Development of a Spatially-Related Phytochemical Database for a Chihuahuan Desert Shrubland

Mary E. Lucero1, Rick Estell1, Ken Ramsey2, Kris Havstad1

1USDA-ARS Jornada Experimental Range, Las Cruces New Mexico (http://jornada-www.nmsu.edu/) 2Jornada Basin LTER, New Mexico State University, Las Cruces, NM (http://jornada-

www.nmsu.edu/)

We examine soil and vegetation dynamics involved in arid land degradation and desertification at multiple scales.

Phytochemistry plays a role in such vegetation dynamics as allelopathy, herbivore deterrence, and plant-plant competition. Our goals are to:

determine the volatile composition of JER plants

define how these volatiles interact with key soil microbes, herbivores, and competitive plants

identify areas where phytochemical interactions contribute to success or failure of a plant species.

Apply findings to development of remediation strategies.

PhytoChemistry Database

Protocols

Chemicals

Datasetsmarker gene sequences

(TBA)

Essential Oil Projects

LTER plant species list

Primary Key ID

Plant species code

Related Dataset ID’s (examples: Oil yield, DMA, GCMS, GCFID)

X, Y, Z GPS coordinates and related details

Scientific Names Code Habit Form PhenologyCyphomeris gypsophiloides CYGY Perennial FORB SUDalea formosa Torr. DAFO Perennial SHRUB SPDasyochloa pulchella (HBK) Hitchci DAPU Perennial GRASS SUDalea scoparia Gray (Legum.) DASC Perennial SHRUB SU

LTER PLANT SPECIES LIST

Steam Distillation of ground plant tissue: Written by Mario Tellez prior to 1998, modified by Mary Lucero August 30, 2000

Reagents 1. Pentane - HPLC grade, Fisher Scientific #p399-1 2. Distilled Water Materials 1. Likens-Nickerson Distillation apparatus, including a West Condenser and a Coiled Condenser

West Condenser Coiled Condenser 2. Water bath 3. Heating Mantle 4. Aluminum foil/ glass wool/ cotton/ paper towels or other appropriate insulating material 5. Round Pear shape flask (about 10 mL, 14/20 joints) 6. 300 mL 3-way flask (though nearly any round-bottom, 300 mL flask will do) 7. Stirring bar (small) 8. Hot plate/stirrer 9. Cold water circulator (I used the Forma Scientific 2160 filled with 60% antifreeze, however, I believe plain water would do. My initial setting of –20oC caused ice to form inside the coils of the upper condenser, so I had to shut the system down. Set up the circulator the night before, to allow the temperature to fall before starting the distillation. A setting of –10oC in the water bath circulator will give you a circulating liquid of about 0oC. ML) 10. Teflon joint liners Procedure 1. Clamp body of Likens-Nickerson apparatus in place. 2. Place the West condenser on top, using a Teflon liner at the joint (use liners at all joints). 3. Place a 10ml pear-shaped recovery flask on the extracting/refluxing (pentane) end of the Likens-Nickerson apparatus with a small magnetic stirring bar. 4. Weigh amount of plant material to be distilled, and record weight (10-20 g good to start, but this will depend on factors like amount of plant available, foaming, etc. These factors may also influence size of flask and amount of water to use) The flask (24/40 joints) can be the usual 3-neck round bottom flask, a 1-neck recovery flask, or just about any other flask that will fit a 24/40 joint. 5. Measure enough distilled water to fill half the flask (This is good for a first run. Conditions, i.e. foaming, might dictate use of much less water or use of a larger flask without a concurrent raise in water volume). 6. Add all but about 20 mL of the measured water to the flask with the plant material and place on the 24/40 joint on the Likens-Nikerson apparatus. 7. From top of West condenser flood bottom part of U-tube in Likens-Nikerson apparatus with remaining water until it starts flowing onto distilling (water/plant) flask. 8. Carefully, from the top of the West condenser, flood top part of U-tube of Likens-Nikerson apparatus with 12-15ml of pentane allowing excess to flow into pear shaped flask [be careful not to let water rise with pentane into pear shaped flask (water level in U-shaped tube should descend enough, once you start adding the pentane, so that pentane has room to go up the elbow and flow into the 10 mL pear-shaped flask)]. 9. Place an Ahlin condenser on top of West condenser (water flow should be from the West condenser into the Ahlin condenser, water temperature preferably 0oC).

Likens and Nickerson Distillation Apparatus

Chemical ID RI Common Name of Chemical

DB-5 RT in Min Structure

Mass Spectrum

508-32-7 927 tricyclene 5.53

Chemicals for which DB-5 column GC retention indices and Mass spectra are available

50 60 70 80 90 100 110 120 130 140

93.00

40

M/Z

GCMS-TIC of essential oil from Dalea scoparia

•Image (*.gif) or ASCII?

•We want to avoid re-creating MS software

•*.CDF files not as universal as we would like