Molecular and phylogenetic analysis of the Salt Creek tiger beetles(Ellipsoptera nevadica lincoliniana)

Innocent Byiringiro Author

04/05/2021 Added

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UCARE 2020-2021 research presentation.

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[00:00:01.490]Hello, my name is Innocent Byiringiro.
[00:00:05.690]I'll be presenting about the research
[00:00:07.610]i'm carrying out, titled Molecular and Phylogenetic Analysis
[00:00:12.490]of the Salt Creek tiger beetles.
[00:00:15.090]I am under the directions of Dr. Thomas Powers.
[00:00:19.960]Salt creek tiger beetle
[00:00:21.700]is one of Nebraska's endangered species.
[00:00:24.760]It is found in the Ellipsoptera genus. a genus
[00:00:28.070]that it shares with 13 other species and 9 subspecies.
[00:00:34.583]Ellipsoptera nevadica lincolniana,
[00:00:36.820]Salt Creek tiger beetle as its common name
[00:00:39.730]is a subspecies in the nevadica species.
[00:00:43.530]Subspecies that have been proven to be closely-related to
[00:00:47.290]Ellipsoptera nevadica lincolniana include:
[00:00:49.940]Ellipsoptera nevadica olmosa
[00:00:53.255]Ellipsoptera nevadica nevadica
[00:00:55.611]and Ellipsoptera nevadica knausii.
[00:00:59.390]Amongst them, only Salt Creek tiger beetle
[00:01:02.800]was listed as a federal endangered subspecies
[00:01:07.290]on November 7th, 2005.
[00:01:11.440]Salt Creek tiger beetle is verily niche specific,
[00:01:15.040]spend most of his life as larvae,
[00:01:18.420]mostly observed as an adult.
[00:01:21.600]It's most common habitats are barren salt flats
[00:01:25.700]and saline stream edges of the saline wetlands.
[00:01:29.730]Due to only surviving on limited conditions,
[00:01:33.350]it has one of the most restricted ranges
[00:01:36.860]of any insect in the United States.
[00:01:41.200]In Nebraska, known-meta populations include
[00:01:45.187]Little Salt Creek Arbor Lake,
[00:01:47.740]Little Salt Creek Roper,
[00:01:49.600]Upper Little Salt Creek - North,
[00:01:51.620]Upper Little Salt Creek - South,
[00:01:53.760]Jack Sinn WMA
[00:01:55.810]and Oak Creek.
[00:01:57.940]Only three meta populations still exist.
[00:02:02.820]On May 6th , 2014,
[00:02:05.930]a final rule designated 449 hectares
[00:02:10.040]for Salt Creek tiger beetle recovery plan.
[00:02:14.270]To protect Salt Creek tiger beetles,
[00:02:16.990]we are interested in learning the evolutionary relationship
[00:02:20.790]between Ellipsoptera nevadica lincolniana
[00:02:23.850]with other species and subspecies
[00:02:26.910]in the genus ellipsoptera genus and beyond.
[00:02:30.090]This study will also allow us to predict
[00:02:33.700]the relatedness of the cultured and wild Salt Creek tiger beetles
[00:02:38.490]to understand the variation
[00:02:40.360]in the Salt Creek tiger beetle populations.
[00:02:43.480]Comparison with other subspecies
[00:02:46.890]will provide more evidence to conserve habitat
[00:02:50.580]that the salt creek tiger beetle depends on.
[00:02:54.600]It will also provide data that can be used
[00:02:58.660]to easily identify this salt creek tiger beetle
[00:03:01.750]and other understudied tiger beetle species.
[00:03:05.430]In addition to that,
[00:03:06.950]it will serve as a basis for further studies
[00:03:09.860]to investigate specific genetic differences
[00:03:13.029]between subspecies and how those influence
[00:03:16.521]their survivability or extinction.
[00:03:21.340]To perform this study, we received 60 samples
[00:03:25.290]from Steve Spomer, including our species of interest,
