Abstract
Tuber melanorufum and T. tomentellum spp. nov. are described and illustrated in this paper based on the morphological and phylogenetic data. Tuber melanorufum is diagnosed by the red-brown to blackish ascomata covered with distinctly fine warts and the ellipsoid to long ellipsoid ascospores ornamented with very crowded spine-reticulations. Tuber tomentellum is characterized by the large ascospores and two types of hairs arising from the outer cells of the peridium. Phylogenetic analyses place T. melanorufum and T. tomentellum in the Rufum and Puberulum clades, respectively.
References
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<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Hillis, D.M. & Bull, J.J. (1993) An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. <em>Systematic Biology</em> 42 (2): 182–192. https://doi.org/10.1093/sysbio/42.2.182 </span></span></span></p>
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<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Ronquist, F. & Huelsenbeck, J.P. (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. <em>Bioinformatics</em> 19 (12): 1572–1574. https://doi.org/10.1093/bioinformatics/btg180 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Song, M.S. (2005) Taxonomic and molecular systematic studies on <em>Tuber</em> in China. PhD thesis, Graduate School of Chinese Academy of Sciences, Beijing. [In Chinese]</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Stamatakis, A. (2014) RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. <em>Bioinformatics</em> 30 (9): 1312–1313. https://doi.org/10.1093/bioinformatics/btu033 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Tao, K. & Liu, B. (1990) Ecology and nutritive value of <em>Tuber sinense</em>. <em>Journal of Shanxi University</em> (Nat Sci Ed) 13 (3): 319–321.</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Trappe, J.M. (1975) Generic synonyms in the Tuberales. <em>Mycotaxon</em> 2: 109–122.</span></span></span></p>
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<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Yan, X.Y., Cao, J.Z. & Fan, L. (2018) Four new <em>Tuber</em> species added to the <em>Rufum</em> group from China based on morphological and molecular evidence. <em>Mycologia</em> 110 (4): 771–779. https://doi.org/10.1080/00275514.2018.1490120 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Bonito, G., Smith, M.E., Nowak, M., Healy, R.A., Guevara, G., Ca´zares, E., Kinoshita, A., Nouhra, E.R., Domý´nguez, L.S., Tedersoo, L., Murat, C., Wang, Y., Moreno, B.A., Pfister, D.H., Nara, K., Zambonelli, A., Trappe, J.M. & Vilgalys, R. (2013) Historical biogeography and diversification of truffles in the <em>Tuberaceae</em> and their newly identified southern hemisphere sister lineage. <em>PLoS One</em> 8 (1): e52765. https://doi.org/10.1371/journal.pone.0052765 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Castresana, J. (2000) Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. <em>Molecular biology and evolution</em> 17 (4): 540–552. https://doi.org/10.1093/oxfordjournals.molbev.a026334 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Courty, P.E., Bucbe, M., Diedhiou, A.G., Frey-Klett, P., Tacon, F.L., Rineau, F., Uroz, S. & Garbaye, J. (2010) The role of ectomycorrhizal communities in forest ecosystem processes: New perspectives and emerging concepts. <em>Soil Biology & Biochemistry</em> 42: 679–698. https://doi.org/10.1016/j.soilbio.2009.12.006</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Deng, X.J., Liu, P.G., Liu, C.Y. & Wang, Y. (2013) A new white truffle species, <em>Tuber panzhihuanense</em> from China. <em>Mycological Progress </em>12 (3): 557–561. https://doi.org/10.1007/s11557-012-0862-6</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Deng, X.J., Yu, F.Q. & Liu, P.G. (2014) Contribution to confirmed & synthesized on mycorrhizae of <em>Tuber indicum</em> s.l. with two dominated & subalpine broad- leaf trees in southwestern China. <em>American Journal of Plant Sciences</em> 5: 3269–3279. https://doi.org/10.4236/ajps.2014.521341</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Dring, D.M. (1971) Chapter III techniques for microscopic preparation. <em>Methods in Microbiology</em> 4: 95–111. https://doi.org/10.1016/S0580-9517(09)70008-X</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Fan, L. & Cao, J.Z. (2012) Two new species of white truffle from China. <em>Mycotaxon</em> 121: 297–305. https://doi.org/10.5248/121.297</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Fan, L., Cao, J.Z. & Hou, C.L. (2013) <em>Tuber subglobosum</em> and <em>T. wenchuanense</em>—two new species with spino-reticulate ascospores. <em>Mycotaxon</em> 123: 95–101. https://doi.org/10.5248/123.95</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Fan, L., Hou, C.L. & Li, Y. (2012) <em>Tuber microverrucosum</em> and <em>T. huizeanum</em> two new species from China with reticulate ascospores. <em>Mycotaxon</em> 