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Accession Number 5004797

Title Assessing the genetic status of Whooping Cranes with AFLP

Project Description The U. S. Fish and Wildlife Service bears full responsibility for the preservation of the whooping

crane and the long-term maintenance of the captive populations in the United States. We need to

know the extant genetic diversity and individual relatedness within the captive whooping crane

populations to adequately manage the captive population and to establish viable wild populations.

The Aransas-Wood Buffalo whooping crane population declined to 15 birds in 1941, the only viable

population in the wild today. Although less than in 1940, the extant genetic diversity in the worlds

whooping crane population is unknown. Studies of genetic diversity in Whooping Cranes to date

have not provided sufficient information to assure long term viability of the species. A newly

developed (and patented) technique Amplified or Anonymous Fragment Length Polymorphism

(AFLP) can provide a robust estimate of diversity. This approach has two major advantages; one is

that a very large number of polymorphisms should be found, and the other is that the startup is

much simpler and less expensive than that of previous approaches. The AFLP technique provides

a better estimate of diversity because it looks at a much broader segment of the nuclear genome

than any of the above techniques. This technique can produce thousands of DNA segments so

adequate polymorphisms to characterize diversity in the cranes should be found. The study will

include samples from other crane populations with robust populations and those of reduced

diversity. A comparison of crane species will give us a better understanding of crane diversity and

the relative state of genetic diversity in the whooping crane. Blood from crane species with robust

populations and those of reduced diversity will be collected and used to prepare DNA. The DNA

will be examined by AFLP for polymorphisms. This will entail determining appropriate reagents

and conditions for 1) endonuclease cleavage of crane DNA, 2) selective PCR of the resulting DNA

pieces, and 3) separation to locate the polymorphisms. The polymorphic DNA segments will be

collected and sequenced. Probes for these polymorphisms will be prepared (via contract) and

used to determined relatedness in Whooping Cranes. We are estimating that between 10 and 50

will be required. After this determination is made the selected DNA segments will be sequenced

and probes for these segments will be prepared (part 2). After the probes are available they will be

utilized to analyze the genetic diversity of all captive Whooping Cranes and of the wild Whooping

Cranes for which samples become available (part 3). These probes should be applicable to all

crane species. Although the use of the AFLP technique requires the use of DNA of the species of

interest without contamination by other DNA (we will utilize crane blood), once the probes are

developed we will apply them to other samples such as feathers and droppings.

Keywords aflp, cranes, genetics, polymorphisms, recovery, restoration, whooping cranes,

Principal Melancon M J., USGS Patuxent Wildlife Research Center: mark_melancon@usgs.gov; Gee G F.,

Investigators Patusent Wildlife Research Center: george_gee@usgs.gov;

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