Marcin Jan Kamiński

Phylogenetics, Biogeography, Entomology, Tenebrionidae, Blaptinae, Platynotini, Sepidiini


National Science Center, OPUS 19 (2020/37/B/NZ8/02496)

High-throughput DNA sequencing on historical collections of beetles:

phylogenetic studies among Platynotini (Tenebrionidae)

PI: dr hab. Marcin Jan Kamiński, Ph.D.

Museum and Institute of Zoology, Polish Academy of Sciences

*Information dedicated for potential PhD candidates is marked as “PhD Project

I. OBJECTIVES

The main aim of this project is to reconstruct the phylogeny of the darkling beetle tribe Platynotini (Coleoptera: Tenebrionidae), primarily based on the high‐throughput DNA sequencing of museum specimens, which was selected as the most time- and cost-effective approach due to the difficulties of collecting platynotoid species caused by geographical (wide distribution), political (inaccessible areas/ambiguous permitting), environmental (habitat degradation), and biological (phenological differences of co-occurring taxa) factors. Specifically, the project will aim to generate DNA sequence data from representatives across the tribe’s known diversity (75 gen., ~590 sp.; Kamiński et al. 2019a) by sequencing 160 historical and ~90 more recently collected specimens (including 30 outgroup taxa). Based on this dataset the following specific problems will be investigated.


  1. Historical biogeography of Platynotini.

This subproject will investigate the origin of transoceanic disjunctions (African, Indomalayan, and New World components) observed within the tribe (Kamiński 2015). The role of long-distance dispersal and vicariance will be assessed by using ancestral area analyses and a time-calibrated phylogeny. In particular, the following opposing biogeographic scenarios will be tested:


  • The transoceanic disjunction in Platynotini was driven by tectonic-driven vicariance resulting from the breakup of the Gondwanan supercontinent;

  • The African-Indomalayan disjunction in Platynotina fits the “boreotropical migration” model (Lavin & Luckow 1993), which assumes that ancestors of the Indomalayan complex dispersed from Africa via northern routes to colonize subtropical or tropical regions, followed by subsequent extinctions in middle-to high-latitude regions of the Northern Hemisphere.


  1. Evolution of ovoviviparity: a case study based on platynotoid darkling beetles.

Ovoviviparity is rare among beetles. This mode of reproduction, where eggs develop within the mother until hatching, has been reported for select carabids, chrysomelids, micromalthids, staphylinids, and tenebrionids (Hagan 1948, 1951, Tschinkel 1978, Liebherr & Kavanaugh 1985, Iwan 2000). Within the latter family, the majority of known cases, besides one (see Dutrillaux et al. 2010), occur in Platynotini (Tschinkel 1978, Iwan 2000). Due to topology uncertainties and limited taxon sampling, currently available phylogenies cannot be used to quantify parallel occurrences of ovoviviparity within the tribe (Kamiński et al. 2019a). Furthermore, as stated by Tschinkel (1978) and Iwan (2000), additional Platynotini species may potentially prove to be ovoviviparous. In order to test this assumption, a comparative study of female terminalia will be conducted within the tribe by dissecting ~1,000 female specimens. The morphological traits associated with ovoviviparity will be analyzed to explore potentially overlooked/unknown ovoviviparous taxa. This data will be compared to the molecular phylogeny (ancestral state reconstruction) to estimate how many times ovoviviparity has evolved across the clade. Moreover, as shifts towards ovoviviparity/viviparity are often associated with colonisation of colder/drier environments (Roth 1970, Pincheira-Donoso et al. 2013), ecological niche modeling will be used to investigate the differences in habitat preference between selected ovovivi- and oviparous species.


