fmicb.2022.1016782
Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41:95–98.
Hosking JE, Mesarich CH, Tarallo M, de la Rosa S, Rivera SA (2024) First Report of Colletotrichum fioriniae and Colletotrichum godetiae causing anthracnose disease of Blueberry (Vaccinium corymbosum L.) Fruit in New Zealand. Australasian Plant Pathology 53:503–506.
https://doi.org/10.1007/s13313-024-01002-5
Huang ZY, Shen HW, Su XJ, Luo ZL (2024) Phyllosticta cangshanensis sp. nov., an endophytic fungus from Rhododendron decorum in Cangshan Mountain, Yunnan Province, China. Phytotaxa 678(3):146-166.
https://doi.org/10.11646/phytotaxa.678.3.1
Katoh K, Rozewicki J, Yamada KD (2019) MAFFT online service: multiple sequence alignment, interactive sequence choice and visualization. Briefings in Bioinformatics 20:1160–1166.
https://doi.org/10.1093/bib/bbx108
Kuraku S, Zmasek CM, Nishimura O, Katoh K (2013) A leaves facilitates on-demand exploration of metazoan gene family trees on MAFFT sequence alignment server with enhanced interactivity. Nucleic Acids Research 41:W22–W28.
https://doi.org/10.1093/nar/gkt389
Kusari S, Pandey SP, Spiteller M (2013) Untapped mutualistic paradigms linking host plant and endophytic fungal production of similar bioactive secondary metabolites. Phytochemistry 91:81–87.
https://doi.org/10.1016/j.phytochem.2012.07.021
Liu JK, Phookamsak R, Doilom M, Wikee S, Li YM, Ariyawansha H, Boonmee S, Chomnunti P, Dai DQ, Bhat JD et al (2012) Towards a natural classification of Botryosphaeriales. Fungal Diversity 57:149–210.
https://doi.org/10.1007/s13225-012-0207-4
Liu YJ, Whelen S, Hall BD (1999) Phylogenetic relationships among ascomycetes: evidence from an RNA polymerse II subunit. Molecular Biology and Evolution 16(12):1799–1808.
https://doi.org/10.1093/oxfordjournals.molbev.a026092
Maharachchikumbura SSN, Guo LD, Cai L, Chukeatirote E, Wu WP, Sun X, Crous PW, Bhat DJ, McKenzie EHC, Bahkali AH et al (2012) A multi-locus backbone tree for Pestalotiopsis, with a polyphasic characterization of 14 new species. Fungal Diversity 56:95–129.
https://doi.org/10.1007/s13225-012-0198-1
Maharachchikumbura SSN, Hyde KD, Groenewald JZ, Xu J, Crous PW (2014) Pestalotiopsis revisited. Studies in Mycology 79(1):121–186.
https://doi.org/10.1016/j.simyco.2014.09.005
Meena H, Hnamte S, Siddhardha B (2019) Secondary metabolites from endophytic fungi: chemical diversity and application. In: Singh BP (ed) Advances in Endophytic Fungal Research: Present Status and Future Challenges. Springer, Cham, pp 145–169.
https://doi.org/10.1007/978-3-030-03589-1_7
Miller MA, Pfeiffer W, Schwartz T (2010) Creating the CIPRES science gateway for inference of large phylogenetic trees. In: Proceedings of the 2010 Gateway Computing Environments Workshop (GCE), November 2010. New Orleans, Louisiana, pp 1–8.
https://doi.org/10.1109/GCE.2010.5676129
Monika G, Benjarong KT (2023) First Detection of Trichothecium roseum Causing Leaf Spots on Tomato in Germany. Plant Dis 107(4):1233.
https://doi.org/10.1094/PDIS-07-22-1588-PDN
Nair DN, Padmavathy S (2014) Impact of endophytic microorganisms on plants, environment and humans. The Scientific World Journal 2014:250693.
https://doi.org/10.1155/2014/250693
Nylander JAA (2004) MrModeltest v2 Program distributed by the author. Evolutionary Biology Centre, Uppsala University: Uppsala, Sweden.
Petrini O, Fisher P (1990) Occurrence of fungal endophytes in twigs of Salix fragilis and Quercus robur. Mycological Research 94:1077–1080.
https://doi.org/10.1016/s0953-7562(09)81336-1
Poveda J, Eugui D, Abril-Urías P, Velasco P (2021) Endophytic fungi as direct plant growth promoters for sustainable agricultural production. Symbiosis 85(1):1–19.
https://doi.org/10.1007/s13199-021-00789-x
Promputtha I, Lumyong S, Dhanasekaran V, McKenzie EH, Hyde KD, Jeewon R (2007) A phylogenetic evaluation of whether endophytes become saprotrophs at host senescence. Microbial Ecology 53(4):579–590.
https://doi.org/10.1007/s00248-006-9117-x
Rannala B, Yang Z (1996) Probability distribution of molecular evolutionary trees: a new method of phylogenetic inference. Journal of Molecular Evolution 43:304–311.
https://doi.org/10.1007/BF02338839
Rathnayaka AR, Tennakoon DS, Jones GE, Wanasinghe DN, Bhat DJ, Priyashantha AKH, Stephenson SL, Tibpromma S, Karunarathna SC (2024) Significance of precise documentation of hosts and geospatial data of fungal collections, with an emphasis on plant-associated fungi. New Zealand Journal of Botany 63(2–3):462–489.
https://doi.org/10.1080/0028825X.2024.2381734
Rojas-Jiménez K, Hernández M, Blanco J, Vargas LD, Acosta-Vargas LG, Tamayo G (2016) Richness of cultivable endophytic fungi along an altitudinal gradient in wet forests of Costa Rica. Fungal Ecology 20:124–131.
https://doi.org/10.1016/j.funeco.2015.12.006
Ronquist F, Huelsenbeck JP (2003) MrBayes3: bayesian phylogenetic inference under mixed models. Bioinformatics 19:1572–1574.
https://doi.org/10.1093/bioinformatics/btg180
Saikkonen K, Faeth SH, Helander M, Sullivan TJ (1998) Fungal endophytes: a continuum of interactions with host plants. Annual Review of Ecology and Systematics 29(1):319–343. https://doi.org/10.1146/annurev.ecolsys.29.1.319
Saikkonen K, Helander M, Faeth SH (2004) Fungal endophytes: hitchhikers of the green world. In: Mukerji KG (ed) Plant Microbiology. Taylor & Francis, London, pp 81–101.
