International Conference on Mathematics and Natural Sciences, 29-30 Nov. 2006, Bandung West Java.
Nengah Dwianita Kuswytasari (1), I Nyoman Pugeg Aryantha (1,2)
1) RG Microbiology, Genetics and Molecular Biology School of Life Sciences and Technology ITB
2) Center for Life Sciences ITB
ABSTRACT
The presence of bacterial endosymbiont on some species of aphids has been studied by using molecular approach. Aphids were collected from BALITSA vegetable field in Lembang West Java. Four methods for DNA isolation were applied, but only modified method of Fukatsu (1999) was successfully isolated the high molecular weight of aphid DNA along with endosymbiont DNA. Polimerase Chain Reaction (PCR) technique was employed to amplify the DNA by using 16SA1 and SB1 primers. The primers (5’-AGAGTTTGATCMTGGCTCAG-‘3[forward], 5’-TACGGYTACCTTGTTACGACTT-‘3[reverse]) were designed to amplify general eubacterial 16S rDNA with a total length 1500 bp of amplification result. Amplification results show that only Aphis gossypii (Glover) give a positive result of bacterial endosymbiont. Other species of aphid [i.e Aphis craccivora (Koch), Myzus ornatus (Laing), Myzus persicae (Sulzer), Lipaphis erysimii (Kaltenbach), and Rhopalosiphum rufiabdominalis (Sasaki)] did not give any positive result of eubacterial endosymbiont. Perhaps there is no endosymbiont exist on these species or they have other kind of endosymbiont rather than eubacterial endosymbiont. It is also found that bacterial endosymbiont DNA can not be well preserved with acetone 100% over 1, 3, 6 and 9 months.
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Keywords : Bacterial endosymbiont, aphid, 16S rDNA
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