Research Article |
Corresponding author: Ning Jiang ( n.jiang@caf.ac.cn ) Academic editor: Huzefa Raja
© 2024 Yaquan Zhu, Lei Ma, Han Xue, Yong Li, Ning Jiang.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Zhu Y, Ma L, Xue H, Li Y, Jiang N (2024) New species of Diaporthe (Diaporthaceae, Diaporthales) from Bauhinia variegata in China. MycoKeys 108: 317-335. https://doi.org/10.3897/mycokeys.108.128983
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Diaporthe species are known as endophytes, saprobes and pathogens infecting a wide range of plants and resulting in important crop diseases. In the present study, four strains of Diaporthe were obtained from diseased leaves of Bauhinia variegata in Guangdong Province, China. Phylogenetic analyses were conducted to identify these strains using five gene regions: internal transcribed spacer (ITS), calmodulin (cal), histone H3 (his3), translation elongation factor 1-α (tef1) and β-tubulin (tub2). The results combined with morphology revealed two new species of Diaporthe named D. bauhiniicola in D. arecae species complex and D. guangzhouensis in D. sojae species complex.
Diaporthales, morphology, multi-gene phylogeny, taxonomy, two new taxa
Diaporthe (syn. Phomopsis) is the type genus of Diaporthaceae in Diaporthales (
The teleomorph of Diaporthe is characterized by aggregated spherical ascomata with tapering necks, unitunicate, 8-spored, elongate to clavate asci, and septate or aseptate, elongated to elliptical, hyaline ascospores with larger guttules at center and smaller ones at the ends (
Diaporthe species are associated with a wide range of plant hosts as pathogens, endophytes and saprobes of crops, forest trees and ornamentals (
Species identification of Diaporthe has traditionally been based on host as well as morphological characters such as the size and shape of fruiting bodies and spores (
Bauhinia variegata is a flowering plant species belonging to Fabaceae. It is native to China and cultivated as an ornamental tree in subtropical and tropical climate for its scented flowers. The aim of the present study was to identify new isolates collected from diseased leaves of Bauhinia variegata in China following the combined approaches of morphology and phylogeny in the genus Diaporthe.
In 2022, a plant disease investigation was conducted in Guangdong Province, China. Small and irregular leaf spots were observed on the leaves of Bauhinia variegata, and 14 leaves were collected for isolation. The leaves were firstly surface-sterilized for 1 min in 75% ethanol, 3 min in 1.25% sodium hypochlorite and 1 min in 75% ethanol, rinsed for 2 min in distilled water and blotted on dry sterile filter paper. Then, the discolored areas were cut into 0.5 × 0.5 cm pieces and transferred to the surface of potato dextrose agar plates (PDA; 200 g potatoes, 20 g dextrose, 20 g agar per litre), incubated at 25 °C to obtain pure cultures. The cultures were deposited in the
China Forestry Culture Collection Center (CFCC; http://cfcc.caf.ac.cn/) and the specimen was deposited in the
Herbarium of the Chinese Academy of Forestry (
The isolates were grown on PDA, MEA and SNA plates, incubated at 25 °C under a 12 h near-ultraviolet light/12 h dark cycle to induce sporulation. Colony characters and pigment production on PDA, MEA and SNA were noted for the 10-day culture. Microscopic structures of the fungi growing on medium were mounted in water and examined under an Axio Imager 2 microscope (Zeiss, Oberkochen, Germany). At least 30 measurements were made for each structure examined.
The genomic DNA was extracted from the fresh mycelium harvested from PDA plates after seven days using a cetyltrimethylammonium bromide (CTAB) method (
Loci | Primers | PCR: Thermal Cycles: (Annealing Temp. in Bold) | Reference |
---|---|---|---|
ITS | ITS1f/ITS4 | (95 °C: 30 s, 48 °C: 30 s, 72 °C: 1 min) × 35 cycles |
|
cal | CAL228F/CAL737R | (95 °C: 15 s, 54 °C: 20 s, 72 °C: 1 min) × 35 cycles |
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his3 | CYLH3F/H3-1b | (95 °C: 30 s, 57 °C: 30 s, 72 °C: 1 min) × 35 cycles |
|
tef1 | EF1-728F/EF1-986R | (95 °C: 15 s, 54 °C: 20 s, 72 °C: 1 min) × 35 cycles |
|
tub2 | T1(Bt2a)/Bt2b | (95 °C: 30 s, 55 °C: 30 s, 72 °C: 1 min) × 35 cycles |
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A PCR reaction was conducted in a 20 µL reaction volume, and the components were as follows: 1 µL DNA template (20 ng/μL), 1 µL forward 10 µM primer, 1 µL reverse 10 µM primer, 10 µL T5 Super PCR Mix (containing Taq polymerase, dNTP and Mg2+, Beijing Tisingke Biotech Co., Ltd., Beijing, China), and 7 µL sterile water. Amplifications were performed using a T100 Thermal Cycler (Bio-Rad, Hercules, CA, USA). All amplified PCR products were evaluated visually with 1.4% agarose gels stained with ethidium bromide and PCR positive products sent to Sangon Biotech (Shanghai) Co., Ltd., (Beijing, China) for sequencing. Strands were sequenced in both directions using PCR primers. The new sequences generated in this study, as well as the reference sequences of all isolates used in the present study, are listed in Table
Isolates and GenBank accession numbers used in the phylogenetic analyses of Diaporthe.
