Higher altitude taxa increasing range and genetic relateness at sub species level during glacial periods or not? The alpine leaf beetle Oreina elongata

My last blog post takes us on a journey to Northern Europe (how exciting!). The taxon of interest this time is the alpine leaf beetle Oreina elongata (Coleoptera: Chrysomelidae). Most phylogeography with regards to short term evolutionary timescales consists of making inferences, based on the Quaternary climate cycles (starting 2.6 mya). Most phylogeographic study is also usually on taxa that are distributed in lowland habitats, which are isolated into refugia (areas of habitat maintained/ not changed or covered in ice). during glacial periods. In Europe these refugia were located in southern Europe, in areas like the Italian, Iberian and Balkan Peninsulas. However this beetle taxa O.elongata is found in high altitudes, and has had its range contracted post last glacial maximum. It is of interest as to weather it expanded its range during glacial periods, and gene flow occurred keeping the taxa similar morphologically and genetically. O.elongata has many isolated populations found on the Alps and Apennines at altitudes of 1200–2500 m above sea level (See Fig 1 below).



Map showing isolated populations on O.elongata in the Alps and Apennines mountain ranges in southern Europe

 Several subspecies have been described for O.elongata allopatrically based on varied morphological characters (male genitalia (the aedeagus) and cuticle microstructure). These beetles are herbivores that feed on hosts from two tribes of Asteraceae. When feeding on the tribe Cirsium (Cynareae) the larvae and adults synthesise cardenolides, where-as individuals feeding on the tribes Adenostyles or Senecio (Senecioneae) encounter pyrrolizidine alkaloid Noxides which are plant produced PAs which they can sequester. These seem to be a form of chemical defence as they also have 3 bright colour morphs (see Fig 2) indicating toxicity to potential predators (blue, green and mixed morphs).

Figure 2. Two colour morphs of the alpine leaf beetle Oriena elongata (It's third morph is a composite green/ blue colour)

 

 



These were the main questions that this paper was trying to answer, using molecular analysis. Males were analysed molecularly as the described sub-species could be identified by the genitalia. For technical aspects about the Molecular analysis refer to the Methods and Results section of this article.

 

1. Did O. elongata, as a representative of the high altitude fauna, survive the cold periods of the Quaternary in situ in the Alps and Apennines?

In order to infer the timescale over which differentiation is likely to have arisen, we make use of an approximate molecular dating method based on a review of published gene-specific mtDNA substitution rates to answer a further question:

2. Was divergence within O. elongata a product of the last glacial cycle or is the differentiation more ancient?

 

The CO1 gene region is commonly used to express polymorphism within genera and species. For this study it has shown that there is 5 distinct clades of O. elongata across its geographic range. This is a complex the authors of this paper prefer to use, compared to the 7 subspecies approach as the genetic differentiation between these different sites and populations is that high. As the polymorphism is higher than expected and 4 of the seven sub-species do not appear to be monophyletic clades (Refer to the phylogenetic tree in the paper as the image is not clear when put onto this blog post) (shared recent common ancestor). Therefore it should be required to sample these populations more intensely and determine more morphological characters that may be variable between these proposed 5 species!

 

It is probable that O.elongata did survive the glacial periods in situ on the Alps and Apennines. Based on the approximate dating method, along with the phylogenetic analysis and molecular clock hypothesis, O.elongata  diverged long before the Last glacial maximum. The general pattern in the phylogeography is that the populations from the central Swiss Alps and northern Italy are basal, with subsequent separation of the eastern, southern (Apennines) and western extremes of the distribution, and finally a second colonization of the Apennines. Clades I and II in the Central Alps separated early and must have survived many glacial cycles in isolation from the rest of the species, perhaps in refuges within the Alps and are the basal group. This structuring is also common within plants in the area. Four refugia are proposed for alpine plants along the southwestern, southern and eastern edges of the Alps, which is what was found similar in this study of O.elongata.

 

This is all interesting, as during glacial periods the habitat that O.elongata could inhabit would have been substantially larger. However most of the populations have remained relatively disjoint since the major divergence of clades about 0.3 to 1.0 million years ago! Therefore it is indicative that this genus of beetles are low dispersers and there geographic range contraction during interglacial periods has lead to speciation. It's good to see the other side of the coin sometimes!

 

Thanks for reading!

 

Article: http://www.sciencedirect.com.ezproxy.lincoln.ac.nz/science/article/pii/S1055790310003519

Phylogeography of two species in the "neglected" taxon of mygalomorph; the Funnel web spiders of Austrailia



Dinner plate for scale!