122: 161–169. https://doi.org/10.5248/122.161 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Fan, L., Li, T., Xu, Y.Y. & Yan, X.Y. (2022) Species diversity, phylogeny, endemism and geography of the truffle genus <em>Tuber</em> in China based on morphological and molecular data. <em>Persoonia</em> 48: 175–202. https://doi.org/10.3767/persoonia.2022.48.05</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Fan, L., Zhang, J.L., Li, T., Sun, H.J., Xiong, W.P. & Li, Y. (2018) Chinese black truffles: <em>Tuber yigongense</em> sp. nov., taxonomic reassessment of <em>T. indicum</em> s.l., and re-examination of the <em>T. sinense</em> isotype. <em>Mycotaxon</em> 133: 183–196. https://doi.org/10.5248/133.185 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Gardes, M. & Bruns, T.D. (1993) ITS primers with enhanced specificity for basidiomycetes—application to the identification of mycorrhizae and rusts. <em>Molecular ecology</em> 2 (2): 113–118. https://doi.org/10.1111/j.1365-294X.1993.tb00005.x </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">He, X.Y., Li, H.M. & Wang, Y. (2004) <em>Tuber zhongdianense </em>sp. nov. from China. <em>Mycotaxon</em> 90 (1): 213–216.</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Hillis, D.M. & Bull, J.J. (1993) An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. <em>Systematic Biology</em> 42 (2): 182–192. https://doi.org/10.1093/sysbio/42.2.182 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Katoh, K. & Frith, M.C. (2012) Adding unaligned sequences into an existing alignment using MAFFT and BLAST. <em>Bioinformatics</em> 28 (22): 3144–3146. https://doi.org/10.1093/bioinformatics/bts578 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Kinoshita, A., Sasaki, H. & Nara, K. (2011) Phylogeny and diversity of Japanese truffles (<em>Tuber</em> spp.) inferred from sequences of four nuclear loci. <em>Mycologia</em> 103 (4): 779–794. https://doi.org/10.3852/10-138 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Kumar, S., Stecher, G. & Tamura, K. (2016) Mega7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. <em>Molecular Biology & Evolution </em>(7): 1870. https://doi.org/10.1093/molbev/msw054</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Nylander, J. (2004) MrModeltest 2.2. Computer software distributed by the University of Uppsala. Evolutionary Biology Centre, Sweden.</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Page, R.D. (2001) TreeView. Glasgow University, Glasgow.</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Pegler, D.N., Spooner, B.M. & Young, T.W.K. (1993) <em>British truffles: A revision of British hypogeous fungi.</em> Royal Botanic Gardens, England. https://doi.org/10.2307/4110224</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Picco, V. (1788) <em>Melethemata Inauguralia</em>. Ioan. Mich. Briolus,Turin.</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Pollini, C. (1816) <em>Tuber rufum</em>. <em>Giornale di Fisica, Chimica, Storia Naturale, Medicina ed Arti</em> 9: 182. </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Qiao, P., Liu, P.G., Hu, H.T. & Wang, Y. (2013) Typification of <em>Tuber formosanum</em> (Tuberaceae, Pezizales, Ascomycota) from Taiwan, China. <em>Mycotaxon</em> 123 (1): 293–299. https://doi.org/10.5248/123.293</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Ronquist, F. & Huelsenbeck, J.P. (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. <em>Bioinformatics</em> 19 (12): 1572–1574. https://doi.org/10.1093/bioinformatics/btg180 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Song, M.S. (2005) Taxonomic and molecular systematic studies on <em>Tuber</em> in China. PhD thesis, Graduate School of Chinese Academy of Sciences, Beijing. [In Chinese]</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Stamatakis, A. (2014) RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. <em>Bioinformatics</em> 30 (9): 1312–1313. https://doi.org/10.1093/bioinformatics/btu033 </span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Tao, K. & Liu, B. (1990) Ecology and nutritive value of <em>Tuber sinense</em>. <em>Journal of Shanxi University</em> (Nat Sci Ed) 13 (3): 319–321.</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Trappe, J.M. (1975) Generic synonyms in the Tuberales. <em>Mycotaxon</em> 2: 109–122.</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Trappe, J.M., Molina, R., Luoma, D.L., Cazares, E., Pilz, D., Smith, J.E., Castellano, M.A., Miller, S.L. & Trappe, M.J. (2009) <em>Diversity, ecology, and conservation of truffle fungi in forests of the Pacific Northwest.</em> Pacific Northwest Research Station. https://doi.org/10.2737/PNW-GTR-772</span></span></span></p>
<p align="justify"><span style="color: #000000;"><span style="font-family: Times New Roman, serif;"><span style="font-size: small;">Vittadini, C. (1831) <em>Monographia tuberacearum</em>. F. Rusconi, Milan, 88 pp. </span></span></span></p>
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