Schardl CL, Craven KD, Speakman S, Stromberg A, Lindstrom A, Yoshida R (2008) A novel test for host-symbiont codivergence indicates ancient origin of fungal endophytes in grasses. Systematic Biology 57(3):483–498.
https://doi.org/10.1080/10635150802172184
Shrestha N, Wang Z, Su X, Xu X, Lyu L, Liu Y, Dimitrov D, Kennedy JD, Wang Q, Tang Z et al (2018) Global patterns of Rhododendron diversity: The role of evolutionary time and diversification rates. Global Ecology and Biogeography 27:913–924.
https://doi.org/10.1111/geb.12750
Stamatakis A (2006) RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics 22:2688–2690.
https://doi.org/10.1093/bioinformatics/btl446
Stamatakis A, Hoover P, Rougemont J (2008) A rapid bootstrap algorithm for the RAxML web-servers. Systematic Biology 57:758–771.
https://doi.org/10.1080/10635150802429642
Sun H, Zhang J, Deng T, Boufford DE (2017) Origins and evolution of plant diversity in the Hengduan Mountains, China. Plant Diversity 39(4):161–166.
https://doi.org/10.1016/j.pld.2017.09.004
Tao G, Liu ZY, Liu F, Gao YH, Cai L (2013) Endophytic Colletotrichum species from Bletilla ochracea (Orchidaceae), with descriptions of seven new speices. Fungal Diversity 61:139–164.
https://doi.org/10.1007/s13225-013-0254-5
Templeton AR, Crandall KA, Sing CF (1992) A cladistic analysis of phenotypic associations with haplotypes inferred from restriction endonuclease mapping and DNA sequence data. III. Cladogram estimation. Genetics 132(2):619–633.
https://doi.org/10.1093/genetics/132.2.619
Tian W, Zhang CQ, Qiao P, Milne R (2011) Diversity of culturable ericoid mycorrhizal fungi of Rhododendron decorum in Yunnan, China. Mycologia 103(4):703–709.
https://doi.org/10.3852/10-296
UNESCO World Heritage Centre (2025) Three Parallel Rivers of Yunnan Protected Areas. UNESCO World Heritage Centre.
https://whc.unesco.org/en/list/1083/.
Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172(8):4238–4246.
https://doi.org/10.1128/jb.172.8.4238-4246.1990
Wang GN, Yu XD, Dong W, Bhat DJ, Boonmee S, Zhang D, Zhang H (2019) Freshwater hyphomycetes in Eurotiomycetes: a new species of Minimelanolocus and a new collection of Thysanorea papuana (Herpotrichiellaceae). Mycological Progress 18:511–522.
https://doi.org/10.1007/s11557-019-01473-7
Wang XQ, Huang Y, Long CL (2013) Assessing the genetic consequences of flower-harvesting in Rhododendron decorum Franchet (Ericaceae) using microsatellite markers. Biochemical Systematics and Ecology 50:296–303.
https://doi.org/10.1016/j.bse.2013.04.009
White TJ, Bruns T, Lee SJWT, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, pp 315–322.
https://doi.org/10.1016/b978-0-12-372180-8.50042-1
Yuan ZL, Verkley GJM (2015) Pezicula neosporulosa sp. nov. (Helotiales, Ascomycota), an endophytic fungus associated with Abies spp. in China and Europe. Mycoscience 56:205-213.
https://doi.org/10.1016/j.myc.2014.06.004
Zhang JH, Li KJ, Liu XF, Yang L, Shen SK (2021) Interspecific Variance of Suitable Habitat Changes for Four Alpine Rhododendron Species under Climate Change: Implications for Their Reintroductions. Forests 12(11):1520.
https://doi.org/10.3390/f12111520
Zhang YM, Maharachchikumbura SSN, Tian Q, Hyde KD (2013) Pestalotiopsis species on ornamental plants in Yunnan province, China. Sydowia 65(1):113–128.
https://doi.org/10.1016/j.riam.2013.01.005
Zhao J, Shan T, Mou Y, Zhou L (2011) Plant-derived bioactive compounds produced by endophytic fungi. Mini Reviews in Medicinal Chemistry 11(2):159–168.
https://doi.org/10.2174/138955711794519492
Zhu YX, Zhang ZX, Zhang HP, Chai LS, Li L, Ma SG, Li Y (2019) A new ascorbic acid derivative and two new terpenoids from the leaves and twigs of Rhododendron decorum. Journal of Asian Natural Products Research 21(6):579–586.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 Authors

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
Articles in this journal are copyrighted by the x may be read and used for academic purposes, such as teaching, research, or citation, with proper credit given to the author and the journal.use or modification of the articles is prohibited without permission.
statements expressed in the articles are solely the opinions of the authors.
authors are fully responsible for the content and accuracy of their articles.
other reuse or republication requires permission from the journal."