Corresponding author: J. Jennifer Luangsa-ard (
Academic editor: T. Lumbsch
Thailand is known to be a part of what is called the Indo-Burma biodiversity hotspot, hosting a vast array of organisms across its diverse ecosystems. This is reflected by the increasing number of new species described over time, especially fungi. However, a very few fungal species from the specialized spider-parasitic genus
Kuephadungphan W, Tasanathai K, Petcharad B, Khonsanit A, Stadler M, Luangsa-ard JJ (2020) Phylogeny- and morphology-based recognition of new species in the spider-parasitic genus
To arthropodologists or even arachnologists, it is surprising that fungal pathogens of spiders seem to be generally neglected when the host can be completely overgrown by the pathogens to be unrecognizable as a spider. Nonetheless, this group of fungi has been known and studied for more than two centuries (
Thailand is one of the most biodiverse countries in Southeast Asia and the BIOTEC culture collection has more than 700
Our continuous survey of invertebrate-pathogenic fungi in Thailand for over two decades has led to the BIOTEC Bangkok Herbarium (
Spiders parasitized by
Microscopic characteristics were studied based on observation of synnemata and perithecia. Each of them was detached from the stroma and mounted on a microscope slide containing a drop of lactophenol cotton blue solution. Shapes and sizes of individual character were determined and measured according to
The mummified spiders were identified based on morphological characteristics. To better understand the host-pathogen relationship, posture of spider at attachment on leaf surface (touching or lifting), position of spider on the leaf (under or upper side), and leaf type (monocots or dicots) were herein recorded.
DNA extraction, PCR amplification of five DNA regions as well as purification of PCR products were conducted according to the protocols previously described by
DNA sequences were checked manually for ambiguous base calls and all sequences were assembled using BioEdit v.7.2.5 (
Phylogenetic relationships were inferred using maximum likelihood (
The combined data set of 43 taxa (Table
List of taxa included in the phylogenetic analysis and their GenBank accession numbers. The isolates representing four new species and other sequences generated in this study are marked in bold.
Species | Code | GenBank accession numbers | References | ||||
---|---|---|---|---|---|---|---|
|
LSU |
|
|
|
|||
|
HUA 772 |
|
|
|
– | – |
|
|
HUA 186145 | – |
|
|
– | – |
|
|
ANDES-F 1014 |
|
|
|
– | – |
|
|
ANDES-F 1024T |
|
|
|
– |
|
|
|
ARSEF 7518 |
|
– |
|
|
|
|
|
ARSEF 1564T |
|
– |
|
|
|
|
|
OSC 93623 |
|
|
|
|
|
|
|
ARSEF 5050 |
|
– |
|
|
|
|
|
CBS 111113T |
|
|
|
|
|
|
|
CBS 134.22T |
|
|
|
|
|
|
|
|
|
|
|
|
||
|
OSC 93609T | – |
|
|
|
|
|
|
OSC 93610 |
|
|
|
|
|
|
|
CBS 309.85 |
|
|
|
|
|
|
|
CBS 658.80 |
|
|
– | – | – |
|
|
CBS 610.80T |
|
|
|
– |
|
|
|
NHJ 4314 | – | – |
|
|
|
|
|
NHJ 11923 | – |
|
|
|
|
|
|
|
|
|
|
|
||
|
NHJ 3510 | – | – |
|
|
|
|
|
NHJ 11679 | – | – |
|
|
– |
|
|
|
|
– |
|
– | – |
|
|
|
|
|
|
|
|
|
|
|
|
– |
|
– | – |
|
|
|
|
|
|
|
|
|
|
ARSEF 1915T | – |
|
|
|
|
|
|
|
|
– | – |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
– |
|
|
|
|
|
|
|
|
|
|
|
– |
|
|
|
– |
|
|
|
NHJ 12014 | – | – |
|
|
|
|
|
NHJ 10808 | – |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
– |
|
|
– |
|
|
|
|
|
|
|
– |
|
|
|
|
|
|
|
– |
|
|
|
|
|
|
|
|
|
|
|
|
– |
|
– | – |
|
|
|
|
– |
|
– | – |
|
|
|
|
|
|
– |
|
|
|
|
|
|
|
|
|
|
Phylogenetic tree inferred from a RAxML search of a concatenated alignment of
Thailand, Nakhon Ratchasima, Khao Yai National Park, Mo Sing To Nature Trail;
Refers to its spider host.
Colonies derived from conidia, on
Thailand, Nakhon Ratchasima, Khao Yai National Park, Kong Kaeo Waterfall;
Thailand, Chiang Mai, Chiang Dao District, Ban Huathung;
Refers to the fusiform part-spores.
Spiders totally covered by the white to cream mycelial mat. A single synnema or synnemata in pairs cream to light brown, often darker than the mycelia covering the host, narrowing toward the apex and terminating in a swollen sterile tip with acute apex (Fig.
Colonies derived from conidia, on
Thailand, Chiang Mai, Chiang Dao District, Ban Huathung;
The sexual morph of
Thailand. Nakhon Ratchasima, Khao Yai National Park, Mo Sing To Nature Trail;
Refers to the capability of the fungus to produce pigmentosins.
Spider host completely covered by white to yellowish-white mycelial mat.
Colonies derived from ascospores, on
Thailand, Nakhon Ratchasima, Khao Yai National Park, Mo Sing To Nature Trail,
Refers to the outer appearance of the fungus resembling the posture of a scorpion.
White to grayish- or brownish-white mycelial mat velvety, completely covering the spider host, firmly attaching the underside of living leaf by the mycelia covering its legs (Fig.
Colonies derived from conidia, on
The morphology of
A torrubiella-like sexual morph is well-known to be connected with
The morphological resemblance between
In nature, a torrubiella-like sexual morph may occur on spider hosts without the presence of
It has been over a half century since host specialization was suggested as one of the taxonomic criteria for parasitic fungi (
According to the effort of putting toward species identification of spider hosts (
It is notable that all seven host individuals used for identification in this study were found under a leaf.
Since
It is remarkable that our finding has revealed the high possibility to incorporate the host specificity in molecular and morphological criteria for classification and identification of
The biggest challenge for molecular phylogeny-based classification of
Despite molecular phylogeny currently being the most powerful approach available in modern fungal classification and taxonomy (
This work was supported by the National Center for Genetic Engineering and Biotechnology (BIOTEC) Platform Technology Management (Grant no. P19-50231), National Science and Technology Development Agency (NSTDA). This project has also received funding from the European Union’s Horizon 2020 Research and Innovation Staff Exchange (RISE) programme under the Marie Skłodowska-Curie grant agreement No 645701, Project acronym “GoMyTri”; lead beneficiaries JJL and MS. The National Park, Wildlife and Plant Conservation Department in Thailand is gratefully acknowledged for permission to conduct a study in the protected area. We are grateful to Dr. Philip James Shaw for the English editing and the reviewers whose comments and suggestions helped improve our manuscript.