Corresponding author: Henrik Wallin (
Academic editor: F. Vitali
A new subspecies of the European cerambycid
Wallin H, Kvamme T, Bergsten J (2017) To be or not to be a subspecies: description of
The tribe
Recently, there have been some taxonomic changes within the genus
Our study focus mainly on the northern populations of
We have not been able to find any attacks on, or specimens reared from, any other
Habitat of
We have also made a comparison with other
All 17 available sequences for
New material of both
DNA from imagines was extracted from adults using 1 leg, 2 legs, thoracic muscle tissue, or head and prothorax. When DNA from larvae was extracted, tissue from tergites or sternites was used. Extraction of DNA was done by using either the Quiagen tissue kit, or a GeneMole robot (Tab.
Ready-ToGo™ PCR beads (Amersham Biosciences) were used in all PCR recations and 2-4ul of DNA. The longer fragments were amplified under the following conditions: 95C for 5min followed by 40 cycles of 95C for 30s, 50C for 30s and 72C for 60s and a final extension period of 72C for 8min. The shorter fragments were amplified under the same conditions or with a shorter extension time (72C 50s). In second trials with samples that failed the first time, the annealing temperature was lowered to 47C. PCR reactions were purified with Exonuclease I and FastAP (Fermentas) and sequenced with a BigDye™ Terminator ver. 1.1 Cycle Sequencing Kit (Applied Biosystems), cleaned with a DyeEx 96 kit (QIAGEN) and ran on an ABI Prism 3100 Genetic Analyzer (Applied Biosystems).
Sequence chromatograms were edited in SEQUENCHER (Gene Codes Corporation). Contigs were created of the forward and reverse reads and of the two or three overlapping fragments for the older material. Sequences were exported in fasta format after primers had been removed and aligned using CLUSTALX 2.0 (
We calculated genetic distances under the Kimura 2-parameter model using MESQUITE (
Our study includes descriptions of the sclerotised parts of the male terminalia: the aedeagus, endophallus with the sclerites inside the median phallomere and the internal sac, tegmen with parameres and median lobe, and tergite VIII. The internal sac of the males was embedded in glycerol and photographed using a regular light microscope. This method is described in detail by
We maintain the use of the internal sac (part of the median phallomere), since it has been frequently used in the past (
Male genitalia photos were taken using an Olympus SZX 10 UC 30 camera attached to a Zeiss microscope and operated via the software ANALYSIS docum and Olympus Soft Imaging Solutions GmbH Version 5.1 (Build 2677). No stacking was used on these images. Habitus photos were taken using a Canon EOS 5D Mark II DSLR camera with a Canon MP-E 65mm f/2.8 1–5× macro lens and a Canon MT-24EX Macro Twin Lite flash with custom-made light diffusors. The camera was mounted on a motorized Stackshot rail (Cognisys) and operated via the software ZERENE STACKER (Zerene Systems) that was also used for stacking the images. Measurement data of body length (
Metadata for specimens included in the molecular analysis. Column four gives GenBank accession numbers.
Species | Extr. ID | Ext method | CO1 Acc | Stage | from | Country, province, locality | Date | Leg. |
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|
JB941 | Qiagen |
|
larva |
|
Norway, Hedmark, Ljørdalen | 27.06.2013 | Torstein Kvamme |
|
JB942 | Qiagen |
|
larva |
|
Norway, Hedmark, Engerdal | 27.06.2013 | Torstein Kvamme |
|
JB946 | Qiagen |
|
larva |
|
Norway, Hedmark, Ljørdalen | 27.06.2013 | Torstein Kvamme |
|
JB949 | Qiagen |
|
larva |
|
Sweden, Lule lappmark, Kiruna | 24.06.2013 | Torstein Kvamme |
|
JB950 | Qiagen |
|
larva |
|
Norway, Hedmark, Trysil | 27.06.2013 | Torstein Kvamme |
|
JB016 | GeneMole | Failed | adult | Sweden, Torne Lappmark, Silkimuotka | 28.VI.1948 | N. Höglund | |
|
JB017 | GeneMole | Failed | adult | Sweden, Torne Lappmark, Silkimuotka | 28.VI.1948 | N. Höglund | |
|
JB021(JB250) | GeneMole |
|
adult | Sweden, Åsele Lappmark, Kittelfjäll | 28.VI.1972 | T-E Leiler | |
|
JB022(JB249) | GeneMole |
|
adult | Sweden, Torne Lappmark, Soppero | 30.VI.1980 | Stig Lundberg | |
|
JB023(JB248) | GeneMole | Failed | adult | Sweden, Torne Lappmark, Soppero | 15.VI.1968 | Stig Lundberg | |
|
JB024(JB251) | GeneMole |
|
adult | Sweden, Lule Lappmark, Messaure | 14.VII.1971 | S. Lundberg & T. Müller | |
|
JB945 | Qiagen |
|
larva |
|
Sweden, Uppland, Uppsala | 07.2013 | Henrik Wallin |
|
JB018(JB247) | GeneMole |
|