[00:03:30.025]Ellipsoptera nevadica lincolniana,
[00:03:33.272]Ellipsoptera nevadica knausii,
[00:03:36.048]and other species in Ellipsoptera genus.
[00:03:40.790]One hind leg was cut from each insect
[00:03:44.710]and DNA was extracted and purified
[00:03:47.990]using Qiagen blood and tissue kit.
[00:03:51.940]The CO1 one region of the mitochondrial genome
[00:03:54.447]which have been proven to be conserved and efficient
[00:03:58.080]for evolutionary analysis among species
[00:04:01.780]was amplified using the polymerase chain reaction.
[00:04:06.640]Successful amplicons were cleaned
[00:04:09.470]and sent to Eton Bioscience lab,
[00:04:11.970]which sent us DNA sequences.
[00:04:15.150]After receiving the DNA from Eton Biosciences,
[00:04:18.359]we edited and formatted the sequences
[00:04:21.160]for phylogenetic analysis.
[00:04:24.040]Our analysis using a maximum likelyhood tree,
[00:04:28.130]showed Ellipsoptera nevadica knausii
[00:04:30.913]and Ellipsoptera nevadica lincolniana
[00:04:33.820]to be closely related with minor differences.
[00:04:38.030]It also showed other species that are closely related
[00:04:42.270]to Ellipsoptera nevadica lincolniana
[00:04:45.240]by falling in the same clade.
[00:04:48.040]These findings support the taxonomical classification
[00:04:52.360]of subspecies in the Ellipsoptera nevadica.
[00:04:55.840]Further analysis of the subspecies
[00:04:58.479]in the Ellipsoptera nevadica
[00:05:00.860]showed that only one polymorphic site was found
[00:05:05.730]between Ellipsoptera nevadica lincolniana
[00:05:08.110]and Ellipsoptera nevadica knausii.
[00:05:10.832]13 polymorphic sites are found
[00:05:13.216]between Ellipsoptera nevadica lincolniana
[00:05:15.902]and Ellipsoptera nevadica nevadica.
[00:05:18.873]12 polymorphic sites were found
[00:05:21.009]between Ellipsoptera nevadica lincolniana
[00:05:23.635]and Ellipsoptera nevadica olmosa.
[00:05:27.190]So what is the meaning of these results?
[00:05:30.910]Originaly , the Ellipsoptera nevadica lincolniana
[00:05:34.400]was described as a separate species,
[00:05:37.370]but more recently,
[00:05:39.150]based on geographical and morphological evidence,
[00:05:42.500]it was re-classified as a subspecies
[00:05:45.580]in the Ellipsoptera nevadica species.
[00:05:48.330]Our study supported that evidence.
[00:05:52.420]Further studies demonstrated
[00:05:54.550]that Ellipsoptera nevadica lincolniana
[00:05:57.230]is a very new specific species
[00:06:00.160]compared to other subspecies in the nevadica species.
[00:06:03.930]Even though it is geographically and morphologically
[00:06:06.453]related to Ellipsoptera nevadica knausii,
[00:06:10.550]the two subspecies have significant ecological
[00:06:14.246]and behavioral differences.
[00:06:16.630]This study forms the basis to further
[00:06:19.820]explore specific genetic differences
[00:06:23.170]between Ellipsoptera nevadica lincolniana
[00:06:25.862]and Ellipsoptera nevadica knausii
[00:06:28.750]in order to identify genes
[00:06:31.370]that may account for the ecological and behavioral
[00:06:34.670]preferences of each subspecies.
[00:06:38.280]This study also provided genetic material information
[00:06:42.107]that will be used to design environmental DNA tests
[00:06:46.120]for the Ellipsoptera nevadica lincolniana.
[00:06:50.950]Thank you.

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Molecular and phylogenetic analysis of the Salt Creek tiger beetles(Ellipsoptera nevadica lincoliniana)

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