Simple sequence repeat SSR polymorphisms STM 3023 of potatoes belonging to Native

1
Outreach on Potato Genome Project
By Tia Beavert Funded by Mellon Foundation,
Yakama Nation, and Potato Genome Project,
National Science Foundation.
COMMUNITY OUTREACH During the visit to Baker Lab
in Albany, California, I was able to present my
research project to the staff at the Lab. Also
later during the week I was able to present and
teach a second grade class about Potatoes and the
Genetic makeup of the potato, as well as the
basic characteristics of the potato. This
presentation was tied into their class lesson
from the book Frog and Toad, which was on growing
a garden.
ABSTRACT At Heritage College, we have a community
evolutionary garden that has a variety of
Solanaceous plants.  The purpose of the garden is
to show evolutionary differences, changes, or
similarities of one plant throughout time.  My
research has a similar purpose, but is focused
upon the potato plant.   The first process
learned was the extraction of potato DNA in order
to trace the history of the potato.  The DNA
extractions were from various potatoes from the
Brown Lab in Prosser, Washington. The research
center was willing to train me to complete the
process using the potato plant as a model.  The
process practiced can be used in the future to
evaluate plants of my choice using similar
methods.  The reason I am interested in this
research is because I would like to learn more
about the natural foods and medicines the Yakama
Indians use traditionally throughout the year.
The main purpose of my research is to gain
knowledge of how to evaluate plants from a
smaller perspective, as in genetic composition.

• METHODS
• Grind leaf tissue in a pre-chilled mortar in
  liquid nitrogen to a fine powder.
• Transfer 100mg powder into an Eppendorf tube. Add
  700µl of fresh 2X CTAB buffer (and 2µl of
  ß-mercaptoethanol and vortex). Place in 65C
  water bath for 45 min, inverting tubes every 15
  min. Cool samples to RT for 2 min.
• Add 700µl of chloroformisoamyl alcohol (241) to
  each tube. Vortex briefly and gently to avoid
  shearing DNA, then turn over several times.
• Spin 5 min at 14,000 rpm in a microcentrifuge.
  Remove aqueous top layer and transfer to a new
  labeled Eppendorf tube. Be careful to avoid
  carrying over interphase material. Dispose
  chloroformisoamyl alcohol waste properly in a
  labeled waste container.
• Add 50µl of 10 CTAB (in 0.7 M NaCl), vortex
  gently, and mix thoroughly.
• Repeat steps 3 and 4.
• Add equal volume of cold (-20C) isopropanol
  (400-500µl) to each tube. Turn over several times
  and let tubes sit at 4C for 30 min (15 min at
  -20C or 5 min at 70C).
• Spin at 14,000 rpm for 20 min. Carefully pour off
  supernatant so as not to lose the DNA pellet
  turn over tubes to dry (1-2 min).
• Wash pellet with 1ml 70 EtOH (at least 1-3 min)
  and spin 30 min at 14,000 rpm. Carefully pour off
  EtOH, wash pellet in 1ml 95 EtOH, spin 30 min at
  14,000 rpm, and carefully pour off EtOH. Turn
  tubes upside down and air-dry overnight or for
  few minutes using a vaccum pump.
• Dissolve DNA in 150µl T10E1 or distilled H2O per
  sample, add 1-2µl 10mg/ml (DNAse-free)-RNAseA,
  vortex briefly and gently, and spin briefly.
  Incubate at 37C for 1 hr.
• DNA concentration is estimated in comparison with
  known concentration of lambda DNA in 1 agarose
  gel.
• Store at 4C, or at -20C for long-term
  storage.

My purpose of attending the workshop was to learn
how to trace the DNA of plants using the potato
as a basis. Hopefully in the future I can use
the same method to trace the DNA origin of our
natural foods and medicines.
CONCLUSION I learned a scientific process that
involved the extraction of DNA from a plant.
This process can be and will be used in the
future to evaluate plants native to my tribe.
After the workshop, I practiced the method of DNA
extraction thoroughly in order to transmit the
information from a scientific level to the
kindergarten through twelfth grade level. This
enabled the students to learn information about
plants at a molecular level and show them that
science is useful in many ways.
Simple sequence repeat (SSR) polymorphisms (STM
3023) of potatoes belonging to Native Americans
living on the Olympic Peninsula of Washington
State (To-Le-Ak Quileute potato , Ozette
(Victoria, Vancouver Island), Haida Queen
Charlotte Islands, Mak 2.2 and Mak 1.2 Ozette
from Neah Bay compared to native cultivars from
South America (365373 Peru, 365345 Peru, and
Yema de Huevo Colombia) and Mexico (307743,
324461, 281033). The Native American potatoes
from the Olympic Peninsula appear be more
similar to Mexican land-races than to Peruvian
cultivars, but more data is needed.
The Makah Potato Workshop participants included
Hazel Denny, Cassandra Smith, Neuee Ward, Tia
Beavert, and Daniel Andrade. Dr. Charles Brown,
Ching Pa Yang, and David Culley assisted by being
the instructors of the workshop. During the
Makah Potato Workshop, the method of DNA
extraction was practiced and completed.
Throughout the workshop we participated in the
extraction of the DNA and the completion of the
gel.