adult | Sweden, Öland, Räpplinge | 03.V.1976 | Bert Gustafsson | |
|
JB019(JB246) | GeneMole |
|
adult | Sweden, Småland, Åseda | 26.XII.1974 | Bert Gustafsson | |
|
JB020(JB245) | GeneMole |
|
adult | Sweden, Uppland, Uppsala | 01.V.1984 | Stig Lundberg | |
|
JB025(JB252) | GeneMole |
|
adult | Sweden, Norrbotten, Kalix | 30.VI.1994 | S. Lundberg & T. Müller | |
|
JB026 | GeneMole | Failed | adult | Sweden, Västerbotten, Umeå | 09.V.1969 | Lars Huggert | |
|
JB027 | GeneMole | Failed | adult | Sweden, Halland, Släp | 02.V.1965 | Lars Huggert | |
|
JB028 | GeneMole | Failed | adult | Sweden, Västergötland, Amundön | 31.12.1968 | Lars Huggert | |
|
JB029 | GeneMole |
|
larva |
|
Sweden, Uppland, Uppsala | 05.2009 | Henrik Wallin |
|
JB030 | GeneMole |
|
adult |
|
Sweden, Uppland, Knutby | 05.2009 | Henrik Wallin |
|
JB938(RB122) | Qiagen |
|
larva |
|
Sweden, Uppland, Uppsala | 07.2013 | Henrik Wallin |
|
JB944 | Qiagen |
|
larva |
|
Sweden, Uppland, Knutby | 07.2013 | Henrik Wallin |
|
JB031 | GeneMole |
|
adult | Sweden, Södermanland, Haninge | 20.IX.2009 | Julio Ferrer | |
|
JB939 | Qiagen | Failed | adult |
|
Canada, Ontario, Ottawa | 07.07.1961 | S.D. Hicks |
|
JB943 | Qiagen | Failed | adult |
|
USA, California, Turlock | 24.05.1955 | R.R. Snelling |
|
JB948 | Qiagen |
|
larva |
|
Sweden, Uppland, Knutby | 07.2013 | Henrik Wallin |
Stems and branches were cut from shrubs of
The species nomenclature follows
Specific information on examined specimens is mentioned under each species in the section “Taxonomy”. The dates and other information were copied from the labels. In some cases, additional information provided by collectors has been added.
There are 69 published and released 5-prime end fragments of
Gene tree from strict clock analysis with Beast of a 5-prime end fragment of mitochondrial cytochrome oxidase subunit I (the animal Barcoding fragment). Numbers at nodes are posterior probability values, only given for nodes >0.5. Scale bar = expected number of substitutions per site.
Amplification of the 3-prime end fragment of
Gene tree from strict clock analysis with Beast of a 3-prime end fragment of mitochondrial cytochrome oxidase subunit I. Numbers at nodes are posterior probability values, only given for nodes >0.5. Scale bar = expected number of substitutions per site.
The genetic distance between
The body length, among the examined specimens, was significantly smaller in
Body lengths of males and females of
The subspecies are not diagnosable based on body length in the sense requiring 75% of individuals of subspecies A to be outside the distribution of 99% of subspecies B (
Body shape measured as the ratio of total body length to maximum body width of males and females of
Type species.
Habitus (dorsal view).
The following specimens are available through Boldsystems Public Data Portal and
A medium-sized and subcylindrical species with body length 9.0–15.0 mm according to e.g.
Habitus (dorsal view).
Morphological characteristics of
Females form a “U-shaped mark” in the bark of
The preferred host tree is
Host tree species of
Host tree species | Reference |
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Habitus (lateral view).
The following specimens collected in Finland and available (through Boldsystems Public Data Portal) for photo examination includes: 1 ♀ COLFA181-10, Lapland, Inari, 1980-07-11, leg. Erkki Laasonen, id
A relatively small to medium-sized and subcylindrical subspecies with body length 9.5–13.0 mm in females and 8.0–12.0 mm in males, according to measurements from the present study. Body 3.1 times longer than wide in females and 3.4 times longer than wide in males (Fig.
Frons.
morphological characteristics are mainly based on type specimens, either collected on, or reared from branches of
Aedeagi (
The name is an adjective used as a substantive in the genitive case derived from the specific name of the host plant
The distribution of
Hind wings.
The attacks are similar to
In addition, parasites including wasps and flies frequently attack
Host tree attacks.
Distribution of records mainly from Fennoscandia. Open circles:
Subspecies of
The new subspecies
We agree with
The remaining species within the subgenus subgenus
The male genitalia of all other Palaearctic species of
The other species synonymised by
That
The trinomial subspecies remain a contentious hierarchical level in zoological taxonomy (
We thank Mikhail Danilevsky (Moscow), Larry Bezark (Sacramento, California) and Dan Heffern (Houston, Texas) for support during various stages of this work. Rasa Bukontaite (