Species | Location | Host | Strain | GenBank Accession Number | ||||
---|---|---|---|---|---|---|---|---|
ITS | tef1 | tub2 | cal | his3 | ||||
Diaporthe absenteum | China | Camellia sinensis | LC3429* | KP267897 | KP267971 | KP293477 | NA | KP293547 |
D. absenteum | China | Camellia sinensis | LC3564 | KP267912 | KP267986 | KP293492 | NA | KP293559 |
D. acaciarum | Tanzania | Acacia tortilis | CBS 138862* | KP004460 | NA | KP004509 | NA | KP004504 |
D. acericola | Italy | Acer negundo | MFLUCC 17-0956* | KY964224 | KY964180 | KY964074 | KY964137 | NA |
D. aceris | Japan | Acer sp. | LC8112 | KY491547 | KY491557 | KY491567 | KY491575 | NA |
D. actinidiae | New Zealand | Actinidia deliciosa | ICMP 13683* | KC145886 | KC145941 | NA | NA | NA |
D. acuta | China | Pyrus pyrifolia | CGMCC 3.19600* | MK626957 | MK654802 | MK691225 | MK691124 | MK726161 |
D. alangii | China | Alangium kurzii | CFCC 52556* | MH121491 | MH121533 | MH121573 | MH121415 | MH121451 |
D. alangii | China | Alangium kurzii | CFCC 52557 | MH121492 | MH121534 | MH121574 | MH121416 | MH121452 |
D. alnea | Netherlands | Alnus sp. | CBS 146.46 | KC343008 | KC343734 | KC343976 | KC343250 | KC343492 |
D. amaranthophila | Japan | Amaranthus tricolor | MAFF 246900 | LC459575 | LC459577 | LC459579 | LC459583 | LC459581 |
D. ambigua | South Africa | Pyrus communis | CBS 114015* | KC343010 | KC343736 | KC343978 | KC343252 | KC343494 |
D. angelicae | Austria | Heracleum sphondylium | CBS 111592* | KC343027 | KC343753 | KC343995 | KC343269 | KC343511 |
D. anhuiensis | China | Cunninghamia lanceolata | CNUCC 201901* | MN219718 | MN224668 | MN227008 | MN224549 | MN224556 |
D. arctii | Austria | Arctium lappa | CBS 139280* | KJ590736 | KJ590776 | KJ610891 | KJ612133 | KJ659218 |
D. arecae | India | Areca catechu | CBS 161.64* | KC343032 | KC343758 | KC344000 | KC343274 | KC343516 |
D. arengae | Hong Kong | Arenga engleri | CBS 114979* | KC343034 | KC343760 | KC344002 | KC343276 | KC343518 |
D. arezzoensis | Italy | Cytisus sp. | MFLUCC 15-0127 | MT185503 | NA | NA | NA | NA |
D. aseana | Thailand | Unidentified dead leaf | MFLUCC 12-0299a* | KT459414 | KT459448 | KT459432 | KT459464 | NA |
D. australiana | Australia | Macadamia | CBS 146457 | MN708222 | MN696522 | MN696530 | NA | NA |
D. bauhiniicola | China | Bauhinia variegata | CFCC 58154* | PP864723 | PP938599 | PP938603 | PP938607 | PP938611 |
D. bauhiniicola | China | Bauhinia variegata | GZ13B | PP864724 | PP938600 | PP938604 | PP938608 | PP938612 |
D. batatas | USA | Ipomoea batatas | CBS 122.21* | KC343040 | KC343766 | KC344008 | KC343282 | KC343524 |
D. beilharziae | Australia | Indigofera australis | BRIP 54792* | JX862529 | JX862535 | KF170921 | NA | NA |
D. biconispora | China | Citrus grandis | ZJUD62 | KJ490597 | KJ490476 | KJ490418 | MT227578 | KJ490539 |
D. biguttulata | China | Citrus limon | ZJUD47* | KJ490582 | KJ490461 | KJ490403 | NA | KJ490524 |
D. brasiliensis | Brazil | Aspidosperma sp. | CBS 133183* | KC343042 | KC343768 | KC344010 | KC343284 | KC343526 |
D. caatingaensis | Brazil | Tacinga inamoena | CBS 141542* | KY085927 | KY115603 | KY115600 | NA | KY115605 |
D. camelliae-oleiferae | China | Camellia oleifera | HNZZ027* | MZ509555 | MZ504707 | MZ504718 | MZ504685 | MZ504696 |
D. caryae | China | Carya illinoensis | CFCC 52563* | MH121498 | MH121540 | MH121580 | MH121422 | MH121458 |
D. caryae | China | Carya illinoensis | CFCC 52564 | MH121499 | MH121541 | MH121581 | MH121423 | MH121459 |
