การพัฒนาเทคนิค Tetra-primer ARMS-PCR เพื่อคัดเลือกลักษณะอะไมโลสสูง ในการปรับปรุงพันธุ์มันสำปะหลัง

Adcharapun Chaicharoen; Suwaluk  Sansanee; Jeeraporn Kansup; Vipavee  Chanroj; Krittaya Petchpoung; Siriwan Soiklom

doi:10.14456/thaidoa-agres.2026.16

Authors

Adcharapun Chaicharoen Biotechnology Research and Development Division, Thanyaburi, Pathum Thani 12110, Thailand
Suwaluk Sansanee Rayong Field Crops Research Center, Huai Pong, Mueang, Rayong 21150, Thailand
Jeeraporn Kansup Biotechnology Research and Development Division, Thanyaburi, Pathum Thani 12110, Thailand
Vipavee Chanroj Biotechnology Research and Development Division, Thanyaburi, Pathum Thani 12110, Thailand
Krittaya Petchpoung Kasetsart University Research and Development Institute, Ngamwongwan Rd., Chatuchak, Bangkok 10900, Thailand
Siriwan Soiklom Kasetsart University Research and Development Institute, Ngamwongwan Rd., Chatuchak, Bangkok 10900, Thailand

DOI:

https://doi.org/10.14456/thaidoa-agres.2026.16

Keywords:

amylose content, molecular marker, SNP, Tetra-primer ARMS-PCR, plant breeding

Abstract

Amylose starch in cassava (Manihot esculenta Crantz) can be widely used in many industries. Rapid selection of cassava varieties with high amylose starch content can be done using advanced techniques. This study aimed to develop molecular markers associated with high amylose content by designing primers for single nucleotide polymorphism (SNP) markers. The results revealed that the SNP marker S2_25293441 on chromosome 2 was significantly associated with high amylose content. Optimal conditions for tetra-primer amplification refractory mutation system polymerase chain reaction (tetra-primer ARMS-PCR) were achieved at an annealing temperature of 60°C. In heterozygous (CT) genotypes, DNA bands of 339, 229, and 166 bp were observed, while homozygous (CC) genotypes produced bands of 339 and 229 bp. Genotypes exhibiting the CT genotype demonstrated phenotypes with high amylose content. In testing with cassava varieties containing amylose content exceeding 18%, the developed SNP marker achieved an accuracy rate of 75.90% in selecting high amylose varieties. The use of this molecular marker reduced the number of plants required for field selection, enabled screening with fairly high accuracy, and shortened the time for selecting cassava varieties with high amylose content.

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