aquilonis within the species complex) and Ceratitis capitata. tryoni(although indistinguishable from B. However, a slightly modified version ( Semeraro and Malipatil 2005) provided below, is recommended instead of Test 1 specifically for the diagnosis of the high-risk pest species B. Test 2 takes slightly longer, the RFLP patterns are sometimes more complex to interpret and the variation in size of the ITS1 region not as easy to detect is in Test 1. Therefore this test is recommended when the potential species is within that listed for this test. Test 1 is more rapid, the RFLP patterns less complex to interpret and the variation in size of the ITS1 region easier to detect than Test 2. Host records for the target taxa may assist in the elimination of possible non-target species. narrow the possible species at the outset for choice of restriction enzymes.tryoni complex from Ceratitis capitata only.Īll tests should be considered in terms of the host fruit (for immature life stage samples) and likely country/place of origin to: ![]() Test 2 can distinguish 24 of 33 target species within the current list of species nine species form four species groups. aquilonis) cannot be distinguished from each other. Test 1 can distinguish 25 of 27 target species within the current list of 59 species all four species within the Bactrocera tryoni group (including B. In the specific circumstance to distinguish just Bactrocera tryoni complex from Ceratitis capitata the enzymes AluI, DdeI, RsaI and SspI are diagnostic. This test can be used for identification of 33 fruit fly species ( Diagnostic methods used to identify fruit flies) details are available in Armstrong and Cameron (1998). The specific suite will depend on the likely species identity if it can be narrowed down by likely country of origin or the fruit. Various combinations of three or four out of 10 restriction enzymes are recommended to produce a species-specific RFLP profile. See diagram and PCR RFLP test 2 for methods. Because of this, the variability in PCR fragment length is not so discernible and is therefore not useful as an additional character. Test 2, originally developed by Armstrong and Cameron (1998), is similar to Test 1 but amplifies a larger 1.5-1.8 kb DNA fragment, encompassing the 18S and the ITS1 gene regions. ![]() Part of the ribosomal RNA operon with the location of primer positions for Tests 1 and 2 Test 2
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