  1. Molecular phylogeny-based revisions on subtribal, generic, and species levels.

Aside from solving specific biological problems described below, this subproject aims to assess the usefulness of museum sequencing in phylogenetic studies at different classification levels.


a. Subtribal level: Phylogeny and taxonomic revision of subtribe Eurynotina (PhD Project). As no phylogenetic hypotheses for this platynotoid subtribe exist, this study will be the first to address this issue. The group currently consists of 16 genera grouping 95 flightless species and subspecies (Kamiński 2016). The molecular-based phylogenetic analysis will aim to include representatives of all Eurynotina genera, while species concepts will be revised using a matrix-based approach. Based on the results a new classification of Eurynotina will be proposed.


b. Generic level: Biogeography of the Ectateus generic group (Platynotina). The Ectateus complex consists of 12 recently revised genera (Kamiński 2015). The majority of genera are restricted either to the Eastern or Western regions of Central Africa. This biogeographic pattern is not reflected in the recently published morphology-based phylogeny, i.e. neither the Eastern or Western components are clustered together. Based on this observation it was previously hypothesized that the natural history of the group fits the turnover-pulse model (see Vrba 1993). The aim of this subproject is to conduct a detailed – Central Africa restricted – biogeographic investigation in order to model the evolution of the distributional ranges of the genera representing the Ectateus generic group.


c. Species level: Phylogenetic revision of the genus Gonopus and related Stenogonopus (Platynotina). Gonopus consists of 21 apterous species and subspecies distributed in extremely arid ecosystems in Southern Africa (Endrödy-Younga 2000). The genus is an interesting model for ethology (parabiosis, parental care) and developmental biology (evolution of aptery) (Rasa 1994, NCN 2016). However, those studies are largely inhibited by the lack of phylogenetic data on the genus. Moreover, the limits of several species within the genus remain blurry, as well as the distinctiveness of Gonopus in relation to Stenogonopus (Endrödy-Younga 2000). The aim of this subproject is to revise the genus by using a total evidence approach based on molecular, morphological, and distributional data.

  1. Phylogenetic potential of selected historic entomological collections.

As the correlation between quality of DNA yield and time since the collection of a particular specimen is not direct (Kanda et al. 2015, Sproul & Maddison 2017), assessment of the phylogenetic potential of a given collection is challenging without conducting a targeted study. The current project will primarily be based on the specimens preserved at the Museum and Institute of Zoology of the Polish Academy of Sciences (MIZ PAS). However, the analysed individuals will originate from collections created by specifically selected entomologists, i.e. Hans Gebien (1874–1947), Witold Eichler (1874–1960), Szymon Tenenbaum (1892–1941), Carl Koch (1904–1970), Zoltán Kaszab (1915–1986), and Sebastian Endrödy-Younga (1934–1999). As an outcome, profiles of their collections, in terms of their usefulness for molecular phylogenetics, will be produced. Results of this subproject will be used to implement specific protocols of specimens preservation in the largest Polish zoological collection (MIZ PAS).


II. SIGNIFICANCE

Platynotini (platynotoid beetles) is a highly diverse tribe of mostly flightless darkling beetles (Tenebrionidae). Morphologically, Platynotini is easily distinguishable from other tribes within the subfamily Tenebrioninae by the presence of stridulatory gula, used for sound production (Koch 1956, Iwan 2002a). Furthermore, monophyly of the tribe was strongly supported in recent molecular phylogenies within Tenebrioninae (Kamiński et al. 2019a, b). Based on morphological and molecular evidence, two subtribes were distinguished within Platynotini (Kamiński et al. 2019a), Eurynotina (16 gen., 92 sp.) and Platynotina (59 gen., ~500 sp.). Platynotina can be further subdivided into the following groupings (See Fig. 1 for details) based on previous studies (Koch 1956, Iwan 2002a, 2010, Kamiński et al. 2019a): anomalipoid generic complex (1 gen., 51 sp.), gonopoid g. c. (2 gen., 16 sp.), melanocratoid g. c. (8 gen., 35 sp.), platynotoid g. c. (32 gen., 197 sp.), and trigonopoid g. c. (16 gen., >200 sp.). Iwan (2002a) conducted a generic level phylogenetic analysis on the subtribe based on morphology. The results provide a foundation for future work (this study); however, the resulting topology was poorly supported, partly not convergent with the preliminary molecular phylogeny (Fig. 2), and thus highlighted the need for further studies. Furthermore, the phylogenetic relations among Eurynotina were never investigated.