Spiders are massively cool creatures! Reasons for this include big pointed (sometimes poisonous) fangs (Chelicerae), or they can get to the size of dinner plates (including leg span). Mygalomorph (Infraorder) is an infamous taxa of spider which includes the likes of trapdoor spiders and Tarantulas. They also include the spider of interest in this paper the Funnel web spider! Most phylogeographic studies look at how varying environmental conditions or dispersal barriers cause speciation, in certain related taxa. The reason this paper caught my attention was due to their reference too ‘neglected taxa’. This was mainly due to the hypothesis they were testing. This was how two niche differentiated spiders in the same taxa that are sympatrically dispersed, can have such variation in evolution and population structure under the same environmental conditions. 

 

Unfortunately with phylogeographic research there is a reliance on large, multilocus datasets. These data sets are usually derived from past research on high profile taxa. Interestingly proposed by the authors of this paper, is that these species do not paint the whole proverbially picture. Taxa like terrestrial invertebrates have characteristics like longevity and persistence in the environment, which can retain and display phylogeographical signals over a longer time period than “obvious” groups (Probably stupid overrated groups like mammals!!). These obvious groups get sucked into the ‘snowball down a hill’ theory (research spawns more research) leading to a noticeable absence of research on possibly more interesting taxa. Initial pioneer studies are unattractive however due to initial ground work required! Collection of understudied taxa is difficult for obvious reasons (Where the heck do I find these things!). Molecular techniques can be highly specialised so for some taxa, development of the methods to extract genetic information can take a long time. With enough determination and perseverance these barriers can be overcome to show some really remarkable phylogeographic patterns.

 

So what’s the deal with these funnel web spiders? Well for one they are distinguished from another common group of spiders the Araneomorphae. Mygalomorphs only respire via booklungs compared to Araneomorphae which respire via booklungs and trachea. The booklungs expose a large surface area to the air making them highly susceptible to desiccation. This leads to a sedentary lifestyle, living in burrows and expressing a low vagility (movement in the environment!). These characteristics mean that the mygalomorphs can retain phylogeographic signal at finer scales over long time periods. Recent work has been completed revising the taxonomy of the Australian funnel web spiders. The two species of interest in this study are from the family Hexathelidae, and are called Hadronyche cerberea (Koch, 1873) and Atrax sutherlandi Gray, 2010. Both species have largely overlapping ranges and distributions in SE Austrailia, also being sympatrically and continuously distributed in the study site region of Tallaganda. These spiders are non-balloners which means the dispersal into their currently large range must have been over a long time. Biological characteristics include both species are of similar size, long lived and construct permanent burrow retreats (homes). Prey items include other invertebrates and small vertebrates that ends up in the funnel of death! Previous research looked at burrow detritus and lab experiments to look for specific prey preference. No definitive preference was found, so it was assumed the diet is based on what is in its microhabitat. These species both share common characters as referenced above, however have a marked variation in where they build their homes. A.sutherlandi burrows exclusively in soil, where H.cerberea is saproxylic meaning it makes its burrows in decomposing logs on the forest floor!

 

The study site Tallaganda, is shown in the above image. The pink areas are soils of metasedimentary segments dating back to the Ordovician period (490-443 Mya). The areas on the above image that are brown are dated from (443-354 Mya), making this landscape geologically stable. This region was subject to 100,000 year glacial interglacial cycles within the Pleistocene, with 20 glacial-interglacial periods in the last 2.4 Myr. As Tallaganda is within both A.sutherlandi and H.cerberea's wider distribution it implies they are not recent arrivals, therefore historical conditions have been the same for them. The alternate hypothesis is that both species show disparate phylogeographical patterns. This means there life histories and niches differences have played a role in there evolution which varies with each other.
The results were consistent with the alternate hypothesis that these two sympatric, niche differentiated species have different underlying population structures!

H.cerberea population structure suggest a recent radiation from a point source or recent colonization of Tallaganda. Low nucleotide diversity and high haplotype diversity is found in H.cerberea  which is consistent with this idea of recent rapid expansion. Either a small population survived in Tallaganda during the last glaciation and radiated, or the more likely idea that they recolonized from the Greater Diving Range (GDR). The GDR overlaps Tallaganda, which had individuals from the Monga forest in the GDR sequenced. The H.cerberea from Tallganda were shallowly nested (within the same clade, or close to) with the Mongo forest individuals meaning this is probably the area they recolonized from. This pattern is not found in any other saproxylic species studied within the Tallganda region, indicating that something about H.cerberea biology or association with its habitat is the causal factor. This pattern is proposed to be due to H.cerberea relying on wood to be present and decompose, so they can make burrows. But during glaciations the treelines are decreased substantially and no forest was left in the Tallaganda region. Therefore glaciation and environmental change will have a greater effect on H.cerberea and have an increased chance of causing local extinction.

