Editing Eastern wolf (section)

====Genetic evidence====
[[Mitochondrial DNA]] (mDNA) passes along the maternal line and can date back thousands of years.<ref name=beeland2013/>

In 1991, a study of the [[mitochondrial DNA]] (mDNA) sequences of wolves and coyotes from across North America found that the wolves of the Minnesota, Ontario and Quebec regions possessed coyote [[genotypes]]. The study proposes that dispersing male gray wolves were mating with coyote females in deforested areas bordering wolf territory. The distribution of coyote genotypes within wolves matched the phenotypic differences between these wolves found in an earlier study, with the larger Great Lakes wolf found in Minnesota, the smaller Algonquin ([[Algonquin Provincial Park|Provincial Park]]) type found in central Ontario, and the smallest and more coyote-like [[Eastern coyote|tweed wolf or eastern coyote]] type occupying sections of southeastern Ontario and southern Quebec.<ref name=lehman1991/>

{{cladogram
|title=Proposed phylogenetic tree of wolf evolution
|cladogram=
  {{clade
  |label1 = Ancestral Canid{{br}}(1-2 million years ago)
  |1 =
    {{clade
    |1 =
      {{clade
      |1 = [[Coyote]]
      |2 =
        {{clade
        |1 = [[Red wolf]]
        |2 = Eastern wolf
        }}
      }}
    |label2 = [[Gray wolf]]
    |grouplabel2 = [[Subspecies of Canis lupus|Gray wolf subspecies]]
    |bar2 = grey
    |2 =
      {{clade
      |1 = {{clade}}
      |2 = {{clade}}
      }}
    }}
  }}
}}

In 2000, a study looked at red wolves and eastern wolves from both eastern Canada and Minnesota. The study agreed that these two wolves readily hybridize with the coyote. The study used 8&nbsp;[[microsatellites]] (genetic markers taken from across the [[genome]] of a specimen). The phylogenetic tree produced from the genetic sequences showed a close relationship among the red wolves and the eastern wolves from Algonquin Park, southern Quebec, and Minnesota such that they all clustered together. These then clustered next closer with the coyote and away from the gray wolf. A further analysis using mDNA sequences indicated the presence of coyote in both of these two wolves, and that these two wolves had diverged from the coyote 150,000–300,000&nbsp;years ago. No gray wolf sequences were detected in the samples. The study proposed that these findings are inconsistent with the two wolves being subspecies of the gray wolf, that red wolves and eastern wolves (eastern Canadian and Minnesota) evolved in North America after having diverged from the coyote, and therefore they are more likely to hybridize with coyotes.<ref name=wilson2000/>

In 2009, a study of eastern Canadian wolves – which was referred to as the "Great Lakes" wolf in this study – using microsatellites, mDNA, and the paternally-inherited [[yDNA]] markers found that the eastern Canadian wolf was a unique [[ecotype]] of the gray wolf that had undergone recent hybridization with other gray wolves and coyotes. It could find no evidence to support the findings of the earlier 2000 study regarding the eastern Canadian wolf. The study did not include the red wolf.<ref name=koblmuller2009/> This study was quickly rebutted on the grounds that it had misinterpreted the findings of earlier studies that it relied upon, nor did it provide a definition for a number of the terms that it used, such as "ecotype".<ref name=cronin2009/>

In 2011, a study compared the genetic sequences of 48,000&nbsp;[[single nucleotide polymorphisms]] ([[mutations]]) taken from the genomes of canids from around the world. The comparison indicated that the red wolf was about 76% coyote and 24% gray wolf with hybridization having occurred 287–430&nbsp;years ago. The eastern wolf – which was referred to as the "Great Lakes" wolf in this study – was 58% gray wolf and 42% coyote with hybridization having occurred 546–963&nbsp;years ago. The study rejected the theory of a common ancestry for the red and eastern wolves.<ref name=vonholdt2011/><ref name=beeland2013/> However the next year, a study reviewed a subset of the 2011 study's [[Single-nucleotide polymorphism]] (SNP) data and proposed that its methodology had skewed the results and that the eastern wolf is not a hybrid but a separate species.<ref name=rutledge2012a/><ref name=beeland2013/> The 2012 study proposed that there are three true canis species in North America – the gray wolf, the western coyote, and red wolf/eastern wolf with the eastern wolf represented by the Algonquin wolf, with the Great Lakes wolf being a hydrid of the eastern wolf and the gray wolf, and the eastern coyote being a hybrid of the western coyote and the eastern (Algonquin) wolf.<ref name=rutledge2012a/>