Ad. 1. While the distribution of Eurynotina seems to be confined to southern Africa (Kamiński 2016), representatives of Platynotina are disjunctly distributed throughout Sub-Saharan Africa, North and South America, and the Indomalayan biogeographic realm (Fig. 1), with no taxa currently shared between regions (Kamiński 2015). However, the current classification needs to be tested, particularly the currently accepted taxonomic interpretations of the three flighted genera, i.e. Alaetrinus (New World), Penthicoides (India), and Zidalus (Africa). As definitions of these taxa are largely based on plesiomorphic features (Iwan 1995, 2002a), it is possible that they represent a monophyletic lineage that might be interpreted as a single genus (Kamiński et al. 2019a). This study will test the generic and biogeographic limits of these taxa using new molecular data and an updated morphological analysis for the tribe (Kamiński et al. 2019a, b). Unlike the three flighted genera, the flightless Platynotina genera display high degrees of morphological distinctiveness between the above-mentioned regions; however their phylogenetic relations remain uninvestigated by the means of molecular systematics.

The origin of transoceanic disjunctions (Africa, Indomalayan, and New World) within Platynotini has never been studied due to essential sampling gaps in the previous phylogenetic project, which was directed towards higher-classification problems (Kamiński et al. 2019a, b). Specifically, due to the lack of ethanol-preserved specimens, no Indomalayan species were included, while the New World fauna was represented only by the flighted Alaetrinus (Fig. 2). Moreover, the inclusion of several endemic forms seems to be crucial for the proper reconstruction of the historical biogeography of the tribe, e.g. Nesopatrum, a monotypic genus known only from Democratic Republic of São Tomé and Príncipe (coast of Central Africa), which displays a high degree of morphological similarity to some New World components. Inclusion of this genus might be crucial for investigating the African-New World disjunction.

As Platynotini occur in many of the most threatened habitats designated as biodiversity hotspots on different continents, such as Borneo Lowland Forests, Central and Eastern Miombo Woodlands, Drakensberg Montane Woodlands, Guinean Moist Forests, Namib-Karoo-Kaokoveld Deserts, Southeastern Conifer and Broadleaf Forests, or Southwestern Ghats Moist Forest (Iwan 1997, Kamiński & Iwan 2013b, Kamiński 2015, 2016), the biogeographic study of the tribe based on dense taxon sampling can provide new insights on the origin and evolution of those essential regions. At the same time, the widespread distribution of Platynotini creates a logistical challenge in terms of collecting material, which can be fully overcome by using museum sequencing.

Although the existence transoceanic disjunctions (Africa, Indomalayan, and New World) have been linked to the Gondwanan breakup (Gvinish & Renner 2004), more recent studies tend to lean towards other scenarios (see Lavin & Luckow 1993, Ye et al. 2017). This project aims to test differing historical distribution models (e.g., Gondwanan breakup versus “boreotropical migration”) in the context of Platynotini, a species-rich and distributionally variable tribe that can offer unique insights into our understanding of global biodiversity patterns.


Ad. 2. Previously, ovoviviparity was reported for ten species representing Platynotini (Tschinkel 1978, Iwan 1996, 2000, 2005, 2010, Iwan & Ferrer 2000). Specifically, fully developed 1st instar larvae were found in the bursa copulatrix of one species of Eurynotina (South African Eurynotus capensis), two from the trigonopoid generic complex (South African Atrocrates medvedevi and Melanopterus marginicollis), and seven from melanocratoid g. c. (Malagasy Clastopus aberlenci, Melanocratus ferreri, Styphacus bartolozzi, decorsei, S. kochi, Sebastianus projectus, and S. simplex). Taking into consideration the available phylogenetic/taxonomic context, it can be initially assumed that ovoviviparity has arisen three times within the tribe (Fig. 2). However, due to limited taxon sampling, previous molecular studies have not challenged the phylogenetic status of the melanocratoid Platynotina (Kamiński et al. 2019a), which currently consists of eight Malagasy genera grouped only by having deeply emarginate clypei (Iwan 2010). As this morphological feature proved to be variable within the other platynotoid generic complexes (Kamiński 2016), the monophyly of the melanocratoid Platynotina remains questionable. Moreover, the relation between the trigonopoid and melanocratoid genetic complexes is poorly understood due to the limited taxon sampling and low clade support separating these complexes in the current phylogeny (Fig. 2).