The genetic analysis on H.cerberea has provided insight into how it has interacted in the Tallganda region, due to its more dynamic niche. A.sutherlandi however are indicative of long-term persistence within the Tallaganda region during glacial periods. H.cerberea had a phylogeny with a low variety of clades showing expansion from one source back into Tallaganda. A.sutherlandi however displayed a phylogeny which was more monophyletic, corresponding to a population that is distinct and has non-overlapping geographical ranges. Phylogeny of A.sutherlandi showing genetically and geographically distinct clades. This pattern is consistent with the idea that small areas of refugia were maintained during the glacial cycle. This means there was areas that the two main different soil types, in areas not covered or impacted by ice, allowing burrows and habitats to be built and sustained. This allows evolution and preference to the soil type of the refugia. These are also barriers to gene flow and is why there is several distinct genetic clades. I don't want to go into excess detail about soil type preference so I'll keep it short and sweet. Certain groups had a preference to either granite or metasediment soils based on what was found at the refugia. Post glacial, these groups began recolonizing the other soil type making paraphyletic groups (groups sourced from granitic soils) to monophyletic  metasediment clades. There is some degree of preference and structuring based on the underlying lithology. (Read the paper if you want a more in-depth look at the structuring of soil preference!).

Their are a few things that this can show us about evolution of invertebrates over time. One is that closely related but niche differentiated species like the funnel web spiders, can evolve differently as climate change affects their particular niche differently (no forest vs soil). This trivial difference of burrowing in wood vs soil shows that even the subtle and overlooked traits/ behaviours or niches can influence evolution drastically. Also as the funnel webs are a neglected taxon, it is important to realise how these taxon can show previously underestimated evolution history cues in certain environments, and on different organisms like invertebrates. I enjoyed this paper as it was on spiders :D. It also shows that phylogeography has a long way to go, but invertebrates are a possible solution to furthering our understanding of organisms distributed in space and time!

Thanks for reading! The link to the paper is posted below

Article: http://apps.webofknowledge.com.ezproxy.lincoln.ac.nz/full_record.do?product=UA&search_mode=GeneralSearch&qid=1&SID=Y13dKoo@2n8B2ldJAfC&page=1&doc=1



 

Habitat and climatic variations impact on the population of the Capr velvet worm on the South African Cape peninsula and surrounding mountain area

A few weeks ago I was sitting in on a Masters Research proposal on genetic variation in invertabrate taxa on Banks Peninsula. By chance a famous New Zealander was also attending this proposal,

Ruud with a weta on his face!

who goes by the name Ruud Kleinpaste! If you don't know he is an iconic Kiwi personality off the T.V. show 'Buggin with Rudd'. He asked the person presenting there research ideas about an interesting group of invertabrates called the Onychophora (Velvet worms) found worldwide, and if he had come across any of them in his sampling. This person had not, as they are a interesting group (it has a crazy biology!) dating back to the Cambrian period 500 million years ago! So once I had decided to do a blog on insect phylogeography, it only seemed right that I investigate this group. This lead me to a paper showing how a velvet worm species biology, has influenced it's crytic evolution in a dynamic geological and climatic landscape.

First I should describe this creature of anicent proportions and what makes it so bizarre!

The mighty velvet worm!

Its a segmented invertabrate, with a flattened cylindrical body. It moves very slowly using its lobopods (stub feet) by expanding and contracting internal muscles in each lobopod. Each foot has a pair of retractable, sclerotised (hardened) chitin claws which are where its scientific name is derived: 'Onycho' 'phora' meaning "claws" and "carry" respectively, and are used to gain firm grips on uneven terrain. They are soft bodied creatures with a fluid-filled body cavity acting as a hydro skeleton, which is similar in unrelated soft bodied animals like worms. It respires via its whole body surface via diffusion. It lives in preferably dark environments with high air humidity and is found in tropical habits in the temperate zone within the South hemisphere. Due to there body make-up and slow movement habits they are night active to avoid desiccation (drying up) during the day time. They have oral papillae (slime glands) on the side of their head which is used to contain, then excrete slime onto approaching predators as defence or prey of interest.

For a more detailed description of velvet worms which I have only summarised above, go to this link: http://en.wikipedia.org/wiki/Onychophora#Conservation_status

The paper of interest is called; Phylogeography of the Cape velvet worm (Onychophora: Peripatopsis capensis) reveals impact of Pliocene and Pleistocene climatic oscillations on Afromontane froest in Western Cape, South Africa

The study site is on the south-western Cape, more specifically Cape Floristic Region (CFR) with a history of an early Miocene period (5-23mya) with a characteristic subtropical rainforest making up most of the vegetation. However the climate changed during the Pliocene era (2-5mya) amplifying the east-west rainfall gradient as mountains began forming in the south-eastern Africa making west CFR more arid winter only rainfall climate. The east contrastingly had year round rainfall. The current topography is shown in the below picture showing the molecular results based on the isolated habitats.

