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Title:	Filogeni molekul subfamili Alysiinae dan Opiinae (Hymenoptera : Braconidae) di Malaysia
Authors:	Ameyra Aman Zuki (P57257)
Supervisor:	Dr Salmah Yaakop
Keywords:	Braconidae Alysiinae Opiinae Subfamili Braconidae-Malaysia.
Issue Date:	10-Sep-2013
Description:	Alysiinae dan Opiinae merupakan dua subfamili di bawah famili Braconidae dan dianggap kumpulan beradik kerana berkongsi ciri biologi iaitu sebagai endoparasitoid ke atas larva Diptera siklorafus. Objektif kajian ini adalah untuk menentukan hubungan filogenetik di antara genus-genus daripada subfamili Alysiinae dan Opiinae, menentukan status monofiletik ke atas setiap klad Alysiinae dan Opiinae serta menentukan keberkesanan penanda genetik 28S dan ND1 dalam menyelesaikan pertalian bagi subfamili Alysiinae dan Opiinae. Sebanyak 48 takson kajian telah berjaya dikumpul dari seluruh Malaysia dan pengekstrakan DNA, tindakan berantai polymerase (PCR), penjujukan serta analisis filogenetik telah dijalankan. Dua penanda genetik yang digunakan adalah gen nuklear 28S D2 (28S) dan gen mitokondria NADH dehydrogenase subunit I (ND1). Tiga kriteria filogenetik telah digunakan iaitu kriteria parsimoni, jarak, dan likelihood. Perisian PAUP* versi 4.0b10 dan MrBayes v 3.1.2 digunakan bagi menjalankan analisis Maksimum Parsimoni (MP), Neighbor-Joining (NJ) dan Inferens Bayesian (BI). Berdasarkan hasil kajian, pohon 50% peraturanmajoriti konsensus data 28S telah memberikan resolusi topologi pohon yang lebih baik ke atas genus berbanding ND1 dengan berjaya mengelompokkan kumpulan genus antaranya Aspilota Foerster 1862, Alysia Latreille 1804, Dapsilarthra Foerster 1862 dan suku Opiini serta Biosterini. Walau bagaimanapun, pengelasan kumpulan genus tersebut mendapat sokongan peratusan butstrap yang rendah (<50) akan tetapi disokong oleh ciri biologi masing-masing. Selain itu, pohon ND1 menunjukkan percampuran genus ke dalam kumpulan genus yang salah. Hasil ujian kehomogenan (Incongruent Length Different Test- ILD) gabungan data 28S dan ND1 memberikan nilai p> 0.05 dan pohon 50% peraturan-majoriti konsensus MP 28S+ND1 memberikan resolusi topologi pohon yang baik bagi status filogeni subfamili Alysiinae dan Opiinae dengan membentuk kumpulan monofiletik dan parafiletik masing-masing. Pemecahan klad tersebut juga disokong oleh ciri morfologi Alysiinae sebagai satu kumpulan monofiletik kerana mempunyai mandibel yang mencapah keluar apabila mulut ditutup, exodont dan Opiinae parafiletik kerana ketidakpastian ciri sinapomorfik disebabkan variasi ciri morfologi yang banyak. Walau bagaimanapun, topologi pohon menunjukkan percampuran genus di dalam kumpulan genus yang salah dan hasil kajian tidak dapat membuktikan hubungan adik beradik antara Alysiinae dan Opiinae. Walaupun tidak mendapat sokongan butstrap dan nilai kebarangkalian akhir yang tinggi, lokus genetik 28S didapati lebih berkesan dalam menyelesaikan hubungan di peringkat kumpulan genus di dalam Alysiinae dan Opiinae berbanding ND1. Hasil kajian berkepentingan di peringkat awalan program pengurusan serangga perosak (Insect Pest Management- IPM) iaitu dapat membantu mengesahkan status spesies dengan menjalankan pengecaman molekul. Ini kerana pengecaman berasaskan ciri-ciri morfologi ke peringkat genus dan spesies ke atas Alysiinae dan Opiinae amat mengelirukan dan tidak stabil sehingga kini.,Alysiinae and Opiinae are two subfamilies under the family Braconidae and assumed to be sister-groups because of their shared biological characterisitcs as endoparasitoids of cyclorrhaphous Dipteran larvae. The objectives of this study were to determine the phylogenetic relationships among genera under the subfamily Alysiinae and Opiinae, to prove the monophyletic status of each clade of Alysiinae and Opiinae, and to determine the effectiveness of 28S and ND1 genetic markers towards revealing relationships in Alysiinae and Opiinae. A total of 48 taxa were collected from all over Malaysia and DNA extraction, polymerase chain reaction (PCR), sequencing analysis and phylogenetic analysis were performed. Two genetic markers were used namely 28S D2 nuclear gene (28S) and NADH dehydrogenase subunit I mitochondrial gene (ND1). The three criteria used in the analyses were parsimony, distance and likelihood. The software of PAUP* version 4.0b10 and MrBayes v 3.1.2 were used to run the Maximum Parsimony (MP), Neighbor-joining (NJ) and Bayesian Inference (BI) analyses. The results of this study showed that the topology of the 50% majority-rule consensus tree based on 28S data using MP analysis provided a better resolution for classifying the genus groups viz. Aspilota Foerster 1862, Alysia Latreille 1804, Dapsilarthra Foerster 1862 and the tribes of Opiini and Biosterini. However, the groupings were supported with very low percentage of bootstrap values (<50), but were supported by their biological characteristics. Besides that, the ND1 tree showed the intermixture of genera in the wrong genus groups. The homogeneity test (Incongruent Length Different test - ILD) showed p> 0.05 from the combined 28S+ND1 data. The 50% majority-rule consensus tree of 28S+ND1 data using MP analysis provided a good resolution in the tree topology for the phylogenetic status of the subfamily by forming a monophyletic group in Alysiinae, while showing a paraphyletic in the Opiinae. Clade divergence was supported by the morphological characteristics of the Alysiinae in having an exodont mandible, where the mandible pointed outwards when the mouth is closed whilst forming a paraphyletic group in the Opiinae because of the undetermined of the synapomorphic character due to large variations in morphological characters. Unfortunately, the tree topology showed the mixing of genera in the wrong genus grouping and the results did not prove the sistergroup relationships of both subfamilies. Even though the bootstrap and the Bayesian posterior probability did not give high support value, however the 28S marker gave results which were more effective than those of ND1 for resolving the relationships of the genus groups in Alysiinae and Opiinae. The results from this study were very significant for the early stage of the Insect Pest Management (IPM) programme as it helps in clarifying the species status through molecular identification. This is because the morphological character based identification system to the genus and species levels are confusing and unstable until now,Master
Pages:	144
Call Number:	QL568.B8 A468 2013
Publisher:	UKM, Bangi
Appears in Collections:	Faculty of Science and Technology / Fakulti Sains dan Teknologi

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