Quantification of parallel occurrences of ovoviviparity within the tribe faces further challenges, as several other (non-trigonopoid and non-melanocratoid) species were listed as potentially ovoviviparous, e.g. Anomalipus variolosus (Tschinkel 1978). This assumption was made based on the discovery of a single large egg within the bursa copulatrix of a dissected female. Reduction of the number of eggs and increased egg size are often viewed as preadaptations to ovoviviparity in insects (Roth 1970, Tschinkel 1978). Similar eggs were later reported for many other species representing Platynotina (Iwan 2010, Kamiński 2013a). In order to define morphological traits associated with ovoviviparity and explore potentially overlooked/unknown ovoviviparous taxa additional dissections of female reproductive tracts are needed.

The biological cause of ovoviviparity within Platynotina has never been comprehensively investigated. Available literature on other beetle groups, specifically rove beetles, links this mode of reproduction with a termitophilous lifestyle (Zilberman et al. 2019). However, this does not seem to be the case in Platynotina, as no termitophilous species have been documented (Kamiński et al. 2019b). On the other hand, several herpetological studies link the evolution of viviparity, and indirectly ovoviviparity, with the colonization of colder habitats (Pincheira-Donoso et al. 2013). The cold-climate hypothesis assumes that low temperatures experienced in colder environments (higher altitudes and/or latitudes) by externally developing eggs compromise successful incubation; and therefore, selection favours the evolution of prolonged retention of eggs within the female (Blackburn 2000, 2015, Shine 2005). A different climate-based assumption was proposed by Iwan (2010), who hypothesised that ovoviviparity in Platynotina evolved as a response to the low humidity and large diurnal amplitude of temperatures. A similar suggestion was also made for ovoviviparous cockroaches (Roth 1970).

The combination of a robust phylogeny for the tribe, along with ecological niche modeling based on climatic and geographic factors performed for selected ovoviviparous and oviparous species, will be used to test climatic-based ovoviviparity hypotheses.


Ad. 3. While the low-coverage whole genome sequencing approach is well suited for the recovery of mitogenomes and nuclear ribosomal DNA, it may not be sufficient to fully resolve phylogenetic relationships at higher-classification levels for which more genetic loci are often needed (Cameron et al. 2004, Sproul & Maddison 2017). To resolve this potential issue and reconstruct a well-supported phylogeny for Platynotini, this project will combine the shotgun sequencing of rarely collected species (museum specimens) with the targeted enrichment of multiple low copy nuclear loci (plus mitogenomes and nuclear ribosomal DNA) on select, phylogenetically crucial modern taxa preserved in ethanol. This strategy will provide a well-resolved backbone topology for the higher-level phylogenetic/biogeographic considerations, as well as a diverse data set for solving more specific problems, i.e. Phylogeny and taxonomic revision of subtribe Eurynotina; Biogeography of the Ectateus generic group (Platynotina); and Phylogenetic revision of the genus Gonopus and related Stenogonopus (Platynotina). The following subprojects were designated to investigate the limits of this combinative approach (i.e. targeted enrichment and genome skimming) at different classification levels.


a. (PhD Project) While several phylogenetic and taxonomic contributions are available for Platynotina (e.g., Koch 1956, Iwan 2002a), little is known about Eurynotina. The most comprehensive papers on the taxonomy of this subtribe were published by Koch (1954a, b, 1955). In relation to Platynotina, Eurynotina is defined by the absence aedeagal clavae (Kamiński 2016). The phylogenetic distinctiveness of this lineage was initially supported by molecular data (Kamiński et. al. 2019a, b). However, representatives of only two genera (Eurynotus, Oncotus) were included in that study (Fig. 2).

