Sains Malaysiana 51(4)(2022): 1017-1026

http://doi.org/10.17576/jsm-2022-5104-06

Clustering of Fifth Generation Mutants of Genetically - Improved Kipas Putih Soybean with High Protein Content

(Pengelompokan Mutan Generasi Kelima Kacang Soya Kipas Putih Terubah Suai Genetik dengan Kandungan Protein Tinggi)

ZUYASNA^1,*, ANDARI RISLIAWATI² & GUNAWAN¹

¹Agrotechnology Study Program, Faculty of Agriculture, Syiah Kuala University, Jl. T.H.Krueng Kalee No3, Banda Aceh, Indonesia

²Indonesian Centre for Agricultural Biotechnology and Genetic Resources Research and Development, Bogor, Indonesia

Received: 28 October 2020/Accepted: 2 September 2021

Abstract

Kipas Putih is one of the local varieties of soybean from Aceh, Indonesia. To improve the productivity of this variety, the irradiation mutation technique was applied. This study aimed to examine the level of genetic diversity in the M₅ generation of Kipas Putih soybean using SSR markers and to identify whether any used markers were associated with the loci that control the protein content in soybean. A total of 18 SSR markers were used to amplify the DNA of 11 mutant lines of Kipas Putih. The total alleles detected were 55 alleles which range from 1 to 7 alleles per locus. The average of major allele’s frequency was 0.71 with the heterogeneity of 0.17. The PIC value ranged from 0.00 to 0.73 with an average of 0.35. Satt308 was identified as the most polymorphic marker among others since it had a PIC value greater than 0.70. The genetic distance analysis showed that the mutant lines B4, B10 and B13 had the furthest genetic distance (0.45) with the Kipas Putih (B0) soybean variety. The phylogenetic analysis showed Kipas Putih soybean and ten mutants of M₅ generation grouped into four main clusters at a similarity coefficient of 0.20. Furthermore, association analysis showed that only two SSR markers, Satt577 and Satt431 were significantly associated with protein content with an R² value of 0.7855 to 0.9643.

Keywords: Genetic diversity; mutant lines; phylogenetic; SSR markers

Abstrak

Kipas Putih adalah sejenis kacang soya tempatan yang berasal dari Aceh, Indonesia. Untuk meningkatkan produktivitinya, teknik mutasi penyinaran telah diaplikasikan. Kajian ini bertujuan untuk mengetahui tahap kepelbagaian genetik mutan kacang soya Kipas Putih generasi M₅ menggunakan penanda SSR dan untuk menentukan sama ada mana-mana penanda yang digunakan berkorelasi dengan lokus yang mengawal ciri kandungan protein dalam kacang soya. Sebanyak 18 penanda SSR digunakan untuk mengamplifikasi 11 titisan mutan kacang soya Kipas Putih. Jumlah alel yang dikesan adalah 55 alel dengan jarak 1 hingga 7 alel setiap lokus. Purata frekuensi alel dominan adalah 0.71 dengan tahap kepelbagaian 0.17. Nilai PIC berjulat antara 0.00 hingga 0.73 dengan purata 0.35. Satt308 dikenal pasti sebagai penanda yang paling polimorfik berbanding penanda SSR lain memandangkan nilai PICnya yang melebihi 0.70. Analisis jarak genetik menunjukkan bahawa titisan mutan B4, B10 dan B13 mempunyai jarak genetik terjauh (0.45) dengan kacang soya Kipas Putih (B0). Analisis filogenetik menunjukkan bahawa kacang soya Kipas Putih dan sepuluh mutan M₅ dikelompokkan menjadi empat kelompok utama dengan nilai pekali 0.20. Seterusnya, analisis perkaitan menunjukkan bahawa hanya dua penanda SSR iaitu Satt577 dan Satt431 berkait secara signifikan dengan kandungan protein dengan nilai R² 0.7855 hingga 0.9643.

Kata kunci: Filogenetik; kepelbagaian genetik; penanda SSR; titisan mutan

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^*Corresponding author; email: zuyasna@unsyiah.ac.id