DNA studies?; New DNA studies; More about Port Famine

Insectifera 2019, Vol 27, (2019), pp 42
Authors

DNA studies?

In 1997 I was wondering whether one could do DNA analysis of Colias ponteni. I then wrote in our magazine "Insectifera":

"If one could do a DNA analysis of the butterflies, this would of course also be extremely interesting. Hopefully, such an analysis might clarify how close the relative "imperialis specimens" are to the "ponteni specimens". Maybe it would even be able to get an idea of how large the population of the species could be, but above all it would be extremely interesting to see if it was possible to form a family tree with the other Colias species in the world. Then one might perhaps get an idea of where this isolated population in Hawaii originated."

A few years after the article was written, I received a letter from Professor Omoto in Japan who was doing just that kind of study. I sent him a small fragment of a leg of Wallengren's specimen of Colias ponteni.

Unfortunately, however, it proved impossible to obtain any useful material from this 150-year-old ponteni leg. Since this analysis took place more than 20 years ago and the technology has developed a lot since then, it would of course be very interesting to try again with this method, not least in order to find out more about Colias ponteni’s relationship with the other American Colias species.

New DNA-studies

Hopefully the time has finally come when it is planned that during this year, 2019, this kind of DNA study will be carried out because the technology has developed considerably over the past twenty years. Now, as this is written, I can only hope that we can get answers to the questions where Colias ponteni was actually found and where it ends up in a Colias family tree! Perhaps the pupa I found at the Evolutionary Museum in Uppsala might also give the crucial answer to the question of Port Famine or Honolulu?

Quite soon after I found the pupa, I consulted retired researcher Roland Moberg at the Botanical Department in Uppsala to see if it would be possible to do a DNA analysis of the stalk on which the pupa was attached. It could be a decisive proof if one could determine which species the stalk belonged to. He then replied that it was unfortunately impossible to get any useful information out of that 150 years old dry plant stalk.

More about Port Famine

Can the areas around Port Famine be the place for the collection of Colias ponteni and how did it look from the beach when the frigate Eugenie arrived in 1852 and also later? An interesting insight into this can be found in a note in the Swedish research traveler Carl Skottberg's book "Båtfärder" (Boat Voyages) which deals with his expedition for example to Tierra del Fuego. He describes the conditions around the Punta Arena, which is only about 30 miles north of Port Famine along the Strait of Magellan, around 50 years after the frigate Eugenie arrived at Port Famine.

Skottberg writes:

“In front of us lies a low yellow sandy beach, from which here and there narrow bridges go out into the strait, and beyond the beach the country rises to forested hills and ridges, where the fire burned black fields in the rich greenery. And between the sea and the forest lies Punta Arenas, the town at the Strait of Magelháes' type for the settler city and the upstart community, with an astoundingly rapid development behind it. In ten years, it has multiplied its population, and 12,000 people now have their home there - Chileans and Spaniards, Germans and Englishmen, French and Italian, Swedes, Norwegians, Danes, Russians Austrians. . . a language blunder, against which Babels almost fades. Stone palace among the metal houses, myriads of hotels and taverns, large storefronts and dirty windows, howling gramophones, slurries of "cocktails", which are brewed - see the first impression. We settle down on the traditional Swedish-traveling place, the hotel cosmos, whose low, white building rests on the sandy beach.”

Based on this description, it seems quite unlikely to me that nobody from any country for almost 170 years would have succeeded in finding this extremely eye-catching orange Colias butterfly. Which in that case of Kinberg and Andersson, who were not even entomologists, on a single day when they were probably completely concentrating on birds and plants, would have collected at least five specimens. Based on this, there is hardly any reason to suggest that the area south of Port Famine down towards Mt Tarn would be the place where Colias ponteni was collected. I have already mentioned the same thing about Mt Tarn based on Charles Darwin's description of the mountain in his book about the voyage on the Beagle.

Dr. Gerardo Lamas, who by many is considered to possess the greatest knowledge of South America's butterflies, wrote in 1981: “Obviously, no one has seen Colias ponteni alive for over 137 years. Hardly anyone, however, has visited the supposed localities and looked for it”. I have no idea how big the research was that actually happened in the area. However, based on Lama's suggestion, I read Skogman's book about which other landings were made on Eugenie's voyage through the Strait of Magellan. At that time, the beach-landing at the mouth of the Bachelor River appeared as the only possible alternative to Port Famine and Mt Tarn.