Most Eurynotina genera have not been studied since their descriptions, e.g. Menederopsis, Phaleriderma, Schyzoschelus, or Stridigula (all described by Koch 1954b); hence the validity of their taxonomic concepts have not been tested. Furthermore, many genus-level taxa are defined by the lack of apomorphies found in other taxa. For example, Stridigula can be differentiated from Psectropus only by having narrow protarsi in males (Kamiński 2016). There is a strong possibility that some poorly defined Eurynotina genera are paraphyletic and should not be maintained. Furthermore, in many cases the diagnostic characters are extremely ambiguous. This disrupts the identification process even at the generic level. Reliable identifications can often only be made by referring to the type material (Kamiński pers. obs.). As a consequence, only a fraction of the globally collected species in museums have been identified. Large amounts of unidentified specimens representing Eurynotina were located in the following institutions: TNHN, Durban Natural Science Museum, Iziko South African Museum, Muséum National d’Histoire Naturelle in Paris, Museum für Naturkunde der Humboldt–Universität in Berlin, Natural History Museum in London, and California Academy of Sciences in San Francisco. Taking into consideration the scarcity of available contributions to the subtribe, museums are potentially a rich and unexplored source of undescribed Eurynotina taxa.

Although, some species of Eurynotina are commonly observed in certain types of environments (e.g., Eurynotus capensis; Kamiński pers. obs.), over 75% of the known species and subspecies are only known from the type series (Kamiński 2016). Data on the biology of Eurynotina is extremely scarce (Schulze 1969, Tschinkel 1978, Kamiński et. al. 2019b), and collecting fresh specimens is difficult. During eight research trips to South Africa and Namibia, conducted by the authors of this application, representatives of only four genera were collected (i.e. Eurynotus, Oncotus, Phylacastus, and Schyzoschelus). In consequence, the application of museum sequencing techniques seems to be a reasonable and cost-effective approach for the phylogenetic study of Eurynotina.


b. Major generic lineages of the subtribe Platynotina are characterised by specific distributional patterns (Fig. 1). For example, the trigonopoid group is strictly confined to the dryer ecosystems of southern Africa (Iwan 2002), the melanocratoid group is endemic to Madagascar (Iwan 2010), while the platynotiod lineage seems to prefer tropical savanna climates (Kamiński & Iwan 2013b). This creates an interesting evolutionary problem, which can be further investigated while achieving the first detailed goal of this application (Ad. 1); however, more specific biogeographic questions can also be formulated within particular generic groups.

The Ectateus generic group (EGG) is a recently redefined lineage within the Afrotropical platynotoid Platynotina that groups 100 species classified in 12 genera (Kamiński 2015). The phylogeny-based revision of this complex resulted in an interesting distributional pattern. Namely, some of the sister clades were found to inhabit opposing, Eastern or Western, parts of Tropical Africa, e.g. Anchophthalmus vs Phallocentrion+Nesopatrum (Fig. 1). Ancestral area analyses implied that ancestors of these clades likely had wider, pancentral African, distributionals, which were later confined (Kamiński 2015). Furthermore, the revealed diversity hotspots of the EGG correspond to the tropical areas of endemism that were designated based on comparative analyses of flora (Linder 2001), especially in the following centres: ‘the East African Coast’, ‘Huilla’, ‘Kivu’, ‘Lower Guinea’, and ‘Upper Guinea’. This picture was also reflected in studies concerning other organisms, e.g. birds (de Klerk et al. 2002), mammals (Turpie & Crowe 1994), leaf beetles (Biondi & D’Alessandro 2006), and sphingids (Ballesteros-Mejia et al. 2013). The higher diversity and local endemism reported in these areas may suggest that they served as ‘refuges’ for several species during climate changes in the past. The congruence between Afrotropical centres of endemism and refuges has been suggested by several authors (Endrödy-Younga 1988, Burgess et al. 1998, Linder 2001, Aldasoro et al. 2004). Based on this evidence, Kamiński (2015) hypothesized that the diversification of the EGG can be explained by the turnover-pulse model (Vrba 1993). Specifically, it was assumed that cyclic paleoclimatic changes, by affecting the plant communities, caused pulsative shrinkage and extensions of the ancestors range/s. During the shrinkage phases the ancestral populations were fragmented, which promoted allopatric evolution (Vrba 1993).