Also in 2011, a scientific [[literature review]] was undertaken to help assess the taxonomy of North American wolves. One of the findings proposed was that the eastern wolf, whose range includes eastern Canada and the Upper Peninsula of Michigan plus Wisconsin and Minnesota is supported as a separate species by morphological and genetic data. Genetic data supports a close relationship between the eastern and red wolves, but not close enough to support these as one species. It was "likely" that these were the separate descendants of a common ancestor shared with coyotes. This review was published in 2012.<ref name=chambers2012/>

Another study of both mDNA and yDNA in wolves and coyotes by the same authors indicates that the eastern wolf is genetically divergent from the gray wolf and is a North American evolved species with a long-standing history. The study could not dismiss the possibility of the eastern wolf having evolved from an ancient hybridization of gray wolf and coyote in the Late Pleistocene or Early Holocene.<ref name=wilson2012/> Another study by the same authors found that eastern wolf mDNA genetic diversity had been lost after their culling in the early 1960s, leading to the invasion of coyotes into their territory and introgression of coyote mDNA.<ref name=rutledge2012b/>

In 2014, the [[National Center for Ecological Analysis and Synthesis]] was invited by the [[United States Fish and Wildlife Service]] to provide an independent review of its proposed rule relating to gray wolves. The Center's panel findings were that the proposed rule was heavily dependent upon the analysis contained in a scientific literature review conducted in 2011 (Chambers et al.), that this work was not universally accepted and that the issue was "not settled", and that the rule does not represent the "best available science".<ref name=dumbacher2014/> Also in 2014, an experiment to hybridize a captive western gray wolf and a captive western coyote was successful, and therefore possible. The study did not assess the likelihood of such hybridization in the wild.<ref name=mech2014/>

In 2015, the [[Committee on the Status of Endangered Wildlife in Canada]] changed its designation of the eastern wolf from ''Canis lupus lycaon'' to ''Canis cf. lycaon'' (''Canis'' species believed to be ''lycaon'')<ref name=COSEWIC2015/> and a species at risk.<ref name=SRPR2015/>

Later that year, a study compared the [[DNA sequences]] using 127,000&nbsp;single-nucleotide polymorphisms (mutations) of wolves and coyotes, but did not include red wolves and used Algonquin wolves as the representative eastern wolf, not wolves from the western Great Lakes states (usually referred to as Great Lakes wolves). The study indicated that Algonquin wolves were a distinct genomic cluster, even distinct from the Great Lakes states' wolves, which it found were actually hybrids of the gray wolf and the Algonquin wolf.  The study's results did not exclude a possibility that the Great Lakes states' wolf (the gray wolf x eastern wolf hybrid (''C.&nbsp;l.&nbsp;lycaon'')) historically inhabited southern Ontario, southern Quebec and the northeastern United States alongside the Algonquin wolf, as there is evidence to suggest both inhabited those areas.<ref name=rutledge2015/>

In 2016, a study of mDNA once again indicated the Eastern wolf as a coyote–wolf hybrid.<ref name=ersmark2016/>

In 2018, a study looked at the [[Y-chromosome]] male lineage of canines. The unexpected finding was that the one Great Lakes wolf specimen included in this study showed a high degree of [[genetic divergence]]. Previous studies propose the Great Lakes wolf to be an ancient ecotype of the gray wolf that had experienced genetic [[introgression]] from other types of gray wolves and coyotes. The study called for further research into the Y-chromosomes of coyotes and wolves to ascertain if this is where this unique genetic male lineage may have originated from.<ref name=oetjens2018/>