D. cercidis | China | Cercis chinensis | CFCC 52565* | MH121500 | MH121542 | MH121582 | MH121424 | MH121460 |
D. cercidis | China | Cercis chinensis | CFCC 52566 | MH121501 | MH121543 | MH121583 | MH121425 | MH121461 |
D. chiangraiensis | Thailand | Bauhinia sp. | MFLUCC 17-1669* | MF190119 | MF377598 | NA | NA | NA |
D. chrysalidocarpi | China | Chrysalidocarpus lutescens | SAUCC194.35 | MT822563 | MT855760 | MT855876 | MT855646 | MT855532 |
D. cichorii | Italy | Cichorium intybus | MFLUCC 17-1023* | KY964220 | KY964176 | KY964104 | KY964133 | NA |
D. cinmomi | China | Cinnamomum sp. | CFCC 52569* | MH121504 | MH121546 | MH121586 | NA | MH121464 |
D. cinmomi | China | Cinnamomum sp. | CFCC 52570 | MH121505 | MH121547 | MH121587 | NA | MH121465 |
D. citriasiana | China | Citrus unshiu | CGMCC 3.15224* | JQ954645 | JQ954663 | KC357459 | KC357491 | KJ490515 |
D. columnaris | USA | Vaccinium vitisidaea | AR3612* | AF439625 | NA | NA | NA | NA |
D. compacta | China | Camellia sinensis | CGMCC 3.17536* | KP267854 | KP267928 | KP293434 | NA | KP293508 |
D. convolvuli | Turkey | Convolvulus arvensis | CBS 124654* | KC343054 | KC343780 | KC344022 | KC343296 | KC343538 |
D. cucurbitae | Canada | Cucumis sp. | DAOM 42078* | KM453210 | KM453211 | KP118848 | NA | KM453212 |
D. cuppatea | South Africa | Aspalathus linearis | CBS 117499* | KC343057 | KC343783 | KC344025 | KC343299 | KC343541 |
D. cyatheae | Taiwan | Cyathea lepifera | YMJ 1364* | JX570889 | KC465406 | KC465403 | KC465410 | NA |
D. discoidispora | China | Citrus unshiu | ZJUD89* | KJ490624 | KJ490503 | KJ490445 | NA | KJ490566 |
D. drenthii | Australia | Macadamia | CBS 146453 | MN708229 | MN696526 | MN696537 | NA | NA |
D. durionigena | Vietnam | Durio zibethinus | VTCC 930005 | MN453530 | MT276157 | MT276159 | NA | NA |
D. endocitricola | China | Citrus maxima | ZHKUCC20-0012* | MT355682 | MT409336 | MT409290 | MT409312 | NA |
D. endophytica | Brazil | Schinus terebinthifolius | CBS 133811* | KC343065 | KC343791 | KC344033 | KC343307 | KC343549 |
D. eucalyptorum | China | Eucalyptus | CBS 132525* | MH305525 | NA | NA | NA | NA |
D. eugeniae | Indonesia | Eugenia aromatica | CBS 444.82* | KC343098 | KC343824 | KC344066 | KC343340 | KC343582 |
D. fraxini-angustifoliae | Australia | Fraxinus angustifolia | BRIP 54781* | JX862528 | JX862534 | KF170920 | NA | NA |
D. fructicola | Japan | Passiflora edulis × P. edulis | MAFF 246408* | LC342734 | LC342735 | LC342736 | LC342738 | LC342737 |
D. fulvicolor | China | Pyrus pyrifolia | CGMCC 3.19601* | MK626859 | MK654806 | MK691236 | MK691132 | MK726163 |
D. ganjae | USA | Cannabis sativa | CBS 180.91* | KC343112 | KC343838 | KC344080 | KC343354 | KC343596 |
D. goulteri | Australia | Helianthus annuus | BRIP 55657a* | KJ197290 | KJ197252 | KJ197270 | NA | NA |
D. guangdongensis | China | Citrus maxima | ZHKUCC20-0014* | MT355684 | MT409338 | MT409292 | MT409314 | NA |
D. guangxiensis | China | Vitis vinifera | JZB320094* | MK335772 | MK523566 | MK500168 | MK736727 | NA |
D. guangzhouensis | China | Bauhinia variegata a | CFCC 58151* | PP864725 | PP938601 | PP938605 | PP938609 | PP938613 |
D. guangzhouensis | China | Bauhinia variegata | GZ13E | PP864726 | PP938602 | PP938606 | PP938610 | PP938614 |
D. gulyae | Australia | Helianthus annuus | BRIP 54025* | JF431299 | JN645803 | KJ197271 | NA | NA |
