Genetic Diversity among Imperata cylindrica Josiah Raymer, Undergraduate Research Assistant Mack Thetford, Associate Professor West Florida Research and.

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Presentation transcript:

Genetic Diversity among Imperata cylindrica Josiah Raymer, Undergraduate Research Assistant Mack Thetford, Associate Professor West Florida Research and Education Center, Milton, FL Institute of Food and Agricultural Sciences University of Florida

Invasive weed No reliable methods of control Obligate outcrosser (can not self fertilize) Lack of genetic diversity (clonal populations) Red leaf variants are sold as an ornamental Limited knowledge of genetic relationship Imperata cylindrica Background Information Imperata cylindrica Cogon Grass

Cranberry Grass Red Baron Grass Red Baron Cranberry Grass Imperata cylindrica rubra Imperata cylindrica ‘Red Baron’ Imperata cylindrica rubra ‘Red Baron’ Imperata cylindrica Variety?Cultivar?Cultivar of a variety? Imperata cylindrica Cogon Grass Variety?Cultivar?Cultivar of a variety?

Objectives Develop a reliable method of extracting amplifiable DNA from both red and green leaf selections of Imperata Determine if diversity exists among red leaf selections of Imperata

Objective 1 Determine a reliable DNA extraction method Evaluate the effectiveness of two alternative DNA extraction procedures compared to the standard CTAB procedure to obtain amplifiable DNA

Qiagen DNeasy® Extraction DNeasy Plant Mini Kits A commercially available Plant DNA extraction kit

Amplification RAPD/PCR reactions Replication of DNA fragments of specific size based on the primer used in the reaction Gel Electrophoresis Separation of DNA fragments of specific size using an electrical current

Amplification (Qiagen) DNA from green leaf samples produced well amplified fragments  DNA from red leaf samples did not produce amplified fragments

DNA Purification (red samples) Centricon YM-100 ultrafilters Membrane based filtration tubes

Results of Amplification Ultrafiltered DNA isolated from red leaf Imperata via Qiagen DNeasy kits, produced amplified fragments

Results DNA of green leaf Imperata extracted using the Qiagen DNeasy kits produced amplifiable DNA DNA of red leaf Imperata extracted using the Qiagen DNeasy kits required an extra ultrafiltration step before amplification occurred

Objective 2 Is There Genetic Diversity Within Selections of Red Leaf Cogongrass? Are Red Leaf Selections similar to Green Leaf Selections?

Samples 29 Samples 13 green leaf (UF Collection) 16 red leaf (multiple industry sources)

Amplification RAPD/PCR 5 Primers 5 Different fingerprints/sample Horizontal gel electrophoresis Digital Images

Imperata cylindrica Gel Images Imperata cylindrica Cogon Grass 1 Kb ladder A02A07 A09 A19 B02 A02 A07 A09A19 B02 Sample #9 Sample SB120

Gel Processing Gel imaging software (Gel Comprar II) 5 fingerprints for each sample were linked to form a composite data set Dendogram was constructed Principal component analysis Jacknife comparison

Dendogram Imperata cylindrica Cogon Grass

Imperata cylindrica Principal Component Analysis Imperata cylindrica Cogon Grass

Imperata cylindrica Jackknife comparison Imperata cylindrica Cogon Grass Results of jackknife comparison of red and green leaf samples of Imperata cylindrica. Actual Sample Morphology Predicted Sample Morphology (%) Green LeafRed Leaf Green Leaf8515 Red Leaf0100

Conclusions Red and green leaf selections of Imperata cylindrica were separated into two distinct groups.

Conclusions Red leaf selections of Imperata cylindrica were further separated into two distinct groups.

Significance to Industry  Genetic diversity is present within populations of red and green leaf selections of cogongrass.  Potential exists for cross pollination between the different selections.