Lokus SSR Berasosiasi Karakter Tahan Penyakit Mati-Pohon Durian Berdasarkan Bulked Pseudo-Segregant Analysis (SSR Loci Associated to Resistance Traits to Durian Die-Back based on Bulked Pseudo-Segregant Analysis)

Panca Jarot Santoso, I Nyoman Pugeg Aryantha, Sony Suhandono, Adi Pancoro

Abstract


Penyakit mati-pohon disebabkan cendawan Pythiaceae khususnya Phytophtora palmivora, Pythium vexans, dan Pythium cucurbitacearum menjadi salah satu kendala utama dalam budidaya durian. Di antara upaya pengendaliannya adalah melalui pemuliaan dan seleksi tanaman tahan berbasis molekuler menggunakan marka SSR. Penelitian untuk mengidentifikasi lokus SSR yang berasosiasi dengan karakter tahan penyakit mati-pohon pada durian telah dilaksanakan di Laboratorium Genetika Tumbuhan SITH-ITB dari bulan April sampai dengan Desember 2014. Penelitian dilaksanakan secara bulked pseudo-segregant analysis dua pool DNA durian tahan dan rentan. Amplifikasi lokus SSR menggunakan 77 pasang primer mikrosatelit berlabel fluorescent. Produk amplifikasi dibaca menggunakan GeneMarker v.2.4.0., setiap puncak pancaran fluorescent yang memiliki nilai intensitas tinggi dipilih sebagai alel. Pembandingan panjang alel dilakukan di antara dua pool dan pembanding aksesi tahan. Lokus yang memiliki alel berbeda antara dua pool tetapi memiliki alel sama dengan pembanding dianggap sebagai marka yang berasosiasi dengan sifat tahan durian terhadap Pythiaceae. Hasil analisis ditemukan tiga lokus mDz03F10, mDz4B2, dan mDz3B1 dengan motif berturut-turut (GAA)3.A(GA)4, (GAGT)2ttGAGT, dan (TTTTATG)2(GCCC)2 teridentifikasi sebagai marka yang berasosiasi dengan karakter tahan Pythiaceae. Hasil analisis ini memerlukan satu langkah validasi untuk meyakinkan keterpautan marka dengan karakter target sebelum digunakan sebagai marka molekuler.

Keywords

Durian; SSR; BpSA; Tahan; Pythiaceae

Abstract

Die-back disease caused by Pythiaceae especially Phytophtora palmivora, Pythium vexans, and Pythium cucurbitacearum is one of the obstacles in durian cultivation. An effort to control this disease is through breeding and selection of resistant plants based on molecular assays such as SSR markers. Research to identify SSR loci associated with durian die-back resistance was done at Plant Genetics Laboratory, SITH-ITB from April to December 2014. The research was conducted through bulked pseudo-segregant analysis of two DNA pools, resistance, and susceptible durians. Amplification of SSR loci was carried out by using 77 fluorescent labeled primers. Amplification products were analyzed using GeneMarker v.2.4.0. Fluorescent peak with high intensity was considered as a selected allele. Comparison of allele length was executed amongst two pools and resistance reference. A locus showed different allele between two pools, while it given the same allele to reference was considered as SSR marker associated with Phytiaceae resistance. The analysis were found three loci, mDz03F10, mDz4B2, and mDz3B1 with motif of (GAA)3.A(GA)4, (GAGT)2ttGAGT, and (TTTTATG)2(GCCC)2 recpectively identified as SSR markers associated to die-back resistance. This result, therefore, requires further validation to convince markers association to target traits before they are used as molecular markers.


Keywords


Durian; SSR; BpSA; Resistance; Pythiaceae

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DOI: http://dx.doi.org/10.21082/jhort.v30n1.2020.p9-20

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