D. guttulata | China | Unknown | CGMCC 3.20100 | MT385950 | MT424685 | MT424705 | MW022470 | MW022491 |
D. helianthi | Serbia | Helianthus annuus | CBS 592.81* | KC343115 | KC343841 | KC344083 | KC343357 | KC343599 |
D. heterostemmatis | China | Heterostemma grandiflorum | SAUCC194.85* | MT822613 | MT855925 | MT855810 | MT855692 | MT855581 |
D. hongkongensis | China | Dichroa febrífuga | CBS 115448* | KC343119 | KC343845 | KC344087 | KC343361 | KC343603 |
D. hordei | Norway | Hordeum vulgare | CBS 481.92* | KC343120 | KC343846 | KC344088 | KC343362 | KC343604 |
D. huangshanensis | China | Camellia oleifera | CNUCC 201903* | MN219729 | MN224670 | MN227010 | NA | MN224558 |
D. hubeiensis | China | Vitis vinifera | JZB320123 | MK335809 | MK523570 | MK500148 | MK500235 | NA |
D. hunanensis | China | Camellia oleifera | HNZZ023* | MZ509550 | MZ504702 | MZ504713 | MZ504680 | MZ504691 |
D. infecunda | Brazil | Schinus sp. | CBS 133812* | KC343126 | KC343852 | KC344094 | KC343368 | KC343610 |
D. infertilis | Suriname | Camellia sinensis | CBS 230.52* | KC343052 | KC343778 | KC344020 | KC343294 | KC343536 |
D. kochmanii | Australia | Helianthus annuus | BRIP 54033* | JF431295 | JN645809 | NA | NA | NA |
D. kongii | Australia | Portulaca grandifla | BRIP 54031* | JF431301 | JN645797 | KJ197272 | NA | NA |
D. krabiensis | Thailand | marine based habitats | MFLUCC 17-2481* | MN047101 | MN433215 | MN431495 | NA | NA |
D. leucospermi | Australia | Leucospermum sp. | CBS 111980* | JN712460 | KY435632 | KY435673 | KY435663 | KY435653 |
D. limonicola | Malta | Citrus limon | CPC 28200* | NR_154980 | MF418501 | MF418582 | MF418256 | MF418342 |
D. litchiicola | Australia | Litchi chinensis | BRIP 54900* | JX862533 | JX862539 | KF170925 | NA | NA |
D. lithocarpi | China | Lithocarpus glabra | CGMCC 3.15175* | KC153104 | KC153095 | KF576311 | KF576235 | NA |
D. longicolla | USA | Glycine max | FAU599* | KJ590728 | KJ590767 | KJ610883 | KJ612124 | KJ659188 |
D. longispora | Canada | Ribes sp. | CBS 194.36* | KC343135 | KC343861 | KC344103 | KC343377 | KC343619 |
D. lusitanicae | Portugal | Foeniculum vulgare | CBS 123212 | KC343136 | KC343862 | KC344104 | KC343378 | KC343620 |
D. lusitanicae | Portugal | Foeniculum vulgare | CBS 123213* | MH863280 | KC343863 | KC344105 | KC343379 | KC343621 |
D. malorum | Portugal | Malus domestica | CAA 734* | KY435638 | KY435627 | KY435668 | KY435658 | KY435648 |
D. manihotia | Rwanda | Manihot utilissima | CBS 505.76 | KC343138 | KC343864 | KC344106 | KC343380 | KC343622 |
D. masirevicii | Australia | Helianthus annuus | BRIP 57892a* | KJ197276 | KJ197239 | KJ197257 | NA | NA |
D. mayteni | Brazil | Maytenus ilicifolia | CBS 133185 | KC343139 | KC343865 | KC344107 | KC343381 | KC343623 |
D. megalospora | Not stated | Sambucus canadensis | CBS 143.27 | KC343140 | KC343866 | KC344108 | KC343382 | KC343624 |
D. melitensis | Malta | Citrus limon | CPC 27873* | MF418424 | MF418503 | MF418584 | MF418258 | MF418344 |
D. melonis | USA | Cucumis melo | CBS 507.78* | KC343142 | KC343868 | KC344110 | KC343384 | KC343626 |
D. melonis | Indonesia | Glycine soja | CBS 435.87 | KC343141 | KC343867 | KC344109 | KC343383 | KC343625 |
D. middletonii | Australia | Rapistrum rugostrum | BRIP 54884e* | KJ197286 | KJ197248 | KJ197266 | NA | NA |
D. millettiae | China | Millettia reticulata | GUCC9167* | MK398674 | MK480609 | MK502089 | MK502086 | NA |
D. minusculata | China | saprobic on decaying wood | CGMCC 3.20098* | MT385957 | MT424692 | MT424712 | MW022475 | MW022499 |