As the investigation conducted by Kamiński (2015) was not time-stratified, it cannot be viewed as an attempt to test the turnover-pulse hypothesis. However, as that paper summarized the results of a long lasting taxonomic project based on investigation of over 13,000 specimens (including types of all the species), which resulted in several taxonomic publications (e.g., Iwan 1995, 1998, 2002a, 2003, 2004, 2005, 2006, Iwan & Kamiński 2012, Kamiński 2012, 2013a, b, c, 2014, Kamiński & Iwan 2013a, b, Kamiński & Ras 2011), it can be concluded that it provided a well-established biogeographic hypothesis, along with reliable tools for testing it, i.e. well-defined taxa, verified distributional records, and preselected geographic regions. The aim of this subproject is to put this data within the framework of molecular phylogenetics.


c. The genus Gonopus likely forms a divergent evolutionary lineage within Platynotina, based on its unique morphology and the available phylogenetic data (Figs 1, 2). The last taxonomic revision of the genus was conducted by Endrödy-Younga (2000), who distinguished 21 species and subspecies divided into two subgenera (Agonopus, Gonopus). While Dr. Endrödy-Younga was working on this manuscript, his health rapidly declined due to terminal illness. He died one year before his revision was published. According to the editorial note, at that time some of the taxonomic problems within the genus remained open (Endrödy-Younga 2000). Nevertheless the manuscript was published and can be treated as a valuable starting point for further investigations.

The identification of Gonopus species is changing due to the high degree of morphological variability observed even between the closely collected specimens (Kaminski & Iwan pres. obs.). This phenomenon is also common among other southern African beetle groups, such as toktokkies (Kamiński et al. 2020), and creates a major challenge for the morphology-based taxonomy. In such cases the search for reliable diagnostic characters forces researchers either to designate multiple monomorphic species (known from relatively few specimens), or to describe a few morphologically diverse and widely distributed ones. Present classification of Gonopus is a result of both those approaches, as currently the genus is subdivided into a few widely distributed species (e.g. G. tibialis), and many highly apomorphic ones known from relatively small type series (Endrödy-Younga 2000), such as G. transvaalensis described from Limpopo (South Africa). However, the morphological characters used by Endrödy-Younga for delimitation of the widely distributed species often break down when specimens from multiple populations are examined. Furthermore, the majority of the currently designated subspecies are sympatric, which logically undermines their status (e.g., G. tibialis tibialis and G. tibialis capensis or G. tibialis alternatus). All of this indicates that Gonopus is badly in need of revision, and suggests that the molecular approach for species delimitation should be used in such investigation.

In addition to reassessing the taxonomy within Gonopus, its relationship to the genus Stenogonopus (currently containing two species) must also be reexamined. Currently this taxon is strictly defined by the lack of apomorphic features (flat collar margin of prosternum, not exposing mouthparts) reported for Gonopus (Endrödy-Younga 2000). Taking into consideration the plesiomorphic definition of Stenogonopus, it can be assumed that Gonopus is probably paraphyletic in regard to that genus. However, detailed morphological and molecular analyses are needed to test this assumption.

Representatives of Gonopus are among the most extreme thermophiles within Platynotini. They seem to prefer sandy soils, and are common faunistic elements of the Namib and Kalahari deserts (Endrödy-Younga 2000; authors pers. obs.). This specific habitat preference seems to be linked to an interesting ethology. Gonopus agrestis was defined as an nocturnal fossorial detritivore that inhabits burrows during the daytime, and emerges at dusk to feed (Rasa 1994). Adults of this species carry food into their tunnels where it is consumed by them and all other burrow inhabitants. This includes their first instar larvae and other species of darkling beetles, such as Herpiscius damaralis (Rasa 1994). Parabiosis, parental care, and burrowing behaviours have not been reported for any other Platynotini genera, and is rare among Tenebrionidae in general. Furthermore, this specific ethology might be linked to the longevity of these beetles, as adults of different species of Gonopus were observed to live for over 8 years (Iwan 2010).

Although potentially interesting, the ethology of Gonopus remains poorly investigated. One of the main reasons for this is the lack of reliable taxonomic and faunistic data. This subproject aims to test generic, species, and subspecies boundaries within Gonopus and Stenogonopus using a total evidence approach based on molecular, morphological (imaginal and larval), and distributional data.


