OBJECTIVES

The primary objectives of this proposal are to 1) identify the minimum number of DNA sequences (barcodes) required to definitively classify plant species and differentiate between native and invasive species; and 2) generate a database of suitable DNA barcodes sequences from 300 species that are a) native to Hawaii, b) known to be invasive in Hawaii or c) suspected to have the potential of becoming invasive in Hawaii. This knowledge will be used to develop a diagnostic method for the detection of invasive species in Hawaii. The ultimate goal is to develop an assay that can positively identify invasive species at any life stage and detect small amounts of contaminating invasive species in bulk material (e.g. seed mimics in imported seed lots). This proposal is not intended to assess the invasiveness of plants - several existing programs already focus on this task (e.g. Hawaii Weed Risk Assessment Group, NatureServe).

APPROACH

1) We will use bioinformatic analysis of existing GenBank data, as well as published literature, to identify the DNA region most suitable for the DNA barcoding of land plants. These regions will be selected from the plastid, mitochondrial and/or nuclear genomes. 2) The DNA barcode region will be amplified via PCR from a number of families containing species that are difficult to tell apart, beginning with the family Poaceae (grasses). 3) The DNA sequence for these amplicons will be obtained and examined for useful sequence polymorphisms. 4) Sequence polymorphisms will be stored in a web-accessible database.