D. miriciae | Australia | Helianthus annuus | BRIP 54736j* | KJ197282 | KJ197244 | KJ197262 | NA | NA |
D. musigena | Australia | Musa sp. | CBS 129519* | KC343143 | KC343869 | KC344111 | KC343385 | KC343267 |
D. myracrodruonis | Brazil | Astronium urundeuva | URM 7972* | MK205289 | MK213408 | MK205291 | MK205290 | 17 |
D. nelumbonis | Taiwan | Nelumbo nucifera | R. Kirschner 4114* | KT821501 | NA | LC086652 | NA | NA |
D. neoarctii | USA | Ambrosia trifi | CBS 109490* | KC343145 | KC343871 | KC344113 | KC343387 | KC343629 |
D. neoraonikayaporum | Thailand | Tectona grandis | MFLUCC 14-1136* | KU712449 | KU749369 | KU743988 | KU749356 | NA |
D. oculi | Japan | Homo sapiens | HHUF 30565* | LC373514 | LC373516 | LC373518 | NA | NA |
D. osmanthi | China | Osmanthus fragrans | GUCC9165* | MK398675 | MK480610 | MK502091 | MK502087 | NA |
D. ovalispora | China | Citrus limon | CGMCC 3.17256* | KJ490628 | KJ490507 | KJ490449 | NA | KJ490570 |
D. oxe | Brazil | Maytenus ilicifolia | CBS 133186* | KC343164 | KC343890 | KC344132 | KC343406 | KC343648 |
D. pandanicola | Thailand | Pandanus sp. | MFLUCC 17-0607* | MG646974 | NA | MG646930 | NA | NA |
D. paranensis | Brazil | Maytenus ilicifolia | CBS 133184* | KC343171 | KC343897 | KC344139 | KC343413 | KC343655 |
D. pascoei | Australia | Persea americana | BRIP 54847* | JX862532 | JX862538 | KF170924 | NA | NA |
D. passiflorae | South America | Passifla edulis | CBS 132527* | JX069860 | KY435633 | KY435674 | KY435664 | KY435654 |
D. passifloricola | Malaysia | Passiflora foetida | CBS 141329* | KX228292 | NA | KX228387 | NA | KX228367 |
D. perseae | Netherlands | Persea gratissima | CBS 151.73* | KC343173 | KC343899 | KC343141 | KC343415 | KC343657 |
D. pescicola | China | Prunus persica | MFLUCC 16-0105* | KU557555 | KU557623 | KU557579 | KU557603 | NA |
D. phaseolorum | USA | Phaseolus vulgaris | AR4203* | KJ590738 | KJ590739 | KJ610893 | KJ612135 | KJ659220 |
D. phoenicicola | India | Areca catechu | CBS 161.64* | MH858400 | GQ250349 | JX275440 | JX197432 | NA |
D. podocarpi-macrophylli | China | Podocarpus macrophyllus | CGMCC 3.18281* | KX986774 | KX999167 | KX999207 | KX999278 | KX999246 |
D. pseudobauhiniae | Thailand | Bauhinia sp. | MFLU 17-1670 | MF190118 | MF377599 | NA | NA | NA |
D. pseudobauhiniae | Thailand | Bauhinia sp. | MFLUCC 17-1669* | MF190119 | MF377598 | NA | NA | NA |
D. pseudolongicolla | Serbia | Glycine max | PL42* | JQ697843 | JQ697856 | NA | NA | NA |
D. pseudolongicolla | Croatia | Glycine max | CBS 127269 | KC343155 | KC343881 | KC344123 | KC343397 | KC343639 |
D. pseudomangiferae | Dominican Republic | Mangifera indica | CBS 101339* | KC343181 | KC343907 | KC344149 | KC343423 | KC343665 |
D. pseudooculi | Japan | Homo sapiens | HHUF 30617* | NR_161019 | LC373517 | LC373519 | NA | NA |
D. pseudophoenicicola | Spain | Phoenix dactylifera | CBS 462.69* | KC343184 | KC343910 | KC344152 | KC343426 | KC343668 |
D. pseudophoenicicola | Iraq | Mangifera indica | CBS 176.77 | KC343183 | KC343909 | KC344151 | KC343425 | KC343667 |
D. pterocarpicola | Thailand | Pterocarpus indicus | MFLUCC 10-0580a* | JQ619887 | JX275403 | JX275441 | JX197433 | NA |
D. pyracanthae | Portugal | Pyracantha coccinea | CBS 142384* | KY435635 | KY435625 | KY435666 | KY435656 | KY435646 |
D. racemosae | South Africa | Euclea racemosa | CPC 26646* | MG600223 | MG600225 | MG600227 | MG600219 | MG600221 |