Ad. 4. As far as the authors of this proposal are aware, no studies have attempted to evaluate the usefulness of the Polish historical-entomological collections for molecular systematics. Due to the complicated history of the collection, and the efforts of present and former curators, the MIZ PAS contains a diverse set of taxonomically important and/or rare specimens (Iwan et al. 1995). This project aims to investigate the ‘molecular potential’ of the entomological collections of the following researchers based on the specimens preserved in the MIZ PAS.

Hans Gebien, a leading specialist in darkling beetles during the first half of the 20th century. Author of many species descriptions within Tenebrionidae, including Platynotini (Gebien 1904, 1910, 1911). The major part of his collection is preserved in the Natural History Museum in Basel (Switzerland), while the specimens hosted by MIZ PAS were historically obtained from the Szczecin National Museum. This part of Gebien’s collection remains unknown to a wider spectrum of entomologists.

Witold Eichler, Polish gynecologist, traveler, and entomologist. He collected a diverse assemblage of Coleoptera (different families). The core of his collection is composed of specimens collected in Northern Rhodesia (present Zambia) between 1941 and 1946. This includes several specimens of rare and undescribed Platynotina species.

Szymon Tenenbaum, one of the most prominent European beetle workers (Daszkiewicz & Bauer 2010) of the 20th century. His entomological collection includes over 250 drawers containing multiple families of Coleoptera. From the historical point of view, this is one of the most interesting collections in the world, as it was involved in the process of rescuing people from the Warsaw Ghetto during the Second World War. This history was described in a book (Ackerman 2007) and movie (Caro 2017) both entitled “The Zookeeper's Wife”. The platynotines in Tenebaum’s collection originate from his trips to Brazil (1923) and Mexico (1926).

Carl Koch, former curator at the Ditsong National Museum of Natural History (TNHN). Koch described the majority of African Platynotini species and genera (see Iwan 2002a, Kamiński 2015, 2016). Aside from Tenebrionidae, his diverse beetle collection contains representatives of many different families, including Carabidae and Scarabaeidae. The major part of this collection is deposited in the TNHM, while the MIZ PAS specimens were obtained by institutional exchange (identification of materials).

Zoltán Kaszab, former researcher at the Hungarian Natural History Museum (HNHM). He revised the Oriental component of Platynotina, describing several species (Kaszab 1975). The main part of his darkling beetle collection is deposited in the HNHM (Matskási 1987). The specimens deposited in MIZ PAS were obtained by institutional exchange (identification of materials).

Sebastian Endrödy-Younga, successor of C. Koch at the TNHN (Bellamy & Jäch 1999). Over the course of his career Endrödy-Younga amassed an extremely diverse beetle collection, which includes several Platynotini species (e.g., Endrödy-Younga 2000). His specimens are especially well-labeled (geographic coordinates, collection methods, additional details), which enables more detailed analyses of the factors influencing the DNA yield. Endrödy-Younga’s specimens deposited in MIZ PAS were obtained by institutional exchange (identification of materials).

Because all of the above-mentioned researchers gathered a wide variety of beetle families from different parts of the world, the results of the hereby proposed investigation would potentially impact many future phylogenetic projects. Furthermore, due to the historical value and general public interest in Szymon Tenenbaum’s entomological collection, the outcome of this study might generate publicity. The experience gathered during the realisation of this subproject will be used to implement detailed protocols for museum sequencing in the largest Polish zoological collection (MIZ PAS).

Fig. 1. Distribution and generic diversity of major evolutionary lineages of Platynotini. Eurynotina, anomalipoid, gonopoid and trigonopoid Platynotina display similar distributional patterns in Southern Africa. (b) Synapomorphic for Platynotini structure of the gular region (presence of stridulatory surface).

Fig. 2. Phylogeny of Platynotini (reanalysed from Kamiński et al. 2019a). Topology obtained in ML and Bayesian analyses of the concatenated 12S, 28S, COII, wg, CAD2, and ArgK matrix (3,892 bp). Posterior probabilities (PP) are displayed above branches, while bootstrap values (BS) are shown below. No value indicates PP=1.00 and/or BS=100. Abbreviations: ovo - indicates potential ovoviviparous clades (Ad. 2); EGG - member of Ectateus generic group (Ad. 3b).