Explores DNA analysis as a tool to determine *Colias ponteni*'s phylogenetic position within the genus and clarify its origin locality. The author describes his 1997 proposal to sequence historical specimens and the failed attempt to extract usable DNA from a 150-year-old butterfly leg; by 2019, improved technology offered renewed hope for testing whether the species originated in Hawaii, Port Famine, or elsewhere, and for constructing a genealogy with other American *Colias* species.
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INSECTIFERA • ÅR 2019 • ÅRGÅNG 11
Men kan San Fransiscoområdet vara lokalen för Colias ponteni? Visst skulle det kunna vara så.
Området hyser ju många unika arter men jag måste ändock ställa mig starkt tvivlande till att denna
spektakulära art skulle ha funnits där och att den oerfarne Samuel Pontén eller Kinberg som alls icke
var någon entomolog skulle ha kunnat insamla minst fem, men kanske så många som åtta exemplar av
en sådan fjäril vid ett enda besök och att arten därefter aldrig återfunnits. Nej San Fransisco-området
kan nog med den största säkerhet avfärdas som möjlig lokal för Colias ponteni med hänsyn till de
många entomologer som redan vid denna tid i mitten av 1800-talet inventerat området runt San
Fransisco.

DNA-studier?

Redan 1997 funderade jag på huruvida man kunde göra en DNA-analys av djuren. Jag skrev då i vår
tidskrift ”Insectifera”:

”Kunde man göra en DNA-analys av djuren vore naturligtvis detta också ytterst intressant.
Förhoppningsvis skulle kanske en sådan analys kunna klargöra hur nära släkt ”imperialisdjuren” är
med ”pontenidjuren”. Den svarta vinkelteckningen vid framkanten är ju något mer framträdande på
”pontenidjuren”. Kanske skulle det t.o.m. gå att få en uppfattning om hur stor populationen av arten
kunde tänkas vara men framför allt vore det ju ytterst intressant att se om det gick att bilda ett släktträd
med de övriga Colias-arterna i världen. Då skulle man ju kanske kunna få en uppfattning om varifrån
denna isolerade population på Hawaii härstammade.”

Några år efter det artikeln skrevs fick jag brev från Professor Omoto i Japan som höll på med just
sådana studier. Jag skickade honom ett litet fragment av ett ben av Wallengrens exemplar av Colias
ponteni.

Det visade sig dock tyvärr omöjligt att få ut något användbart material ur detta då 150 år gamla ponteni-
ben. Några nya studier utifrån denna analysmetod har inte varit aktuellt för mig. Eftersom denna analys
skedde för mer än 20 år sedan och tekniken utvecklats mycket sedan dess vore det naturligtvis mycket
intressant att åter göra försök med denna metod, inte minst då för att närmare utröna Colias pontenis
släktskap med övriga Amerikanska Colias-arter.

Nya DNA-studier

Förhoppningsvis har denna tid nu äntligen kommit då man planerar att nu i år 2019 företa just
denna form av DNA-studier sedan tekniken avsevärt utvecklats de senaste 20 åren. Jag kan nu
när detta skrivs i maj bara hoppas att vi kan få svar på frågorna var Colias ponteni faktiskt påträffades
och var den hamnar i ett Colias-släktträd! Kanske kan då också den puppa jag fann hos Evolutions-
muséet i Uppsala ge de avgörande svaret på frågan Port Famine eller Honolulu? Ganska snart efter det
att jag hittat puppan rådfrågade jag för övrigt Roland Moberg på Botaniska institutionen i Uppsala om
det skulle vara möjligt att göra en DNA-analys av stjälken på vilken puppan satt fastgördlad. Det skulle
ju kunna bli ett avgörande bevis om man kunde fastställa vilken art stjälken tillhörde. Han svarade då
att det tyvärr var omöjligt att få fram något ur denna 150 år gamla torra stjälkstump.

Mer om Port Famine

Kan själva samhället Port Famine vara platsen för insamlandet av Colias ponteni och hur såg det ut
vid denna strandremsa vid fregatten Eugenies ankomst 1852 och tiden därefter? En intressant inblick
om detta kan man kanske få i en notis i den svenske forskningsresanden Carl Skottbergs bok
”Båtfärder” som handlar om hans expedition till bl.a. Eldslandet. Han beskriver där förhållandena runt
Punta Arena som ju endast ligger några mil norr om Port Famine längs Magellans sund runt 50 år efter
det att Eugenie anlöpte Port Famine.