D. raonikayaporum | Brazil | Spondias mombin | CBS 133182* | KC343188 | KC343914 | KC344156 | KC343430 | KC343672 |
D. rhodomyrti | China | Rhodomyrtus tomentosa | CFCC 53101 | MK432643 | MK578119 | MK578046 | MK442965 | MK442990 |
D. rhodomyrti | China | Rhodomyrtus tomentosa | CFCC 53102 | MK432644 | MK578120 | MK578047 | MK442966 | MK442991 |
D. rosae | Thailand | Rosa sp. | MFLUCC 17-2658* | MG828894 | NA | MG843878 | MG829273 | NA |
D. rosiphthora | Brazil | Rosa sp. | COAD 2914* | MT311197 | MT313693 | NA | MT313691 | NA |
D. rossmaniae | Portugal | Vaccinium corymbosum | CAA762* | MK792290 | MK828063 | MK837914 | MK883822 | MK871432 |
D. sackstonii | Australia | Helianthus annuus | BRIP 54669b* | KJ197287 | KJ197249 | KJ197267 | NA | NA |
D. salinicola | Thailand | Xylocarpus sp. | MFLU 18-0553* | MN047098 | MN077073 | NA | NA | NA |
D. sambucusii | China | Sambucus williamsii | CFCC 51986* | KY852495 | KY852507 | KY852511 | KY852499 | KY852503 |
D. sambucusii | China | Sambucus williamsii | CFCC 51987 | KY852496 | KY852508 | KY852512 | KY852500 | KY852504 |
D. schimae | China | Schima superba | CFCC 53103* | MK432640 | MK578116 | MK578043 | MK442962 | MK442987 |
D. schimae | China | Schima superba | CFCC 53104 | MK432641 | MK578117 | MK578044 | MK442963 | MK442988 |
D. schini | Brazil | Schinus terebinthifolius | CBS 133181* | KC343191 | KC343917 | KC344159 | KC343433 | KC343675 |
D. schoeni | Italy | Schoenus nigricans | MFLU 15-1279* | KY964226 | KY964182 | KY964109 | KY964139 | |
D. sclerotioides | Netherlands | Cucumis sativus | CBS 296.67* | KC343193 | KC343919 | KC344161 | KC343435 | KC343677 |
D. searlei | Australia | Macadamia | CBS 146456* | MN708231 | NA | MN696540 | NA | NA |
D. sennae | China | Senna bicapsularis | CFCC 51636* | KY203724 | KY228885 | KY228891 | KY228875 | NA |
D. sennae | China | Senna bicapsularis | CFCC 51637 | KY203725 | KY228886 | KY228892 | KY228876 | NA |
D. serafiniae | Australia | Helianthus annuus | BRIP 55665a* | KJ197274 | KJ197236 | KJ197254 | NA | NA |
D. siamensis | Thailand | Dasymaschalon sp. | MFLUCC 10-0573a* | JQ619879 | JX275393 | JX275429 | JX197423 | NA |
D. sinensis | China | Amaranthus sp. | ZJUP0033-4* | MK637451 | MK660449 | MK660447 | NA | MK660451 |
D. sojae | USA | Glycine max | FAU635* | KJ590719 | KJ590762 | KJ610875 | KJ612116 | KJ659208 |
D. spinosa | China | Pyrus pyrifolia | CGMCC 3.19602* | MK626849 | MK654811 | MK691234 | MK691129 | MK726156 |
D. stewartii | Not stated | Cosmos bipinnatus | CBS 193.36* | MH867279 | GQ250324 | JX275421 | JX197415 | NA |
D. subellipicola | China | On dead wood | KUMCC 17-0153* | MG746632 | MG746633 | MG746634 | NA | NA |
D. subordinaria | New Zealand | Plantago lanceolata | CBS 464.90* | KC343214 | KC343940 | KC344182 | KC343456 | KC343698 |
D. taiwanensis | Taiwan | Ixora chinensis | NTUCC 18-105-1* | MT241257 | MT251199 | MT251202 | MT251196 | NA |
D. taoicola | China | Prunus persica | MFLUCC 16-0117* | KU557567 | KU557635 | KU557591 | NA | NA |
D. tarchonanthi | South Africa | Tarchonanthus littoralis | CBS 146073* | MT223794 | NA | MT223733 | NA | MT223759 |
D. tecomae | Brazil | Tabebuia sp. | CBS 100547* | KC343215 | KC343941 | KC344183 | KC343457 | KC343699 |
D. tectonae | Thailand | Tectona grandis | MFLUCC 12-0777* | KU712430 | KU749359 | KU743977 | KU749345 | NA |
D. tectonendophytica | Thailand | Tectona grandis | MFLUCC 13-0471* | KU712439 | KU749367 | KU743986 | KU749354 | NA |