The afromontane forest was once one large connected forest. But aridification that occurred during the Miocene/ Pliocene separated this forest in discontinuous fragmented forests patches.With the two main types consisting of southern afromontane forest and southern coastal forest. This lead them to 3 main hypothesise (1) P.capensis will show a genetic history mirroring the paleogeography (isolation of habitat over time) due to it's specificity to habitat/ low dispersal; (2) The physical barriers of low lying coastal plains and lack of forest connectivity lead to this isolation also and (3) Increasing genetic differentiation should be observed moving along a west-to-south-easterly trajectory as the south-eastern part of the Cape Fold mountains have historically had a larger annual rainfall with larger forest patches creating more habitat heterogeneity (intra forest fragment variation/ ecotypes).

Specimens of P.capensis were collected from many sites over the Southern Cape. They were collected from forest floor litter and decaying plant material (there preferred habitat). A minimum of one specimen to a maximum of 10 were collected from 21 different sites in the western and southwestern cape regions of the western Cape Province of South Africa. Conventional DNA analysis methods were used to extract, amplify, sequence and analyse the sample DNA data. Refer to the paper for in-depth methods of DNA analysis if sort of thing interests you!

The results produced clear findings which eluded to the fact that their has been genetic differentiation. The phylogenetic tree produced showed that there were 3 genetically variable and distinct clades of P.capensis. Clade A is the Cape Peninsula population, which is separated by the Cape Flats and mountains from Clade C, the Theewaterskloof-Overstrand population. Furthermore Clade C is separated from Clade B which is the Overberg population, by the Breede River-valley and adjacent mountain ranges. These separations are in the form of habitat isolation due to geological change over time. The Cape flats is a shrub dominated and nutrient poor ecosystem, which has been shown in past studies to also limit gene flow between other invertebrate groups! In the case of P.capensis this habitat is less desirable to live in as there is less woody detritus and leaf litter to live in. It is also susceptible to desiccation in this habitat as it is more arid than the afromontane forest of the Cape Peninsula. The Breede River-valley produces unique abiotic conditions creating a semi-arid environment, with a multitude of variable soils (e.g. sandy, aeolian, acidic, alluvial, clay and loam) with only 270mm rainfall per annum. These unique condition are not suited for forest growth and therefore there is no dispersal pathway between Clades B and C, leading to this genetic differentiation. These results are consistent with the first and second hypothesis of the authors!

This is a phylogenetic tree showing the three clades of velvet worm based on variation in the sequences on the CO1 gene

Within habitat heterogeneity was different between all three clades. This means there is a larger amount varying habitats in certain clades, leading to a greater population structure and genetic variability. Clade A shown in red on the above Fig 3 had the lowest haplotypic diversity (inter clade genetic variation), as the habitat in the region was relatively continuous and was relatively consist (no large variation in topography or climate). Clade B had the highest level of haplotypic diversity as the Overberg region had many fragmented areas of forest habitat meaning dispersal by P.capensis is greatly limited. Clade C was the largest in area with a varying climate. Rainfall varied from only in winter on the western part of Theewaterskloof-Overstrand to year round in the south east. Also as the Cape mountains (where a large proportion of afromontane forest grows) lies in this area there is a high amount of topographic heterogeneity. It was suggested that this area has had a long evolutionary history with 3 distinct haplotypes arising between the most variable and extreme environments due to evolution at the meta population level. The level of habitat heterogeneity varying from low in the west (Clade A) to highest in the south east (Clade C)! The authors again were able to validate their third hypothesis.

This paper is a really cool way to show how habitat can influence invertebrate species interactions in a dynamic environment. It provides insight that allows people to realise that; hey what we thought was one well dispersed species is possibly 2 or even 3! This could impact the taxonomic classification of species as many areas in the world will have similar patterns of evolution and population isolation occurring.

Thanks for reading my first blog post!!! (I hope you could follow most of it!)

This is the link to the paper:http://onlinelibrary.wiley.com.ezproxy.lincoln.ac.nz/doi/10.1111/j.1420-9101.2012.02482.x/pdf

 
If you wanna find out what the bug man is all about i suggest you check out the links below :D

http://www.youtube.com/watch?v=UefNF-65hTA&list=PL3562ECF090AF7264 

(Just copy and paste the link this stupid blog is messing with my formatting :O)