Polymorphisms flanking the mariner integration sites in the rice gall midge (Orseolia oryzae Wood-Mason) genome are biotype-specific

Citation
S. Behura et al., Polymorphisms flanking the mariner integration sites in the rice gall midge (Orseolia oryzae Wood-Mason) genome are biotype-specific, GENOME, 44(6), 2001, pp. 947-954
Citations number
30
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology,"Molecular Biology & Genetics
Journal title
GENOME
ISSN journal
0831-2796 → ACNP
Volume
44
Issue
6
Year of publication
2001
Pages
947 - 954
Database
ISI
SICI code
0831-2796(200112)44:6<947:PFTMIS>2.0.ZU;2-U
Abstract
In an effort to study genome diversity within and between the Indian biotyp es of the Asian rice gall midge, Orseolia oryzae, a major insect pest of ri ce, we made use of mariner transposable element integration site polymorphi sms. Using degenerate primers, the design of which is based on mariner sequ ences, we amplified a ca. 450 bp mariner sequence from the rice gall midge. The mariner sequence showed homology with that of a mariner element isolat ed from the Hessian fly, Mayetiola destructor, a major dipteran pest of whe at. Southern hybridization, using this mariner fragment as a probe, reveale d that the mariner elements are moderately to highly repetitive in the rice gall midge genome. Based on the sequence information of this 450-bp PCR-am plified fragment, outward-directed primers were designed and used in an inv erse PCR (iPCR) to amplify the DNA flanking the conserved regions. To study the regions flanking the mariner integration sites, we employed a novel PC R-based approach: a combination of sequence specific amplification polymorp hism (SSAP) and amplified fragment length polymorphism (AFLP). The outward- directed mariner-specific primer was used in combination with adapter-speci fic primers with 1-3 selective nucleotides at their 3' ends. The amplificat ion products were resolved on an agarose gel, Southern-transferred onto nyl on membranes, and probed with the iPCR fragment. Results revealed biotype-s pecific polymorphisms in the regions flanking the mariner integration sites , suggesting that mariner elements in the rice gall midge may be fixed in a biotype-specific manner. The implications of these results are discussed i n the context of biotype differentiation.