D. tectonigena | China | Tectona grandis | MFLUCC 12-0767* | KU712429 | KU749371 | KU743976 | KU749358 | NA |
D. tectonigena | China | Camellia sinensis | LC6512 | KX986782 | KX999174 | KX999214 | KX999284 | KX999254 |
D. terebinthifolii | Brazil | Schinus terebinthifolius | CBS 133180* | KC343216 | KC343942 | KC344184 | KC343458 | KC343700 |
D. thunbergiicola | Thailand | Thunbergia laurifolia | MFLUCC 12-0033* | KP715097 | KP715098 | NA | NA | NA |
D. tulliensis | Australia | Theobroma cacao | BRIP 62248a* | KR936130 | KR936133 | KR936132 | NA | NA |
D. ueckeri | USA | Cucumis melo | FAU656* | KJ590726 | KJ590747 | KJ610881 | KJ612122 | KJ659215 |
D. unshiuensis | China | Fortunella margarita | CGMCC 3.17566* | KJ490584 | KJ490463 | KJ490405 | NA | KJ490526 |
D. unshiuensis | China | Carya illinoensis | CFCC 52594 | MH121529 | MH121571 | MH121606 | MH121447 | MH121487 |
D. unshiuensis | China | Carya illinoensis | CFCC 52595 | MH121530 | MH121572 | MH121607 | MH121448 | MH121488 |
D. vawdreyi | Australia | Psidium guajava | BRIP 57887a | KR936126 | KR936129 | KR936128 | NA | NA |
D. vexans | USA | Solanum melongena | CBS 127.14 | KC343229 | KC343955 | KC344197 | KC343471 | KC343713 |
D. viniferae | China | Vitis vinifera | JZB320071* | MK341550 | MK500107 | MK500112 | MK500119 | NA |
D. vochysiae | Brazil | Vochysia divergens | LGMF1583* | MG976391 | MK007526 | MK007527 | MK007528 | MK033323 |
D. xishuangbanica | China | Camellia sinensis | CGMCC 3.18283* | KX986784 | KX999176 | KX999217 | NA | NA |
D. xishuangbanica | China | Camellia sinensis | LC6707 | KX986783 | KX999175 | KX999216 | NA | KX999255 |
For the phylogenetic analysis, sequences of reference Diaporthe species and related taxa were downloaded from NCBI GenBank based on recent publications on the genus Diaporthe (
In the present study, we inferred a genus tree of Diaporthe covering a large proportion of sequence data available as last summarized by
In the D. arecae species complex, the combined sequence alignments comprised 61 strains, with D. eucalyptorum (CBS 13252), D. biconispora (ZJUD62) and D. vawdreyi (BRIP 57887a) as the outgroup taxa. The dataset comprised 2662 characters including alignment gaps (590 for ITS, 499 for cal, 485 for his3, 375 for tef1 and 713 for tub2). CFCC 58154 and GZ13B from Bauhinia variegata formed a distinct clade close to D. sennae (Fig.
Phylogram of Diaporthe arecae species complex resulting from a maximum likelihood analysis based on a combined matrix of ITS, cal, his3, tef1 and tub2 loci. Numbers above the branches indicate ML bootstrap values (left, ML BS ≥ 50%) and Bayesian posterior probabilities (right, BPP ≥ 0.9). Isolates from the present study are in bold and ex-type strains are marked with *.
Phylogram of Diaporthe sojae species complex resulting from a maximum likelihood analysis based on a combined matrix of ITS, cal, his3, tef1 and tub2 loci. Numbers above the branches indicate ML bootstrap values (left, ML BS ≥ 50%) and Bayesian posterior probabilities (right, BPP ≥ 0.9). Isolates from the present study are in bold and ex-type strains are marked with *.
China • Guangdong Province, Guangzhou City, Luhu Park, 23°9'11.15"N, 113°16'46.01"E, 92 m asl, on diseased leaves of Bauhinia variegata, 8 Aug 2022, Yong Li, Chengbin Wang & Yaquan Zhu, (holotype: CAF800094; ex-type culture: CFCC 58154).
Named after the host genus, Bauhinia.
Conidiomata formed on PDA pycnidial, scattered to aggregated, black, erumpent, raising above surface of culture medium, subglobose, 150–300 μm diam., exuding white or yellowish creamy conidial droplets from central ostioles after 30 days at 25 °C. Conidiophores reduced to conidiogenous cells. Conidiogenous cells hyaline, unbranched, septate, straight, slightly tapering towards the apex, 6.0–15.0 × 1.5–4.0 μm. Alpha conidia hyaline, aseptate, ellipsoidal to spindle-shaped, biguttulate or with one guttulate, 4.5–7.0 × 2.0–3.0 μm. Beta conidia and gamma conidia not observed. Teleomorph not observed.
Colonies covering entire plate after 2 weeks. On PDA with profuse aerial mycelium, white surface, reverse fulvous. On MEA with fluffy aerial mycelium, dirty white surface, reverse ochreous. On SNA white sparse aerial mycelium, surface and reverse white.
China • Guangdong Province, Guangzhou City, Luhu Park, 23°9'11.15"N, 113°16'46.01"E, 92 m asl, on diseased leaves of Bauhinia variegata, 8 Aug 2022, Yong Li, Chengbin Wang & Yaquan Zhu, living culture GZ13B.
Two strains representing Diaporthe bauhiniicola clustered in a clade distinct from its closest phylogenetic neighbour, D. sennae (Fig.
Named after the collection site of the type specimen, Guangzhou City.
China • Guangdong Province, Guangzhou City, Longdong straight street, 23°11'41.02"N, 113°22'8.33"E, 46 m asl, on diseased leaves of Bauhinia variegata, 8 Aug 2022, Yong Li, Chengbin Wang & Yaquan Zhu, (holotype: CAF800095; ex-type culture: CFCC 58151).
Conidiomata pycnidial, scattered to aggregated, black, erumpent, raising above surface of culture medium, subglobose, 150–450 µm diam, exuding white or yellowish creamy conidial droplets from central ostioles after 30 days at 25 °C. Conidiophores 12.5–24.5 × 1–2.5 μm, cylindrical, hyaline, unbranched, straight to sinuous. Conidiogenous cells densely aggregated, phiailidic, unbranched, straight or slightly curved, 5.5–10 × 2.0–7.5 μm. Beta conidia filiform, hyaline, straight or slightly curved, aseptate, 17.0–29.5 × 1.0–2.0 μm. Alpha conidia and gamma conidia not observed. Teleomorph not observed.
Colonies covering entire plate after 2 weeks. On PDA with profuse aerial mycelium, white surface, reverse amber. On MEA with fluffy aerial mycelium, dirty white surface, reverse ochreous. On SNA white sparse aerial mycelium, surface and reverse white.
China • Guangdong Province, Guangzhou City, Longdong straight street, 23°11'41.02"N, 113°22'8.33"E, 46 m asl, on diseased leaves of Bauhinia variegata, 8 Aug 2022, Yong Li, Chengbin Wang & Yaquan Zhu, living culture GZ13E.
Diaporthe guangzhouensis from the present study is phylogenetically close to D. tulliensis (Fig.
In the current study, phylogenetic analyses based on five combined loci (ITS, cal, his3, tef1 and tub2), as well as morphological characters of the anamorph obtained in culture, revealed D. bauhiniicola and D. guangzhouensis spp. nov. from Bauhinia variegata, which contributed to our knowledge of the diversity of Diaporthe species in China.
Diaporthe pseudobauhiniae (syn. D. chiangraiensis, Chiangraiomyces bauhiniae) was described as a saprobic fungus on branches of Bauhinia sp. in Thailand (
The initial species concept of Diaporthe based on the assumption of host-specificity, resulted in the introduction of more than 1000 taxa (http://www.indexfungorum.org/). However, more than one species of Diaporthe have been often discovered from the same host (
Diaporthe is considered as a species-rich genus. Nevertheless, an emerging perspective posits that the quantity of recognized Diaporthe species may have been substantially overestimated. The D. amygdali species complex has been proven a single species evidenced from the genealogical concordance phylogenetic species recognition principle (GCPSR) and coalescence-based models: general mixed yule-coalescent (GMYC) and poisson tree processes (PTP), with several species becoming synonyms (
The authors have declared that no competing interests exist.
No ethical statement was reported.
This study was supported by Fundamental Research Funds of
Conceptualization: LM, YL, NJ. Methodology: YZ. Formal analysis: HX. Investigation: YL. Data Curation: LM, HX. Writing - Original draft: YZ. Writing - Review and Editing: NJ. Visualization: NJ.
Yaquan Zhu https://orcid.org/0000-0002-3296-239X
Han Xue https://orcid.org/0000-0003-0414-6237
Yong Li https://orcid.org/0000-0002-4406-1329
Ning Jiang https://orcid.org/0000-0002-9656-8500
All of the data that support the findings of this study are available in the main text.